CN116685576A

CN116685576A - Cysteine protease inhibitors and methods of use thereof

Info

Publication number: CN116685576A
Application number: CN202180056069.4A
Authority: CN
Inventors: 李·D·阿诺德; 安迪·珍妮丝; 瓦特·姜
Original assignee: American Co Paddis Biosciences
Current assignee: American Co Paddis Biosciences
Priority date: 2020-06-09
Filing date: 2021-06-09
Publication date: 2023-09-01

Abstract

The present disclosure provides compounds of formula II having warheads and their use in the treatment of medical diseases or conditions, such as viral infections. Pharmaceutical compositions and methods of preparing various warhead-bearing compounds are provided. The compounds are expected to inhibit proteases, such as 3C, CL-like or 3 CL-like proteases. II (II)

Description

Cysteine protease inhibitors and methods of use thereof

Cross Reference to Related Applications

The application requires U.S. S. N.63/036,866 submitted on 6/9 of 2020; U.S. S. N.63/039,297 submitted on 6/15 of 2020; U.S. S. N.63/067,669 submitted on day 19, 8, 2020; U.S. S. N.63/091,630 submitted on day 14 of 10 of 2020; U.S. S. N.63/129,018 submitted on 12/22 2020; U.S. S. N.63/171,675 submitted at 4/7 of 2021; U.S. S. N.63/172,478 submitted at 4/8 of 2021; U.S. S. N.63/173,146 submitted at 4/9 of 2021; U.S. S. N.63/179,128 submitted at month 23 of 2021; and U.S. S. N.63/195,460, filed on 1,6, 2021, the contents of each of which are hereby incorporated by reference in their entirety.

Brief description of the sequence Listing

The present application contains a sequence listing submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. ASCII copy name created at month 6 and 3 of 2021 is PARB-004wo_sl. Txt and is 3,285 bytes in size.

Background

The coronaviridae (Coronaviridae family) in viruses are enveloped, single-stranded, sense RNA viruses and include 141 species, which are classified into four genera according to their systemic relationship: alpha-, beta-, gamma-and delta-coronaviruses. Coronaviruses (covs) are zoo viruses that infect a variety of animals ranging from whales to birds, bats, cats and humans. Generally, coV infection causes mild to moderate respiratory tract infections; however, some CoV species are extremely toxic and can cause extensive death. Severe acute respiratory syndrome coronavirus (Severe acute respiratory syndrome coronavirus; SARS-CoV) is human CoV, which causes the first pandemic in the 21 st century, infects over 8,000 people and has a mortality rate of 10%. Coronavirus of the middle east respiratory syndrome (Middle East respiratory syndrome coronavirus; MERS-CoV) was identified in 11 in 2012 and was from this infected more than 1,600 people in 26 countries and had a mortality rate of 36%. It is therefore important to identify coronavirus drug targets that can be used to develop broad-spectrum anti-coronavirus therapeutics to combat existing and emerging infections with coronaviruses.

All covs express >800kDa replicase polymeric proteins comprising two or three cysteine proteases, papain-like proteases (PLPpro, or PLP1 and PLP 2), and 3C-like proteases (3 CLpro, nsp5 or Mpro). These proteases treat the CoV replicase polymeric protein by cleaving the polymeric protein into 16 nonstructural proteins that are responsible for various aspects of CoV replication. CoV 3CLpro is responsible for handling 11 cleavage sites within replicase polymeric proteins and is critical for CoV replication, making it a very valuable therapeutic development target. In all CoV 3CLpro, the overall active site architecture and substrate recognition pocket are conserved in structure, increasing their attractiveness as targets for the development of broad-spectrum anti-CoV therapeutics. Furthermore, the high sequence conservation near the active site in CoV 3CLpro from different coronavirus subclasses makes it an excellent target for the development of broad-spectrum therapeutics against coronavirus infections. Thus, the development of CoV 3CLpro inhibitors is a promising approach for the treatment of respiratory infections and related diseases.

Extensive research by targeting a direct animal epidemic host of coronaviruses with small molecule inhibitors (zoonotic reservoirs) helps to learn a structure-based design strategy aimed at creating molecular scaffolds that can help develop therapeutics for coronavirus infection; however, small molecule antiviral agents or commercially available effective broad-spectrum therapeutic agents have not been identified. There is an urgent need to develop broad spectrum CoV therapeutics to overcome the challenges of traditional anti-CoV therapeutic development, as broad spectrum therapeutics can be rapidly applied in the outbreak of human and animal comorbidities.

Disclosure of Invention

The present disclosure relates in part to viral protease inhibitors. Also disclosed herein are pharmaceutical compositions comprising at least one disclosed compound and a pharmaceutically acceptable carrier.

In one embodiment, disclosed herein is an antiviral compound comprising a warhead covalently bound to a 3C or 3CL protease inhibitor, wherein the antiviral compound is covalently bound to a Cys residue of a protease, and wherein the antiviral compound has activity against one or more viruses.

Also disclosed herein are compounds represented by formula II:

or a pharmaceutically acceptable salt, stereoisomer, ester, or prodrug thereof, wherein:

R ^3a Selected from the group consisting ofAnd 4-10 membered heterocycles, wherein the heterocycles may optionally be one, two or three are each independently selected from the group consisting of hydroxy, C ₁ -C ₈ Alkoxy, oxo (oxo) and warhead a; r is R ^3b Selected from hydrogen and C ₁ -C ₈ An alkyl group; wherein R is ^3a And R is ^3b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring, wherein the heterocyclic ring can optionally be selected from C independently from one, two or three ₆ -C ₁₄ Aryl and warhead a substituents; r is R ^1a Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl; r is R ^1b Selected from hydrogen and C ₁ -C ₈ An alkyl group; or R is ^1a And R is ^1b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl; r is R ¹ Selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ¹ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a); r is R ^A Independently at each occurrence selected from the group consisting of: halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、-CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-C(O)-OC(CH ₃ ) ₃ 、C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocyclyl, wherein R ^B The alkyl, heterocyclyl, heteroaryl, or aryl may be optionally substituted with one, two, or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Hydroxyl and oxo; r is R ² Selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 、-O-(C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl), 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ^B Or R is ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a); or R is ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered mono-or bicyclic heterocycle having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a); r is R ³ Selected from 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a); r is R ^B Independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₁₆ Cycloalkyl, fluorenylmethyloxy, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; r is R ^C Independently at each occurrence selected from hydrogen, halogen and C ₁ -C ₈ An alkyl group; r is R ^x Independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano, -O- (R) ^xx )-OCH ₃ 、-OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, -O-C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; wherein two are geminal C ₁ -C ₈ Alkyl groups may be joined together to form, together with the carbon to which they are attached, C optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy and oxo ₃ -C ₆ Cycloalkyl; and wherein alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each independently selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl; r is R ^G Selected from the group consisting of: hydrogen, optionally by one, two or three R ^gg Substituted C _1-6 Alkyl, optionally substituted with one, two or three R ^hh substituted-C (=o) -C _1-6 Alkyl, -C (=o) -C _3-6 Cycloalkyl, -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl), -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ Aryl), -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocyclyl) and-C (O) - (4-10 membered heterocyclyloxy); wherein the aryl, heterocyclyl or heteroaryl group may optionally be substituted with one,Two or three R' s ^jj Substitution; r is R ^gg Independently at each occurrence selected from the group consisting of: -C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ；R ^hh Independently at each occurrence selected from the group consisting of: halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocyclyl and C ₁ -C ₆ An alkoxy group; r is R ^jj Independently at each occurrence selected from the group consisting of: halo, oxo, hydroxy, cyano, C ₁ -C ₆ Alkyl, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, C ₁ -C ₆ Alkoxy, C _3-6 Cycloalkyl, SF ₅ And NH ₂ ；R ^m Independently at each occurrence selected from the group consisting of: hydrogen, C _1-3 Alkyl, phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl; wherein C is _1-3 Alkyl, phenyl and C _3-6 Cycloalkyl groups may be optionally substituted with one, two or three halo groups; r is R ^xx Is- (OCH) ₂ CH ₂ ) _nn -wherein nn is selected from 1, 2, 3, 4, 5 and 6; r is R ^y Independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CF ₃ 、-CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH; a is a warhead; and X is selected from C (R) ^xy ) And N, wherein R is ^xy Selected from the group consisting of: H. d, -OH, -NH ₂ Halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl and C ₁ -C ₈ An alkoxy group.

In some embodiments, disclosed herein are compounds represented by formula II-A:

in some embodiments, disclosed herein are compounds represented by formula II-B:

in some embodiments, disclosed herein are compounds represented by formulas II-C:

in some embodiments, disclosed herein are compounds represented by formula II-D-A or formula II-D-B:

in some embodiments, disclosed herein are compounds represented by formula II-E-A or formula II-E-B:

in some embodiments, disclosed herein are compounds represented by formulas II-F:

in some embodiments, disclosed herein are compounds represented by formulas II-G:

in some embodiments, disclosed herein are compounds represented by formula II-H-A or formula II-H-B:

wherein pp is selected from 0, 1, 2 and 3.

In some embodiments, disclosed herein are compounds represented by formulas II-E:

wherein ss is selected from 0, 1, 2 and 3 and mm is selected from 1, 2 and 3.

In some embodiments, disclosed herein are compounds represented by formulas II-I:

or a pharmaceutically acceptable salt thereof, wherein:

R ³ is thatR ^t Independently at each occurrence H or methyl; or each R ^t Can form a cyclopropyl group together with the carbon to which it is attached; r is R ^B Selected from the group consisting of: 9-10 membered bicyclic heteroaryl having one ring nitrogen, C ₁ -C ₆ Alkyl and C ₂ -C ₃ Alkenyl groups; wherein R is ^B Optionally substituted with one, two or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₃ Alkoxy, NHR ^m And phenyl (optionally substituted with one or two halogens); and R is ^m Is C _1- C ₃ Alkyl or-C (O) -C _1-3 Alkyl, wherein each C _1- C ₃ Alkyl is independently optionally substituted with one, two or three halogens.

In certain embodiments, disclosed herein are conjugates represented by formula III:

wherein Cys is ₁₄₅ Is cysteine at position 145 or an equivalent active site on a CL or 3CL protease; IR is a viral protease inhibitor; and wherein the conjugate-forming compound comprises-CN warhead.

Detailed Description

Features and other details of the present disclosure will now be described more particularly. Before further describing the present disclosure, certain terms used in the specification, examples, and appended claims are collected here. These definitions should be read in light of the remainder of the present disclosure and as understood by those skilled in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

Definition of the definition

The term "treatment" includes any symptom-independent effect that causes an improvement (e.g., alleviation, reduction, modulation, or elimination) of a condition, disease, disorder, or the like, including reduction of detoxification in asymptomatic individuals and prevention of exposed individuals.

The term "alkyl" as used herein refers to a saturated straight or branched chain hydrocarbon. Exemplary alkyl groups include, but are not limited to, straight or branched chain hydrocarbons having 1-6, 1-4, or 1-3 carbon atoms, respectively referred to herein as C _1-6 Alkyl, C _1-4 Alkyl and C _1-3 An alkyl group. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-butyl, 3-methyl-2-butyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-dimethyl-1-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, and the like.

As used herein, the term "alkynyl" refers to an unsaturated straight or branched hydrocarbon having at least one carbon-carbon triple bond. Exemplary alkynyl groups include, but are not limited to, straight or branched chain groups having 2-6 or 3-6 carbon atomsRespectively referred to herein as C _2-6 Alkynyl and C _3-6 Alkynyl groups. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, and the like.

As used herein, the term "alkenyl" refers to an unsaturated straight or branched hydrocarbon having at least one carbon-carbon double bond. Exemplary alkenyl groups include, but are not limited to, straight or branched chain groups having 2-6 or 3-4 carbon atoms, respectively referred to herein as C ₁ -C ₅ Alkenyl, C ₂ -C ₆ Alkenyl and C ₃ -C ₄ Alkenyl groups. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, and the like.

As used herein, the term "alkoxy" refers to a straight or branched chain alkyl group (alkyl-O-) linked to oxygen. Exemplary alkoxy groups include, but are not limited to, alkoxy groups having 1 to 6 or 2 to 6 carbon atoms, respectively referred to herein as C ₁ -C ₅ Alkoxy, C ₁ -C ₆ Alkoxy and C ₂ -C ₆ An alkoxy group. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

The term "aryl" refers to a group of a mono-or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., sharing 6, 10, or 14 pi electrons in a ring array) provided with 6 to 14 ring carbon atoms and zero heteroatoms ("C) _6-14 Aryl "). In some embodiments, the aryl group has six ring carbon atoms ("C ₆ Aryl "; such as phenyl). In some embodiments, aryl groups have ten ring carbon atoms ("C ₁₀ Aryl "; for example naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has fourteen ring carbon atoms ("C ₁₄ Aryl "; such as anthracenyl). "aryl" also includes ring systems in which an aryl ring as defined above is fused with one or more carbocyclyl or heterocyclyl groups, wherein the linking group or point of attachment is on the aryl ring, and in such cases the number of carbon atoms continues to indicate the number of carbon atoms in the aryl ring system. Typical aryl groups include, but are not limited to, groups derived from: acetocene, acenaphthylene, acephenanthrene, anthracene, azulene, benzene,Coronene, fluoranthene, fluorene, hexaacene (hexacene), hexacene (hexacene), indanadiene (as-indacene), s-indenodiene (s-indacene), indane, indene, naphthalene, octacene (octacene), ovobenzene (oxacene), penta-2, 4-diene, pentacene (pentacene), pentacene (pentalene), pentalene (pentacene), perylene, phenalene, phenanthrene, picene, heptacene, pyrene, pyran, shavings (rubicene), triphenylene, and binaphthene (trinaphthalene). Specifically, aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl.

Examples of representative substituted aryl groups include the following:

wherein R is ⁵⁶ And R is ⁵⁷ One of which may be hydrogen and R ⁵⁶ And R is ⁵⁷ At least one of which is independently selected from halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl, 4-10 membered heterocyclyl, alkanoyl, C ₁ -C ₈ Alkoxy, heteroaryloxy, alkylamino, arylamino, heteroarylamino, NR ⁵⁸ COR ⁵⁹ 、NR ⁵⁸ SOR ⁵⁹ 、NR ⁵⁸ SO ₂ R ⁵⁹ COO-alkyl, COO-aryl, CONR ⁵⁸ R ⁵⁹ 、CONR ⁵⁸ OR ⁵⁹ 、NR ⁵⁸ R ⁵⁹ 、SO ₂ NR ⁵⁸ R ⁵⁹ S-alkyl, SO ₂ -alkyl, S-aryl, SO-aryl and SO ₂ -an aryl group; or R is ⁵⁶ And R is ⁵⁷ A cyclic ring (saturated or unsaturated) having 5 to 8 atoms and optionally containing one or more heteroatoms selected from the group consisting of N, O and S; r is R ⁶⁰ And R is ⁶¹ Independently hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₄ Haloalkyl, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ Aryl, substituted C ₆ -C ₁₀ Aryl, 5-10 membered heteroaryl, or substituted 5-10 membered heteroaryl.

As used herein, the term "carbonyl" refers to the group-C (O) -.

As used herein, the term "cyano" refers to the group-CN.

As used herein, the term "cycloalkoxy" refers to cycloalkyl (cycloalkyl-O-) linked to oxygen. Exemplary cycloalkoxy groups include, but are not limited to, cycloalkoxy groups having 3 to 6 carbon atoms, referred to herein as C _3-6 A cycloalkoxy group. Exemplary cycloalkoxy groups include, but are not limited to, cyclopropoxy, cyclobutoxy, cyclohexyloxy, and the like.

The term "cycloalkyl" or "carbocyclyl" as used herein refers to a saturated or partially unsaturated hydrocarbon group having, for example, 3-6 or 4-6 carbons, respectively referred to herein as C ₃ -C ₁₀ Cycloalkyl, C _3-6 Cycloalkyl or C _4-6 Cycloalkyl groups. Exemplary cycloalkyl groups include, but are not limited to, cyclohexyl, cyclopentyl, cyclopentenyl, cyclobutyl, or cyclopropyl.

As used herein, the term "halo" or "halogen" refers to F, cl, br or I.

As used herein, the term "haloalkyl" refers to an alkyl group in which the alkyl group is substituted with one or more halogens. Typical haloalkyl groups include, but are not limited to, trifluoromethyl (i.e., CF ₃ ) Difluoromethyl, fluoromethyl, chloromethyl, dichloromethyl, dibromoethyl, tribromomethyl, tetrafluoroethyl, and the like. Exemplary haloalkyl groups include, but are not limited to, straight or branched chain hydrocarbons having 1-6, 1-4, or 1-3 carbon atoms substituted with halogen (i.e., cl, F, br, and I), referred to herein as C, respectively _1-6 Haloalkyl, C _1-4 Haloalkyl and C _1-3 Haloalkyl.

When used to describe a compound or a group on a compound, the term "hetero" means that one or more carbon atoms in the compound or group have been replaced with nitrogen, oxygen, or sulfur heteroatoms. Hetero may be applied to any of the hydrocarbon groups described above, such as alkyl groups (e.g., heteroalkyl groups), cycloalkyl groups (e.g., heterocyclyl groups), aryl groups (e.g., heteroaryl groups), cycloalkenyl groups (e.g., cycloheteroalkenyl groups), and the like having from 1 to 5, and specifically, from 1 to 3 heteroatoms.

As used herein, the term "heteroaryl" or "heteroaromatic group" refers to an aromatic 5-10 membered ring system containing one or more heteroatoms (e.g., one to three heteroatoms such as nitrogen, oxygen, and sulfur). The term may also be used to refer to 5-7 membered monocyclic heteroaryl or 8-10 membered bicyclic heteroaryl. Where possible, the heteroaryl ring may be attached to the adjacent group through carbon or nitrogen. Examples of heteroaryl rings include, but are not limited to, furan, thiophene, pyrrole, pyrrolopyridine, indole, thiazole, oxazole, isothiazole, isoxazole, imidazole, benzimidazole, imidazopyridine, pyrazole, triazole, pyridine, pyrimidine, or the like.

Examples of representative heteroaryl groups include the following:

wherein each Z is selected from carbonyl, N, NR ⁶⁵ O and S; and R is ⁶⁵ Each independently is hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₁₀ Cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ Aryl and 5-10 membered heteroaryl.

The term "heterocyclyl", "heterocycle" or "heterocyclic group" is art-recognized and refers to a saturated or partially unsaturated 4-10 membered ring structure, which includes one to three heteroatoms, such as nitrogen, oxygen and sulfur. Where possible, the heterocyclyl ring may be attached to an adjacent group through carbon or nitrogen. The term may also be used to refer to 4-10 membered saturated or partially unsaturated ring structures, which are bridged, fused or spiro ring structures, the ring structure of which includes one to three heteroatoms, such as nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to, pyrrolidine, piperidine, morpholine, thiomorpholine, piperazine, oxetane, azetidine, tetrahydrofuran, dihydrofuran, dihydropyran, tetrahydropyran, and the like. In some embodiments, the heterocycle is a spiroheterocycle (e.g., 2, 8-diazaspiro [4.5 ] ]Decane). In some embodiments, the heterocycle is a bridged heterocycle (e.g., octahydro-1H-4, 7-methanoisoindole). "A spiroheterocyclyl "or" spiroheterocycle "refers to a polycyclic heterocyclyl having rings connected by one common atom (referred to as the spiro atom), wherein the rings have one or more heteroatoms selected from the group consisting of: n, O and S (O) _m (wherein m is an integer of 0 to 2). Representative examples of heterocyclyl groups include, for example:

as used herein, the term "heterocyclyloxy" refers to a heterocyclyl group (heterocyclyl-O-) attached to oxygen.

As used herein, the term "heteroaryloxy" refers to a heteroaryl group (heteroaryl-O-) attached to oxygen.

As used herein, the term "hydroxy" refers to the group-OH.

As used herein, the term "oxo" refers to the group = O.

"pharmaceutically or pharmacologically acceptable" includes molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or human as desired. For human administration, the formulation should meet sterility, pyrogenicity, and general safety and purity standards as required by the FDA biological formulation standard office (FDA Office of Biologics standards).

As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may also contain other active compounds that provide supplemental, additional, or enhanced therapeutic functions.

As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one compound as disclosed herein formulated with one or more pharmaceutically acceptable carriers.

"individual," "patient," or "subject" are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, and most preferably humans. The compounds of the present disclosure may be administered to mammals, such as humans, but may also be administered to other mammals, such as animals in need of veterinary treatment, e.g., livestock (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). "modulation" includes antagonism (e.g., inhibition), agonism, partial antagonism and/or partial agonism.

In this specification, the term "therapeutically effective amount" refers to the amount of the subject compound that will elicit the biological or medical response of a tissue, system or animal (e.g., a mammal or human) that is being sought by the researcher, veterinarian, medical doctor or other clinician. The compounds of the present disclosure are administered in a therapeutically effective amount to treat a disease. Alternatively, a therapeutically effective amount of a compound is that amount necessary to achieve the desired therapeutic and/or prophylactic effect.

As used herein, the term "pharmaceutically acceptable salt" refers to salts of acidic or basic groups that may be present in the compounds used in the composition. The essentially basic compounds included in the compositions of the present invention are capable of forming a wide variety of salts with a wide variety of inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts (e.g., salts containing pharmacologically acceptable anions) including, but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate (tannate), pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, sucrate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1' -methylene-bis- (2-hydroxy-3-naphthalene)). The intrinsically acidic compounds included in the compositions of the present invention are capable of forming base salts with a variety of pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, in particular, calcium salts, magnesium salts, sodium salts, lithium salts, zinc salts, potassium salts and iron salts. The compounds comprising basic or acidic moieties contained in the compositions of the present invention may also form pharmaceutically acceptable salts with various amino acids. The compounds of the present disclosure may contain both acidic and basic groups; for example, an amino group and a carboxylic acid group. In such cases, the compounds may exist in the form of acid addition salts, zwitterionic or basic salts.

The compounds of the present disclosure may contain one or more chiral centers and thus exist as stereoisomers. As used herein, the term "stereoisomer" consists of all enantiomers or diastereomers. Depending on the configuration of substituents around the sterically derived carbon atom, these compounds may be designated by the symbols "(+)", "(-)", "R" or "S", but those skilled in the art will recognize that the structure may implicitly represent a chiral center. The present disclosure encompasses various stereoisomers of these compounds and mixtures thereof. In the nomenclature, a mixture of enantiomers or diastereomers may be designated "(±)", but those skilled in the art will recognize that the structure may implicitly represent a chiral center.

The compounds of the present disclosure may contain one or more double bonds and thus exist as geometric isomers arising from the arrangement of substituents around the carbon-carbon double bond. Sign symbolRepresents a bond, which may be a single bond, a double bond, or a triple bond as described herein. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration, wherein the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass the "E" and "Z" isomers. Substituents around a carbon-carbon double bond may alternatively be referred to as "cis" or "trans", where "cis" represents the same side of the double bond The substituents on the above, and "trans" represents substituents on the opposite side of the double bond.

The compounds of the present disclosure may contain carbocycles or heterocycles and thus exist as geometric isomers arising from the arrangement of substituents around the ring. The arrangement of substituents around a carbocycle or heterocycle is designated as being in the "Z" or "E" configuration, wherein the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise specified, structures depicting carbocycles or heterocycles encompass both the "Z" and "E" isomers. Substituents around a carbocycle or heterocycle may also be referred to as "cis" or "trans", where the term "cis" refers to substituents on the same side of the ring plane and the term "trans" refers to substituents on the opposite side of the ring plane. Mixtures of compounds in which substituents are disposed on the same and opposite sides of the ring plane are designated "cis/trans".

Individual enantiomers and diastereomers of the compounds of the present disclosure may be prepared synthetically from commercially available starting materials containing asymmetric or stereogenic centers or by preparing racemic mixtures followed by analytical methods well known to those of ordinary skill in the art. These analytical methods are exemplified by: (1) attaching the mixture of enantiomers to a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and releasing the optically pure product from the auxiliary, (2) salt formation using an optically active resolving agent, (3) direct separation of the mixture of optical enantiomers on a chiral liquid chromatography column, or (4) kinetic resolution using a stereoselective chemical or enzymatic reagent. The racemic mixture may also be resolved into its constituent enantiomers by well-known methods such as chiral phase liquid chromatography or crystallization of the compounds in chiral solvents. Stereoselective synthesis, a chemical or enzymatic reaction in which a single reactant forms a heterogeneous mixture of stereoisomers during the creation of a new stereogenic center or during the conversion of a pre-existing one, is well known in the art. Stereoselective synthesis encompasses both enantioselective and diastereoselective transformations, and may involve the use of chiral auxiliary. For examples, see Carreira and Kvaerno, classics in Stereoselective Synthesis, wiley-VCH: weinheim,2009.

The compounds disclosed herein may exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents (such as water, ethanol, and the like), and the disclosure is intended to cover solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in crystalline form.

The present disclosure also contemplates isotopically-labeled compounds of the present disclosure as those recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as respectively ² H、 ³ H、 ¹³ C、 ¹⁴ C、 ¹⁵ N、 ¹⁸ O、 ¹⁷ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F and F ³⁶ Cl. For example, one or more H atoms in the compounds of the present disclosure may be replaced with deuterium.

Certain isotopically-labeled disclosed compounds (e.g., with ³ H or ¹⁴ Those compounds labeled with C) are suitable for use in compound and/or substrate tissue distribution assays. Tritiated (i.e., ³ H) And carbon-14 (i.e., ¹⁴ c) Isotopes are particularly preferred for their ease of preparation and detectability. In addition, the use of heavier isotopes (such as deuterium (i.e., ² h) Substitution) may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., extended in vivo half-life or reduced dosage requirements), and thus may be preferred in some cases. Isotopically-labeled compounds of the present disclosure can generally be prepared by following procedures analogous to those disclosed in the examples herein by substituting a non-isotopically-labeled reagent with an isotopically-labeled reagent.

The term "prodrug" refers to a compound that is converted in vivo to the disclosed compound or a pharmaceutically acceptable salt, hydrate, or solvate of the compound. Transformation can occur at various locations (such as in the intestinal lumen or while traversing the intestine, blood, or liver) by various mechanisms (such as by esterases, amidases, phosphatases, oxidative and/or reductive metabolism). Prodrugs are well known in the art (see, e.g., rautio, kumpulain et al, nature Reviews Drug Discovery 2008,7,255). For example, if a compound or a pharmaceutically acceptable salt, hydrate, or solvate of a compound of the present disclosure contains a carboxylic acid functional group, a prodrug may comprise an ester formed by replacing a hydrogen atom of an acid group with a group such as: (C) _1-8 ) Alkyl, (C) _2-12 ) Alkylcarbonyloxymethyl, 1- (alkylcarbonyloxy) ethyl having 4 to 9 carbon atoms, 1-methyl-1- (alkylcarbonyloxy) -ethyl having 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having 3 to 6 carbon atoms, 1- (alkoxycarbonyloxy) ethyl having 4 to 7 carbon atoms, 1-methyl-1- (alkoxycarbonyloxy) ethyl having 5 to 8 carbon atoms, N- (alkoxycarbonyl) aminomethyl having 3 to 9 carbon atoms, 1- (N- (alkoxycarbonyl) amino) ethyl having 4 to 10 carbon atoms, 3-phthaloyl, 4-crotonoyl lactone, gamma-butyrolactone-4-yl, di-N, N- (C) _1-2 ) Alkylamino (C) _2-3 ) Alkyl (such as beta-dimethylaminoethyl), carbamoyl- (C) _1-2 ) Alkyl, N-di (C) _1-2 ) Alkylcarbamoyl- (C) _1-2 ) Alkyl and piperidinyl- (C) _2-3 ) Alkyl, pyrrolidinyl- (C) _2-3 ) Alkyl or morpholino (C) _2-3 ) An alkyl group.

Similarly, if a compound of the present disclosure contains an alcohol functional group, a prodrug may be formed by replacing a hydrogen atom of the alcohol group with a group such as: (C) _1-6 ) Alkylcarbonyloxymethyl, 1- ((C) _1-6 ) Alkylcarbonyloxy) ethyl, 1-methyl-1- ((C) _1-6 ) Alkylcarbonyloxy) ethyl (C _1-6 ) Alkoxycarbonyloxymethyl, N- (C) _1-6 ) Alkoxycarbonylaminomethyl group, succinyl group, (C) _1-6 ) Alkylcarbonyl group,Alpha-amino (C) _1-4 ) Alkylcarbonyl, arylalkylcarbonyl or α -aminoalkylcarbonyl- α -aminoalkylcarbonyl, wherein each α -aminoalkylcarbonyl is independently selected from the group consisting of naturally occurring L-amino acid, P (O) (OH) ₂ 、-P(O)(O(C _1-6 ) Alkyl group ₂ Or a glycosyl group (a group produced by removing a hydroxyl group in the form of a hemiacetal of a carbohydrate).

If a compound of the present disclosure incorporates an amine functionality, a prodrug may be formed, for example, by producing an amide or carbamate, an N-alkylcarbonyloxyalkyl derivative, (oxo-dioxolyl) methyl derivative, an N-Mannich base (N-Mannich base), an imine, or an enamine. In addition, secondary amines can be metabolically cleaved to produce bioactive primary amines, or tertiary amines can be metabolically cleaved to produce bioactive primary or secondary amines. See, for example, simplii cio et al, molecules 2008,13,519 and references therein.

As used herein, the term "warhead" or "warhead group" refers to a functional group present on a compound, wherein the functional group is capable of participating in a reaction with a protein (e.g., a 3C or 3CL protease (e.g., cysteine on a protease such as Cys 145) in a reversible or irreversible manner. For example, the warhead may form a covalent bond with the protein, or may create a stable transition state, or be a reversible or irreversible alkylating agent. For example, the warhead moiety may be a functional group on the inhibitor that may participate in a bond formation reaction in which a new covalent bond is formed between a portion of the warhead and the donor (e.g., amino acid residue of a protein). In embodiments, the warhead is an electrophile and the "donor" is a nucleophile, such as a side chain of a cysteine residue. As provided herein, the warhead may comprise a nitrile or a halogen group. As also provided herein, warheads can include aldehydes, ketoamides, hydroxyl bisulfites, heterocyclic moieties, aziridines, oxiranes, epoxyketones, halomethyl ketones, hydroxymethyl ketones, electrophilic ketones (e.g., trifluoromethyl ketones), acyloxymethyl ketones, benzothiazolyl ketones, and Michael acceptors. For example, the nitrile may be a reversible covalent warhead for cysteine protease inhibition, e.g., where the mechanism of action may involve the formation of a reversible covalent bond between the nitrile and the active cysteine for the formation of a thioiminoester adduct. The reaction of glutathione or cysteine of other proteins is typically reversible, while the reaction with cysteine or aminoethylthiol typically irreversibly forms a thiazolidine adduct. It will be appreciated that the compounds contemplated herein may be reversible or irreversible inhibitors.

Examples of exemplary warheads include, but are not limited to, moieties having cyano groups, halomethyl groups, aldehydes, ketoamides, hydroxy bisulfites, heterocycles, epoxyketones, halomethyl ketones, hydroxymethyl ketones, electrophilic ketones, acyloxymethyl ketones, benzothiazolyl ketones, or michael acceptors, such as:

/>

and-CN.

In some embodiments, the warhead is a moiety having a cyanohydrin or cyanoacrylate moiety. Examples of exemplary cyanohydrins and cyanoacrylate warheads include, but are not limited to:

/>

wherein R is ^13bb Selected from the group consisting of: halogen, C ₁ -C ₆ Alkyl, C ₁ -C ₆ Haloalkyl, C ₁ -C ₆ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, -N (R) ^e R ^f ) and-C (O) -N (R) ^e R ^f )；R ^e And R is ^f Each selected from hydrogen and C ₁ -C ₆ Alkyl groups; or R is ^e And R is ^f Can form a 4-6 membered heterocyclic ring together with the nitrogen to which it is attached; and where the valency permits, p is 0, 1, 2, 3 or 4.

In some embodiments, the warhead is a moiety having a cyanoamine or cyanoamide moiety. Examples of exemplary cyanoamine warheads include, but are not limited to:

/>

In some embodiments, the warhead is a moiety having an imino-oxazolidone moiety. Examples of exemplary imino-oxazolidinone warheads include, but are not limited to:

in some embodiments, the warhead is a moiety having an iminoimidazolidone. Examples of exemplary iminoimidazolidone warheads include, but are not limited to:

wherein each R is ^ccc And R is ^ccc Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) and C ₆ -C ₁₄ Aryl groups. In some embodiments, the warhead is selected from the group consisting of: />

Other examples of exemplary warheads include, but are not limited to:

wherein R is ^cc Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl group, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), 5-10 membered heterocycle and-N (R) ^b R ^c ) Wherein R is ^b And R is ^c Are each selected from hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl group, or R ^b And R is ^c Can be joined together to form a 5-10 membered heterocyclic ring together with the nitrogen to which it is attached.

Some other examples of exemplary warheads include, but are not limited to:

Wherein R is ^cd Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl groups.

Other examples of exemplary warheads include, but are not limited to:

wherein R is ^c Selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl group, wherein C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl;

wherein X is ² Selected from the group consisting of NH, O, and S; x is X ³ Independently at each occurrence selected from N and CH; r is R ^D Independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl group,R ^E Independently at each occurrence selected from the group consisting of: halogen, hydroxy, C ₁ -C ₈ Alkyl and C ₁ -C ₈ An alkoxy group; p is selected from 0, 1 and 2; and q is selected from 0, 1 and 2; />

Wherein X is ² Selected from NH, NR ^P O and S, wherein R ^P Is C ₁ -C ₈ An alkyl group;

wherein R is ^D Selected from the group consisting of: c (C) ₃ -C ₆ Cycloalkyl, C ₁ -C ₈ Alkyl groupX ⁴ Independently at each occurrence selected from CH and N; r is R ^E Independently at each occurrenceA group consisting of: halogen, -CH ₃ 、-OCH ₃ 、-OCH ₂ CH ₃ 、-OCH(CH ₃ ) ₂ 、-CN、-CF ₃ 、-OCF ₃ and-SCF ₃ The method comprises the steps of carrying out a first treatment on the surface of the And p is selected from 0, 1 and 2; -C (O) R ^D Wherein R is ^D Selected from the group consisting of: hydrogen, -CH ₂ OH、-CH ₂ OR' and-CH _x F _y Wherein x is 0, 1 or 2; y is 1, 2 or 3; and the sum of x and y is 3, wherein R' is selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered aryl), C ₁ -C ₈ Heteroalkyl, C ₃ -C ₆ Cycloalkyl and 5-10 membered aryl; - (CH=CH) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

Those skilled in the art will appreciate that the compounds disclosed herein including the above warhead also encompass precursors of those compounds, e.g., the cyano moiety involved in the warhead may be replaced with, e.g., a halo moiety.

Those skilled in the art will appreciate that the compounds disclosed herein may also bind irreversibly or may otherwise inhibit, for example, viral proteins by any other mechanism of action.

As used herein, the term "inhibitor" refers to a compound that binds to and/or inhibits a protease of interest with a measurable affinity.

As used herein, the term "reversible" or "reversible inhibitor" refers to a protease inhibitor that associates with a protease in a manner that inhibits the activity of the protease when the protease is bound to the inhibitor, but does not associate with the protease in a manner that inhibits the activity of the protease when the protease is no longer bound to the inhibitor. Reversible inhibitors can be inhibited by: competing with the substrate for binding to the active site of the protease (competitive reversible inhibitor), or associating with the protease bound to its substrate in a manner that renders the complex inactive (non-competitive reversible inhibitor), or associating with the protease and/or protease-substrate complex in a manner that inhibits the activity of either and/or both of the protease and/or protease-substrate complex.

As used herein, the term "irreversible" or "irreversible inhibitor" refers to an inhibitor (i.e., a compound) capable of covalently bonding to a protease of interest in a substantially irreversible manner. After covalent bond formation has occurred, the irreversible inhibitor will remain substantially bound to the target protease. Irreversible inhibitors generally show a time dependence, whereby the extent of inhibition increases with the time the inhibitor is contacted with the enzyme. In certain embodiments, after covalent bond formation occurs, the irreversible inhibitor will remain substantially bound to the target protease and will remain bound for a period of time longer than the lifetime of the protein.

I. Reversible or irreversible viral protease inhibitor compounds

The present invention relates in part to compounds that inhibit viral proteases. Examples of viral proteases include, but are not limited to, cathepsin K, coronavirus main protease (Mpro), caspase 3, calpain 1 and cathepsin S. Thus, in various embodiments, the compounds of the present disclosure (e.g., compounds of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-a or IV-B) are viral protease inhibitors, wherein the viral protease is selected from the group consisting of: cathepsin K, coronavirus main protease (Mpro), caspase 3, calpain 1 and cathepsin S. In certain embodiments, the viral protease is a coronavirus major protease (Mpro). In some embodiments, the viral protease is cathepsin K. In some embodiments, the viral protease is caspase 3. In some embodiments, the viral protease is calpain 1. In some embodiments, the viral protease is cathepsin S.

Also provided herein are compounds represented by:

R ^3a selected from the group consisting ofAnd 4-10 membered heterocycles, wherein the heterocycles may be optionally substituted with one, two or three substituents each independently selected from the group consisting of: hydroxy, C ₁ -C ₈ Alkoxy, oxo, and warhead a; r is R ^3b Selected from hydrogen and C ₁ -C ₈ An alkyl group; wherein R is ^3a And R is ^3b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring, wherein the heterocyclic ring can optionally be selected from C independently from one, two or three ₆ -C ₁₄ Aryl and warhead a substituents; r is R ^1a Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl; r is R ^1b Selected from hydrogen and C ₁ -C ₈ An alkyl group; or R is ^1a And R is ^1b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl; r is R ¹ Selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ¹ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a); r is R ^A Independently at each occurrence selected from the group consisting of: halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、-CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-C(O)-OC(CH ₃ ) ₃ 、C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocyclyl, wherein R ^B The alkyl, heterocyclyl, heteroaryl, or aryl may be optionally substituted with one, two, or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Hydroxyl and oxo; r is R ² Selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 、-O-(C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl), 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ^B Or R is ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a); or R is ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered mono-or bicyclic heterocycle having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a); r is R ³ Selected from 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);R ^B independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₆ Cycloalkyl, fluorenylmethyloxy, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; r is R ^C Independently at each occurrence selected from hydrogen, halogen and C ₁ -C ₈ An alkyl group; r is R ^x Independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano, -O- (R) ^xx )-OCH ₃ 、-OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, -O-C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; wherein two are geminal C ₁ -C ₈ Alkyl groups may be joined together to form, together with the carbon to which they are attached, C optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy and oxo ₃ -C ₆ Cycloalkyl; and wherein the alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl; r is R ^G Selected from the group consisting of: hydrogen, optionally by one, two or three R ^gg Substituted C _1-6 Alkyl, optionally substituted with one, two or three R ^hh substituted-C (=o) -C _1-6 Alkyl, -C (=o) -C _3-6 Cycloalkyl, -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl), -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ Aryl), -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocyclyl) and-C (O) - (4-10 membered heterocyclyloxy); wherein aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj Substitution; r is R ^gg Independently at each occurrence selected from the group consisting of: -C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ；R ^hh Independently at each occurrence selected from the group consisting of: halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocyclyl and C ₁ -C ₆ An alkoxy group; r is R ^jj Independently at each occurrence selected from the group consisting of: halo, oxo, hydroxy, cyano, C ₁ -C ₆ Alkyl, C _1-6 Haloalkyl, C ₁ -C ₆ Alkoxy, C _1-6 Haloalkoxy, C _3-6 Cycloalkyl, SF ₅ And NH ₂ ；R ^m Independently at each occurrence selected from the group consisting of: hydrogen, C _1-3 Alkyl, phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl; wherein C is _1-3 Alkyl, phenyl and C _3-6 Cycloalkyl groups may be optionally substituted with one, two or three halo groups; r is R ^xx Is- (OCH) ₂ CH ₂ ) _nn -wherein nn is selected from 1, 2, 3, 4, 5 and 6; r is R ^y Independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CF ₃ 、-CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH; a is a warhead; and X is selected from C (R) ^xy ) And N, wherein R is ^xy Selected from the group consisting of: H. d, -OH, -NH ₂ Halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl and C ₁ -C ₈ An alkoxy group.

In some embodiments, R ^3b Is hydrogen.

In certain embodiments, disclosed herein are compounds represented by formula II-a:

in certain embodiments, disclosed herein are compounds represented by formula II-B:

in various embodiments, disclosed herein are compounds represented by formulas II-C:

in some embodiments, provided herein are compounds represented by formulas II-F:

in some embodiments, provided herein are compounds represented by formulas II-G:

/>

wherein pp is selected from 0, 1, 2 and 3.

Wherein ss is selected from 0, 1, 2 and 3 and mm is selected from 1, 2 and 3.

In other embodiments, disclosed herein are compounds represented by formula II-E-II:

wherein ss is selected from 0, 1, 2 and 3 and mm is selected from 1, 2 and 3.

or a pharmaceutically acceptable salt thereof, wherein:

R ³ is thatR ^t Independently at each occurrence H or methyl; or each R ^t Can form a cyclopropyl group together with the carbon to which it is attached; r is R ^B Selected from the group consisting of: 9-10 membered bicyclic heteroaryl having one ring nitrogen, C ₁ -C ₆ Alkyl and C ₂ -C ₃ Alkenyl groups; wherein R is ^B Optionally substituted with one, two or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₃ Alkoxy, NHR ^m And phenyl (optionally substituted with one or two halogens); and R is ^m Is C _1- C ₃ Alkyl or-C (O) -C _1-3 Alkyl, wherein C _1- C ₃ Alkyl is independently optionally substituted with one, two or three halogens.

In certain embodiments, R ^3a Is thatWherein R is ^xy Selected from the group consisting of: H. d, OH, NH ₂ Halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl and C ₁ -C ₈ An alkoxy group. In embodiments, R ^xy Selected from the group consisting of: H. d, CH ₃ 、CH ₂ CH ₃ F and CF ₃ . In some embodiments, R ^xy F. In some embodiments, R ^xy Is CF (CF) ₃ . In some embodiments, R ^xy Is CH ₃ . In some embodiments, R ^xy H.

In various embodiments, X is selected from the group consisting of: CH. CD, C (CH) ₃ )、C(CH ₂ CH ₃ )、N、CF、CCl、CBr、C(CHF ₂ )、C(CH ₂ F) And C (CF) ₃ ). In some embodiments, X is CH. In some embodiments, X is CD. In some embodiments, X is C (CH ₃ ). In some embodiments, X is C (CF ₃ ). In some embodiments, X is CF. In some embodiments, X is N.

In some embodiments, a is selected from the group consisting of: cyano, -C (O) R ^D 、-C(O)CH ₂ N(R ^b R ^c )、-C(O)CH ₂ OC(O)R ^D 、-C(O)C(O)R ^D 、-(CH＝CH)C(O)OR ^D 、-(CH＝CCN)C(O)OR ^D 、-(CH＝CCN)C(O)(NH)R ^D 、-CH(CN)(OH)、-CH(CN)(NR ^b R ^c )、Wherein R is ^D Selected from the group consisting of: hydrogen, hydroxy, -OR ^bb -N(R ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; wherein R is ^D Can be optionally selected from one, two or three of halogen, hydroxy and R ^E Substituent groups of the group are substituted; r is R ^E Selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally substituted with one, two or three substituents each independently selected from the group consisting of: halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ An alkoxy group; r is R ^bb Selected from the group consisting of: c (C) ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, - (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; r is R ^cc Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl group, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), 5-10 membered heterocycle and-N (R) ^b R ^c ) Wherein R is ^b And R is ^c Each independently selected from hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl group, or R ^b And R is ^c Can be joined together to form, together with the nitrogen to which they are attached, a 5-10 membered heterocyclic ring; r is R ^cd Selected from hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl groups; and R is ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl group, wherein C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl.

In embodiments, a is selected from the group consisting of: the number of the-CN,/>

in embodiments, R ^1a Selected from the group consisting of:

in embodiments, R ^1a Is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

In embodiments, R ^1b Is hydrogen.

In certain embodiments, R ^1a And R is ^1b Joined together to form

In certain embodiments, R ^3a Is a 4-10 membered heterocycle.

In some embodiments, R ^3a Selected from the group consisting of:

/>

in some embodiments, R ³ Is a 4-10 membered heterocycle.

In some embodiments, R ³ Selected from the group consisting ofIn some embodiments, R ³ Is that(e.g.)>) Wherein R is ^x3 Independently at each occurrence selected from the group consisting of: hydrogen, halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl, C ₃ -C ₆ Cycloalkyl and C ₁ -C ₈ An alkoxy group; and pp is selected from 0, 1, 2 and 3. In some embodiments, R ³ Is->In some embodiments, R ³ Is->In some embodiments, R ³ Is thatIn some embodiments, R ³ Is->In some embodiments, R ³ Is thatAnd R is ^t Independently at each occurrence H or methyl; or each R ^t Can form a cyclopropyl group together with the carbon to which it is attached.

In some embodiments, R ³ Selected from the group consisting of:

in embodiments, R ³ Is a substituted bicyclic heterocycle, a substituted monocyclic heterocycle, a substituted bicyclic heteroaryl, or a substituted monocyclic heteroaryl. />

In some embodiments, R ³ Selected from the group consisting of:

in some embodiments, R ³ Selected from the group consisting of:

in various embodiments, R ² is-NHC (O) R ^B . In various embodiments, R ^B Is a 5-10 membered heteroaryl. In various embodiments, R ^B Is a bicyclic heteroaryl (e.g., 9 membered heteroaryl). In various embodiments, R ^B Is substituted. In various embodiments, R ^B Is unsubstituted. In various embodiments, R ^B Substituted with halogen. In various embodiments, R ^B is-OCH ₃ And (3) substitution. At the position ofIn various embodiments, R ^B Substituted with-OH. In various embodiments, R ^B Quilt C ₁ -C ₈ Alkyl substitution. In various embodiments, R ^B Quilt C ₁ -C ₈ Alkoxy substitution. In various embodiments, R ² Is substituted. In various embodiments, R ² Is unsubstituted. In various embodiments, R ² Substituted with halogen. In various embodiments, R ² is-OCH ₃ And (3) substitution. In various embodiments, R ² Substituted with-OH. In various embodiments, R ² Quilt C ₁ -C ₈ Alkyl substitution. In various embodiments, R ² Quilt C ₁ -C ₈ Alkoxy substitution.

In some embodiments, R ² Selected from the group consisting of:

/>

in some embodiments, R ^1a And R is ² Joined together to form a heterocycle selected from the group consisting of:

wherein R is ^1b H.

/>

Wherein R is ^1b H.

/>

wherein R is ^1b H.

/>

wherein R is ^1b H.

In some embodiments, R ^G Selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh substituted-C (=o) -C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl; -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ An aryl group); -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ An aryl group); -C (O) - (5-10 membered heteroaryl); -C (O) - (4-10 membered heterocyclyl) -C (O) - (4-10 membered heterocyclyloxy); wherein aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj And (3) substitution.

In some embodiments, R ^G Selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh substituted-C (=o) -C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl groups.

In other embodiments, R ^G Selected from the group consisting of: -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl), -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ Aryl), -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocyclyl) and-C (O) - (4-10 membered heterocyclyloxy); wherein aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj And (3) substitution.

In some embodiments, R ^gg Independently at each occurrence selected from the group consisting ofThe group consisting of: -C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m . In other embodiments, R ^hh Independently at each occurrence selected from the group consisting of: halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocyclyl and C ₁ -C ₆ An alkoxy group. In other embodiments, R ^jj Independently at each occurrence selected from the group consisting of: halo, oxo, hydroxy, cyano, C ₁ -C ₆ Alkyl, C _1-6 Haloalkyl, C ₁ -C ₆ Alkoxy, C ₁ -C ₆ Haloalkoxy, C _3-6 Cycloalkyl, SF ₅ And NH ₂ 。

In certain embodiments, R ^m Independently at each occurrence selected from the group consisting of: hydrogen, C _1-3 Alkyl (optionally substituted by one, two or three F), phenyl (optionally substituted by halo), -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl (optionally substituted with one, two or three F) and 5-6 membered heteroaryl.

In some embodiments, R ^G Selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl and heterocycle, wherein R ^m Selected at each occurrence from H and C _1-3 Alkyl (optionally substituted with one, two or three halogens (e.g., F)), or C ₃ -C ₆ Cycloalkyl (optionally substituted with one, two or three F).

In some embodiments, R ^G Selected from the group consisting of: 5-6 membered monocyclic-C (O) -heteroaryl or 8-10 membered bicyclic-C (O) -heteroaryl, each having at leastOne ring nitrogen and optionally substituted with one, two or three substituents each selected from halo, methoxy, cyano and hydroxy; -C (O) -C (R) ⁵⁵ R ⁵⁶ )-NH-C(O)-R ⁵⁷ Wherein R is ⁵⁵ Is H and R ⁵⁶ Is straight-chain or branched C _1-5 Alkyl (optionally substituted with halo), or R ⁵⁵ And R is ⁵⁶ Together with the carbon to which it is attached form C _3-5 Cycloalkyl (optionally substituted by halo) and wherein R ⁵⁷ Is C _1-3 Alkyl (optionally substituted with one, two or three halo).

In some embodiments, R ^G Selected from the group consisting of:

/>

in some embodiments, R ^G Is that

In some embodiments, the disclosed compounds are represented by the formula: For example->Wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl, C _3-6 Cycloalkyl (e.g., t-butyl, propyl, cyclopropyl), phenyl, and heterocyclyl; and R is ^G2 is-NH (C=O) R ^m Wherein R is ^m Selected at each occurrence from H, methyl and CF ₃ 。

In some embodiments, the disclosed compounds are represented by:wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl, C _3-6 Cycloalkyl, phenyl and heterocyclyl; and R is ^G2 is-NH (C=O) R ^m Wherein R is ^m Selected at each occurrence from H, methyl and CF ₃ 。

In some embodiments, the disclosed compounds are represented by:wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted by one, two or three C ₁ -C ₆ Alkoxy substitution), C _3-6 Cycloalkyl, phenyl and heterocyclyl; and R is ^G2 Selected from the group consisting of: -NH (C) _1-3 Alkyl) (optionally substituted with one, two or three substituents each independently selected from the group consisting of: halo, optionally substituted phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl) and-NH (c=o) R ^m Wherein R is ^m At each occurrence selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally one, two or three independently selected from halo, cyano and C ₁ -C ₆ Substituted with a substituent selected from the group consisting of alkoxy), CHF ₂ 、CF ₃ A 5-6 membered heteroaryl (optionally substituted with halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy, CHF ₂ Or CF (CF) ₃ Substitution).

In some embodiments, R ^G3 Selected from the group consisting of:

in some embodiments, R ^G2 Selected from the group consisting of:

wherein R is ^F Selected from the group consisting of: c (C) _1-6 Alkyl, C _3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, wherein R ^F Optionally one, two or three are each selected from halogen, cyano, hydroxyGroup C ₁ -C ₆ Substituent substitution of the group consisting of alkoxy; and X is ^F Selected from the group consisting of: H. halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy and C _1-6 Haloalkyl.

/>

also disclosed herein are compounds represented by formulSup>A IV-Sup>A or formulSup>A IV-B:

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ^1a selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

or R is ^1a And R is ^1b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl;

R ¹ Selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ¹ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);

R ^A independently at each occurrence selected from the group consisting of: halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、-CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-C(O)-OC(CH ₃ ) ₃ 、C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heterogeniesAryl group, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocyclyl, wherein R ^B The alkyl, heterocyclyl, heteroaryl, or aryl may be optionally substituted with one, two, or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Hydroxyl and oxo;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)OR ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 、-O-(C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl), 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ^B Or R is ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

or R is ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered mono-or bicyclic heterocycle having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a);

R ^3b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^B independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₃ -C ₆ Cycloalkyl, fluorenylmethyloxy, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

R ^C independently at each occurrence selected from hydrogen, halogen and C ₁ -C ₈ An alkyl group;

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano group、-O-(R ^xx )-OCH ₃ 、-OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, -O-C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

wherein two are geminal C ₁ -C ₈ Alkyl groups may be joined together to form, together with the carbon to which they are attached, C optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy and oxo ₃ -C ₆ Cycloalkyl; and is also provided with

Wherein alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each independently selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl;

R ^G selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh substituted-C (=o) -C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl; -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ An aryl group); -C (O) - (5-10 membered heteroaryl); -C (O) - (4-10 membered heterocyclyl); and-C (O) - (4-10 membered heterocyclyloxy); wherein aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj Substitution;

R ^gg independently at each occurrence selected from the group consisting of: -C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ；

R ^hh Independently at each occurrence selected from the group consisting of: halogen radical, cyanogenRadical, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocyclyl and C ₁ -C ₆ An alkoxy group;

R ^jj independently at each occurrence selected from the group consisting of: halo, oxo, hydroxy, cyano, C ₁ -C ₆ Alkyl, C _1-6 Haloalkyl, C ₁ -C ₆ Alkoxy, C _3-6 Cycloalkyl, SF ₅ And NH ₂ ；

R ^m Independently at each occurrence selected from the group consisting of: hydrogen, C _1-3 Alkyl, phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl; wherein C is _1-3 Alkyl, phenyl and C _3-6 Cycloalkyl groups may be optionally substituted with one, two or three halo groups;

R ^xx Is- (OCH) ₂ CH ₂ ) _nn -wherein nn is selected from 1, 2, 3, 4, 5 and 6; and is also provided with

R ^y Independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CF ₃ 、-CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), 5-10 membered heteroaryl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH.

In some embodiments, the disclosed compounds are selected from the group consisting of the compounds identified in tables 1 and 2 below:

table 1. Exemplary compounds.

/>

Table 2 exemplary compounds.

/>

II method

Another aspect of the present disclosure provides a method of treating a patient suffering from a viral infection, such as a coronavirus infection. In particular, in certain embodiments, the present disclosure provides a method of treating a covered medical indication comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein, such as a compound of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-a or IV-B.

In certain embodiments, the present disclosure provides a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds described herein. In some embodiments, the viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses (e.g., enterovirus 71 (EV 71)), orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses. In certain embodiments, the viral infection is a coronavirus infection. In some embodiments, the viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19). In embodiments, the viral infection is SARS-CoV-2.

In some embodiments, the viral infection is caused by a virus selected from the group consisting of: calicivirus, MD145, murine norovirus, porcine herpesvirus, rabbit hemorrhagic disease virus, porcine teschovirus, bovine coronavirus, feline infectious peritonitis virus, EV-68 virus, EV-71 virus, poliovirus, norovirus, human Rhinovirus (HRV), hepatitis A Virus (HAV), and Foot and Mouth Disease Virus (FMDV).

In embodiments, the viral infection is an arenavirus infection. In some embodiments, the arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus. In some embodiments, the viral infection is an influenza infection. In some embodiments, the influenza is influenza H1N1, H3N2, or H5N1.

Another aspect of the present disclosure provides a method of treating a patient suffering from a viral infection, such as a norovirus infection. In some embodiments, the present disclosure provides a method of treating a viral infection caused by norovirus in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of any of the compounds described herein.

In certain embodiments, provided herein is also a method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying a virus a therapeutically effective amount of a compound described herein, and/or contacting an effective amount of a compound described herein with a virus-infected cell. In some embodiments, the method further comprises administering another therapeutic agent. In some embodiments, the method further comprises administering an additional antiviral therapeutic agent. In embodiments, the antiviral therapeutic agent is selected from the group consisting of: ribavirin (ribavirin), fapiravir (favirapivir), ST-193, oseltamivir (oseltamivir), zanamivir (zanamivir), peramivir (peramivir), danoprevir (danoprevir), ritonavir (ritonavir), adeciclovir (remdesivir), cobicistat (cobicistat), etipravir (elvitegravir), emtricitabine (emtricitabine), tenofovir (tenofovir), tenofovir disoproxil (tenofovir disoproxil), tenofovir alafenamide hemi-fumarate (tenofovir alafenamide hemifumarate), abacavir (abaavir), doluteprevir (dotevir), efavirenz (aviranz), elbavir, ledipasvaravir (glaprasuvir), capeprevir (glapravir) sofosbuvir (sofosbuvir), bitravir (bicegravir), dasabavir (dasabavir), lamivudine (lamivudine), atazanavir (atazanavir), obetavir (ombetavir), lamivudine, stavudine (stavudine), nevirapine (nevirapine), rilpivirine (rilpivirine), parliaprevir, cimaprevir (simeprevir), dacarbatavir (daclatasvir), glazoprevir (grazoprevir), pirenz (pibrentprevir), adefovir), amprenavir (amprenavir), an Puli near (amplien), alavuc (caprine), anti-antibodies, bafuvirr (bavirapine), carbotevir (cabtevir), cytarabine, ecalcine, epigallocatechin gallate (epigallocatechin gallate), itravirine (etravirine), fostemavir Sha Wei (fostemavir), gemcitabine (gemcitabine), griffinine (griffithiin), isoprinosine (imunovir), indinavir (indinavir), maraviroc, metiran Sha Zong (methisazone), MK-2048, nelfinavir (nelfmavir), nevirapine (nevirapine), nitazoxanide (nitazoxanide), nomavir (noravervir), plexaformer (plectadine), PRO 140, telangavir (raltegavir), prazidine (pravastatin), saquinadine (saquinadine), teudine (tenadine), valadidine (X-355), and fluzafidine (valacillin-355). In some embodiments, the other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir (aciclovir), acyclovir (acyclovir), protease inhibitors, arbidol (arbidol), atazanavir, nitroprussian (atrippla), boceprevir, cidofovir (cidofovir), bivariate (combimir), darunavir (darunavir), doconavir (docosanol), edexuridine (edoxudine), entry inhibitors, entecavir (entecavir), famciclovir (famciclovir), fomivir (fomivirsen), fosamprenavir (fosamprenavir), foscarnet (foscarnet) phosphine ethanol (fosfonet), ganciclovir (ganciclovir), ibacitabine (ibalitabine), amplupulin (immuoxovir), idoxillin (idoxidine), imiquimod (imiquimod), inosine (inosine), integrase inhibitors, interferons, lopinavir (lopinavir), lovir amine (loviride), moroxydine (moroxydine), ne Sha Wei (nexavir), nucleoside analogs, penciclovir (penciclovir), plica (pleconaril), podophyllotoxin (podophyllotoxin), ribavirin, telangavir (tipranavir), trifluouridine (trizidine), tricxivir (trizivir), qu Jingang amine (amandine), teluvalida), valacyclovir (valacivir), valaciclovir (valaciclovir), valaciclovir (valacivir), vickers Wei Luoke (vicriloc), vidarabine (vidarabine), verapamil (viramidine) and zidovudine (zodovudine). In embodiments, the additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibody, enfuvirtide, amantadine, rimantadine, pramipexole (pleconaril), acyclovir, zidovudine (zidovudine), zidovudine, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampicin (rifampicin), zanamivir, oseltamivir Weidan noprevir, ritonavir, radevir, cobicistat, eptitavir, emtricitabine, tenofovir disoproxil, valproamide hemifumarate, abacavir, dolutegravir, efavirenz, dipivoxil, ganciclovir, sofalcavir, bitalavir, dacarbazine, lamivudine atazanavir, obitaconvir, lamivudine, stavudine, nevirapine, rilpivirine, palirivir, cimiravir, dacarbazine, glazopivir, pirenz-tavir, adefovir, amprenavir, an Puli, alavirrol, anti-goat antibodies, balafrevir, cabazithromycin, cytarabine, elciclovir, epigallocatechin gallate, itravirlin, formoterol Sha Wei, gemcitabine, grifefosine, isoprinosine, indinavir, valavir, metisazone, MK-2048, nelfinavir, nevirapine, nifedipine, noolovir, prazifovir, PRO 140, raltegafur, prazidine, saquinavir, telbivudine, TNX-355, valacyclovir, VIR-576 and cebine.

The patients encompassed include not only humans, but also other animals such as companion animals (e.g., dogs, cats), livestock (e.g., cows, pigs) and wild animals (e.g., monkeys, bats, snakes).

Thus, in one embodiment, described herein is a method of ameliorating or treating a viral infection in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., a compound of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-a or IV-B as described herein) or a pharmaceutically acceptable salt thereof.

Other methods of treatment contemplated include a method of treating or ameliorating a viral infection condition or co-disease by administering to a subject in need thereof an effective amount of a compound disclosed herein.

Exemplary co-diseases include lung disease, heart disorders, endocrine disorders, respiratory disorders, liver disorders, skeletal disorders, psychiatric disorders, metabolic disorders, and reproductive disorders.

In some embodiments, the viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses. In some embodiments, the viral infection is a coronavirus infection. In some embodiments, the viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19). In some embodiments, the viral infection is SARS-CoV-2. In some embodiments, the viral infection is an arenavirus infection. In some embodiments, the arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus. In some embodiments, the viral infection is an influenza infection. In some embodiments, the influenza is influenza H1N1, H3N2, or H5N1. In some embodiments, the viral infection is a respiratory viral infection. In some embodiments, the viral infection is an upper respiratory tract viral infection or a lower respiratory tract viral infection. In some embodiments, the method further comprises administering another therapeutic agent.

In certain embodiments, the virus is selected from the group consisting of: retroviruses (e.g., human Immunodeficiency Virus (HIV), simian Immunodeficiency Virus (SIV), human T cell lymphotropic virus (HTLV) -1, HTLV-2, HTLV-3, HTLV-4), ebola virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, herpes Simplex Virus (HSV) (e.g., HSV-1, HSV-2, varicella-zoster virus, cytomegalovirus), adenoviruses, orthomyxoviruses (e.g., influenza A, influenza B, influenza C, influenza D, togaviruses)), flaviviruses (e.g., dengue virus (dengue virus), zika virus (Zika virus), west Nile virus, rift valley fever virus (Rift Valley fever virus), arenavirus, crimea-oka hemorrhagic fever virus (Crimean-Congo hemorrhagic fever virus), epstein-Barr virus (echo virus), rhinovirus, coxsackie virus (coxsackie virus), coronavirus (e.g., severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), coronavirus disease 2019 (COVID-19), respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus (e.g., adeno-associated virus), vaccinia virus, smallpox virus, molluscum virus, bovine leukemia virus, bovine diarrhea virus, poliovirus, st.Louis encephalitis virus, japanese encephalitis virus (Japanese encephalitis virus), tick-borne encephalitis virus, mo Ruigu virus (Murray Valley virus), powassan virus, luo Xiao virus (Rocio virus), jumping-ill virus (jumping-ill virus), banzi virus, ilheus virus, kekobera virus (Kokobera virus), kunjin virus, alfuy virus, rabies virus (rabies virus), polyoma virus (e.g., JC virus, BK virus), alphavirus, and rubella virus (rubella virus) (e.g., rubella virus).

In certain embodiments, the disease or disorder is a viral infection, e.g., a disease or disorder selected from the group consisting of: acquired immunodeficiency syndrome (AIDS), HTLV-1 associated spinal cord disease/tropical spastic paraplegia, ebola virus disease, hepatitis a, hepatitis b, hepatitis c, herpes, shingles, acute varicella, mononucleosis, respiratory tract infection, pneumonia, influenza, dengue fever, encephalitis (e.g., japanese encephalitis, san lewy encephalitis or tick-borne encephalitis such as boltzfeldt-jakob encephalitis (Powassan encephalitis)), west Nile fever (West Nile fever), rift valley fever (Rift Valley fever), crimia-oka hemorrhagic fever, kosal forest disease (Kyasanur Forest disease), yellow fever, zhai ka fever (Zika fever), aseptic meningitis, myocarditis, cold, pulmonary infection, molluscum contagiosum (molluscum), regional bovine leukemia, coronavirus disease 2019 (COVID-19), mumps, gastroenteritis, measles, rubella, palmoplasm disease (applied-ek disease), smallpox, warts (e.g., warts), warts, condyloma, molluscum contagiosum, and pityriasis (pita).

In some embodiments, the virus is an RNA virus (having a genome comprised of RNA). The RNA virus may be single stranded RNA (ssRNA) or double stranded RNA (dsRNA). RNA viruses have a high mutation rate compared to DNA viruses because RNA polymerase lacks proofreading ability (see, e.g., steinhauer DA, holland JJ (1987), "Rapid evolution of RNA viruses". Annu. Rev. Microbiol. 41:409-33). In some embodiments, the RNA virus is a positive-stranded RNA virus (e.g., SARS-CoV virus, polio virus, coxsackievirus, enterovirus, human rhinovirus, foot/mouth disease virus, encephalomyocarditis virus, dengue virus, zika virus, hepatitis c virus, or newcastle disease virus (New Castle Disease virus)).

RNA viruses are classified by type of genome (double stranded, negative (-) or positive (+) single stranded). Double stranded RNA viruses contain a number of different RNA molecules, each encoding one or more viral proteins. Sense ssRNA viruses use their genome directly as mRNA; ribosomes within the host cell translate the mRNA into a single protein, which is then modified to form the various proteins required for viral replication. One such protein is an RNA-dependent RNA polymerase (RNA replicase), which replicates viral RNA to form a double-stranded replicative form. The antisense ssRNA virus has a genome replicated by RNA replicase to produce sense RNA for replication. Thus, the virus comprises an RNA replicase. The resulting sense RNA then acts as a viral mRNA and is translated by the host ribosome. In some embodiments, the virus is a dsRNA virus. In some embodiments, the virus is a negative ssRNA virus. In some embodiments, the virus is a positive ssRNA virus. In some embodiments, the positive ssRNA virus is a coronavirus.

SARS-CoV2, sometimes also referred to as 2019 novel coronavirus or 2019-nCoV, is a positive sense single stranded RNA virus. SARS-CoV-2 has four structural proteins, called S (spike), E (envelope), M (membrane) and N (nucleocapsid) proteins. The N protein holds the RNA genome together; s, E and M proteins form the viral envelope. The spike protein allows the virus to attach to the membrane of a host cell, such as the ACE2 receptor in a human cell. F1000Research, 9:72). SARS-CoV2 is a highly contagious, pathogenic agent of the globally prevalent coronavirus disease 2019 (COVID 19).

In some embodiments, the virus is a DNA virus (having a genome comprised of DNA). Exemplary DNA viruses include, but are not limited to, parvoviruses (e.g., adeno-associated viruses), adenoviruses, african swine fever viruses (asfarviruses), herpes viruses (e.g., herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), epstein-Barr viruses (EBV), cytomegalovirus (CMV)), papillomaviruses (e.g., HPV), polyomaviruses (e.g., simian vacuole virus 40 (SV 40)), and poxviruses (e.g., vaccinia virus, smallpox virus, fowl pox virus, capripoxvirus, myxoma virus). Exemplary RNA viruses include, but are not limited to, bunyaviruses (e.g., hantavirus), coronaviruses, flaviviruses (e.g., yellow fever virus, west nile virus, dengue virus), hepatitis viruses (e.g., hepatitis a virus, hepatitis C virus, hepatitis e virus), influenza viruses (e.g., influenza a virus, influenza B virus, influenza C virus), measles virus, mumps virus, calicivirus, norovirus (e.g., norwalk virus)), poliovirus, respiratory Syncytial Virus (RSV), retroviruses (e.g., human immunodeficiency virus-1 (HIV-1)), and toroviruses.

The methods described herein can inhibit viral replication transmission, replication, assembly, or release, or minimize expression of viral proteins. In one embodiment, described herein is a method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release comprising administering to a patient carrying a virus a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable salt thereof, and/or contacting an effective amount of a compound described herein or a pharmaceutically acceptable salt thereof with a virally infected cell.

Also described herein is a method of treating a respiratory disorder in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., a compound of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-a or IV-B, etc., described herein) or a pharmaceutically acceptable salt thereof. In certain embodiments, the respiratory disorder is selected from the group consisting of: chronic Obstructive Pulmonary Disease (COPD), asthma, fibrosis, chronic asthma, acute asthma, pulmonary disease secondary to environmental exposure, acute pulmonary infection, chronic pulmonary infection, a1 antitrypsin disease, cystic fibrosis, and autoimmune disease. In some embodiments, the respiratory disorder is associated with a heart attack.

Also described herein is a method of treating a disorder associated with a cathepsin (e.g., cathepsin K) in a subject in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., a compound of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-a or IV-B, etc.), or a pharmaceutically acceptable salt thereof, as described herein. In some embodiments, the disorder is a cathepsin-dependent condition or disease. In embodiments, the disorder is selected from the group consisting of: breast cancer, compact osteogenesis imperfecta, glioblastoma, bone sclerosis, osteoporosis, glucocorticoid-induced osteoporosis, paget's disease, abnormally increased bone turnover, periodontal disease, tooth loss, bone fracture, rheumatoid arthritis, osteoarthritis, peri-prosthetic osteolysis, osteogenesis imperfecta, atherosclerosis, obesity, glaucoma, chronic obstructive pulmonary disease, metastatic bone disease, hypercalcemia of malignancy, and multiple myeloma.

The compounds described herein, e.g., compounds of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-A, IV-B, and the like, as defined herein, may be administered in combination with one or more additional therapeutic agents to treat conditions described herein, such as infections caused by a pathogen (e.g., virus, fungus, or protozoa) described herein. For clarity, fixed compositions comprising the disclosed compounds and another therapeutic agent, such as disclosed herein, are contemplated herein, as well as methods of separately administering the disclosed compounds and the disclosed therapeutic agents. For example, provided in the present disclosure is a pharmaceutical composition comprising a compound described herein (e.g., a compound of formula I as defined herein), one or more additional therapeutic agents, and a pharmaceutically acceptable excipient. In some embodiments, a compound of formula I as defined herein and one additional therapeutic agent are administered. In some embodiments, the disclosed compounds as defined herein and two additional therapeutic agents are administered. In some embodiments, the disclosed compounds as defined herein and three additional therapeutic agents are administered. Combination therapy may be achieved by administering two or more therapeutic agents, each of which is formulated and administered separately. For example, compounds of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-A, IV-B, and the like, as defined herein, and additional therapeutic agents, may be formulated and administered alone. Combination therapy may also be achieved by administering two or more therapeutic agents in a single formulation, e.g., a pharmaceutical composition comprising a compound of formula I as one therapeutic agent and one or more additional therapeutic agents, such as an antibiotic, viral protease inhibitor, or antiviral nucleoside antimetabolite. For example, a compound of formula I as defined herein and an additional therapeutic agent may be administered in a single formulation. Combination therapies also encompass other combinations. Although two or more agents in combination therapy may be administered simultaneously, they need not be. For example, the administration of a first dose (or combination of doses) may be several minutes, hours, days, or weeks earlier than the administration of a second dose (or combination of doses). Thus, administration of two or more agents may be within minutes of each other, or within 1, 2, 3, 6, 9, 12, 15, 18, or 24 hours of each other, or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14 days of each other, or within 2, 3, 4, 5, 6, 7, 8, 9 weeks, or weeks of each other. In some cases, even longer intervals are possible. Although in many cases it is desirable that two or more agents used in combination therapy be present simultaneously within the patient's body, this need not be the case.

Combination therapy may also include two or more administrations of one or more of the agents used in combination performed using different sequences of the component agents. For example, if agent X and agent Y are used in combination, they may be administered sequentially one or more times in any combination, e.g., in the order X-Y-X, X-X-Y, Y-X-Y, Y-Y-X, X-X-Y-Y, etc.

In some embodiments, the one or more additional therapeutic agents that may be administered in combination with the compounds provided herein may be antibiotics, viral protease inhibitors, antiviral antimetabolites, lysosomal agents, M2 proton channel blockers, polymerase inhibitors (e.g., EIDD-2801, which is also known as molin upirair), neuraminidase inhibitors, reverse transcriptase inhibitors, viral entry inhibitors, integrase inhibitors, interferons (e.g., types I, II and III), or nucleoside analogs. In some embodiments, the one or more additional therapeutic agents that may be administered in combination with the compounds provided herein may be a steroid (e.g., a corticosteroid such as betamethasone, prednisone, prednisolone, triamcinolone, methylprednisolone, dexamethasone; mineralocorticoids such as fludrocortisone; glucocorticoids such as hydrocortisone (hydrocortisone), cortisone, dexamethasone (ethabiosorb), prednisone, prednisolone, triamcinolone, dexamethasone, vitamin D such as dihydrofast stay alcohol (dihydroachyurol), androgens such as Ai Bo homo (apoptone), oxaandrone (oxanolone), oxybolone (oxabolone), testosterone, nandrolone (nandrolone) (also known as anabolic steroid), estrogens such as diethylstilbestrol (diethylstilbestrol), progestins such as danazol (danazol), norethindrone (norethindrone), medroxyprogesterone acetate (medroxyprogesterone acetate), caproic acid 17-hydroxyprogestrel, and progestins such as mifepristone (mifepristone) and triesterone (gestrone) or immunomodulators such as 6 mercaptopurine, 6MP, aflurone, N (Alston), abalon (oxabolone), testosterone (Avermentain), avermentain, basta (Tar-12, tar-Tar), bastone (Tar-12, tar-Tar (Tar), basil (Tar-A), and (Tabana. RTM) may be used as a pharmaceutical composition for the treatment of the oral and a pharmaceutical composition for the oral administration Recombinant C1 esterase inhibitors, human C1 inhibitors, crigz (Cinryz), copazone, dimethyl fumarate, dittany Luo Ximei (diroximel fumarate), ai Kala peptide (ecallantide), epratuzumab (emapalumab), epratuzumab-lzsg, itavey (Extavia), fingolimod (fingolimod), phenanthrazyer (Firazyr), gamifant Mi Fante (Gamifant), ji Lunya (gillenya), glamer (glatirer), glatopa (Glatopa), hagarda (Haegardera), atibant (icarbant), fukand (Intergen), interferon alpha n3, interferon alfacon 1, interferon beta 1a, interferon beta 1b, interferon beta Carlbit (Kalbittor), remeziram (Kineret), mercaptopurine, monomethyl fumarate, pegylated interferon beta-1 a, pralerdidi (Plegridy), pralethol (Purinethol), pralentan (Purixan), ribifer (Rebif), ribifida-dox (Rebif rebidse), ruimestemcel-L, li Naxi Pralept, luo Peigen interferon alpha 2b (ropeginterfer alfa b), russert (Ruconest), renilicil (Ryoncil), siltuximab (siltuximab), su Timo monoclonal antibody (Sutimimab), sawytem (Sylvant), talbira (Tecfidera) and Wu Mirui t (Vumety)). In some embodiments, the one or more additional therapeutic agents is cathepsin L. In some embodiments, the one or more additional therapeutic agents is dehydromembrane ecteinascidin B (also known as pritidine (plitidpsin) or aplidine) or zotefene (Zotatifin) (eFT 226).

In some embodiments, the methods described herein further comprise administering an additional antiviral therapeutic agent. In some embodiments, the antiviral therapeutic agent is selected from the group consisting of: ribavirin, famprivir, ST-193, oseltamivir, zanamivir, pamil, danoprevir, ritonavir, radenpyr, bivalirudin, mitsubishi, etila, emtricitabine, tenofovir disoproxil, tenofovir alafenamide hemi-fumarate, abacavir, dolutegravir, efavirenz, elbavir, ledipavir, ganciclovir, sofosbuvir, bitaravir, dasabacavir, lamivudine, atazanavir, obetavir, lamivudine, stavudine, nevirapine, rilpivirine, ziprairivir, cimiravir, dacarbavir Grazopicir, pirenzvir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafavir, cabazitaxel, cytarabine, ecalciline, epigallocatechin gallate, itravir, fotif Sha Wei, gemcitabine, grifficine, isoprinosine, indinavir, malaevir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, nootkatole, pulefafur, PRO 140, raltegafur, prazidine, saquinavir, tebuvir, TNX-355, valacyclovir, VIR-576, and zalcitabine. In some embodiments, the other therapeutic agent is selected from the group consisting of: protease inhibitors (e.g., nafamostat, camostat, gabexate, epothilone (epsilon) -aminocaproic acid and aprotinin), fusion inhibitors (e.g., BMY-27709, CL 61917, and CL 62554), M2 proton channel blockers (e.g., amantadine and rimantadine), polymerase inhibitors (e.g., 2-deoxy-2 'fluoroguanosine (2' -fluoroGuo), 6-endonuclease inhibitors (e.g., L-735,822 and flutamide), neuraminidase inhibitors (e.g., zanamivir (renavir), oseltamivir, peramivir, and ABT-675 (a-315675)), reverse transcriptase inhibitors (e.g., such as, abacavir, adefovir, delavirdine, didanosine, efavirenz, emtricitabine, lamivudine, nevirapine, stavudine, tenofovir disoproxil and zalcitabine), acyclovir, aclarven, protease inhibitors (e.g., amprenavir, indinavir, nelfinavir (nelfinavir), ritonavir and saquinavir), arbidol, atazanavir, rituximab, boceprevir, clobetavir, darunavir, duo Sha Nuo, ideuridine, entry inhibitors (e.g., env Wei De and malavirrol), entecavir, famciclovir, foscarnet, phosphazene, fludroxide, ciclovir, valacil, famciclovir, valproir, valproic acid, and the like, ganciclovir, ibacitabine, imperforate, idoxuridine, imiquimod, inosine, integrase inhibitors (e.g., raltegravir), interferons (e.g., I, II and form III), lopina Weiluo valamine, moroxydine, naphthalene Sha Wei, nucleoside analogs (e.g., acyclovir), penciclovir, praecoraline, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricyclovir, triamcinolone, telavadade, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine, and zidovudine. In some embodiments, the additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibody, enfu Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivir, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampicin, zanamivir, oseltamivir Weidan nopuvir, ritonavir, radciclovir, cobalastat, eptifibatide, emtricitabine, tenofovir disoproxil, tenofovir, valavermectin, abacavir, dulutegravir, efavirenz, ganciclovir, sofosbuvir, bitorubivir, dasabavir, lamivudine, atazanavir, obuwei, obutyvir, emtricitabine, tenofovir, valacil, valvulafir, and valvulafir lamivudine, stavudine, nevirapine, rilpivirine, palivir, cimlopivir, dacarbazine, pezopivir, pirenz-tavir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafrevir, cabavir, cytarabine, eleganciclovir, epigallocatechin gallate, itraprine, ferti Sha Wei, gemcitabine, grifeis, isoprinosine, indinavir, malavir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, norelvir, praxafovir, PRO 140, raltegafur, prazidine, quinavir, telbivudine, TNX-355, valacyclovir, pir-576 and zanciclovir. In some embodiments, the other therapeutic agent is selected from the group consisting of: quinine (optionally in combination with clindamycin), chloroquine (chloroquine), amodiaquine (amodiaquine), artemisinin (artemesinin) and derivatives thereof (e.g., artemether (artemether), artesunate (artesuline), dihydroartemisinin (dihydroartemesinin), arteether (artemether)), doxycycline (doxycycline), pyrimethamine (pyrimethamine), mefloquine (mefloquine), halofantrine (halofantrine), hydroxychloroquine (hydroxychloroquine), epothilone (efloquine), azazoxanide (azazonidine), ornidazole (oxanidazole), paromomycin (paromomycin), pentadine (pentazane), primaquine (priquine), pyrimethamine (guanamine), chlormequine (protamine), and optionally in combination with a sulfonamide (ppfluquine.g., 1, 05), and a sulfonamide (ppfluquine.g., fluquine). In some embodiments, the other therapeutic agent is an antibiotic. In some embodiments, the antibiotic is a penicillin (penicillin) antibiotic, a quinolone antibiotic, a tetracycline antibiotic, a macrolide antibiotic, a lincosamide antibiotic (lincosamide antibiotic), a cephalosporin antibiotic, or an RNA synthetase inhibitor. In some embodiments, the antibiotic is selected from the group consisting of: azithromycin, vancomycin, metronidazole, gentamicin, colistin, fidaxomicin, telavancin, oritavancin, dalbavancin, daptomycin, cephalexin, cefalexin, and cefuroxime, cefadroxil, cefazolin, cephalosporin, cefaclor cefamandole, cefoxitin, cefprozil, ceftopirane, ciprofloxacin, cefprozil, ceftriaxone, and the like levofloxacin (levaquilin), fluxoxin (floxin), tequinine (tequin), moxifloxacin (avelox), norfloxacin (norflolox), tetracycline, minocycline (minocycline), oxytetracycline (oxyetracyline), doxycycline, amoxicillin (amoxicillin), ampicillin (ampicillin), penicillin V, bischlorocilin (dicycloacillin), carbenicillin (carbicillin), methicillin (methicillin), ertapenem (ertapene), doripenem (doripeem), imipenem (imipenem)/cilastatin (cilatin), meropenem (meropenem), amikacin (kanamicin), kanamycin (kanamicin), neomycin (neomycin), neticin (kamicin), tobramycin (tobramycin), paromomycin, cefixime (cefdinir), cefdinir (cefditoren), cefoperazone (cefperazone), cefotaxime (ceftaxime), ceftazidime (ceftazidime), ceftibuten (ceftibuten), ceftizoxime (ceftizoxime), ceftriaxone (ceftriaxone), cefoxitin (ceftiotin) and streptomycin (streetmycin). In some embodiments, the antibiotic is azithromycin.

In some embodiments, the one or more additional therapeutic agents that may be administered in combination with the compounds provided herein may be selected from the group consisting of: ribavirin, famprivir, ST-193, oseltamivir, zanamivir, pamil, danoprevir, ritonavir, radenpyr, bivalirudin, mitsubishi, etila, emtricitabine, tenofovir disoproxil, tenofovir alafenamide hemi-fumarate, abacavir, dolutegravir, efavirenz, elbavir, ledipavir, ganciclovir, sofosbuvir, bitaravir, dasabacavir, lamivudine, atazanavir, obetavir, lamivudine, stavudine, nevirapine, rilpivirine, ziprairivir, cimiravir, dacarbavir Grazopicir, pirenzvir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafavir, cabazitaxel, cytarabine, ecalciline, epigallocatechin gallate, itravir, fotif Sha Wei, gemcitabine, grifficine, isoprinosine, indinavir, malaevir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, nootkatole, pulefafur, PRO 140, raltegafur, prazidine, saquinavir, tebuvir, TNX-355, valacyclovir, VIR-576, and zalcitabine.

In some embodiments, the compounds described herein (e.g., compounds of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-A, IV-B, etc.) and pharmaceutically acceptable salts thereof can be used in combination with: for example, one or more other agents that may be useful in the prevention or treatment of respiratory diseases, inflammatory diseases, autoimmune diseases; antihistamines, corticosteroids (e.g., fluticasone propionate (fluticasone propionate), fluticasone furoate, beclomethasone dipropionate (beclomethasone dipropionate), budesonide (budesonide), ciclesonide (ciclesonide), mometasone furoate (mometasone furoate), triamcinolone, flunisolide), NSAIDs, leukotriene modulators (e.g., montelukast (montelukast), zafirlukast, profluust (pranlukast)), tryptase inhibitors, IKK2 inhibitors, p38 inhibitors, syk inhibitors, protease inhibitors (such as elastase inhibitors), integrin antagonists (e.g., beta-2 integrin antagonists), adenosine A2a agonists, mediator release inhibitors (such as cromolyn sodium), 5-lipoxygenase inhibitors (Ji Fuluo (zyflo)), DP1 antagonists, DP2 antagonists, PI3K delta inhibitors, ITK inhibitors, LP (lysophosphatidic acid inhibitors) or FLAP (5-lipoxygenase activating protein) inhibitors (e.g., 3- (3- (tert-butylsulfanyl) -1- (4- (6-ethoxypyridin-3-yl) benzyl) -5- ((5-ethylpyridin-2-yl) methoxy) -1H-indol-2-yl) -2, 2-dimethylpropionic acid sodium), bronchodilators (e.g., muscarinic antagonists, beta-2 agonists), methotrexate and similar agents; monoclonal antibody therapies such as anti-lgE, anti-TNF, anti-IL-5, anti-IL-6, anti-IL-12, anti-IL-1 and the like; cytokine receptor therapies, such as etanercept (etanercept) and the like; antigen non-specific immunotherapy (e.g., interferons or other cytokines/chemokines, chemokine receptor modulators (such as CCR3, CCR4 or CXCR2 antagonists), other cytokine/chemokine agonists or antagonists, TLR agonists and the like), suitable anti-infective agents, including antibiotics, antifungals, anthelmintics, antimalarials, antiprotozoal agents, and antitubercular agents.

In some embodiments, the additional therapeutic agent may be a kinase inhibitor, including but not limited to erlotinib, gefitinib, lenacitinib, afatinib, aoxitinib, lapatinib, crizotinib, britinib, ceritinib, aletinib, everatinib, and moxib, and ibratinib.

In some embodiments, the additional therapeutic agent may be a therapeutic antiviral vaccine.

In some embodiments, the additional therapeutic agent may be an immunomodulatory agent, including, but not limited to, an anti-PD-1 or anti-PDL-1 therapeutic agent, including pamphlezumab (pembrolizumab), nivolumab (nivolumab), alemtuzumab (atezolizumab), devaluzumab (durvalumab), BMS-936559, or avermectin (avelumab); anti-TIM 3 (anti-HAVcr 2) therapeutic agents, including but not limited to TSR-022 or MBG453; anti-LAG 3 therapeutic agents, including but not limited to, rilatumab (rellatlimab), LAG525, or TSR-033; anti-4-1 BB (anti-CD 37, anti-TNFRSF 9); CD40 agonist therapeutic agents, including but not limited to SGN-40, CP-870, CP-893 or RO7009789; anti-CD 47 therapeutic agents including, but not limited to, hu5F9-G4; an anti-CD 20 therapeutic agent; an anti-CD 38 therapeutic agent; STING agonists, including but not limited to ADU-S100, MK-1454, ASA404, or amidobenzimidazole; anthracyclines, including but not limited to doxorubicin (doxorubicin) or mitoxantrone (mitoxantrone); hypomethylating agents including, but not limited to, azacytidine (azacytidine) or decitabine (decetabine); other immunomodulatory therapeutic agents include, but are not limited to, epidermal growth factor inhibitors, statins, metformin (meta), angiotensin receptor blockers, thalidomide (thalidomide), lenalidomide, pomalidomide (pomalidomide), prednisone, or dexamethasone. In some embodiments, the additional therapeutic agent is a p 2-adrenoreceptor agonist, including, but not limited to, vilanaterol (vilanaterol), salmeterol (salmeterol), salbutamol (salbutamol), formoterol (formoterol), salmeterol (salmeterol), fenoterol Luo Kamo terol (fenoterol carmoterol), etaterol (etanterol), natto (namiterol), clenbuterol (clenbuterol), pirbuterol (pirbuterol), fluorobutanol (flinbuterol), rapoterol (reproterol), buterol (bambuterol), indacaterol (terbutamol), terbutaline (terbutaline), and salts thereof, such as salmeterol's (1-hydroxy-2-naphthate), salbutamol's sulfate, or formoterol's fumarate. In some embodiments, the additional therapeutic agent is an anticholinergic agent, including, but not limited to, turnip ammonium bromide (e.g., in bromide form), ipratropium (e.g., in bromide form), oxitropium (e.g., in bromide form), and tiotropium (e.g., in bromide form).

In particular, in certain embodiments, the present disclosure provides a method of treating the above medical indications comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein, such as the disclosed compounds.

The term "booster amount" or "booster dose" is the amount of compound required to improve the pharmacokinetics (or increase availability or exposure) of the second compound. The boosting amount or dose may improve the pharmacokinetics of the second compound (or increase availability or exposure) to the extent that it reaches therapeutic levels in the subject.

In one embodiment, the present disclosure provides a disclosed compound that is co-administered with an antiviral therapeutic agent such as disclosed herein and, for example, thereby boosting the dose of one or more antiviral therapeutic agents. Such a boosting combination may be used, for example, as a prophylactic or therapeutic treatment of a viral infection in a subject in need thereof. In one embodiment, the protease inhibitor is a compound described herein (e.g., a compound of formula II, II-A, II-B, II-C, II-D-A, II-D-B, II-E-A, II-E-B, II-F, II-G, II-H-A, II-H-B, II-E, II-I, IV-A, IV-B, etc.).

Reversible or irreversible conjugates

For example, disclosed herein is an engineered CL or 3CL viral protease, wherein:

the cysteine at position 145 of the CL or 3CL protease has a non-naturally occurring covalent modification resulting from the reaction between an exogenous nitrile modifier having a nitrile function and the cysteine at position 145 of the CL or 3CL protease, and

wherein the sulfur atom at the cysteine residue and the nitrile of the exogenous nitrile modifier undergo a reaction to form a thioimidate adduct, and wherein the engineered SARS-protease does not retain the protease activity of the unmodified CL or 2CL protease.

In some embodiments, the engineered viral protease substantially prevents viral replication of SARS-COV 2. In some embodiments, the CL or 3CL protease is represented by SEQ ID NO: 1. In other embodiments, the exogenous nitrile modifier of SEQ ID NO. 1 enzymatically inhibits IC ₅₀ Less than 20 micromoles.

In some embodiments, the thioimidate adducts resulting from in vivo reactions between an exogenous nitrile modifier and a cysteine at position 145 of SEQ ID NO:1 are represented by:

wherein the method comprises the steps of

IR is an exogenous nitrile modifier after undergoing a reaction.

For example, disclosed herein is an engineered 3CL or 3C protease, such as a SARS-COV2-3CL viral protease represented by SEQ ID No. 1, wherein the cysteine at position 145 of SEQ ID No. 1 has a non-naturally occurring covalent modification resulting from a reaction (e.g., an in vivo reaction) between an exogenous nitrile modifier having a nitrile function and the cysteine at position 145 of SEQ ID No. 1, and wherein the sulfur atom at the cysteine residue and the nitrile of the exogenous nitrile modifier undergo a reaction to form a thioiminoester adduct, and wherein the engineered-3 CL protease does not retain the protease activity of the unmodified-3 CL or 3C protease.

In some embodiments, the engineered SARS-COV2-3CL viral protease substantially prevents viral replication of SARS-COV 2. In other embodiments, the exogenous nitrile modifier of SEQ ID NO. 1 enzymatically inhibits IC ₅₀ Less than, for example, 20 micromoles.

In further embodiments, the thioimidate adducts resulting from the reaction between an exogenous nitrile modifier and a cysteine at position 145 of SEQ ID NO:1 may be represented, for example, by:

wherein IR is an exogenous nitrile modifier after undergoing a reaction.

Also disclosed herein is an engineered SARS-COV2-3CL viral protease represented by SEQ ID No. 1, wherein the cysteine at position 145 of SEQ ID No. 1 has a non-naturally occurring covalent modification resulting from a reaction between an exogenous nitrile modifier and the cysteine at position 145 of SEQ ID No. 1, wherein the exogenous nitrile modifier is represented by:

wherein the sulfur atom at the cysteine residue and the-C.ident.N of the exogenous nitrile modifier undergo a reaction to form a thioiminoester adduct, and wherein

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

R ^B Is C ₁ -C ₆ Alkyl (optionally one, two or three independently selected from halogen, -NR ^m R ^m and-NR ^m (C＝O)R ^m Substituent of the group consisting of R ^m At each occurrence selected from H or C _1-3 Alkyl (optionally substituted with one, two or three halo)); or an 8-10 membered bicyclic heteroaryl (optionally substituted with one, two or three substituents each independently selected from halo or methoxy);

R ^t Independently at each occurrence H or methyl; or each R ^t Can form a cyclopropyl group together with the carbon to which it is attached;

R ^1a is H; or (b)

R ¹ And R is ^1a Together with the nitrogen and carbon to which they are attached form a group which is optionally substituted on the free carbon by one or two groups each selected from methyl, halo or CF ₃ Substituent of (2)Substituted 4-10 membered monocyclic, bicyclic or spiro heterocycles.

Also disclosed herein is a compound represented by:

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

R ^1a is H; or (b)

R ¹ And R is ^1a Together with the nitrogen and carbon to which they are attached form a group which is optionally substituted on the free carbon by one or two groups each selected from methyl, halo or CF ₃ 4-10 membered monocyclic, bicyclic or spiro heterocycle substituted with substituents.

Also disclosed herein is an engineered SARS-COV2-3CL viral protease represented by SEQ ID No. 1, wherein the cysteine at position 145 of SEQ ID No. 1 has a non-naturally occurring covalent modification resulting from an in vivo reaction between an exogenous-c≡n modifier and the cysteine at position 145 of SEQ ID No. 1, wherein the exogenous-c≡n modifier is represented by:

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

R ^B Is C ₁ -C ₆ Alkyl or 8-10 membered bicyclic heteroaryl; wherein C is ₁ -C ₆ Alkyl groups optionally being substituted by one, two or three R ^B1 Substitution; and wherein heteroaryl may be optionally substituted with one, two or three halo;

R ^B1 independently at each occurrence selected from the group consisting of: halo, -NR ^m R ^m and-NR ^m (C＝O)R ^m ；

R ^m Independently at each occurrence selected from hydrogen or C _1-3 Alkyl (optionally substituted with one, two or three halo groups);

n is 1 or 2;

R ^1a is hydrogen; or (b)

R ¹ And R is ^1a Together with the nitrogen and carbon to which they are attached form a group which is optionally selected from CH on free carbon, one or two of which are each independently selected from CH ₃ Halo and CF ₃ A 4-10 membered monocyclic or bicyclic heterocycle substituted with substituents of the group consisting of.

In another embodiment, disclosed herein is a compound represented by:

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

n is 1 or 2;

R ^1a is hydrogen; or (b)

A detailed description of the sequence of SEQ ID NO. 1 is given below.

IV pharmaceutical composition and kit

Another aspect of the present disclosure provides a pharmaceutical composition comprising a compound as disclosed herein formulated with a pharmaceutically acceptable carrier. In particular, the present disclosure provides pharmaceutical compositions comprising a compound as disclosed herein formulated with one or more pharmaceutically acceptable carriers. These formulations include those suitable for oral, rectal, topical, buccal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), rectal, vaginal, or aerosol administration, but in any given case the most suitable form of administration will depend on the extent and severity of the condition being treated and the nature of the particular compound being used. For example, the disclosed compositions may be formulated in unit dosage form, and/or may be formulated for oral or subcutaneous administration.

Exemplary pharmaceutical compositions of the present disclosure may be used in the form of pharmaceutical formulations, e.g., in solid, semi-solid or liquid form, comprising as an active ingredient a mixture of one or more compounds of the present disclosure with an organic or inorganic carrier or excipient suitable for external, enteral or parenteral application. The active ingredient may be compounded, for example, with conventional non-toxic, pharmaceutically acceptable carriers for tablets, pills, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. The active target compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the course or condition of the disease.

For preparing solid compositions such as tablets, the primary active ingredient may be mixed with a pharmaceutical carrier, e.g., conventional tableting ingredients (such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums), and other pharmaceutical diluents, e.g., water, to form a solid preformulation composition containing a homogeneous mixture of a compound of the present disclosure or a non-toxic pharmaceutically acceptable salt thereof. When referring to these preformulation compositions as homogeneous compositions, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

In solid dosage forms (capsules, tablets, pills, dragees, powders, granules, etc.) for oral administration, the subject compositions are admixed with one or more pharmaceutically acceptable carriers such as sodium citrate or dicalcium phosphate and/or any of the following: (1) Fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and/or silicic acid; (2) Binders such as carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and/or acacia (acacia); (3) humectants, such as glycerin; (4) Disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarders such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) Humectants such as acetyl alcohol and glycerol monostearate; (8) adsorbents such as kaolin and bentonite; (9) Lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) a colorant. In the case of capsules, tablets and pills, the compositions may also comprise buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose/milk sugar, as well as high molecular weight polyethylene glycols and the like.

Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropyl methylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or croscarmellose sodium), surfactants or dispersants. Molded tablets may be made by molding in a suitable machine a mixture of the subject composition moistened with an inert liquid diluent. Tablets and other solid dosage forms such as sugar-coated pills, capsules, pills, and granules may optionally be scored or prepared with coatings and shells, such as enteric and other coatings well known in pharmaceutical formulations.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the subject compositions, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents; solubilizing agents and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, cyclodextrins, and mixtures thereof.

Suspensions, in addition to the subject compositions, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as suppositories, which may be prepared by mixing the subject compositions with one or more suitable non-irritating excipients or carriers, including for example cocoa butter, polyethylene glycols, a suppository wax or a salicylate, and which are solid at room temperature but liquid at body temperature, and therefore will melt in the body cavity and release the active agent.

Dosage forms of the subject compositions for transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active ingredient may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers or propellants which may be required.

Ointments, pastes, creams and gels may contain, in addition to the subject compositions, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the subject compositions, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. The spray may additionally contain customary propellants, such as chlorofluorohydrocarbons and unsubstituted volatile hydrocarbons, such as butane and propane.

The compositions and compounds of the present disclosure may alternatively be administered by aerosol. This is achieved by preparing an aqueous aerosol, a liposome formulation or solid particles containing the compound. Non-aqueous (e.g., fluorocarbon propellant) suspensions may be used. Sonic sprayers may be used because they minimize exposure of the agent to shear forces that may cause degradation of the compounds contained in the subject compositions. Generally, aqueous aerosols are prepared by formulating an aqueous solution or suspension of the subject compositions with conventional pharmaceutically acceptable carriers and stabilizers. The carrier and stabilizer will vary with the requirements of the particular subject composition, but typically includes a nonionic surfactant (Tween, pluronic or polyethylene glycol); harmless proteins such as serum albumin; sorbitan esters; oleic acid; lecithin; amino acids such as glycine; a buffering agent; a salt; sugar or sugar alcohol. Aerosols are typically prepared from isotonic solutions.

Pharmaceutical compositions of the present disclosure suitable for parenteral administration comprise a subject composition in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders (which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or thickening agents) which can be reconstituted into a sterile injectable solution or dispersion immediately prior to use.

Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like) and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate and cyclodextrins). Proper fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.

In another aspect, the present disclosure provides an enteral pharmaceutical preparation comprising the disclosed compound and an enteric material; and pharmaceutically acceptable carriers or excipients thereof. An enteric material refers to a polymer that is substantially insoluble in the acidic environment of the stomach and is primarily soluble in the intestinal fluid at a particular pH. The small intestine is a part of the gastrointestinal tract (the intestine) between the stomach and the large intestine, and includes the duodenum, jejunum, and ileum. The pH of the duodenum is about 5.5, the pH of the jejunum is about 6.5 and the pH of the distal ileum is about 7.5. Thus, the enteric material is insoluble, for example, up to a pH of about 5.0, about 5.2, about 5.4, about 5.6, about 5.8, about 6.0, about 6.2, about 6.4, about 6.6, about 6.8, about 7.0, about 7.2, about 7.4, about 7.6, about 7.8, about 8.0, about 8.2, about 8.4, about 8.6, about 8.8, about 9.0, about 9.2, about 9.4, about 9.6, about 9.8, or about 10.0. Exemplary enteric materials include Cellulose Acetate Phthalate (CAP); hydroxypropyl methylcellulose phthalate (HPMCP); polyvinyl acetate phthalate (PVAP); hydroxypropyl methyl cellulose acetate succinate (HPMCAS); cellulose acetate trimellitate; hydroxypropyl methylcellulose succinate; cellulose acetate succinate; cellulose acetate hexahydrophthalate; cellulose propionate phthalate; cellulose acetate maleate; cellulose acetate butyrate; cellulose acetate propionate; copolymers of methacrylic acid and methyl methacrylate; copolymers of methyl acrylate, methyl methacrylate and methacrylic acid; copolymers of methyl vinyl ether with maleic anhydride (Gantrez ES series); ethyl methacrylate-methyl methacrylate-chlorotrimethylammonium ethyl acrylate copolymer; natural resins such as zein, shellac, and cabazitaxel (copal colophorium); and several commercially available enteric dispersion systems (e.g., eudragit L30D55, eudragit FS30D, eudragit L100, eudragit S100, kollicoat EMM30D, estacryl D, coateric, and aquaeric). The solubility of each of the above materials is known or can be readily determined in vitro. The foregoing is a list of possible materials, but those skilled in the art having the benefit of this disclosure will recognize that it is not comprehensive and that other enteric materials exist that are consistent with the objects of this disclosure.

Advantageously, the present disclosure also provides kits for use by, for example, consumers in need of 3CL inhibitors. Such kits include suitable dosage forms, such as those described above, and instructions describing methods of using such dosage forms to mediate, reduce or prevent inflammation. The instructions will instruct the consumer or medical personnel to administer the dosage form according to a mode of administration known to those skilled in the art. Such kits may advantageously be packaged and sold in single or multiple kit units. An example of such a kit is a so-called blister pack. Blister packages are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packages are typically composed of a sheet of relatively rigid material covered with a foil of a preferably transparent plastic material. During the encapsulation process, grooves are formed in the plastic foil. The recess has the size and shape of the tablet or capsule to be packaged. Next, the tablet or capsule is placed in the groove and a sheet of relatively rigid material is sealed against the plastic foil on the side of the foil opposite the direction in which the groove is formed. Thus, the tablet or capsule is sealed in the groove between the plastic foil and the sheet. Preferably, the strength of the sheet is such that the tablet or capsule can be removed from the blister package by manually applying pressure on the groove, thereby forming an opening in the sheet where the groove is placed. The tablet or capsule may then be removed through the opening.

It may be desirable to provide memory assistance on the kit, for example in the form of a number immediately adjacent to the tablet or capsule, where the number corresponds to the number of days in the regimen that the tablet or capsule so specified should be ingested. Another example of such memory assistance is a calendar printed on the card, such as the following "first week, monday, tuesday … …, etc. … … second week, monday, tuesday … …, etc. Other variations of memory assistance will be apparent. A "daily dose" may be a single tablet or capsule, or several pills or capsules, to be taken on a given day. In addition, the daily dose of the first compound may consist of one tablet or capsule, while the daily dose of the second compound may consist of several tablets or capsules, and vice versa. The memory assistance should reflect this.

Also contemplated herein are methods and compositions comprising or administering a second active agent. For example, in addition to suffering from a viral infection, a subject or patient may also suffer from a viral infection or a viral-related co-occurrence disease, i.e., a disease or other unhealthy condition associated with, exacerbated by, or facilitated by a viral infection. Combinations of the disclosed compounds with at least one other agent that has previously been demonstrated to treat these virus-related conditions are contemplated herein.

V. further embodiments of the present disclosure

1. Covered embodiments

In one aspect, the compositions, compounds, and methods of the present disclosure can be described in one embodiment as follows:

1. a viral protease inhibitor compound represented by:

wherein:

R ¹ selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein R ¹ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);

R ^A independently at each occurrence selected from halogen, cyano, hydroxy, oxo, -NH ₂ 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy and C ₃ -C ₆ Cycloalkyl;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B A 5-10 membered heterocycle, a 5-10 membered aryl, and a 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

R ^B independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, 5-10 membered aryl, 5-10 membered heteroaryl, and 5-10 membered heterocycle;

R ^C independently at each occurrence selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, cyano, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with oneOr a plurality of groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

R ^y independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl) and C ₃ -C ₆ Cycloalkyl;

a is a reversible or irreversible warhead;

R ³ selected from 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

A is a reversible or irreversible warhead selected from the group consisting of: cyano, -C (O) R ^D 、-C(O)CH ₂ N(R ^b R ^c )、-C(O)CH ₂ OC(O)R ^D 、-C(O)C(O)R ^D And- (ch=ch) C (O) OR ^D Wherein

R ^D Selected from the group consisting of: hydrogen, -N (R) ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, and 5-10 membered heterocycle; wherein R is ^D Can be optionally selected from one, two or three of halogen, hydroxy and R ^E Substituent groups of the group are substituted;

R ^E selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy and 5-10 membered aryl and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group; and is also provided with

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl groups may optionally be substituted with one or more groups each selected from halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl.

A is reversible warheadWherein R is ^c Selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl.

4.R ^c Is thatWherein X is ¹ Independently at each occurrence selected from N and CH.

A is a reversible warhead selected from the group consisting of:/>

a is reversible warheadWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,

R ^E Independently at each occurrence selected from the group consisting of: halogen, hydroxy, C ₁ -C ₈ Alkyl and C ₁ -C ₈ An alkoxy group;

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

a is reversible warheadWherein X is ² Selected from the group consisting of: NH, NR ^P O and S, wherein R ^P Is C ₁ -C ₈ An alkyl group.

9.A reversible warhead

A is irreversible warhead-C (O) CH ₂ OC(O)R ^D Wherein

R ^D Selected from the group consisting of:C ₁ -C ₈ alkyl and C ₃ -C ₆ Cycloalkyl;

X ⁴ independently at each occurrence selected from CH and N;

R ^E independently at each occurrence selected from the group consisting of: halogen, -CN, -CH ₃ 、-CH ₂ CH ₃ 、-CH(CH ₃ ) ₂ 、-OCH ₃ 、-CF ₃ 、-OCF ₃ and-SCF ₃ The method comprises the steps of carrying out a first treatment on the surface of the And is also provided with

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

/>

a is an irreversible warhead selected from the group consisting of:

a is reversible or irreversible warhead-C (O) R ^D Wherein R is ^D Selected from the group consisting of: hydrogen, -CH ₂ OH、-CH ₂ OR' and-CH _x F _y Wherein R' is selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered aryl), C ₁ -C ₈ Heteroalkyl, C ₃ -C ₆ Cycloalkyl and 5-10 membered aryl, wherein x is 0, 1 or 2; y is 1, 2 or 3; and the sum of x and y is 3.

A is a reversible or irreversible warhead selected from the group consisting of:

a is reversible OR irreversible warhead- (ch=ch) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

A is an irreversible warhead selected from the group consisting of:

a is reversible or irreversible warhead-C (O) CH ₂ N(R ^b R ^c )。

A is selected fromReversible or irreversible warheads:

a is a reversible or irreversible warhead

Wherein M is selected from Na and K.

A is cyano.

22.R ¹ Selected from the group consisting of:

23.R ² selected from the group consisting of:

/>

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

R ⁵ independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

R ⁶ is C ₁ -C ₈ An alkyl group;

R ⁷ at each occurrenceIndependently selected from the group consisting of: halogen, hydroxy, oxo, -N (R) ^y ) ₂ 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

R ⁸ selected from the group consisting of: a 5-10 membered aryl, a 5-10 membered heteroaryl, and a 5-10 membered heterocycle;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH and S;

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

R ⁹ selected from the group consisting of: halogen, hydroxy, oxo, -NH ₂ 、-N(CH ₃ ) ₂ 、-N(CH ₂ CH ₃ ) ₂ 、-CH ₃ 、-CH ₂ CH ₃ 、-OCH ₃ and-OCH ₂ CH ₃ 。

26.R ³ Selected from the group consisting of:

27. the viral protease inhibitor compounds are represented by:

wherein:

R ^y independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl) and C ₃ -C ₆ Cycloalkyl; and is also provided with

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The viral protease inhibitor compounds are represented by:

wherein the method comprises the steps of

R ^x Independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, -N (R) ^y ) ₂ 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

R ^y independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy and C ₃ -C ₆ Cycloalkyl;

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

30.R ^x is-OCH ₃ 。

31. A viral protease inhibitor compound selected from the group consisting of:

/>

32. a viral protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, cyano, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroarylAnd a 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

a is a reversible or irreversible warhead;

R ³ selected from 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a); and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group)、-C(O)-(C ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl groups may optionally be substituted with one or more groups each selected from halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl.

34.R ¹ Selected from the group consisting of:

35.R ³ is a 5-10 membered heterocycle.

36.R ³ Selected from the group consisting of:

37.R ² selected from the group consisting of:

/>

38. a reversible conjugate represented by:

wherein the method comprises the steps of

Cys ₁₄₅ Is cysteine at position 145 or an equivalent active site on a CL or 3CL protease;

IR is a viral protease inhibitor;

B is selected from the group consisting of: -R ^D 、-C(O)R ^D and-CH ₂ OR ^D Wherein

R ^E selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy and 5-10 membered aryl and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group; and is also provided with

39. An irreversible conjugate represented by:

wherein the method comprises the steps of

IR is a viral protease inhibitor;

R ^D selected from the group consisting of: hydrogen, -N (R) ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered arylA 5-10 membered heteroaryl and a 5-10 membered heterocycle; wherein R is ^D Can be optionally selected from one, two or three of halogen, hydroxy and R ^E Substituent groups of the group are substituted;

40. A method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

41. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

42. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus (feline calicivirus), MD145, murine norovirus (murine norovirus), swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus (coxsackievirus), foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

43. The viral infection is a coronavirus infection.

44. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

45. The viral infection is SARS-CoV-2.

46. The viral infection is an arenavirus infection.

48. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

48. The viral infection is an influenza infection.

49. The influenza is influenza H1N1, H3N2 or H5N1.

50. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of a compound of the embodiments, and/or contacting an effective amount of a compound of the embodiments with a virally infected cell.

51. The method of embodiments further comprising administering another therapeutic agent.

52. The method of embodiments further comprising administering an additional antiviral therapeutic agent.

53. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, fampicvir, ST-193, oseltamivir, zanamivir, peramivir, danoprevir, ritona Wei Herui, and darunavir.

54. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

55. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibodies, env Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivirgine, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampin, zanamivir, oselta Weidan noprevir, ritona Wei Herui desivir.

56. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of any of the compounds of the embodiments.

57. The compounds are administered prior to viral exposure.

58. The compounds are administered after viral exposure.

2. Covered embodiments

In another aspect, the compositions, compounds, and methods of the present disclosure can be described in another embodiment as follows:

1. a viral protease inhibitor compound represented by:

wherein:

R ² selected from the group consisting of: of-NH bound via carbon or nitrogen atoms ₂ 、-NHC(O)R ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B A 5-10 membered heterocycle, a 5-10 membered aryl, and a 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, cyano, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

R ^y independently at each occurrence selected from the group consisting ofThe group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl) and C ₃ -C ₆ Cycloalkyl;

a is a reversible or irreversible warhead;

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

A is reversible warhead Wherein R is ^c Selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl.

A is a reversible warhead selected from the group consisting of:

a is reversible warheadWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,R ^E Independently at each occurrence selected from the group consisting of: halogen, hydroxy, C ₁ -C ₈ Alkyl and C ₁ -C ₈ An alkoxy group;

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

9.A reversible warhead

A is irreversible warhead-C (O) CH ₂ OC(O)R ^D Wherein

X ⁴ independently at each occurrence selected from CH and N;

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

a is an irreversible warhead selected from the group consisting of:

A is an irreversible warhead selected from the group consisting of:

a is reversible or irreversible warhead-C (O) CH ₂ N(R ^b R ^c )。

a is a reversible or irreversible warheadWherein M is selected from Na and K.

A is cyano.

22.R ¹ Selected from the group consisting of:

23.R ² selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

R ⁶ is C ₁ -C ₈ An alkyl group;

R ⁷ independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, -N (R) ^y ) ₂ 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH and S;

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

26.R ³ Selected from the group consisting of:

27. the compounds are represented by:

wherein:

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The compound is selected from the group consisting of:

30. the compounds are represented by:

wherein the method comprises the steps of

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

31.R ^x is-OCH ₃ 。

32. A viral protease inhibitor compound selected from the group consisting of:

/>

33. a viral protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, cyano, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl；

a is a reversible or irreversible warhead;

x is selected from CH and N;

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

33. The compounds are represented by:

wherein the method comprises the steps of

a is a reversible or irreversible warhead;

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

R ^D Selected from the group consisting of: hydrogen, -N (R) ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, and 5-10 membered heterocycle; wherein R is ^D Can optionally be oneTwo or three are each selected from halogen, hydroxy and R ^E Substituent groups of the group are substituted;

35.R ¹ Selected from the group consisting of:

36.R ³ is a 5-10 membered heterocycle.

37.R ³ Selected from the group consisting of:

38.R ² selected from the group consisting of:

/>

39. the compound is selected from the group consisting of:

42. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

43. The viral infection is a coronavirus infection.

45. The viral infection is SARS-CoV-2.

46. The viral infection is an arenavirus infection.

47. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

48. The viral infection is an influenza infection.

49. The influenza is influenza H1N1, H3N2 or H5N1.

50. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

51. The method further comprises administering another therapeutic agent.

52. The method further comprises administering an additional antiviral therapeutic agent.

57. The compounds are administered prior to viral exposure.

58. The compounds are administered after viral exposure.

3. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

R ^A Independently at each occurrence selected from halogen, cyano, hydroxy, oxo, SF ₅ 、-NH ₂ 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy and C ₃ -C ₆ Cycloalkyl;

a is a warhead;

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

A is selected from the group consisting of: cyano, -C (O) R ^D 、-C(O)CH ₂ N(R ^b R ^c )、-C(O)CH ₂ OC(O)R ^D 、-C(O)C(O)R ^D 、-(CH＝CH)C(O)OR ^D 、-(CH＝CCN)C(O)OR ^D 、-(CH＝CCN)C(O)(NH)R ^D 、-CH(CN)(OH)、-CH(CN)(NR ^b R ^c )、Wherein the method comprises the steps of

R ^D Selected from the group consisting of: hydrogen, hydroxy, -OR ^bb 、-N(R ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; wherein R is ^D Can be optionally selected from one, two or three of halogen, hydroxy and R ^E Substituent groups of the group are substituted;

R ^E selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy and C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group;

R ^bb selected from the group consisting of: c (C) ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, - (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

R ^cc selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl group, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), 5-10 membered heterocycle and-N (R) ^b R ^c ) Wherein R is ^b And R is ^c Are each selected from hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl group, or R ^b And R is ^c Can be joined together to form, together with the nitrogen to which they are attached, a 5-10 membered heterocyclic ring;

R ^cd Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl; and is also provided with

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl.

A is a warhead represented by:wherein R is ^c Selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl.

A is selected from the group consisting of:

a isWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

a isWherein X is ² Selected from the group consisting of: NH, NR ^P O and S, wherein R ^P Is C ₁ -C ₈ An alkyl group.

9.A is selected from the group consisting of:

a is-C (O) CH ₂ OC(O)R ^D Wherein

X ⁴ independently at each occurrence selected from CH and N;

R ^E independently at each occurrenceA group consisting of: halogen, -CN, -CH ₃ 、-CH ₂ CH ₃ 、-CH(CH ₃ ) ₂ 、-OCH ₃ 、-CF ₃ 、-OCF ₃ and-SCF ₃ The method comprises the steps of carrying out a first treatment on the surface of the And is also provided with

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

a is selected from the group consisting of:

a is-C (O) R ^D Wherein R is ^D Selected from the group consisting of: hydrogen, -CH ₂ OH、-CH ₂ OR' and-CH _x F _y Wherein R' is selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered aryl), C ₁ -C ₈ Heteroalkyl, C ₃ -C ₆ Cycloalkyl and 5-10 membered aryl, wherein x is 0, 1 or 2; y is 1, 2 or 3; and the sum of x and y is 3.

A is selected from the group consisting of:

/>

a is- (ch=ch) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

A is selected from

A is-C (O) CH ₂ N(R ^b R ^c )。

A warhead selected from the group consisting of:

a isWherein M is selected from Na and K.

A is cyano.

22.R ¹ Selected from the group consisting of: 23.R ² Selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

R ⁶ is C ₁ -C ₈ An alkyl group;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH and S;

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ selected from the group consisting of:

/>

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

26.R ³ Selected from the group consisting of:

27. the compounds are represented by:

wherein:

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The compound is selected from the group consisting of:

30. the compounds are represented by:

wherein the method comprises the steps of

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

31.R ^x is-OCH ₃ 。

32. A protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^3a Selected from the group consisting ofAnd a 4-10 membered heterocycle, wherein the heterocycle may be optionally substituted with one, two or three substituents each selected from the group consisting of: hydroxy, C ₁ -C ₈ Alkoxy, oxo, and warhead a;

R ^3b selected from hydrogen and C ₁ -C ₈ An alkyl group; wherein R is ^3a And R is ^3b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring, wherein the heterocyclic ring can optionally be substituted with one, two or three groups each selected from C ₆ -C ₁₄ Aryl and warhead a substituents;

R ^1a selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^1a and R is ^1b Can be joined together to form a 4-10 membered heterocyclic ring or C together with the carbon to which it is attached ₃ -C ₁₀ Cycloalkyl;

R ^A at each timeIndependently at each occurrence selected from halogen, cyano, hydroxy, oxo, SF ₅ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocycle), -C (O) -O- (4-10 membered heterocycle), -C (O) -OC (CH) ₃ ) ₃ 、-C(O)-(C ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl group, C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle, wherein the heterocycle, heteroaryl, or aryl may optionally be substituted with one, two, or three halogens, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Substituents for hydroxy or oxo;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

R ^1a and R is ² Can be joined together to form a 4-10 membered heterocyclic ring or C together with the carbon to which it is attached ₃ -C ₁₀ Cycloalkyl wherein the cycloalkyl or heterocycle may optionally be substituted with one, two or three groups each selected from R ^A Is substituted by a substituent of (a);

R ³ Selected from 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);

R ^B independently at each occurrence selected from the group consisting ofGroup: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, SF ₅ Cyano, -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein the aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

a is a warhead;

x is selected from CH, C (CH) ₃ ) And N; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

33. The compounds are represented by:

34. the compounds are represented by:

A is selected from the group consisting of: -CN,

37.R ^1a Selected from the group consisting of:

38.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

39.R ^1b Is hydrogen.

40.R ^1a And R is ^1b Joined together to form

41.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

42.R ^3a Selected from the group consisting of:

43.R ³ is a 4-10 membered heterocycle.

44.R ³ Selected from the group consisting of:

45.R ² selected from the group consisting of:

/>

46.R ^1a and R is ² Joined together to form a heterocycle selected from the group consisting of:

47. the compound is selected from the group consisting of:

/>

48. a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

49. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

50. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

51. The viral infection is a coronavirus infection.

52. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

53. The viral infection is SARS-CoV-2.

54. The viral infection is an arenavirus infection.

55. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

56. The viral infection is an influenza infection.

57. The influenza is influenza H1N1, H3N2 or H5N1.

58. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

59. The method further comprises administering another therapeutic agent.

60. The method further comprises administering an additional antiviral therapeutic agent.

61. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, fampicvir, ST-193, oseltamivir, zanamivir, peramivir, danoprevir, ritona Wei Herui, and darunavir.

62. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

63. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibodies, env Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivirgine, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampin, zanamivir, oselta Weidan noprevir, ritona Wei Herui desivir.

64. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of any of the compounds of the embodiments.

65. The compounds are administered prior to viral exposure.

66. The compounds are administered after viral exposure.

4. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

R ^3a selected from the group consisting ofAnd 4-10 membered heterocyclic ring, wherein the heterocyclic ring may optionally beOne, two or three substituents each selected from the group consisting of: hydroxy, C ₁ -C ₈ Alkoxy, oxo, and warhead a;

R ^1a selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^A independently at each occurrence selected from halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-NHC(O)O(C ₁ -C ₈ Alkyl) R ^B -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocycle), -C (O) -O- (4-10 membered heterocycle), -C (O) -OC (CH) ₃ ) ₃ 、-C(O)-(C ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl group, C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle, wherein R is ^B The alkyl, heterocycle, heteroaryl or aryl groups may optionally be substituted with one, two or three halogens, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Substituents for hydroxy or oxo;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 、-O-(C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl), 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

R ³ selected from 5-10 membered heteroaryl and4-10 membered heterocycle wherein R ³ Can optionally be one, two or three of each selected from R ^A Is substituted by a substituent of (a);

R ^B independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano, -OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein the alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl;

R ^y independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH;

a is a warhead;

x is selected from the group consisting of: CH. C (CH) ₃ ) And N; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

2. The compounds are represented by:

3. the compounds are represented by:

R ^E selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group;

R ^bb selected from the group consisting of: c (C) ₃ -C ₆ Cycloalkyl radicals、C ₆ -C ₁₄ Aryl, - (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

A is selected from the group consisting of: -CN,

6.R ^1a Selected from the group consisting of:

7.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

8.R ^1b Is hydrogen.

9.R ^1a And R is ^1b Joined together to form

10.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

11.R ^3a Selected from the group consisting of:

12.R ³ is a 4-10 membered heterocycle.

13.R ³ Selected from the group consisting of:

14.R ² selected from the group consisting of:

/>

15.R ^1a and R is ² Joined together to form a heterocycle selected from the group consisting of:

/>

16. the compound is selected from the group consisting of:

/>

17. a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

18. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

19. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

20. The viral infection is a coronavirus infection.

21. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

22. The viral infection is SARS-CoV-2.

23. The viral infection is an arenavirus infection.

24. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

25. The viral infection is an influenza infection.

26. The influenza is influenza H1N1, H3N2 or H5N1.

27. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

28. The method further comprises administering another therapeutic agent.

29. The method further comprises administering an additional antiviral therapeutic agent.

30. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, famprivir, ST-193, oseltamivir, zanamivir, pamil, danoprevir, ritonavir, radenpyr, bivalirudin, mitsubishi, etila, emtricitabine, tenofovir disoproxil, tenofovir alafenamide hemi-fumarate, abacavir, dolutegravir, efavirenz, elbavir, ledipavir, ganciclovir, sofosbuvir, bitaravir, dasabacavir, lamivudine, atazanavir, obetavir, lamivudine, stavudine, nevirapine, rilpivirine, ziprairivir, cimiravir, dacarbavir Grazopicir, pirenzvir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafavir, cabazitaxel, cytarabine, ecalciline, epigallocatechin gallate, itravir, fotif Sha Wei, gemcitabine, grifficine, isoprinosine, indinavir, malaevir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, nootkatole, pulefafur, PRO 140, raltegafur, prazidine, saquinavir, tebuvir, TNX-355, valacyclovir, VIR-576, and zalcitabine.

31. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

32. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibody, enfu Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivir, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampicin, zanamivir, oseltamivir Weidan nopuvir, ritonavir, radciclovir, cobalastat, eptifibatide, emtricitabine, tenofovir disoproxil, tenofovir, valavermectin, abacavir, dulutegravir, efavirenz, ganciclovir, sofosbuvir, bitorubivir, dasabavir, lamivudine, atazanavir, obuwei, obutyvir, emtricitabine, tenofovir, valacil, valvulafir, and valvulafir lamivudine, stavudine, nevirapine, rilpivirine, palivir, cimlopivir, dacarbazine, pezopivir, pirenz-tavir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafrevir, cabavir, cytarabine, eleganciclovir, epigallocatechin gallate, itraprine, ferti Sha Wei, gemcitabine, grifeis, isoprinosine, indinavir, malavir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, norelvir, praxafovir, PRO 140, raltegafur, prazidine, quinavir, telbivudine, TNX-355, valacyclovir, pir-576 and zanciclovir.

33. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of any of the compounds of the embodiments.

34. The compounds are administered prior to viral exposure.

35. The compounds are administered after viral exposure.

5. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

a is a warhead;

x is selected from C (R) ^xy ) And N, wherein R is ^xy Selected from the group consisting of: H. d, -OH, -NH ₂ Halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl and C ₁ -C ₈ An alkoxy group; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

2. The compounds are represented by:

3. the compounds are represented by:

A is selected from the group consisting of: -CN,

6.R ^1a Selected from the group consisting of:

7.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

8.R ^1b Is hydrogen.

9.R ^1a And R is ^1b Joined together to form

10.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

11.R ^3a Selected from the group consisting of:

12.R ³ is a 4-10 membered heterocycle.

13.R ³ Selected from the group consisting of:

14.R ² Selected from the group consisting of:

/>

16. a compound selected from the group consisting of:

/>

20. The viral infection is a coronavirus infection.

22. The viral infection is SARS-CoV-2.

23. The viral infection is an arenavirus infection.

25. The viral infection is an influenza infection.

26. The influenza is influenza H1N1, H3N2 or H5N1.

28. The method further comprises administering another therapeutic agent.

34. The compounds are administered prior to viral exposure.

35. The compounds are administered after viral exposure.

6. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

a is a warhead;

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

A is selected from the group consisting of:

a isWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

9.A is selected from the group consisting of:

a is-C (O) CH ₂ OC(O)R ^D Wherein

X ⁴ independently at each occurrence selected from CH and N;

R ^E at each occurrence Independently selected from the group consisting of: halogen, -CN, -CH ₃ 、-CH ₂ CH ₃ 、-CH(CH ₃ ) ₂ 、-OCH ₃ 、-CF ₃ 、-OCF ₃ and-SCF ₃ The method comprises the steps of carrying out a first treatment on the surface of the And is also provided with

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

a is selected from the group consisting of:

A is selected from the group consisting of:

a is- (ch=ch) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

A is selected from/>

A is-C (O) CH ₂ N(R ^b R ^c )。

A warhead selected from the group consisting of:

a isWherein M is selected from Na and K.

A is cyano.

22.R ¹ Selected from the group consisting of:23.R ² selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

R ⁶ is C ₁ -C ₈ An alkyl group;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH and S;

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

26.R ³ Selected from the group consisting of:

27. the compounds are represented by:

wherein:

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The compound is selected from the group consisting of:

30. the compounds are represented by:

wherein the method comprises the steps of

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

31.R ^x is-OCH ₃ 。

32. A protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^3a Selected from the group consisting of And a 4-10 membered heterocycle, wherein the heterocycle may be optionally substituted with one, two or three substituents each selected from the group consisting of: hydroxy, C ₁ -C ₈ Alkoxy, oxo, and warhead a;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group; or (b)

R ^A independently at each occurrence selected from halogen, cyano, hydroxy, oxo, SF ₅ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocycle), -C (O) -O- (4-10 membered heterocycle), -C (O) -OC (CH) ₃ ) ₃ 、-C(O)-(C ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl group, C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein the alkyl, cycloalkyl, heterocycle, heteroaryl or aryl may optionally be substituted with one, two or three halogens, C ₁ -C ₆ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Substituents for hydroxy or oxo;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a); or (b)

R ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered mono-or bicyclic heterocycle having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein the cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a);

R ^B independently at each occurrence selected from the group consisting of：C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

R ^G selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl and heterocycle, wherein R ^m Selected at each occurrence from H and C _1-3 Alkyl (optionally substituted with one, two or three fluoro), or C ₃ -C ₆ Cycloalkyl (optionally substituted with one, two or three fluoro);

a is a warhead;

x is selected from CH, C (CH) ₃ ) And N;and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

33. The compounds are represented by:

34. the compounds are represented by:

R ^E selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy and C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, cyano、C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group;

A is selected from the group consisting of: -CN,

37.R ^1a Selected from the group consisting of:

38.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

39.R ^1b Is hydrogen.

40.R ^1a And R is ^1b Joined together to form

41.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

42.R ^3a Selected from the group consisting of:

43.R ³ is a 4-10 membered heterocycle.

44.R ³ Selected from the group consisting of:

45.R ² selected from the group consisting of:

/>

and R is ^1b H.

48. The compounds are represented by:

wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl, C _3-6 Cycloalkyl, phenyl and heterocycle; and R is ^G2 is-NH (C=O) R ^m Wherein R is ^m Selected at each occurrence from H, methyl or CF ₃ 。

49. The compounds are represented by:

50. The compound is selected from the group consisting of:

/>

51. a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

52. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

53. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

54. The viral infection is a coronavirus infection.

55. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

56. The viral infection is SARS-CoV-2.

57. The viral infection is an arenavirus infection.

58. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

59. The viral infection is an influenza infection.

60. The influenza is influenza H1N1, H3N2 or H5N1.

61. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

62. The method further comprises administering another therapeutic agent.

63. The method further comprises administering an additional antiviral therapeutic agent.

64. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, fampicvir, ST-193, oseltamivir, zanamivir, peramivir, danoprevir, ritona Wei Herui, and darunavir.

65. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

66. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibodies, env Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivirgine, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampin, zanamivir, oselta Weidan noprevir, ritona Wei Herui desivir.

67. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of any of the compounds of the embodiments.

68. The compounds are administered prior to viral exposure.

69. The compounds are administered after viral exposure.

7. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

a is a warhead;

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

A is a warhead represented by:wherein R is ^c Selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, and 5-10 membered heteroaryl. />

A is selected from the group consisting of:

a isWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

9.A is selected from the group consisting of:

a is-C (O) CH ₂ OC(O)R ^D Wherein

X ⁴ independently at each occurrence selected from CH and N;

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

a is selected from the group consisting of:

A is selected from the group consisting of:

a is- (ch=ch) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

A is selected from

A is-C (O) CH ₂ N(R ^b R ^c )。

A warhead selected from the group consisting of:

a isWherein M is selected from Na and K.

A is cyano.

wherein->

Represents a bond, which may be a single bond or a double bond;

R ⁶ is C ₁ -C ₈ An alkyl group;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH andS；

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ Selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

26.R ³ Selected from the group consisting of:

27. the compounds are represented by:

wherein:

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The compound is selected from the group consisting of:

30. the compounds are represented by:

wherein the method comprises the steps of

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

31.R ^x is-OCH ₃ 。

32. A protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^1a selected from the group consisting ofIs set of (3): c (C) ₁ -C ₈ Alkyl, - (C) ₁ -C ₈ Alkyl) -R ¹ 、-(C ₁ -C ₈ Alkyl) -CN, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group; or (b)

R ^G selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocycle, C ₁ -C ₆ Alkoxy, wherein R is ^m At each occurrence selected from H, C _1-3 Alkyl (optionally covered with oneTwo or three substituents each independently selected from the group consisting of: halo, optionally substituted phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl), C (=o) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano and C ₁ -C ₆ Alkoxy), C (=o) -C _3-6 Cycloalkyl or C (=O) - (5-6 membered heteroaryl) (optionally substituted with halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy and C _1-6 Haloalkyl substitution);

a is a warhead;

x is selected from CH, C (CH) ₃ ) And N; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

33. The compounds are represented by:

34. the compounds are represented by:

A is selected from the group consisting of: -CN,

37.R ^1a Selected from the group consisting of:

38.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

39.R ^1b Is hydrogen.

40.R ^1a And R is ^1b Joined together to form

41.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

42.R ^3a Selected from the group consisting of:

43.R ³ is a 4-10 membered heterocycle.

44.R ³ Selected from the group consisting of:

/>

45.R ² selected from the group consisting of:

/>

and R is ^1b H.

48. The compounds are represented by:

49. The compounds are represented by:

50. The compounds are represented by:

wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted by one, two or three C ₁ -C ₆ Alkoxy substitution), C _3-6 Cycloalkyl, phenyl and heterocycle; and R is ^G2 Selected from the group consisting of: -NH (C) _1-3 Alkyl) (optionally substituted with one, two or three substituents each independently selected from the group consisting of: halo, optionally substituted phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl) and-NH (c=o) R ^m Wherein R is ^m At each occurrence selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally one, two or three independently selected from halo, cyano and C ₁ -C ₆ Substituted with a substituent selected from the group consisting of alkoxy), CHF ₂ 、CF ₃ And 5-6 membered heteroaryl (optionally substituted with halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy, CHF ₂ Or CF (CF) ₃ Substitution).

51. The compounds are represented by:

52.R ^G3 Selected from the group consisting of:

53.R ^G2 selected from the group consisting of:

wherein R is ^F Selected from the group consisting of: c (C) _1-6 Alkyl, C _3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, wherein R ^F Optionally one, two or three groups selected from halogen, cyano, hydroxy and C ₁ -C ₆ Substituent substitution of the group consisting of alkoxy; and X is ^F Selected from the group consisting of: H. halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy and C _1-6 Haloalkyl.

54. The compound is selected from the group consisting of:

/>

55. a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

56. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

57. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

58. The viral infection is a coronavirus infection.

59. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

60. The viral infection is SARS-CoV-2.

61. The viral infection is an arenavirus infection.

62. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

63. The viral infection is an influenza infection.

64. The influenza is influenza H1N1, H3N2 or H5N1.

65. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

66. The method further comprises administering another therapeutic agent.

67. The method further comprises administering an additional antiviral therapeutic agent.

68. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, fampicvir, ST-193, oseltamivir, zanamivir, peramivir, danoprevir, ritona Wei Herui, and darunavir.

69. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

70. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibodies, env Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivirgine, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampin, zanamivir, oselta Weidan noprevir, ritona Wei Herui desivir.

71. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of any of the compounds of the embodiments.

72. The compounds are administered prior to viral exposure.

73. The compounds are administered after viral exposure.

8. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

R ² selected from the group consisting of: of-NH bound via carbon or nitrogen atoms ₂ 、-NHC(O)R ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 5-10 yuanHeterocycles, 5-10 membered aryl and 5-10 membered heteroaryl, wherein R ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

a is a warhead;

m is 1 or 2; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

R ^cd Selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ NaphtheneA base; and is also provided with

A is selected from the group consisting of:

/>

a isWherein the method comprises the steps of

X ² Selected from the group consisting of: NH, O, and S;

X ³ independently at each occurrence selected from N and CH;

R ^D independently at each occurrence selected from the group consisting of:

C ₁ -C ₈ alkyl group,

p is selected from 0, 1 and 2; and is also provided with

q is selected from 0, 1 and 2.

A is selected from the group consisting of:

9.A is selected from the group consisting of:

a is-C (O) CH ₂ OC(O)R ^D Wherein

X ⁴ independently at each occurrence selected from CH and N;

p is selected from 0, 1 and 2.

11.R ^D Selected from the group consisting of:

a is selected from the group consisting of:

A is selected from the group consisting of:

/>

a is- (ch=ch) C (O) OR ^D Wherein R is ^D Is C ₁ -C ₈ An alkyl group.

A is selected from

A is-C (O) CH ₂ N(R ^b R ^c )。

A warhead selected from the group consisting of:

a isWherein M is selected from Na and K.

A is cyano.

22.R ¹ Selected from the group consisting of:

23.R ² Selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

R ⁵ independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, -N (R) ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, wherein the heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl；

R ⁶ Is C ₁ -C ₈ An alkyl group;

W ¹ selected from CH and N;

W ² selected from the group consisting of: CH (CH) ₂ O, NH and S;

w is selected from W ¹ And W is ² ；

s is selected from 1 and 2; and is also provided with

t is selected from 0, 1, 2 and 3.

24.R ² Selected from the group consisting of:

/>

25.R ³ Selected from the group consisting of:

wherein the method comprises the steps of

Represents a bond, which may be a single bond or a double bond;

Y ¹ selected from the group consisting of: CH. CH (CH) ₂ N, NH, O and S;

26.R ³ Selected from the group consisting of:

27. the compounds are represented by:

wherein:

m is selected from 1 and 2.

28.R ^y Selected from the group consisting of: hydrogen, hydrogen,

29. The compound is selected from the group consisting of:

30. the compounds are represented by:

wherein the method comprises the steps of

w is CH or N;

m is selected from 1 and 2; and is also provided with

r is selected from 0, 1, 2 and 3.

31.R ^x is-OCH ₃ 。

32. A protease inhibitor compound represented by:

wherein the method comprises the steps of

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group; or (b)

R ^G selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocycle, C ₁ -C ₆ Alkoxy, wherein R is ^m At each occurrence selected from H, C _1-3 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, optionally substituted phenyl, -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl), C (=o) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano and C ₁ -C ₆ Alkoxy), C (=o) -C _3-6 Cycloalkyl or C (=O) - (5-6 membered heteroaryl) (optionally substituted with halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy and C _1-6 Haloalkyl substitution);

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, SF ₅ Cyano, -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle,wherein the aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo and C ₁ -C ₈ Substituent substitution of alkyl;

a is a warhead;

x is selected from CH, C (CH) ₃ ) And N; and

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

33. The compounds are represented by:

34. the compounds are represented by:

R ^D Selected from the group consisting of: hydrogen, hydroxy, -OR ^bb 、-N(R ^b R ^c )、C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle; wherein R is ^D Optionally one, two or three of each selected from halogen Plain, hydroxy and R ^E Substituent groups of the group are substituted;

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, wherein said C ₁ -C ₈ The alkyl groups may optionally be substituted with one or more groups each selected fromA substituent substituted by the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl.

A is selected from the group consisting of: -CN,

37.R ^1a Selected from the group consisting of:

38.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

39.R ^1b Is hydrogen.

40.R ^1a And R is ^1b Joined together to form

41.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

42.R ^3a Selected from the group consisting of:

43.R ³ is a 4-10 membered heterocycle.

44.R ³ Selected from the group consisting of:

/>

45.R ² selected from the group consisting of:

/>

and R is ^1b H.48. The compounds are represented by: />

49. The compounds are represented by:

wherein R is ^G3 Selected from the group consisting of: H. c (C) _1-6 Alkyl, C _3-6 Cycloalkyl, phenyl and heterocycle; and R is ^G2 is-NH (C=O) R ^m Wherein R is ^m Selected at each occurrence from H, methyl or CF ₃ For example R ^G2 Is that

50. The compounds are represented by:

51. The compounds are represented by:

52.R ^G3 Selected from the group consisting of:

53.R ^G2 selected from the group consisting of:

54. The compound is selected from the group consisting of:

/>

58. The viral infection is a coronavirus infection.

60. The viral infection is SARS-CoV-2.

61. The viral infection is an arenavirus infection.

63. The viral infection is an influenza infection.

64. The influenza is influenza H1N1, H3N2 or H5N1.

66. The method further comprises administering another therapeutic agent.

72. The compounds are administered prior to viral exposure.

73. The compounds are administered after viral exposure.

9. Covered embodiments

1. a protease inhibitor compound represented by:

wherein:

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^A independently at each occurrence selected from halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-NHC(O)O(C ₁ -C ₈ Alkyl) R ^B -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocycle), -C (O) -O- (4-10 membered heterocycle), -C (O) - (4-10 membered heteroepoxy), -C (O) -OC (CH) ₃ ) ₃ 、-C(O)-(C ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl group, C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle, wherein R is ^B The alkyl, heterocycle, heteroaryl or aryl groups may optionally be substituted with one, two or three halogens, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Substituents for hydroxy or oxo;

R ² selected from the group consisting of: -NHC (O) R bound through a carbon or nitrogen atom ^B 、-NHC(O)N(R ^B ) ₂ 、-NHC(O)C(R ^C ) ₂ R ^B 、-NHS(O) ₂ R ^B 、-O-(C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl), 4-10 membered heterocycle, C ₆ -C ₁₄ Aryl and 5-10 membered heteroaryl, wherein R ^B Or R is ² Can optionally be one, two or three of each selected from R ^x Is substituted by a substituent of (a);

or R is ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered mono-or bicyclic heterocycle having a ring nitrogen, NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein the cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a);

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano, -O- (R) ^xx )-OCH ₃ 、-OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl and 4-10 membered heterocycle, wherein the alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl;

R ^G selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocycle, C ₁ -C ₆ Alkoxy, wherein R is ^m At each occurrence selected from H, C _1-3 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo), phenyl (optionally substituted with halo), -S (O) ₂ -CH ₃ 、C _3-6 Cycloalkyl and 5-6 membered heteroaryl), -C (=o) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano and C ₁ -C ₆ Alkoxy), C (=o) -C _3-6 Cycloalkyl and C (=O) - (5-6 membered heteroaryl) (optionally substituted with halo, cyano, hydroxy, NH ₂ 、C _1-6 Alkyl, C _3-6 Cycloalkyl, C ₁ -C ₆ Alkoxy and C _1-6 Haloalkyl substitution);

R ^xx is- (OCH) ₂ CH ₂ ) _nn -wherein nn is selected from 1, 2, 3, 4, 5 and 6;

R ^y independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl group,C ₁ -C ₈ Heteroalkyl, -CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH;

a is a warhead;

pharmaceutically acceptable salts, stereoisomers, esters and prodrugs thereof.

2. The compounds are represented by:

3. the compounds are represented by:

4. the compounds are represented by:

5. the compounds are represented by:

6. the compounds are represented by:

7. the compounds are represented by:

8. the compounds are represented by:

9. The compounds are represented by:

wherein pp is selected from 0, 1, 2 and 3.

10. The compounds are represented by:

wherein ss is selected from 0, 1, 2 and 3 and mm is selected from 1, 2 and 3.

R ^E selected from the group consisting ofIs set of (3): c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally one, two or three are each selected from halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ Substitution of the substituent of the alkoxy group;

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 memberedHeterocycles and 5-10 membered heteroaryl groups.

A is selected from the group consisting of: -CN,

/>

13.R ^1a Selected from the group consisting of:

/>

14.R ^1a is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

15.R ^1b Is hydrogen.

16.R ^1a And R is ^1b Joined together to form

17.R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

18.R ^3a Selected from the group consisting of:

19.R ³ is a 4-10 membered heterocycle.

20.R ³ Selected from the group consisting of:

/>

21.R ² selected from the group consisting of:

/>

22.R ^1a and R is ² Joined together to form a heterocycle selected from the group consisting of:

/>

and R is ^1b H.

23.R ^G Selected from the group consisting of: H. c (C) _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of-C (=o), halo, cyano, -NR ^m R ^m and-NH (c=o) R ^m ) And C (=O) -C _1-6 Alkyl (optionally substituted with one, two or three substituents each independently selected from the group consisting of halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl and heterocycle, wherein R ^m Selected at each occurrence from H and C _1-3 Alkyl (optionally substituted with one, two or three halogens (e.g., F)), or C ₃ -C ₆ Cycloalkyl (optionally substituted with one, two or three F).

24.R ^G Selected from the group consisting of: -C (O) -monocyclic 5-6 membered or-C (O) -bicyclic heteroaryl, each having at least one ring nitrogen and optionally substituted with one, two or three substituents each selected from halo, methoxy, cyano and hydroxy; -C (O) -C (R) ⁵⁵ R ⁵⁶ )-NH-C(O)-R ⁵⁷ Wherein R is ⁵⁵ Is H and R ⁵⁶ Is straight-chain or branched C _1-5 Alkyl (optionally substituted with halo), or R ⁵⁵ And R is ⁵⁶ Together with the carbon to which it is attached form C ₃ -C ₅ Cycloalkyl (optionally substituted by halo) and wherein R ⁵⁷ Is C _1-3 Alkyl (optionally substituted with one, two or three halo).

25.R ^G Selected from the group consisting of:

26. a compound selected from the group consisting of:

/>

27. a method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of any of the compounds of the embodiments.

28. The viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

29. The viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

30. The viral infection is a coronavirus infection.

31. The viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

32. The viral infection is SARS-CoV-2.

33. The viral infection is an arenavirus infection.

34. The arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

35. The viral infection is an influenza infection.

36. The influenza is influenza H1N1, H3N2 or H5N1.

37. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release, comprising administering to a patient carrying the virus a therapeutically effective amount of any of the compounds of embodiments, and/or contacting an effective amount of any of the compounds of embodiments with a virus-infected cell.

38. The method further comprises administering another therapeutic agent.

39. The method further comprises administering an additional antiviral therapeutic agent.

40. The antiviral therapeutic agent is selected from the group consisting of: ribavirin, famprivir, ST-193, oseltamivir, zanamivir, pamil, danoprevir, ritonavir, radenpyr, bivalirudin, mitsubishi, etila, emtricitabine, tenofovir disoproxil, tenofovir alafenamide hemi-fumarate, abacavir, dolutegravir, efavirenz, elbavir, ledipavir, ganciclovir, sofosbuvir, bitaravir, dasabacavir, lamivudine, atazanavir, obetavir, lamivudine, stavudine, nevirapine, rilpivirine, ziprairivir, cimiravir, dacarbavir Grazopicir, pirenzvir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafavir, cabazitaxel, cytarabine, ecalciline, epigallocatechin gallate, itravir, fotif Sha Wei, gemcitabine, grifficine, isoprinosine, indinavir, malaevir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, nootkatole, pulefafur, PRO 140, raltegafur, prazidine, saquinavir, tebuvir, TNX-355, valacyclovir, VIR-576, and zalcitabine.

41. The other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

42. The additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibody, enfu Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivir, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampicin, zanamivir, oseltamivir Weidan nopuvir, ritonavir, radciclovir, cobalastat, eptifibatide, emtricitabine, tenofovir disoproxil, tenofovir, valavermectin, abacavir, dulutegravir, efavirenz, ganciclovir, sofosbuvir, bitorubivir, dasabavir, lamivudine, atazanavir, obuwei, obutyvir, emtricitabine, tenofovir, valacil, valvulafir, and valvulafir lamivudine, stavudine, nevirapine, rilpivirine, palivir, cimlopivir, dacarbazine, pezopivir, pirenz-tavir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafrevir, cabavir, cytarabine, eleganciclovir, epigallocatechin gallate, itraprine, ferti Sha Wei, gemcitabine, grifeis, isoprinosine, indinavir, malavir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, norelvir, praxafovir, PRO 140, raltegafur, prazidine, quinavir, telbivudine, TNX-355, valacyclovir, pir-576 and zanciclovir.

43. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of a compound of the embodiment.

44. The compounds are administered prior to viral exposure.

45. The compounds are administered after viral exposure.

Examples

The compounds described herein may be prepared in a variety of ways based on the teachings contained herein and synthetic procedures known in the art. In the description of the synthetic methods described below, it is to be understood that all of the reaction conditions (including the choice of solvents, reaction atmospheres, reaction temperatures, experimental durations, and treatment procedures) set forth may be selected as standard conditions for the reaction, unless otherwise indicated. Those skilled in the art of organic synthesis will appreciate that the functional groups present on the various parts of the molecule should be compatible with the reagents and reactants presented. Substituents that are not compatible with the reaction conditions will be apparent to those skilled in the art and thus indicate alternative methods. The starting materials in the examples are commercially available or are readily prepared from known materials by standard methods.

At least some of the compounds identified herein as "intermediates" are contemplated as compounds of the present disclosure.

Recording is carried out at ambient temperature using, for example, a Varian Unity Inova (400 MHz) spectrometer (for example compounds) and Bruker Avance DRX (400 MHz) spectrometer or Bruker Avance DPX (300 MHz) spectrometer (for intermediate compounds) with triple resonance 5mm probe ¹ H NMR spectrum. Chemical shifts are expressed in ppm relative to tetramethylsilane. The following abbreviations are used: br=broad signal, s=singlet, d=doublet, dd=doublet, dt=doublet, and dt=doubletDdd=doublet doublet, t=triplet, td=doublet triplet, q=quartet, m=multiplet.

Abbreviations:

AcOH acetic acid

Boc protecting group

CbzCl benzoic acid benzyl ester

DCE dichloroethane

DCM dichloromethane

DDQ 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone

DIEA N, N-diisopropylethylamine

DIPEA N, N-diisopropylethylamine

DMA dimethylacetamide

DMAP 4-dimethylaminopyridine

DMF dimethylformamide

EA ethyl acetate

EtOAc ethyl acetate

EDCI 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide

EDTA ethylenediamine tetraacetic acid

EtOH ethanol

FA formic acid

HATU 3-oxidized hexafluorophosphoric acid (1- [ bis (dimethylamino) methylene ] -1H-1,2, 3-triazolo [4,5-b ] pyridinium

HOBt hydroxybenzotriazole

LiHMDS lithium bis (trimethylsilyl) amide

MTBA 1-4- (3-methyltriazene) benzoic acid

MTBE methyl tert-butyl ether

MeOH methanol

MeCN acetonitrile

MS mass spectrum

NMR nuclear magnetic resonance

PE Petroleum ether

PMA phosphomolybdic acid

PMBCl p-methoxybenzyl chloride

Pht phthalyl group

PyBOP (benzotriazol-1-yloxy tripyrrolidinylphosphonium hexafluorophosphate)

t-BuLi tert-butyllithium

T ₃ P-propane phosphoric anhydride

TEA triethylamine

TFA trifluoroacetic acid

TFAA trifluoroacetic anhydride

THF tetrahydrofuran

TMSCN trimethylcyanosilane

General chemical method

Exemplary compounds described herein, including the preparation of intermediates and the preparation of the accompanying examples, can be obtained by the general synthetic methods illustrated in scheme 1.

Synthetic scheme

Scheme 1

Scheme 1 illustrates an exemplary preparation of C-1. Allowing a solution of amine A-1 and acid B-1 to react with a coupling agent (such as T ₃ P, EDCI/HOBt) in the presence of a base (such as TEA, DMAP and DIEA) and a solvent (such as DMF and DCM) to give C-1.

In scheme 1, examples of a include substituted or unsubstituted alkyl groups and substituted or unsubstituted cycloalkyl groups, examples of B include warhead moieties such as cyano, aldehyde, hydroxymethyl ketone, ketoamide, heteroaryl ketone, ketene, and Michael acceptor (Michael accepter) warheads, examples of C include alkyl substituted 4-, 5-, or 6-membered lactams, and examples of D include substituted or unsubstituted bicyclic heteroaryl moieties. In scheme 1, an exemplary preparation of the cyano moiety at B includes dehydrating the amide using a dehydrating agent, such as a bergss reagent, to produce a nitrile.

The compounds of tables 1 and 2 have been prepared according to general scheme 1 following the examples described below (such as examples 19, 25, 27, 32, 39 and 41).

Example 1 Synthesis of viral protease inhibitor Compound 103

Step 1: (2S) -2- [ [ (2S) -2- (1H-benzimidazole-2-carbonylamino) -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (200 mg,483.81 mol,1 eq, TFA) and 1H-benzimidazole-2-carboxylic acid (94.14 mg,580.57 mol,1.2 eq) in DCM (2 mL) was added EDCI (185.49 mg,967.61 mol,2 eq) and DMAP (118.21 mg,967.61 mol,2 eq). DMF (1 mL) was added to the mixture and stirred at 25℃for 4h. The mixture obtained is treated with H ₂ O (20 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM/meoh=5/1) to give (2S) -2- [ [ (2S) -2- (1H-benzimidazole-2-carbonylamino) -4-methyl-pentanoyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (150 mg,338.22 umol).

Step 2: n- [ (1S) -3-methyl-1- [ [ (1S) -1- (nitrosomethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] butyl ] -1H-benzimidazole-2-carboxamide

To (2S) -2- [ [ (2S) -2- (1H-benzimidazole-2-carbonylamino) -4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to methyl propionate (150 mg,338.22umol,1 eq) ₃ MeOH (7M, 5mL,103.48 eq.). The mixture was stirred in a sealed tube at 80℃for 16h. The reaction was concentrated to dryness in vacuo to give the compound N- [ (1S) -3-methyl-1- [ [ (1S) -1- (nitrosomethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]Carbamoyl group]Butyl group]-1H-benzimidazole-2-carboxamide (140 mg, crude). The crude product was used directly in the next step.

Step 3: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -1H-benzimidazole-2-carboxamide

To N- [ (1S) -3-methyl-1- [ [ (1S) -1- (nitrosomethyl) -2- [ (3S) -2-oxo-pyrrolidin-3-yl]Ethyl group]Carbamoyl group]Butyl group]To DCM (5 mL) of 1H-benzimidazole-2-carboxamide (120.00 mg,280.06umol,1 eq.) was added the Bungeus reagent (150 mg,629.45umol,2.25 eq.). The mixture was stirred at 25℃for 4h. At N ₂ Blow-drying the reaction. By preparative HPLC (column: waters Xbridge Prep OBD C: 150 x 40mm x 10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -40%,8 min) to give N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-1H-benzimidazole-2-carboxamide (40 mg,97.45 umol). MS (ESI) m/z 411.1[ M+H ]] ⁺ , ¹ H NMR(400MHz,DMSO-d ₆ )δppm 13.11(br s,1H),8.97-8.81(m,2H),7.90-7.64(m,2H),7.54(br s,1H),7.31(br s,2H),5.08-4.93(m,1H),4.62-4.43(m,1H),3.19-3.05(m,2H),2.44-2.29(m,1H),2.23-2.05(m,2H),1.91-1.50(m,5H),0.91(dd,J＝6.3,8.9Hz,6H)。

EXAMPLE 2 Synthesis of viral protease inhibitor Compound 105

Step 1: (2S) -2- [ [ (2S) -4-methyl-2- (2-naphthylsulfonylamino) pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (150 mg,501.06umol,1 eq.) in DMF (5 mL) was added naphthalene-2-sulfonyl chloride (227.16 mg,1.00mmol,2 eq.) and DMAP (155.35 mg,1.27mmol,2.54 eq.) and stirred at 25 ℃. The reaction was then stirred at 80℃for 16h. H for the reaction mixture ₂ O (20 mL) was diluted and extracted with EtOAc (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM/meoh=10/1) purification residue. To obtain (2S) -2- [ [ (2S) -4-methyl-2- (2-naphthylsulfonylamino) pentanoyl as an oil]Amino group]-3-[(3S) -2-oxo-pyrrolidin-3-yl]Methyl propionate (70 mg,142.98 umol).

Step 2: (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -4-methyl-2- (2-naphthylsulfonylamino) pentanamide

To (2S) -2- [ [ (2S) -4-methyl-2- (2-naphthylsulfonylamino) pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to a mixture of methyl propionate (50 mg,102.13umol,1 eq) ₃ MeOH (7M, 10mL,685.42 eq.) and stirred at 80℃for 16h. The reaction was concentrated to dryness in vacuo to give the crude (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as an oil]Methyl group]Ethyl group]-4-methyl-2- (2-naphthylsulfonylamino) valeramide (50 mg, crude).

Step 3: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4-methyl-2- (2-naphthylsulfonylamino) pentanamide

To (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To DCM (0.5 mL) of-4-methyl-2- (2-naphthylsulfonylamino) valeramide (70 mg, 147.50. Mu. Mol,1 eq.) was added a Buerger's reagent (79.00 mg, 331.52. Mu. Mol,2.25 eq.). The mixture was stirred at 25℃for 4h. At N ₂ Blow-drying the reaction. The residue was purified by preparative HPLC: column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55% and 8min to obtain solid compound (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-pyrrolidin-3-yl]Ethyl group]-4-methyl-2- (2-naphthylsulfonylamino) pentanoamide (30 mg,65.71 umol). MS (ESI) m/z 457.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 8.81(br d,J＝7.5Hz,1H),8.38(s,1H),8.21(br s,1H),8.12-8.03(m,2H),8.00(d,J＝7.7Hz,1H),7.82-7.72(m,1H),7.71-7.56(m,3H),4.64(q,J＝7.6Hz,1H),3.78-3.67(m,1H),3.09-3.01(m,1H),3.00-2.89(m,1H),2.08-1.96(m,1H),1.90-1.78(m,1H),1.71-1.60(m,1H),1.58-1.33(m,4H),1.31-1.19(m,1H),0.78(d,J＝6.6Hz,3H),0.63(d,J＝6.6Hz,3H)。

EXAMPLE 3 Synthesis of benzyl N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] carbamate

Step 1: (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (300 mg,1.05mmol,1 eq.) was added to a mixture of TFA (4.62 g,40.52mmol,3mL,38.67 eq.) in DCM (5 mL), and the mixture was stirred at 25℃for 2h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give a residue and used in the next step. The compound (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a colorless oil]Methyl propionate (180 mg,918.33 umol). MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (189.47 mg,966.66 mol) and (2S) -2- (benzyloxycarbonylamino) -4-methyl-pentanoic acid (256.46 mg,966.66 mol,1 eq.) in DCM (2 mL) was added DMAP (236.19 mg,1.93mmol,2 eq.) and EDCI (370.62 mg,1.93mmol,2 eq.). DMF (1 mL) was added to the mixture and stirred at 25℃for 14h. After the reaction was completed, the reaction mixture was treated with H ₂ O (50 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Purification of the residue by column chromatography (SiO ₂ Petroleum ether/EtOAc=3/1 to 0/1) to give the compound (2S) -2- [ [ (2S) -2- (benzyloxycarbonylamino) -4-methyl-pentanoyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,461.36 umol). MS (ESI) m/z 434.3[ M+H ]] ⁺

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] carbamic acid phenylmethyl ester

To (2S) -2- [ [ (2S) -2- (benzyloxycarbonylamino) -4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]NH was added to methyl propionate (200 mg,369.09umol,1 eq) ₃ MeOH (7M, 58.14mL,1102.58 eq.). The mixture was stirred at 80℃for 16h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give a residue which was used directly in the next step. The compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl was obtained as a colorless oil]Methyl group]Ethyl group]Carbamoyl group]-3

Benzyl methyl-butyl ] carbamate (150 mg,322.59 umol).

Step 4: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] carbamic acid phenylmethyl ester

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of benzyl carbamate (150 mg,179.22umol,1 eq.) in DCM (5 mL) was added the Buerger reagent (42.71 mg,179.22umol,1 eq.). The mixture was stirred at 25℃for 1h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a residue. By neutral prep HPLC (column: waters Xbridge BEH C18 100. Times.30 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the compound N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as solid ]Ethyl group]Carbamoyl group]-3-methyl-butyl]Benzyl carbamate (28 mg,69.92 umol). MS (ESI) m/z 401.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 8.84(br d,J＝7.9Hz,1H),7.70(s,1H),7.54(br d,J＝7.8Hz,1H),7.41-7.24(m,5H),5.02(s,2H),4.97-4.88(m,1H),4.07-3.91(m,1H),3.20-2.94(m,2H),2.38-2.22(m,1H),2.22-1.98(m,2H),1.85-1.26(m,5H),0.87(br dd,J＝6.5,11.2Hz,6H)

EXAMPLE 4 Synthesis of viral protease inhibitor Compound 131

Step 1: (2S) -2- [ [ (2S) -2- (1H-imidazo [4,5-b ] pyridine-2-carbonylamino) -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,604.76umol,1 eq. TFA) and 1H-imidazo [4,5-b]To a mixture of pyridine-2-carboxylic acid (118.39 mg,725.71umol,1.2 eq.) in DCM (4 mL) was added EDCI (231.86 mg,1.21mmol,2 eq.) and DMAP (147.77 mg,1.21mmol,2 eq.). DMF (2 mL) was added to the mixture and stirred at 25℃for 4h. H for the reaction mixture ₂ O (20 mL) was diluted and extracted with DCM (30 mL). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM/meoh=5/1) to give the compound (2S) -2- [ [ (2S) -2- (1H-imidazo [4, 5-b) as a solid]Pyridine-2-carbonylamino) -4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (100 mg,224.98 umol).

Step 2: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -1H-imidazo [4,5-b ] pyridine-2-carboxamide

To (2S) -2- [ [ (2S) -2- (1H-imidazo [4, 5-b)]Pyridine-2-carbonylamino) -4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to a mixture of methyl propionate (100 mg,224.98umol,1 eq) ₃ MeOH (7M, 27.54mL,856.77 eq.) and stirred at 80℃for 16h. The reaction was concentrated to dryness in vacuo to give the crude N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as an oil]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-1H-imidazo [4,5-b]Pyridine-2-carboxamide (90 mg, crude material).

Step 3: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -1H-imidazo [4,5-b ] pyridine-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-1H-imidazo [4,5-b]To DCM (3 mL) of pyridine-2-carboxamide (80 mg,186.28umol,1 eq.) was added a Buerger's reagent (100.00 mg,419.62umol,2.25 eq.). The mixture was stirred at 25℃for 4h. At N ₂ Blow-drying the reaction. By preparative HPLC (column: waters Xbridge Prep OBD C: 150 x 40mm x 10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:10% -35%,8 min) to give N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-1H-imidazo [4,5-b]Pyridine-2-carboxamide (25 mg,60.76 umol). MS (ESI) m/z 412.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 13.58(br s,1H),9.29-8.96(m,1H),8.89(d,J＝7.9Hz,1H),8.49(br s,1H),8.28-7.84(m,1H),7.71(s,1H),7.36(dd,J＝4.6,8.2Hz,1H),5.06-4.93(m,1H),4.61-4.44(m,1H),3.20-3.06(m,2H),2.43-2.31(m,1H),2.20-2.07(m,2H),1.90-1.53(m,5H),0.92(dd,J＝6.4,9.5Hz,6H)。

EXAMPLE 5 Synthesis of viral protease inhibitor Compound 121

Step 1: (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.2 g,3.77 mmol) in THF (3 mL), ACN (3 mL) and H ₂ LiOH.H was added to the mixture in O (3 mL) ₂ O (158.29 mg,3.77mmol,1 eq.). The mixture was stirred at 25℃for 2h. After the reaction was completed, the solution was concentrated to give a residue, and then the residue was adjusted to pH-4 with HCl. The resulting residue was extracted with EtOAc (20 mL x 3) and brine (20 mL) and then concentrated to give the residual compound (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl as an oil]Propionic acid (1 g,3.31 mmol). MS (ESI) m/z 217.1[ M+H-56 ] ] ⁺ 。

Step 2: n- [ (1S) -2- [ methoxy (methyl) amino ] -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamic acid tert-butyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of propionic acid (1.0 g,3.31 mmol) in DCM (20 mL) was added CDI (535.94 mg,3.31mmol,1 eq). The mixture was stirred at 0deg.C for 30min, followed by the addition of DIEA (512.61 mg,3.97mmol,690.85uL,1.2 eq.) and N, O-dimethylhydroxylamine hydrochloride (322.40 mg,3.31mmol,1 eq.). The resulting mixture was stirred at 25℃for 3h. After the reaction was completed, the reaction mixture was treated with H ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=5/1 to 0/1 to give the compound N- [ (1S) -2- [ methoxy (methyl) amino ] as an oil]-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Tert-butyl carbamate (0.9 g,2.57 mmol). MS (ESI) m/z 316.2[ M+H ]] ⁺

Step 3: n- [ (1S) -2- (1, 3-benzothiazol-2-yl) -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamic acid tert-butyl ester

at-78deg.C, at N ₂ To a mixture of 2-bromo-1, 3-benzothiazole (458.22 mg,2.14mmol,1.5 eq.) in THF (20 mL) was added n-BuLi (2.5 m,684.92 μl,1.2 eq.) in one portion. The mixture was stirred at-78 ℃ for 30min, and then N- [ (1S) -2- [ methoxy (methyl) amino ] was added at-78 ℃]-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Tert-butyl carbamate (500 mg,1.43 mmol). The resulting mixture was stirred for 1 hour and then at 0 ℃ by adding NH ₄ Cl (10 mL) quench the reaction mixture and then stir at 0deg.C for 10min. The resulting mixture was diluted with water (100 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By MPLC (SiO) ₂ StoneThe residue was purified with oleyl ether/EtOAc-meoh=10/1 to 0/1 to give the compound N- [ (1S) -2- (1, 3-benzothiazol-2-yl) -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]Tert-butyl carbamate (150 mg,346.63 umol). MS (ESI) m/z 390.1[ M+H ]] ⁺

Step 4: (3S) -3- [ (2S) -2-amino-3- (1, 3-benzothiazol-2-yl) -3-oxo-propyl ] pyrrolidin-2-one

To N- [ (1S) -2- (1, 3-benzothiazol-2-yl) -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a mixture of tert-butyl carbamate (150 mg,346.63 mol) was added HCl/EtOAc (4M, 86.66uL,1 eq). The resulting mixture was stirred at 20 ℃ for 2h and then concentrated under reduced pressure to give the residue (3S) -3- [ (2S) -2-amino-3- (1, 3-benzothiazol-2-yl) -3-oxo-propyl) as an oil]Pyrrolidin-2-one (100 mg, crude) was used directly in the next step. MS (ESI) m/z 290.1[ M+H ]] ⁺

Step 5: n- [ (1S) -1- [ [ (1S) -2- (1, 3-benzothiazol-2-yl) -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 0℃to (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (18.93 mg, 62.21. Mu. Mol,1 eq.) in DMF (1 mL) was added 1-methylimidazole (25.54 mg, 311.04. Mu. Mol, 24.79. Mu.L, 5 eq.) and hexafluorophosphoric acid [ chloro (dimethylamino) methylene ]]Dimethyl-ammonium (20.95 mg,74.65umol,1.2 eq.). The resulting mixture was stirred at 0deg.C for 30min and then (3S) -3- [ (2S) -2-amino-3- (1, 3-benzothiazol-2-yl) -3-oxo-propyl was added ]Pyrrolidin-2-one (18 mg,62.21umol,1 eq). The resulting mixture was stirred at 25℃for 2h. After the reaction was completed, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By neutral preparative HPLC (column: waters Xbridge BEH C18 100. Times.30 mm. Times.10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) and SFC (column: DAICEL CHIRALCEL OX (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ OMEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,12 min) to obtain solid crude materialThe compound N- [ (1S) -1- [ [ (1S) -2- (1, 3-benzothiazol-2-yl) -2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (8 mg,13.48 umol). MS (ESI) m/z 576.3[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69(s,1H),8.75-8.51(m,2H),8.08(d,J＝7.9Hz,1H),7.95(d,J＝8.2Hz,1H),7.68(s,1H),7.50(t,J＝7.4Hz,1H),7.44-7.37(m,1H),7.19-7.07(m,4H),6.93(d,J＝8.2Hz,1H),6.49(d,J＝7.7Hz,1H),3.89(s,3H),3.15-2.99(m,2H),2.46-2.30(m,1H),2.21-1.94(m,4H),1.93-1.74(m,1H),1.57-1.40(m,2H),0.83-0.71(m,6H)。

EXAMPLE 6 Synthesis of viral protease inhibitor Compound 185

Step 1: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclohexylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (170 mg,763.47 mol,1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionic acid (207.17 mg,763.47 mol,1 eq) in DMF (2 mL) were added DMAP (186.55 mg,1.53mmol,2 eq) and EDCI (292.71 mg,1.53mmol,2 eq). DCM (3 mL) was added to the mixture and stirred at 25 ℃ for 2h. LCMS showed the reaction was complete and the desired MS was observed. By adding H at 0 ℃ ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/etoac=0/1) to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,568.77umol,74.50% yield). MS (ESI) m/z 440.3[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2-amino-3-cyclohexylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionyl at 25 ℃]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (200 mg,455.02umol,1 eq.) in EtOAc (0.5 mL) was added dropwise HCl/EtOAc (4M, 2.00mL,17.58 eq.). The mixture was stirred at 25℃for 1h. Concentrating the reaction mixture under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-3-cyclohexyl-propionyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, crude, HCl) and was used directly in the next step. MS (ESI) m/z 340.1[ M+H ] ] ⁺

Step 3: ((S) -2- ((S) -3-cyclohexyl-2- (4-methoxy-1H-indole-2-carboxamide) propionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (99.18 mg,518.77 mol,1.3 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclohexyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (150 mg, 399.05. Mu. Mol,1 eq., HCl) in DMF (2 mL) was added DMAP (97.50 mg, 798.11. Mu. Mol,2.0 eq.) and EDCI (153.00 mg, 798.11. Mu. Mol,2 eq.). DCM (4 mL) was added to the mixture and stirred at 25 ℃ for 2h. By adding H at 0 ℃ ₂ O (20 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give the product (2S) -2- [ [ (2S) -3-cyclohexyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg,292.63umol,73.33% yield). ¹ H NMR (methanol-d) ₄ ,400MHz)：δppm 7.26(s,1H),7.09-7.20(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.6Hz,1H),4.66(br dd,J＝9.0,6.3Hz,1H),4.52-4.58(m,1H),3.93(s,3H),3.72(s,3H),3.22-3.29(m,2H),2.54-2.62(m,1H),2.26-2.33(m,1H),2.15-2.23(m,1H),1.66-1.87(m,9H),1.47-1.54(m,1H),1.25-1.40(m,3H),0.96-1.06(m,2H)

Step 4: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclohexyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

In a sealed tube, (2S) -2- [ [ (2S) -3-cyclohexyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (150 mg,292.63 mmole, 1 eq.) in ammonia (15.30 g,898.39mmol,15.00mL,3070.07 eq.) was heated at 80℃for 12 hours. The reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (140 mg, crude). MS (ESI) m/z 498.2[ M+H ]] ⁺

¹ H NMR (methanol-d) ₄ ,400MHz)：δppm 7.27-7.34(m,1H),7.13-7.20(m,1H),7.05(d,J＝8.3Hz,1H),6.53(d,J＝7.7Hz,1H),4.62(t,J＝7.6Hz,1H),4.42-4.51(m,1H),3.95(s,3H),3.22-3.30(m,2H),2.53(td,J＝9.2,4.5Hz,1H),2.33(ddd,J＝9.2,6.4,3.4Hz,1H),2.17(ddd,J＝14.1,11.4,4.6Hz,1H),1.71-1.88(m,9H),1.46-1.53(m,1H),1.21-1.32(m,3H),0.97-1.09(m,2H)

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclohexyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (80 mg,160.78umol,1 eq.) in DCM (3 mL) was added the berg reagent (114.94 mg,482.33umol,3 eq.) and the resulting mixture was then stirred at 25 ℃ for 3H. The reaction mixture was concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gives the product N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid ]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (20.02 mg,41.75 umol). MS (ESI) m/z 480.1[M+H] ⁺ 。

Preparative HPLC conditions: column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-60％，10min

¹ H NMR (methanol-d) ₄ ,400MHz)：δppm 7.28(s,1H),7.11-7.18(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.6Hz,1H),5.05(dd,J＝10.1,5.9Hz,1H),4.56-4.61(m,1H),3.93(s,3H),3.22-3.30(m,2H),2.55-2.66(m,1H),2.23-2.40(m,2H),1.65-1.94(m,9H),1.41-1.52(m,1H),1.17-1.36(m,3H),0.94-1.10(m,2H)。

EXAMPLE 7 Synthesis of viral protease inhibitor Compound 101

Step 1: methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate; hydrochloride salt

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl at 25 ℃]To methyl propionate (500 mg,1.75mmol,1 eq.) was added HCl/EtOAc (4M, 10mL,22.91 eq.). The mixture was stirred at 25℃for 0.5h. Concentrating the resulting mixture under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl propionate; hydrochloride (300 mg,1.28mmol,73.29% yield, 95% purity) and used directly in the next step. MS (ESI) m/z 187.1[ M+H ]] ⁺

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate; EDCI (258.28 mg,1.35mmol,2 eq.) and DMAP (164.60 mg,1.35mmol,2 eq.) were added to a mixture of hydrochloride (157.89 mg,673.65umol,95% purity, 1 eq.) and (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoic acid (155.81 mg,673.65umol,1 eq.) in DMF (2 mL). DCM (3 mL) was added to the mixture and stirred at 25 ℃ for 14h. The mixture obtained is treated with H ₂ Dilute with O (50 mL) and useDCM (30 ml×3) was extracted. The combined organic layers were washed with brine, dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2, petroleum ether/etoac=3/1 to 1/1) to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,500.65umol,74.32% yield, 80% purity). MS (ESI) m/z 400.3[ M+H ]] ⁺

Step 3: (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4-methyl-pentanoic acid amide

To N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl group at 25 ℃C]Ethyl group]Carbamoyl group]-3-methyl-butyl]To DCM (5 mL) of tert-butyl carbamate (200 mg,491.19umol,90% purity, 1 eq.) was added TFA (770.00 mg,6.75mmol,0.5mL,13.75 eq.). The mixture was stirred at 25℃for 1h. The resulting mixture was concentrated under reduced pressure to give the product (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as an oil]Ethyl group]-4-methyl-pentanamide (120 mg,405.50umol,82.55% yield, 90% purity) and used directly in the next step. MS (ESI) M/z300.2[ M+H ] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (120 mg,627.67umol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (208.78 mg,627.67umol,90% purity, 1 eq.) in DCM (1 mL) was added EDCI (240.65 mg,1.26mmol,2 eq.) and DMAP (153.36 mg,1.26mmol,2 eq.). DMF (0.5 mL) was added to the mixture and stirred at 25℃for 14h. The mixture obtained is treated with H ₂ O (50 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2, petroleum ether/etoac=3/1 to 0/1) to give the residue as a solidThe solid compound (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (160 mg,304.74umol,48.55% yield, 90% purity). MS (ESI) m/z 473.3[ M+H ]] ⁺

Step 5: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]3- [ (3S) -2-Oxopyrrolidin-3-yl]NH was added to methyl propionate (180 mg,342.83umol,90% purity, 1 eq) ₃ MeOH (7M, 54.00mL,1102.58 eq.) and the mixture was stirred at 80℃for 16h. The resulting mixture was concentrated under reduced pressure to give the residue N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as an oil]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (130 mg,255.73umol,74.59% yield, 90% purity). MS (ESI) m/z 458.3[ M+H ]] ⁺

1H NMR (400 MHz, methanol-d 4) delta ppm 0.97-1.02 (dd, J=14.55, 6.11Hz, 6H) 1.74-1.82 (m, 5H) 2.15 (ddd, J=14.03, 11.34,4.58Hz, 1H) 2.25-2.37 (m, 1H) 2.52 (ddt, J=13.82, 9.41,4.71 Hz, 1H) 3.17-3.29 (m, 2H) 3.90 (s, 3H) 4.46 (dd, J=11.25, 4.16Hz, 1H) 4.60 (dd, J=9.66, 5.01Hz, 1H) 6.50-6.52 (d, J=7.70 Hz, 1H) 7.02-7.04 (d, J=8.31 Hz, 1H) 7.15-7.17 (m, 1.90 (s, 3H) 4.46 (dd, J=11.25, 4.01 Hz, 1H) 4.60 (dd, J=9.70 Hz, 7.01H)

Step 6: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl-3-methyl-butyl-4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (100 mg,196.71 mol,90% purity, 1 eq.) in DCM (4 mL) was added the berg reagent (93.75 mg,393.42 mol,2 eq.). The mixture was stirred at 25℃for 1h. The resulting mixture was concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC to giveTo the product N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23 mg,49.50umol,25.16% yield, 94.59% purity). MS (ESI) m/z 440.1[ M+H ]] ⁺ 。

Preparative HPLC conditions:

column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:27% -57%,10min

1H NMR(400MHz,DMSO-d6)δppm 0.88-0.94(m,6H)1.67-1.74(m,5H)2.11-2.13(m,2H)2.14-2.34(m,1H)3.09-3.14(m,2H)3.88(s,3H)4.36-4.57(m,1H)4.90-5.00(m,1H)6.49-6.51(d,J＝7.58Hz,1H)6.99-7.01(m,2H)7.38(s,1H)7.70(s,1H)8.45-8.47(br d,J＝7.70Hz,1H)8.89-8.91(br d,J＝7.95Hz,1H)11.57(br s,1H)

EXAMPLE 8 Synthesis of viral protease inhibitor Compounds 593

Step 1: (2S) -2-amino-3- (1H-imidazol-5-yl) propionic acid methyl ester

To a solution of (2S) -2- (tert-butoxycarbonylamino) -3- (1H-imidazol-5-yl) propionic acid (0.5 g,1.96mmol,1 eq.) in MeOH (0.6 mL) was added HCl/MeOH (4M, 4.90mL,10 eq.) at 25 ℃. The reaction mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give the product. Methyl (2S) -2-amino-3- (1H-imidazol-5-yl) propionate (400 mg, crude material, HCl) was obtained as a solid and used directly in the next step. MS (ESI) m/z 170.1[ M+H ] ] ⁺

Step 2: (2S) -3- (1H-imidazol-5-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] propionic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoic acid (741.86 mg,1.77mmol,1 eq. TFA) and methyl (2S) -2-amino-3- (1H-imidazol-5-yl) propionate (0.3 g,1.77mmol,1 eq. HCl), DIPEA (1.15 g,8.87mmol,1.54mL,5 eq.) in THF (0.3 mL) and DCM (0.3 mL)T3P (1.69 g,2.66mmol,1.58mL,50% purity, 1.5 eq.) was added to the mixture. The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was washed with brine (3 ml x 3) and dried over anhydrous sodium sulfate and concentrated to give the crude product. Obtaining (2S) -3- (1H-imidazol-5-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]-4-methyl-pentanoyl]Amino group]Methyl propionate (300 mg, crude) and was used directly in the next step. MS (ESI) m/z 456.2[ M+H ]] ⁺

1H NMR (400 MHz, methanol-d) ₄ )δppm 7.48(s,1H),7.27(s,1H),7.11-7.18(m,1H),7.02(d,J＝8.16Hz,1H),6.85(s,1H),6.51(d,J＝7.72Hz,1H),4.60-4.71(m,2H),3.93(s,3H),3.68(s,3H),3.00-3.17(m,3H),1.62-1.78(m,3H),0.97(dd,J＝13.78,6.06Hz,6H)

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-1- (1H-imidazol-5-ylmethyl) -2-oxo-ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward (2S) -3- (1H-imidazol-5-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]-4-methyl-pentanoyl]Amino group]NH was added to methyl propionate (200 mg,439.07umol,1 eq.) at a time ₃ MeOH (7M, 11.76mL,187.56 eq.). The mixture was stirred at 80 ℃ and for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the crude product. Obtaining N- [ (1S) -1- [ [ (1S) -2-amino-1- (1H-imidazol-5-ylmethyl) -2-oxo-ethyl ] as a solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (170 mg,378.83 mol,86.28% yield, 98.16% purity) and used directly in the next step. MS (ESI) m/z 441.2[ M+H ]] ⁺

Step 4: n- [ (1S) -1- [ [ (1S) -1-cyano-2- (1H-imidazol-5-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ (1S) -2-amino-1- (1H-imidazol-5-ylmethyl) -2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (140 mg,317.82 u)To a mixture of DCM (2 mL) was added TFAA (133.51 mg, 635.65. Mu.L, 88.41. Mu.L, 2 eq.). The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated to give a crude product. After 36h of storage, the crude product became compound 593. The residue was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ (1S) -1-cyano-2- (1H-imidazol-5-yl) ethyl in solid form ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23.89 mg,56.31umol,17.72% yield, 99.581% purity). MS (ESI) m/z 423.2[ M+H ]] ⁺

Preparative HPLC conditions:

column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-55％，10min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.58(s,1H),7.30(s,1H),7.12-7.21(m,1H),6.99-7.09(m,2H),6.52(d,J＝7.72Hz,1H),5.05(t,J＝7.06Hz,1H),4.61(br dd,J＝9.70,4.85Hz,1H),3.94(s,3H),3.06-3.21(m,2H),1.60-1.83(m,3H),0.99(dd,J＝13.89,6.17Hz,6H)

Step 5: (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoic acid tert-butyl ester

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (5 g,26.15mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (5.88 g,31.38mmol,1.2 eq., HCl), EDCI (6.52 g,34.00mmol,1.3 eq.), HOBt (4.59 g,34.00mmol,1.3 eq.) in DMF (30 mL) was added TEA (7.94 g,78.46mmol,10.92mL,3 eq.) in one portion. The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (90 mL) and extracted with EtOAc (25 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (25 mL) and 5% aqueous sodium bicarbonate (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether etoac=30:1 to 10:1) purification residue. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid ]-4-methyl-pentanoic acid tert-butyl ester (5.93 g,16.45mmol,62.91% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

¹ H NMR (400 MHz, chloroform-d) delta ppm 9.25 (br s, 1H), 7.10-7.16 (m, 1H), 6.93-7.00 (m, 2H), 6.56 (br d, J=8.31 Hz, 1H), 6.44 (d, J=7.70 Hz, 1H), 4.66 (td, J=8.50, 5.14Hz, 1H), 3.88 (s, 3H), 1.62-1.75 (m, 2H), 1.57-1.62 (m, 1H), 1.42 (s, 9H), 0.92 (dd, J=6.17, 3.85Hz, 6H).

Step 6: (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-valerate (2.00 g,5.55mmol,1 eq.) in DCM (8 mL) was added TFA (10.27 g,90.04mmol,6.67mL,16.23 eq.) and H in one portion ₂ O (666.67 mg,37.01mmol, 666.67. Mu.L, 6.67 eq.). The mixture was stirred at 25 ℃ and for 4h. The reaction mixture was concentrated to give a crude product. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoic acid (2.24 g,5.35mmol,96.50% yield, TFA) and used directly in the next step. MS (ESI) M/z305.1[ M+H] ⁺

Example 9 Synthesis of viral protease inhibitor Compounds 135, 595 and 136

Step 1: n- [ (1S) -1- [ [ (1S) -1- (hydroxymethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (1.5 g,2.86mmol,90% purity, 1 eq.) in THF (20 mL) was added LiBH ₄ (124.45 mg,5.71mmol,2 eq.). The mixture was stirred at 25℃for 2h. After the reaction was completed, H was added at 0deg.C ₂ O (10 mL) quench the reaction mixture and extract with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the residual compound N- [ (1S) -1- [ [ (1) as a solidS) -1- (hydroxymethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (1.0 g,2.25mmol,78.74% yield). MS (ESI) m/z 445.1[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.27(s,1H),7.19-7.10(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.7Hz,1H),4.65-4.53(m,1H),4.05-3.97(m,1H),3.93(s,3H),3.60-3.43(m,2H),3.27-3.10(m,2H),2.59-2.43(m,1H),2.39-2.19(m,1H),2.08-1.89(m,1H),1.85-1.63(m,4H),1.60-1.46(m,1H),1.00(dd,J＝6.1,12.5Hz,6H)。

Step 2: n- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1- (hydroxymethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (674 mg,1.52mmol,1 eq.) in DMSO (25 mL) was added IBX (849.14 mg,3.03mmol,2 eq.). The mixture was stirred at 25℃for 15h. After the reaction was completed, the reaction mixture was treated with H ₂ O (30 mL) was diluted and extracted with EtOAc (30 mL x 2). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Ethyl acetate (10 mL) was added to the residue and filtered to give the product N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (420 mg,759.31umol,50.08% yield, 80% purity). MS (ESI) m/z 443.1[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.27(s,1H),7.20-7.09(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.7Hz,1H),4.60(dt,J＝5.5,9.9Hz,1.5H),4.47(dd,J＝1.4,4.1Hz,0.5H),4.02-3.94(m,1H),3.93(s,3H),3.28-3.15(m,2H),2.54-2.39(m,1H),2.37-2.21(m,1H),2.10-1.93(m,1H),1.89-1.49(m,5H),1.17-0.91(m,6H)。

Step 3: n- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (400 mg,723.15umol,80% purity, 1 eq.) in DCM (10 mL) was added saturated NaHSO ₃ (301.01 mg,2.89mmol, 203.38. Mu.L, 4 eq.). The mixture was stirred at 25℃for 30min, and then KCN (42 mg,644.96umol, 27.63. Mu.L, 8.92e-1 eq.) was added to H ₂ O (0.8 mL). The mixture was stirred at 25℃for 3h. After the reaction was complete, the organic phase was collected and the aqueous layer was extracted with DCM (30 ml x 3). The combined organic phases were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Drying and concentration gave the crude material. NaOH was added to the liquid to ph=9, followed by quenching by adding aqueous NaCl solution, followed by NaOH to pH>14. Purification of the crude material by HCl preparative HPLC gave 120mg of a mixture and SFC isolation gave the compound N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (34 mg,70.96umol,9.81% yield, 97.99% purity) and the compound N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl)]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (64 mg,131.75umol,18.22% yield, 96.66% purity). MS (ESI) m/z 470.2[ M+H ]] ⁺ 。

Preparative HPLC conditions: column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ Water (0.04% HCl) -ACN ]; b%:26% -50%,7min

SFC conditions: column: REGIS (R, R) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-IPA ]; b%:35% -35%,11min

Compound 134 isomer 1: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.57(d,J＝1.8Hz,1H),8.40(d,J＝7.9Hz,1H),8.13(d,J＝9.3Hz,1H),7.57(s,1H),7.36(d,J＝1.5Hz,1H),7.13-7.06(m,1H),7.03-6.97(m,1H),6.69(d,J＝7.3Hz,1H),6.50(d,J＝7.7Hz,1H),4.50-4.40(m,1H),4.33(t,J＝7.8Hz,1H),4.10-3.97(m,1H),3.88(s,3H),3.16-2.98(m,2H),2.39-2.26(m,1H),2.15-2.01(m,1H),1.92-1.80(m,1H),1.80-1.63(m,2H),1.62-1.40(m,3H),0.90(dd,J＝6.3,15.5Hz,6H)。

compound 134 isomer 2: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(br d,J＝1.5Hz,1H),8.35(d,J＝7.9Hz,1H),8.21(d,J＝8.6Hz,1H),7.60(s,1H),7.34(d,J＝1.8Hz,1H),7.12-7.06(m,1H),7.03-6.97(m,1H),6.64(d,J＝6.0Hz,1H),6.50(d,J＝7.5Hz,1H),4.60-4.49(m,2H),4.12-3.96(m,1H),3.88(s,3H),3.19-2.98(m,2H),2.41-2.26(m,1H),2.16-1.95(m,2H),1.92-1.35(m,5H),0.98-0.82(m,6H)。

step 4: [ (2S) -1-hydroxy-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propyl ] sulfonyloxy sodium salt

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (50 mg, 112.99. Mu. Mol,1 eq.) in EtOH (0.4 mL), etOAc (0.2 mL) and H ₂ NaHSO was added to the mixture in O (0.1 mL) ₃ (11.76 mg, 112.99. Mu.L, 7.94. Mu.L, 1 eq.). The mixture was stirred at 80℃for 16h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a residue. DCM (3 mL) and ACN (3 mL) were added to the residue and filtered to give the compound [ (2S) -1-hydroxy-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Sodium sulfonyloxy (5 mg,5.26umol,4.66% yield, 57.5% purity). (ESI) M/z525.1[ M+H] ⁺

¹ H NMR(400MHz,DMSO-d6)δ＝11.67-11.44(m,1H),9.42(s,0.02H),8.52-8.27(m,1H),7.74-7.59(m,1H),7.43(s,1H),7.32(dd,J＝1.8,4.9Hz,1H),7.15-6.93(m,2H),6.50(d,J＝7.7Hz,1H),5.40-5.24(m,1H),4.61-4.33(m,1H),4.31-4.15(m,0.5H),4.11-3.96(m,0.5H),3.94(dd,J＝2.4,5.7Hz,0.5H),3.88(s,3H),3.85-3.81(m,0.5H),3.19-2.94(m,2H),2.27-1.87(m,3H),1.85-1.42(m,5H),0.99-0.79(m,6H)

Step 5: 4-methoxy-N- [ (1S) -3-methyl-1- [ [ (E, 1S) -3-methylsulfonyl-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] allyl ] carbamoyl ] butyl ] -1H-indole-2-carboxamide

At 0℃under N ₂ Down to 1- [ ethoxy (methylsulfonylmethyl) phosphoryl group]To a mixture of oxyethane (130.06 mg,564.96umol,5 eq.) in THF (2 mL) was added n-BuLi (2.5 m,180.79 μl,4 eq.). The mixture was stirred at-75deg.C for 30min, followed by the addition of N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,112.99umol,1 eq). The mixture was stirred at-75℃for 2h. After the reaction was completed, H was added at 0deg.C ₂ O (10 mL) to quench the reaction mixture, and then concentrated under reduced pressure to give a residue. Purification of the residue by preparative HPLC gives the compound 4-methoxy-N- [ (1S) -3-methyl-1- [ [ (E, 1S) -3-methylsulfonyl-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Methyl group]Allyl group]Carbamoyl group]Butyl group]-1H-indole-2-carboxamide (15 mg,28.82umol,25.50% yield, 99.638% purity). (ESI) m/z 519.1[ M+H ]] ⁺ Column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ Water (0.04% HCl) -ACN]；B％：26％-52％，7min

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.33-7.26(m,1H),7.20-7.10(m,1H),7.03(d,J＝8.3Hz,1H),6.85(dd,J＝4.8,15.3Hz,1H),6.68(dd,J＝1.6,15.3Hz,1H),6.52(d,J＝7.7Hz,1H),4.77-4.67(m,1H),4.61-4.50(m,1H),3.99-3.83(m,3H),3.28-3.18(m,2H),3.01-2.88(m,3H),2.65-2.50(m,1H),2.39-2.22(m,1H),2.15-1.97(m,1H),1.91-1.62(m,5H),1.09-0.92(m,6H)

EXAMPLE 10 Synthesis of viral protease inhibitor Compounds 740 and 741

Step 1: (S) -4-chloro-3-oxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) carbamic acid tert-butyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (0.6 g,2.10mmol,1 eq.) in THF (24 mL) was added chloromethane (1.48 g,8.38mmol,608.42uL,4 asThe amount) was then cooled to-70 ℃ and LDA (2 m,6.29ml,6 eq.) was added dropwise. The reaction was stirred at-70℃for 1h. After completion, the reaction mixture was quenched by adding a mixture of AcOH (4.5 mL) and THF (22 mL) at-70 ℃ and then diluted with ethyl acetate (50 mL) and with water (30 mL x 2), saturated NaHCO ₃ (30 mL) extraction. The organic layer was washed with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether the residue was purified with etoac=2:1 to 0:1 and then wet-triturated with methyl tert-butyl ether:petroleum ether=4:1 (3 mL) to give N- [ (1S) -3-chloro-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Propyl group]Tert-butyl carbamate (0.35 g,1.03mmol,49.32% yield, 90% purity). MS (ESI) m/z 308.0[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.66(br s,1H),7.53(br d,J＝7.7Hz,1H),4.61(d,J＝2.2Hz,2H),4.22-4.10(m,1H),3.21-3.11(m,2H),2.34-2.06(m,2H),1.93-1.80(m,1H),1.73-1.54(m,2H),1.39(s,9H)。

Step 2: (S) -3- ((S) -2-amino-4-chloro-3-oxobutyl) pyrrolidin-2-one

N- [ (1S) -3-chloro-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Propyl group]A solution of tert-butyl carbamate (0.33 g,1.08mmol,1 eq.) in HCl/EtOAc (4M, 5mL,18.47 eq.) was stirred at 0deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (3S) -3- [ (2S) -2-amino-4-chloro-3-oxo-butyl as an oil]Pyrrolidin-2-one (0.3 g, crude material, HCl). MS (ESI) m/z 205.0[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.75(br s,3H),7.97(br s,1H),4.96-4.91(m,1H),4.77(s,1H),4.37-4.23(m,1H),3.26-3.07(m,2H),2.60(br d,J＝8.6Hz,1H),2.37-2.27(m,1H),1.96-1.90(m,1H),1.79-1.66(m,1H)。

Step 3: n- ((S) -1- (((S) -4-chloro-3-oxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

Direction (2)S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a solution of 4-methyl-pentanoic acid (416.53 mg,1.37mmol,1.1 eq.) in DMF (5 mL) were added HATU (946.18 mg,2.49mmol,2 eq.) and NMM (251.71 mg,2.49mmol, 273.59. Mu.L, 2 eq.) and the solution stirred at 0deg.C for 0.5h. Next, (3S) -3- [ (2S) -2-amino-4-chloro-3-oxo-butyl was added dropwise at 0 ℃]A solution of pyrrolidin-2-one (0.3 g,1.24mmol,1 eq. HCl) in DMF (5 mL). The reaction was stirred at 25℃for 0.5h. After completion, the reaction mixture was diluted dropwise with water (50 mL) at 0 ℃ and extracted with EtOAc (20 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether etoac=2:1 to 0:1) purification residue. To give N- [ (1S) -1- [ [ (1S) -3-chloro-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Methyl group]Propyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (0.3 g,549.92umol,44.20% yield, 90% purity). MS (ESI) m/z 491.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.58(br s,1H),8.74-8.57(m,1H),8.44(br d,J＝5.0Hz,1H),7.65(br d,J＝4.5Hz,1H),7.37(br s,1H),7.15-7.06(m,1H),7.01(br d,J＝8.1Hz,1H),6.50(br d,J＝7.6Hz,1H),4.75-4.60(m,1H),4.59-4.55(m,1H),4.44(br d,J＝9.2Hz,2H),3.88(s,3H),3.13-3.01(m,2H),2.34-2.18(m,1H),2.09(br dd,J＝2.5,3.9Hz,1H),1.99-1.90(m,1H),1.78-1.49(m,5H),0.97-0.81(m,6H)。

Step 4: 2-oxo-2-phenylacetic acid (S) -3- ((S) -2- (4-methoxy-1H-indole-2-carboxamide) -4-methylpentanoylamino) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butyl ester

To N- [ (1S) -1- [ [ (1S) -3-chloro-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Propyl group]Carbamoyl group]-3-methyl-butyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (0.25 g, 509.19. Mu.L, 1 eq.) in DMF (6 mL) was added benzoylformic acid (99.38 mg, 661.94. Mu.L, 1.3 eq.) and CsF (177.89 mg,1.17mmol, 43.18. Mu.L, 2.3 eq.). At N ₂ The reaction was stirred for 4h at 65℃under an atmosphere. After completion, the reaction mixture was usedDilute with water (20 mL) and extract with EtOAc (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives [ (3S) -3- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as an oil]-4-methyl-pentanoyl]Amino group]-2-oxo-4- [ (3S) -2-oxopyrrolidin-3-yl]Butyl group]2-oxo-2-phenyl-acetate (0.3 g, crude material). MS (ESI) m/z 605.2[ M+H ]] ⁺ 。

Steps 5 and 6: n- [ (1R) -1- [ [ (1S) -3-hydroxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] propyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide, N- [ (1S) -1- [ [ (1S) -3-hydroxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] propyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To [ (3S) -3- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-2-oxo-4- [ (3S) -2-oxopyrrolidin-3-yl]Butyl group]To a solution of 2-oxo-2-phenyl-acetate (0.3 g,496.16 mol,1 eq.) in MeOH (10 mL) was added K ₂ CO ₃ (3.43 mg,24.81umol,0.05 eq.). The reaction was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) to give the product.

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.58(s,1H),8.50(d,J＝7.8Hz,1H),8.41(d,J＝7.9Hz,1H),7.63(s,1H),7.35(d,J＝1.5Hz,1H),7.14-7.05(m,1H),7.04-6.94(m,1H),6.50(d,J＝7.7Hz,1H),5.05-4.98(m,1H),4.57-4.46(m,1H),4.41(ddd,J＝4.0,7.7,11.2Hz,1H),4.34-4.25(m,1H),4.22-4.13(m,1H),3.88(s,3H),3.18-3.01(m,2H),2.25-2.14(m,1H),2.13-2.04(m,1H),1.99-1.84(m,1H),1.77-1.48(m,5H),0.93(br d,J＝6.2Hz,3H),0.89(br d,J＝6.4Hz,3H)。

To give N- [ (1S) -1- [ [ (1S) -3-hydroxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Methyl group]Propyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23.86 mg,49.08 mol,9.89% yield, 97.2% purity). MS (ESI) m/z 473.2[ M+H ]] ⁺ . By chiral SFC (column: DAICEL CHIRALCE L OJ (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-MeOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -20%,15 min) to obtain solid N- [ (1R) -1- [ [ (1S) -3-hydroxy-2-oxo-1- [ [ (3S) -2-oxo-pyrrolidin-3-yl]Methyl group]Propyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (15.43 mg,31.22umol,6.29% yield, 95.6% purity). MS (ESI) m/z 473.2[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.57(s,1H),8.45(br d,J＝8.1Hz,1H),8.41(br d,J＝7.8Hz,1H),7.62(s,1H),7.36(d,J＝1.3Hz,1H),7.14-7.05(m,1H),7.04-6.97(m,1H),6.50(d,J＝7.6Hz,1H),5.06(br s,1H),4.62-4.38(m,2H),4.30-4.19(m,1H),4.19-4.09(m,1H),3.88(s,3H),3.19-3.01(m,2H),2.37-2.22(m,1H),2.09(br dd,J＝3.2,6.2Hz,1H),1.99-1.86(m,1H),1.80-1.43(m,5H),0.94(d,J＝6.2Hz,3H),0.89(d,J＝6.2Hz,3H)。

EXAMPLE 11 Synthesis of viral protease inhibitor Compound 143

Step 1: (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl at 25 ℃C]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (250 mg,625.81umol,1 eq.) was added HCl/EtOAc (8 mL) for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue, which gave the product (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl as an oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (230 mg, crude material). MS (ESI) m/z 300.0[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl ] amino ] pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (230 m)To a mixture of g,684.88umol,1 equivalent, HCl) and (E) -3-phenylpropa-2-enoic acid (202.94 mg,1.37mmol, 162.35. Mu.L, 2 equivalent) in DMF (2 mL) and DCM (4 mL) was added EDCI (262.59 mg,1.37mmol,2 equivalent) and DMAP (167.34 mg,1.37mmol,2 equivalent). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By chromatography on silica gel (SiO ₂ Petroleum ether etoac=1:1) the residue was purified to give the product (2S) -2- [ [ (2S) -4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl as an oil]Amino group]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,465.65umol,67.99% yield). MS (ESI) M/z430.1[ M+H] ⁺ 。

Step 3: (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl ] amino ] pentanamide

In a sealed tube, (2S) -2- [ [ (2S) -4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl]Amino group]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,465.65umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 7mL,97% purity, 105.23 eq.) was heated to 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue, which gave the product (2S) -N- [ (1S) -2-amino-2-oxo-1-oxo-pyrrolidin-3-yl as an oil]Methyl group]Ethyl group ]-4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl]Amino group]Pentanamide (200 mg, crude material). MS (ESI) m/z 415.1[ M+H ]] ⁺ 。

Step 4: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl ] amino ] pentanamide

To (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl]Amino group]To a mixture of valeramide (200 mg,482.51umol,1 eq.) in DCM (2 mL) was added methoxycarbonyl- (triethylammonio) sulfonyl-imide574.93mg,2.41mmol,5 eq.) and the mixture stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give the product (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]-4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl]Amino group]Pentanamide (23.1 mg,58.26umol,12.07% yield, 100% purity). MS (ESI) M/z397.2[ M+H] ⁺ 。

¹ H NMR(400MHz,CDCl ₃ )＝8.70(br d,J＝6.6Hz,1H),7.66-7.55(m,1H),7.54-7.44(m,2H),7.35(br s,3H),6.72-6.52(m,2H),6.47(d,J＝15.7Hz,1H),5.02-4.67(m,2H),3.49-3.22(m,2H),2.56-2.27(m,3H),2.02-1.88(m,1H),1.88-1.80(m,1H),1.75-1.61(m,3H),1.07-0.87(m,6H)

EXAMPLE 12 Synthesis of viral protease inhibitor Compound 598

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl at 25 ℃C]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (300 mg,750.98umol,1 eq.) was added HCl/EtOAc (4M, 6mL,31.96 eq.) for 1h. After completion, use N ₂ Drying the product directly to obtain the oily product (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (260 mg, crude material). MS (ESI) m/z 300.1[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ] amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3-[(3S) -2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (250 mg,744.43umol,1 eq., HCl) and (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid (298.66 mg,1.49mmol, 81.96. Mu.L, 2 eq.) in DMF (2 mL) and DCM (4 mL) was added EDCI (285.42 mg,1.49mmol,2 eq.) and DMAP (181.89 mg,1.49mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Purification of the residue by chromatography on silica gel (SiO ₂ Petroleum ether: etoac=0:1) to give the product (2S) -2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as an oil]Amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (80mg,165.99umo l,22.30% yield). MS (ESI) m/z 482.1[ M+H ]] ⁺ 。

Step 3: (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ] amino ] -4-methyl-pentanamide

Stirring (2S) -2- [ [ (2S) -2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl at 80 ℃]Amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (70 mg,145.25umol,1 eq.) in NH ₃ A mixture in MeOH (7M, 6mL,97% purity, 289.17 eq.) for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as an oil]Methyl group ]Ethyl group]-2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Amino group]-4-methyl-pentanamide (70 mg, crude). MS (ESI) m/z 467.1[ M+H ]] ⁺ 。

Step 4: (2S) -2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ] amino ] -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4-methyl-pentanamide

To (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- [ [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Amino group]-4-methyl-pentanamide (70 mg, 1)49.91 mol,1 eq.) to a mixture of methoxycarbonyl- (triethylammonium) sulfonyl-imide (160.77 mg,674.62 mol,4.5 eq.) in DCM (1.5 mL) and the mixture stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as solid]Ethyl group]-4-methyl-2- [ [ (E) -3-phenylprop-2-enoyl]Amino group]Pentanamide (13.4 mg,58.26umol,12.07% yield, 100% purity). MS (ESI) m/z 449.1[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,CDCl ₃ )δ＝8.67(br d,J＝5.7Hz,1H),7.63(d,J＝15.7Hz,1H),7.42(t,J＝8.3Hz,1H),7.19-7.06(m,2H),6.55(d,J＝15.7Hz,1H),6.34(br s,1H),6.19(br s,1H),4.83-4.67(m,2H),3.47-3.33(m,2H),2.58-2.28(m,3H),2.04(br s,1H),1.95-1.82(m,1H),1.81-1.62(m,3H),0.99(d,J＝6.0Hz,6H)

EXAMPLE 13 Synthesis of viral protease inhibitor Compounds 149

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.75mmol,1 eq.) in HCl/EtOAc (4M, 20 mL). The mixture was stirred at 25 ℃ and for 1h. After the reaction is completed, the reaction mixture is concentrated to give crude (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg, crude) (oil). The crude product was used without further purification. MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -2-inden-2-yl-acetyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxoSubstituted pyrrolidin-3-yl]Methyl propionate (190 mg,1.02mmol,1 eq.) and (2S) -2- (tert-butoxycarbonylamino) -2-inden-2-yl-acetic acid (297.27 mg,1.02mmol,1 eq.) were added DMAP (249.31 mg,2.04mmol,2 eq.) and EDCI (391.21 mg,2.04mmol,2 eq.) in DCM (9 mL) and DMF (3 mL). The mixture was stirred at 25℃for 2h. After completion of the reaction, the reaction was poured into ice water (30 mL) and extracted with EtOAc (20 mL x 3). The combined organic phases were dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250mm, diameter: 100mm,100-200 mesh silica gel, petroleum ether/etoac=1/1, 0/1), giving (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -2-inden-2-yl-acetyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (300 mg,522.27umol,51.18% yield, 80% purity) (solid). MS (ESI) m/z 460.3[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -2-inden-2-yl-acetyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

A mixture of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (400 mg,870.4umol,1 eq) in HCl/EtOAc (4M, 20 mL). The mixture was stirred at 25℃for 2h. After the reaction was completed, the reaction mixture was concentrated to give the product (2S) -2- [ [ (2S) -2- (t-butoxycarbonylamino) -2-inden-2-yl-acetyl group as an oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (330 mg, crude) and was used directly in the next step. MS (ESI) m/z 360.2[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -2-amino-2-inden-2-yl-acetyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -2-inden-2-yl-acetyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (300 mg,652.84 mol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (149.77 mg,783.40 mol,1.2 eq.) in DCM (6 mL) and DMF (2 mL) was added DMAP (159.51 mg,1.31mmol,2 eq.) and EDCI (250.30 mg,1.31mmol,2 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. After the reaction is completedAfter this time, the reaction was poured into ice water (30 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250mm, diameter: 100mm,100-200 mesh silica gel, petroleum ether/ethyl acetate=1/1, 0/1), giving (2S) -2- [ [ (2S) -2-amino-2-inden-2-yl-acetyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (300 mg,506.96umol,77.66% yield, 90% purity) (solid). MS (ESI) m/z 533.2[ M+H ]] ⁺

Step 5: n- [ (1S) -1- [ [ (1S) -2-amino-1- [ (3-methylimidazol-4-yl) methyl ] -2-oxo-ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To a mixture of methyl (S) -2- ((S) -2- (2, 3-dihydro-1H-inden-2-yl) -2- (4-methoxy-1H-indole-2-carboxamido) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (100 mg,187.76umol,1 eq.) was added ammonia (3.20 mg,187.76umol,3.13 μl,1 eq.). The mixture was stirred at 80 ℃ and for 16h. After completion of the reaction, the reaction was concentrated to give crude N- ((S) -2- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1- (2, 3-dihydro-1H-inden-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide (70 mg,108.20umol,57.62% yield, 80% purity) as a solid. The crude product was used without further purification. MS (ESI) m/z 518.2[ M+H ] ] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1-inden-2-yl-2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1-inden-2-yl-2-oxo-ethyl]A mixture of 4-methoxy-1H-indole-2-carboxamide (60 mg,115.93umol,1 eq.) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (55.25 mg,231.85umol,2 eq.) in DCM (0.5 mL). The mixture was stirred at 25 ℃ and stirred for 2h. After completion of the reaction, the reaction was poured into ice water (30 mL) and extracted with DCM (20 mL x 3). The combined organic phasesAnhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mM. Times.3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Amino group]-1-inden-2-yl-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (23.83 mg,47.70umol,41.15% yield, 100% purity) (solid). MS (ESI) m/z 500.3[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.26(s,1H),7.13-7.17(m,2H),7.11-7.12(m,3H),7.03(s,1H),6.55-6.52(d,J＝12.4Hz,1H),5.05-5.01(m,1H),4.85-5.00(m,1H),3.92(s,3H),3.25-3.26(m,3H),3.21-3.24(m,2H),2.90-3.01(m,2H),2.88-2.89(m,1H),2.31-3.33(m,2H),1.81-1.92(m,2H)

EXAMPLE 14 Synthesis of viral protease inhibitor Compounds 165

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (250 mg,873.14umol,1 eq.) was added HCl/EtOAc (4M, 30 mL) at 25 ℃. The mixture was stirred at 25℃for 1h. Concentrating the reaction mixture under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate as a solid; hydrochloride (200 mg, crude) and used directly in the next step.

Step 2:

(2S, 4R) - (9H-fluoren-9-yl) methyl-4- (tert-butoxy) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (190 mg,853.29umol,1 eq., HCl), (2S, 4R) -4-tert-butoxy-1- (9H-fluoren-9-ylmethoxycarbonyl) pyrrolidineA mixture of 2-carboxylic acid (349.40 mg,853.29umol,1 eq), EDCI (327.15 mg,1.71mmol,2 eq), DMAP (208.49 mg,1.71mmol,2 eq), DMF (3 mL) and DCM (6 mL) was stirred at 25℃for 1h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the product (2S, 4 r) - (9H-fluoren-9-yl) methyl-4- (tert-butoxy) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid ester (230 mg,319.96umol,37.50% yield, 80.36% purity) as an oil. MS (ESI) m/z 578.2[ M+H ]] ⁺

Step 3: (S) -methyl-2- ((2S, 4R) -4- (tert-butoxy) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate

A mixture of (2S, 4R) - (9H-fluoren-9-yl) methyl-4- (tert-butoxy) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid ester (170 mg,294.29 mol,1 eq), piperidine (3.76 g,8.83mmol,4.36mL,20% purity, 30 eq) and DMF (1 mL) was stirred at 25℃for 1H. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (DCM/meoh=10/1) to give the product (S) -methyl-2- ((2S, 4 r) -4- (tert-butoxy) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (40 mg,112.54umol,38.24% yield) as an oil.

Step 4: (S) -methyl-2- ((2S, 4R) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate

A mixture of (S) -methyl-2- ((2S, 4R) -4- (tert-butoxy) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (40 mg, 112.54. Mu. Mol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (21.52 mg, 112.54. Mu. Mol,1 eq), EDCI (43.15 mg, 225.08. Mu. Mol,2 eq), DMAP (27.50 mg, 225.08. Mu. Mol,2 eq), DMF (0.5 mL) and DCM (1 mL) was stirred at 25℃for 1H.H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the compound (S) -methyl-2- ((2S, 4 r) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (30 mg,22.33umol,19.84% yield) as an oil.

Step 5: (2S, 4R) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

(S) -methyl-2- ((2S, 4R) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (27 mg,20.10umol,39.35% purity, 1 eq.) and NH ₃ The mixture of MeOH (7M, 3 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give the product (2S, 4 r) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (22 mg, crude material) as a solid. MS (ESI) m/z 514.2[ M+H ]] ⁺

Step 6: (2S, 4R) -4- (tert-butoxy) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

A mixture of (2S, 4R) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butoxy) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (20 mg,38.94umol,1 eq), bogis reagent (27.84 mg,116.83umol,3 eq) and DCM (1 mL) was stirred at 25℃for 4H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -40%,8 min) to obtain (2S, 4R) -4- (tert-butyl acetate as solid productButoxy) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (5 mg,10.09umol,25.91% yield, 100% purity). MS (ESI) m/z 496.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.73-11.43(m,1H),9.26-8.84(m,1H),7.84-7.49(m,1H),7.19-7.07(m,1H),7.05-6.96(m,1H),6.94-6.65(m,1H),6.57-6.41(m,1H),5.08-4.92(m,1H),4.85-4.40(m,2H),4.34-4.08(m,1H),3.98-3.75(m,3H),3.74-3.50(m,1H),3.22-2.80(m,2H),2.47-2.37(m,1H),2.27-2.04(m,3H),2.03-1.87(m,1H),1.86-1.36(m,2H),1.15(s,9H)

EXAMPLE 15 Synthesis of viral protease inhibitor Compound 167

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

Methyl (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg,1.05mmol,1 eq.) in HCl/EtOAc (4M, 5mL,19.09 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (hydrochloride, 210mg, crude material) as a solid.

Step 2: 4-cyclohexyl-2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid (2S, 4S) - (9H-fluoren-9-yl) methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-pyrrolidin-3-yl]Methyl propionate (200 mg,1.07mmol,1 eq.) and (2S, 4S) -4-cyclohexyl-1- (9H fluoren-9-ylmethoxycarbonyl) pyrrolidine-2-carboxylic acid (450.58 mg,1.07mmol,1 eq.) in DMF (1 mL) and DCM (2 mL) were added DMAP (262.43 mg,2.15mmol,2 eq.) and EDCI (411.80 mg,2.15mmol,2 eq.). The mixture was stirred at 25℃for 4h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL) washing with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether the residue is purified with EtOAc=5:1 to 1:1 to give the product (2S, 4S) -4-cyclohexyl-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a solid]Methyl group]Ethyl group]Carbamoyl group]Pyrrolidine-1-carboxylic acid 9H-fluoren-9-ylmethyl ester (500 mg,850.77umol,79.21% yield). MS (ESI) m/z 588.3[ M+H ]] ⁺

Step 3: pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid (S) -methyl ester 2- ((2S, 4S) -4-cyclohexyl ester

Containing (2S, 4S) -4-cyclohexyl-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Pyrrolidine-1-carboxylic acid 9H-fluoren-9-ylmethyl ester (480 mg,816.74umol,1 eq.) in DMF (4 mL) and PIPERIDINE (PIPERIDINE) (862.20 mg,10.13mmol,1mL,12.40 eq.) were stirred at 25℃for 0.5H. After completion, the mixture was treated with N ₂ Dried and then diluted with DCM (10 mL) and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give the product (2S) -2- [ [ (2S, 4S) -4-cyclohexylpyrrolidine-2-carbonyl as a solid ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (210 mg,574.61umol,70.35% yield).

Step 4: (S) -2- ((2S, 4S) -4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S, 4S) -4-cyclohexylpyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg, 547.25. Mu. Mol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (104.62 mg, 547.25. Mu. Mol,1 eq.) in DMF (2 mL) and DCM (3 mL) were added DMAP (133.71 mg,1.09mmol,2 eq.) and EDCI (209.82 mg,1.09mmol,2 eq.). The mixture was stirred at 25℃for 4h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to obtainTo residue. By preparative TLC (SiO) ₂ Petroleum ether EtOAc=0:1) the residue was purified to give the product (2S) -2- [ [ (2S, 4S) -4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (210 mg,389.88umol,71.24% yield). MS (ESI) m/z 539.2[ M+H ] ] ⁺

Step 5: (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

(2S) -2- [ [ (2S, 4S) -4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,371.31umol,1 eq.) was placed in NH ₃ In MeOH (7M, 10mL,188.52 eq.). The mixture was stirred at 80℃for 16h. After completion, the mixture was concentrated under reduced pressure to give the product (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a solid]Methyl group]Ethyl group]-4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (110 mg, crude). MS (ESI) m/z 524.2[ M+H ]] ⁺

Step 6: (2S, 4S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

To (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To DCM (1 mL) of-4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (100 mg, 190.98. Mu. Mol,1 eq.) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (227.55 mg, 954.89. Mu. Mol,5 eq.). The mixture was stirred at 25℃for 3h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product (2S, 4S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a solid]Ethyl group]-4-cyclohexyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (30.7mg,60.17 mol,31.51% yield, 99.1% purity). MS (ESI) m/z 506.3[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23-6.82(m,3H),6.60-6.36(m,1H),5.21-4.96(m,1H),4.72-4.56(m,1H),4.34-4.07(m,1H),4.00-3.80(m,3H),3.57(br t,J＝9.4Hz,1H),3.02-2.54(m,1H),2.46-0.92(m,20H)

EXAMPLE 16 Synthesis of viral protease inhibitor Compound 209

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (0.55 g,1.92mmol,1 eq.) and HCl/EtOAc (4M, 10mL,20.82 eq.) was stirred at 25℃for 0.5h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (0.35 g, crude material) as an oil.

Step 2: (2S, 4S) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester

A mixture of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (0.15 g, 805.55. Mu. Mol,1 eq), (2S, 4S) -1-tert-butoxycarbonyl-4-phenyl-pyrrolidine-2-carboxylic acid (234.69 mg, 805.55. Mu. Mol,1 eq), DMAP (196.83 mg,1.61mmol,2 eq), EDCI (308.85 mg,1.61mmol,2 eq) in DMF (1 mL) and DCM (2 mL) was stirred at 25℃for 0.5h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether etoac=2:1 to 0:1) to give (2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]Carbamoyl group]-4-phenyl-pyrrolidine-1-carboxylic acid tert-butyl ester(0.25 g,500.51umol,62.13% yield, 92% purity). MS (ESI) m/z 460.1[ M+H ]] ⁺ 。

Step 3: (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((2S, 4S) -4-phenylpyrrolidine-2-carboxamido) propanoic acid methyl ester

(2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A mixture of tert-butyl 4-phenyl-pyrrolidine-1-carboxylate (0.25 g,544.03umol,1 eq.) and HCl/EtOAc (4M, 10mL,73.53 eq.) was stirred at 25℃for 0.5h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl as an oil]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]Methyl propionate (0.2 g, crude material). MS (ESI) m/z 360.1[ M+H ]] ⁺ 。

Step 4: (S) -2- ((2S, 4S) -1- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) -4-phenylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

(2S) -3- [ (3S) -2-oxopyrrolidin-3-yl at 25 ℃C]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]A mixture of methyl propionate (0.17 g,472.99umol,1 eq), (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid (94.88 mg,472.99umol,1 eq), T3P (451.48 mg,709.48umol,421.95uL,50% purity, 1.5 eq) and TEA (143.58 mg,1.42mmol,197.50uL,3 eq) in DMF (4 mL) was stirred for 0.5h with degassing. After completion, the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether etoac=2:1 to 0:1) to give (2S) -2- [ (2S, 4S) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl in solid form]-4-phenyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.11 g,162.36umol,34.33% yield, 80% purity). MS (ESI) m/z 542.1[ M+H ]] ⁺ 。

Step 5: (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) -4-phenylpyrrolidine-2-carboxamide

In a sealed tube, (2S) -2- [ [ (2S, 4S) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]-4-phenyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.1 g,184.50umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 3 mL) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow oil]Methyl group]Ethyl group]-1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]-4-phenyl-pyrrolidine-2-carboxamide (0.09 g, crude material). MS (ESI) m/z 527.0[ M+H ]] ⁺ 。

Step 6: (2S, 4S) -1- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -4-phenylpyrrolidine-2-carboxamide

To (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]To a solution of 4-phenyl-pyrrolidine-2-carboxamide (0.09 g,170.78umol,1 eq.) in DCM (1 mL) was added the Buerger's reagent (203.50 mg,853.91umol,5 eq.) and the solution was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to obtain (2S, 4S) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl in solid form]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-4-phenyl-pyrrolidine-2-carboxamide (29.73 mg,56.89umol,33.31% yield, 97.4% purity). MS (ESI) m/z 509.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.17-8.86(m,1H),8.07-7.75(m,1H),7.75-7.65(m,1H),7.62-7.49(m,2H),7.48-7.30(m,5H),7.26(tt,J＝3.0,5.6Hz,1H),7.22-6.73(m,1H),5.09-4.83(m,1H),4.69-4.47(m,1H),4.40-4.01(m,1H),3.77-3.50(m,3H),3.19-3.04(m,2H),2.44-2.31(m,2H),2.22-2.09(m,2H),1.88-1.59(m,2H)。

EXAMPLE 17 Synthesis of viral protease inhibitor Compounds 183

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg,1.05mmol,1 eq.) was added HCl/EtOAc (4M, 30 mL) at 25 ℃. The mixture was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate as an oil, HCl (230 mg, crude material) and used directly in the next step.

Step 2:

(S) -5- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [2.5] octane-6-carboxylic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (230 mg,1.03mmol,1 eq. HCl), (7S) -6-tert-butoxycarbonyl-6-azaspiro [2.5] ]Octane-7-carboxylic acid (263.72 mg,1.03mmol,1 eq.) T ₃ P (657.31 mg,2.07mmol,614.31uL,2 eq.) Et ₃ A mixture of N (522.60 mg,5.16mmol,718.85uL,5 eq.) and DMF (5 mL) was stirred at 25℃for 1h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the product (S) -5- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [2.5] as a yellow oil]Tert-butyl octane-6-carboxylate (300 mg,708.38umol,68.58% yield). MS (ESI) m/z 424.1[ M+H ]] ⁺

Step 3: (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((S) -6-azaspiro [2.5] octane-5-carboxamido) propanoic acid methyl ester

A mixture of (S) -5- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [2.5] octane-6-carboxylic acid tert-butyl ester (290 mg,684.77umol,1 eq) and HCl/EtOAc (4M, 30 mL) was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((S) -6-azaspiro [2.5] octane-5-carboxamido) propionate (240 mg, crude material, HCl) as an oil and was used directly in the next step.

Step 4: (S) -2- ((S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5] octane-5-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

(S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((S) -6-azaspiro [2.5]A mixture of methyl octane-5-carboxamido) propionate (240 mg, 666.95. Mu. Mol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (127.51 mg, 666.95. Mu. Mol,1 eq.), DMAP (162.96 mg,1.33mmol,2 eq.), EDCI (255.71 mg,1.33mmol,2 eq.), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the compound (S) -2- ((S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5] as an oil]Octane-5-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (150 mg,275.74umol,41.34% yield, 91.28% purity). MS (ESI) m/z 495.2[ M-H ]] ^-

Step 5: (S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5] octane-5-carboxylic acid

(S) -2- ((S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [ 2.5)]Octane-5-carboxamide) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester, liOH (24.12 mg,1.01mmol,5 eq.) H ₂ A mixture of O (1 mL) and THF (4 mL) was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to give the product (S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5 ] as a solid]Octane-5-carboxylic acid (65 mg, crude material).MS(ESI)m/z 327.1[M-H] ^-

Step 6: (S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamic acid tert-butyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg,1.40mmol,1 eq.) and NH ₃ The mixture of MeOH (7M, 10 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give tert-butyl ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamate (380 mg, crude material) as a solid.

Step 7: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propanamide

A mixture of tert-butyl ((S) -1-amino-1-oxo-3- ((S) -2-oxo-pyrrolidin-3-yl) propan-2-yl) carbamate (300 mg,1.11mmol,1 eq.) and HCl/EtOAc (4M, 15mL,54.26 eq.) was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give the product (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionamide (190 mg, crude) as a solid and used directly in the next step.

Step 8: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5] octane-5-carboxamide

(S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5]A solution of octane-5-carboxylic acid (65 mg,197.95umol,1 eq), (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionamide (33.89 mg,197.95umol,1 eq), DMAP (48.37 mg,395.91umol,2 eq), EDCI (75.90 mg,395.91umol,2 eq), DMF (1 mL) and DCM (3 mL) was stirred at 25℃for 16h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:10% -40%,8 min) purifying the residue to obtain a solid(S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [ 2.5)]Octane-5-carboxamide (45 mg,79.43umol,40.13% yield, 85% purity). MS (ESI) m/z 480.2[ M-H ] ^-

Step 9: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5] octane-5-carboxamide

(S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [2.5]A mixture of octane-5-carboxamide (40 mg,83.07umol,1 eq), prague reagent (237.55 mg,996.80umol,12 eq) and DCM (20 mL) was stirred at 25℃for 8h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemi ni-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH4HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -40%,8 min) to give the product (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [ 2.5) as a solid]Octane-5-carboxamide (17mg,34.79um ol,41.89% yield, 94.87% purity). MS (ESI) m/z 462.2[ M-H ]] ^- 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.64(s,1H),9.26-8.52(m,1H),7.87-7.61(m,1H),7.18-7.07(m,1H),7.06-6.96(m,1H),6.85-6.60(m,1H),6.51(d,1H),5.30-4.93(m,2H),4.61-4.41(m,1H),3.85(s,3H),3.21-2.96(m,2H),2.39-2.03(m,5H),1.96-1.56(m,4H),0.99(d,1H),0.45-0.15(m,4H)

EXAMPLE 18 Synthesis of viral protease inhibitor Compound 185

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidine-3-yl]To a solution of methyl propionate (170 mg,763.47 mol,1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionic acid (207.17 mg,763.47 mol,1 eq) in DMF (2 mL) were added DMAP (186.55 mg,1.53mmol,2 eq) and EDCI (292.71 mg,1.53mmol,2 eq). DCM (3 mL) was added to the mixture and stirred at 25 ℃ for 2h. By adding H at 0 ℃ ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/etoac=0/1) to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,568.77umol,74.50% yield). MS (ESI) m/z 440.3[ M+H ]] ⁺

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclohexyl-propionyl at 25 ℃C]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a solution of methyl propionate (200 mg,455.02umol,1 eq.) in EtOAc (0.5 mL) was added dropwise HCl/EtOAc (4M, 2.00mL,17.58 eq.). The mixture was stirred at 25℃for 1h. Concentrating the reaction mixture under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-3-cyclohexyl-propionyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, crude, HCl) and was used directly in the next step. MS (ESI) m/z 340.1[ M+H ]] ⁺

To 4-methoxy-1H-indole-2-carboxylic acid (99.18 mg,518.77 mol,1.3 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclohexyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (150 mg,399.05umol,1 equivalent, HCl) in DMF (2 mL) was added DMAP (97.50 mg,798.11umol,2.0 equivalent) and EDCI (153.00 mg,798.11umol,2 equivalents). DCM (4 mL) was added to the mixture and stirred at 25 ℃ for 2h. By adding H at 0 ℃ ₂ O (20 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give the product (2S) -2- [ [ (2S) -3-cyclohexyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg,292.63umol,73.33% yield).

¹ H NMR (methanol-d) ₄ ,400MHz)：δppm 7.26(s,1H),7.09-7.20(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.6Hz,1H),4.66(br dd,J＝9.0,6.3Hz,1H),4.52-4.58(m,1H),3.93(s,3H),3.72(s,3H),3.22-3.29(m,2H),2.54-2.62(m,1H),2.26-2.33(m,1H),2.15-2.23(m,1H),1.66-1.87(m,9H),1.47-1.54(m,1H),1.25-1.40(m,3H),0.96-1.06(m,2H)

In a sealed tube, (2S) -2- [ [ (2S) -3-cyclohexyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (150 mg,292.63 mmole, 1 eq.) in ammonia (15.30 g,898.39mmol,15.00mL,3070.07 eq.) was heated to 80℃for 12h. The reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (140 mg, crude). MS (ESI) m/z 498.2[ M+H ] ] ⁺

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (80 mg,160.78umol,1 eq.) in DCM (3 mL) was added the Buerger's reagent (114.94 mg,482.33umol,3 eq.) followed by stirring the mixture at 25℃for 3H. The reaction mixture was concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gives the product N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]Amino group]-1- (cyclohexylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (20.02 mg,41.75umol,25.97% yield, 100% purity). MS (ESI) m/z 480.1[ M+H ]] ⁺ 。

EXAMPLE 19 Synthesis of viral protease inhibitor Compound 197

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (500 mg,1.75mmol,1 eq.) inHCl/EtOAc (4M, 20 mL). The mixture was stirred at 25 ℃ and for 1h. After the reaction is completed, the reaction mixture is concentrated to give crude (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg, crude material, oil). The crude product was used without further purification. MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (2S, 5S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-3-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (230 mg,1.24mmol,1 eq.) and (2S, 5S) -3-tert-butoxycarbonyl-6, 6-dimethyl-3-azabicyclo [3.1.0]]To a mixture of hexane-2-carboxylic acid (315.35 mg,1.24mmol,1 eq.) in DCM (4.5 mL) and DMF (1.5 mL) was added EDCI (473.57 mg,2.47mmol,2 eq.) and DMAP (301.80 mg,2.47mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion of the reaction, the reaction was concentrated and purified by preparative HPLC (column Waters Xbridge BEH C18 100 x 30mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give (2S, 5S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-3-carboxylic acid tert-butyl ester (200 mg,425.03umol,34.41% yield, 90% purity) (solid). MS (ESI) m/z 424.1[ M+H ]] ⁺

Step 3: (S) -2- ((1S, 2S, 5S) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

(1S, 2S, 5S) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6, 6-dimethyl-3-azabicyclo [ 3.1.0)]A mixture of tert-butyl hexane-3-carboxylate (200 mg,236.13umol,50% purity, 1 eq.) in HCl/EtOAc (4M, 20 mL). The mixture was stirred at 25 ℃ and stirred for 2h. After the reaction is complete, the reaction mixture is concentrated to give crude (S) -2- ((1S, 2S, 5S) -6, 6-dimethyl-3-azabicyclo [ 3.1.0)]Hexane-2-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (150 mg, crudeMass, oil). The crude product was used without further purification. MS (ESI) m/z 324.1[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S, 5S) -3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (S) -2- ((1S, 2S, 5S) -6, 6-dimethyl-3-azabicyclo [ 3.1.0)]To a mixture of methyl hexane-2-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (150 mg,463.84umol,1 eq) and 4-methoxy-1H-indole-2-carboxylic acid (88.68 mg,463.84umol,1 eq) in DCM (3 mL) and DMF (1 mL) was added EDCI (177.84 mg,927.68umol,2 eq) and DMAP (113.33 mg,927.68umol,2 eq). The mixture was stirred at 25 ℃ and stirred for 14h. After completion of the reaction, the mixture was poured into water (50 mL) and extracted with DCM (20 mL x 3). The combined organic phases were washed with brine (60 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250mm, diameter: 100mm,100-200 mesh silica gel, petroleum ether/ethyl acetate=1/1, 0/1) to give (2S) -2- [ [ (2S, 5S) -3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] as a solid]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (50 mg,80.56umol,17.37% yield, 80% purity). MS (ESI) m/z 497.2[ M+H ]] ⁺

Step 5: (2S, 5S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

(2S) -2- [ [ (2S, 5S) -3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (100 mg,201.39umol,1 eq.) in ammonia (5.10 g,299.46mmol,5mL,1486.99 eq.). The mixture was stirred at 80 ℃ and for 16h. After the reaction is completed, the reaction mixture is concentrated to give crude (2S, 5S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-di-Methyl-3-azabicyclo [3.1.0]]Hexane-2-carboxamide (100 mg, crude) (solid). The crude product was used without further purification. MS (ESI) m/z 482.3[ M+H ]] ⁺

Step 6: (2S, 5S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

To (2S, 5S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0]To a mixture of hexane-2-carboxamide (50 mg,103.83umol,1 eq) in DCM (3 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (49.49 mg,207.67umol,2 eq). The mixture was stirred at 25℃for 2h. After completion of the reaction, the reaction was concentrated and purified by preparative HPLC (column: phenomenex Gemini-NX 80X 40mm X3 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -40%,8 min) to give (2S, 5S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide (14.44 mg,31.15umol,30.00% yield, 100% purity). MS (ESI) m/z 464.2[ M+H ]] ⁺ 。

1H NMR (400 MHz, methanol-d 4): delta ppm 7.16-7.18 (m, 1H), 7.11-7.14 (m, 2H), 6.4-6.88 (m, 1H), 5.05-5.08 (m, 0.5H), 4.06 (s, 2H), 3.94-3.98 (m, 0.5H), 3.77-3.86 (m, 4H), 3.28 (s, 2H), 2.61-3.69 (m, 1H), 2.27-2.32 (m, 1H), 2.25-2.26 (m, 1H), 1.78-2.00 (m, 1H), 1.74-1.75 (m, 1H) 1.35-1.64 (m, 2H), 0.97-1.15 (m, 6H)

EXAMPLE 20 Synthesis of viral protease inhibitor Compound 213

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

Methyl (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (501 mg,1.75mmol,1 eq.) in HCl/EtOAc (4M, 10.02mL,22.91 eq.) was stirred at 25℃for 1h. After completion, the solution was concentrated. The crude material was used directly in the next step without further purification. Methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg, crude material) was obtained as a yellow oil.

Step 2: (S) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) tetrahydropyridazine-1 (2H) -carboxylic acid benzyl ester

Containing (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (295.93 mg,1.59mmol,1.4 eq.) and DCM (2 mL)/THF (2 mL) of (3S) -1-benzyloxycarbonyl hexahydropyridazine-3-carboxylic acid (300 mg,1.14mmol,1 eq.) were cooled to 0deg.C, then T3P (1.08 g,1.70mmol,1.01mL,50% purity, 1.5 eq.) and DIEA (440.14 mg,3.41mmol, 593.18. Mu.L, 3 eq.) were added and the solution stirred at 25deg.C for 13h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with ethyl acetate (30 mL. Times.3), and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The crude material was used directly in the next step without further purification. Obtaining (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]Hexahydropyridazine-1-carboxylic acid benzyl ester (455 mg, crude material). MS (ESI) m/z 433.1[ M+H ]] ⁺ 。

Step 3: (S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) tetrahydropyridazine-1 (2H) -carboxylic acid benzyl ester

To (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-pyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]To benzyl hexahydropyridazine-1-carboxylate (200 mg,462.46 mol,1 eq) in DCM (2 mL) was added DIEA (119.54 mg,924.92 mol, 161.10. Mu.L, 2 eq), and (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl chloride (121.56 mg,554.95 mol,1.2 eq) was added and the solution stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (10 mL) was diluted, extracted with DCM (20 mL. Times.3) and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtering and concentrating to obtainTo a crude material. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. Obtaining (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a yellow oil]-3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Hexahydropyridazine-1-carboxylic acid benzyl ester (160 mg,248.88umol,53.82% yield, 95.67% purity). MS (ESI) m/z 433.1[ M+H ]] ⁺ 。

Step 4: (S) -2- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) hexahydropyridazin-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

Containing (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ]-3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Benzyl hexahydropyridazine-1-carboxylate (160 mg,260.14umol,1 eq.) of TFA (5 mL) was stirred at 75deg.C for 1h. After completion, the solution was concentrated to remove TFA, with NaHCO ₃ The solution was diluted, extracted with EtOAc (20 ml x 3) and the combined organic phases were taken up over Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The crude material was used directly in the next step without further purification. Obtaining (2S) -2- [ [ (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl in solid form]Hexahydropyridazine-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (80 mg, crude material). MS (ESI) m/z 481.0[ M+H ]] ⁺ 。

Step 5: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) hexahydropyridazine-3-carboxamide

Containing (2S) -2- [ [ (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Hexahydropyridazine-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH of methyl propionate (80 mg,166.35umol,1 eq) ₃ MeOH (7M, 4.00mL,168.32 eq.) was stirred at 80℃for 17h. After completion, the solution was concentrated to remove MeOH. The crude material was used directly in the next step without further purification. Obtaining (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow oil ]Methyl group]Ethyl group]-2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Hexahydropyridazine-3-carboxamide (75 mg, crude material). MS (ESI) m/z 481.0[ M+H ]] ⁺ 。

Step 6: (S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acryloyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) hexahydropyridazine-3-carboxamide

To (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]To DCM (0.5 mL) of hexahydropyridazine-3-carboxamide (75 mg,160.98umol,1 eq.) was added the Bunges reagent (76.72 mg,321.95umol,2 eq.) and the solution stirred at 25℃for 2h. After completion, the solution was concentrated to remove DCM. The residue was purified by preparative HPLC (neutral conditions). Column: phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ Water (10 mM NH4HCO 3) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8min. Obtaining (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Hexahydropyridazine-3-carboxamide (20 mg,44.65umol,27.74% yield, 100% purity). ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.79-7.60(m,3H),7.32-7.22(m,2H),5.17(dd,J＝2.2,6.0Hz,1H),5.07(dd,J＝6.4,9.7Hz,1H),3.38-3.32(m,2H),3.12(br d,J＝13.7Hz,1H),2.90-2.74(m,1H),2.56(dq,J＝5.8,9.0Hz,1H),2.44-2.14(m,3H),2.08-1.79(m,3H),1.75-1.53(m,2H)。MS(ESI)m/z 448.2[M+H] ⁺ 。

Step 7: (E) -3- (4-chloro-2-fluorophenyl) acryloyl chloride

To DCM (0.5 mL) containing (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid (120 mg, 598.22. Mu.mol, 1 eq.) was added DMF (437.26. Mu.g, 5.98. Mu.L, 0.46. Mu.L, 0.01 eq.) and cooled to 0deg.C followed by addition of (COCl) ₂ (151.86 mg,1.20mmol, 104.73. Mu.L, 2 eq.) and the solution stirred at 25℃for 1h. After completion, the solution was concentrated to remove DCM and to give the crude material. The crude material was used directly in the next step without further purification. (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl chloride (125 mg, crude material) was obtained as a solid.

EXAMPLE 21 Synthesis of viral protease inhibitor Compound 201

Step 2: (S) -methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) pent-4-ynoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate; hydrochloride (180 mg, 808.38. Mu.L, 1 eq), (2S) -2- (tert-butoxycarbonylamino) pent-4-ynoic acid (172.37 mg, 808.38. Mu.L, 1 eq), TEA (572.59 mg,5.66mmol, 787.61. Mu.L, 7 eq), T ₃ A mixture of P (1.03 g,1.62mmol, 961.53. Mu.L, 50% purity, 2 eq.) and DMF (3 mL) was stirred at 25℃for 1h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the product (S) -methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) pent-4-ynoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (150 mg,393.26umol,48.65% yield) as an oil. MS (ESI) M/z382.1[ M+H ]] ⁺

Step 3: (S) -2- ((S) -2-aminopent-4-alkynoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of (S) -methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) pent-4-alkynoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (140 mg,367.05umol,1 eq) and HCl/EtOAc (4M, 30 mL) was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (S) -2- ((S) -2-aminopent-4-alkynylamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (120 mg, crude material, HCl) as an oil and used directly in the next step.

Step 4: (S) -methyl-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamide) pent-4-yn-lamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid ester

A mixture of methyl (S) -2- ((S) -2-aminopentan-4-ynylamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (120 mg, 377.63. Mu. Mol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (72.20 mg, 377.63. Mu. Mol,1 eq.), EDCI (144.78 mg, 755.27. Mu. Mol,2 eq.), DMAP (92.27 mg, 755.27. Mu. Mol,2 eq.), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the compound (S) -methyl-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) pent-4-ynamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (90 mg,160.56umol,42.52% yield, 81.08% purity) as an oil. MS (ESI) m/z 455.1[ M+H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1-oxopent-4-yn-2-yl) -4-methoxy-1H-indole-2-carboxamide

(S) -methyl-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) pent-4-ynamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (85 mg,187.03umol,1 eq.) and NH ₃ The mixture of MeOH (7M, 10 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give the product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1-oxopent-4-yn-2-yl) -4-methoxy-1H-indole-2-carboxamide (85 mg, crude) as a solid. MS (ESI) m/z 440.2[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -1-oxopent-4-yn-2-yl) -4-methoxy-1H-indole-2-carboxamide

A mixture of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1-oxopent-4-yn-2-yl) -4-methoxy-1H-indole-2-carboxamide (80 mg,182.04umol,1 eq), pragus reagent (216.91 mg,910.20umol,5 eq) and DCM (5 mL) was stirred at 25℃for 4H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Waters Xbridge BEH C, 100X 30mm X10 um; mobile phase: [ water (0.04% NH) ₃ H ₂ O+10mM NH4HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified for 20% -50%,10 min) to give the product N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -1-oxopent-4-yn-2-yl) -4-methoxy-1H-indole-2-carboxamide (20 mg,47.46umol,26.07% yield, 100% purity) as a solid. MS (ESI) m/z 422.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.61(d,J＝1.8Hz,1H),9.18-8.93(m,1H),8.74-8.58(m,1H),7.78-7.62(m,1H),7.37-7.29(m,1H),7.15-7.07(m,1H),7.05-6.97(m,1H),6.51(d,J＝7.5Hz,1H),5.03-4.91(m,1H),4.65-4.50(m,1H),3.89(s,3H),3.20-3.05(m,2H),2.91-2.85(m,1H),2.78-2.59(m,2H),2.43-2.29(m,1H),2.21-2.06(m,2H),1.88-1.59(m,2H)

EXAMPLE 22 Synthesis of viral protease inhibitor Compound 205

Step 1: (S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclopropylpropionic acid

To (2S) -2-amino-3-cyclopropyl-propionic acid (1 g,7.74mmol,1 eq.) in THF (5 mL) and H ₂ K was added to the solution in O (5 mL) ₂ CO ₃ (3.75 g,27.10mmol,3.5 eq.) and (Boc) ₂ O (2.20 g,10.07mmol,2.31mL,1.3 eq.). Additional water was added to the mixture, and the mixture was then stirred at 25 ℃ for 16h. The organic solvent is then evaporated and the aqueous solution is petroleumEther (10 mL) and acidified to pH-3 with aqueous 1N citric acid (30 mL). The solution was extracted with DCM (30 ml x 3) and concentrated in vacuo to give (S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionic acid (1.8 g, crude material) as an oil.

Step 2: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (500 mg,1.75mmol,1 eq.) was added HCl/EtOAc (4M, 5 mL) at 25deg.C. The mixture was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (350 mg, hcl, crude) as a yellow gum and was used directly in the next step.

Step 3: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (250 mg,1.12mmol,1 eq. HCl) and (S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionic acid (386.12 mg,1.68mmol,1.5 eq.) in DCM (5 mL) was added TEA (568.05 mg,5.61mmol, 781.36. Mu.L, 5 eq.) at 0deg.C, T3P (2.14 g,3.37mmol,2.00mL,50% purity, 3 eq.) was added to the mixture, followed by stirring the mixture at 25deg.C for 2h. The reaction mixture was quenched with water (10 mL) and extracted with DCM (5 mL x 3). The obtained solution was treated with Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give a residue. By column chromatography (SiO ₂ Petroleum ether the residue was purified with etoac=1:0 to 0:1 to give the gummy product methyl (S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (400 mg,905.74umol,80.67% yield, 90% purity).

¹ H NMR(400MHz,CDCl ₃ )δppm 7.60(d,J＝5.6Hz,1H),5.96(s,1H),5.24(d,J＝7.5Hz,1H),4.65-4.47(m,1H),4.24(d,J＝6.6Hz,1H),3.73(s,3H),3.44-3.27(m,2H),2.51-2.36(m,2H),2.25-2.13(m,1H),1.98-1.82(m,1H),1.66-1.58(m,1H),1.44(s,9H),1.30-1.21(m,1H),0.86-0.71(m,1H),0.49(d,J＝7.9Hz,2H),0.13(d,J＝4.4Hz,2H)。

Step 4: (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate in HCl/EtOAc (4M, 4 mL) was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (330 mg, crude material, HCl) as a yellow gum and used directly in the next step.

1H NMR(400MHz,MeOD-d ₄ )δppm 4.57(dd,J＝4.1,11.0Hz,1H),3.94(t,J＝6.7Hz,1H),3.73(s,3H),3.40-3.33(m,2H),2.55-2.33(m,2H),2.19-2.07(m,1H),2.03-2.00(m,1H),1.93-1.84(m,2H),1.24(t,J＝7.1Hz,1H),0.89-0.79(m,1H),0.59(dd,J＝4.5,7.9Hz,2H),0.26-0.17(m,2H)。

Step 5: (S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (257.73 mg,1.35mmol,1.5 eq.) and methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (300 mg,898.71 mol,1 eq., HCl) in DCM (8 mL) were added EDCI (861.43 mg,4.49mmol,5 eq.) and DMAP (329.38 mg,2.70mmol,3 eq.) followed by stirring the mixture at 25 ℃ for 2H. The combined organic layers were quenched with water (10 mL) and extracted with DCM (4 mL x 3). The obtained solution is treated by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ EtOAc) to give the compound (S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (250 mg,425.06umol,47.30% yield, 80% purity) as a yellow oil. MS (ESI) m/z 471.1[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To methyl (S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (250 mg,531.33umol,1 eq.) NH was added ₃ MeOH (7M, 6.00 mL). The mixture was stirred at 80℃for 16h. The resulting mixture was concentrated under reduced pressure to give the residue N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (200 mg, crude material) as a solid. MS (ESI) m/z 456.1[ M+H ]] ⁺

Step 7: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (100 mg, crude material) in DCM (4 mL) was added the berg reagent (104.63 mg,439.07umol,2 eq). The mixture was stirred at 25℃for 16h. The reaction mixture was quenched with water (0.5 mL) and purified by blowing N ₂ To be dried. The residue was purified by neutral prep HPLC to give the product N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (15 mg,34.29umol,15.62% yield, 100% purity) as a solid. MS (ESI) m/z 438.2[ M+H ] ] ⁺ 。

Preparative HPLC conditions:

column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：20％-50％，10min。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.57(d,J＝1.8Hz,1H),8.90(d,J＝8.2Hz,1H),8.50(d,J＝7.5Hz,1H),7.78-7.65(m,1H),7.36(d,J＝1.5Hz,1H),7.13-7.04(m,1H),7.03-6.96(m,1H),6.50(d,J＝7.8Hz,1H),5.04-4.94(m,1H),4.54-4.38(m,1H),3.89(s,3H),3.19-3.06(m,2H),2.44-2.33(m,1H),2.22-2.07(m,2H),1.90-1.75(m,2H),1.74-1.63(m,1H),1.54-1.41(m,1H),0.87-0.73(m,1H),0.47-0.34(m,2H),0.25-0.15(m,1H),0.14-0.04(m,1H)。

EXAMPLE 23 Synthesis of viral protease inhibitor Compound 401

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

Methyl (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (400 mg,1.40mmol,1 eq.) in HCl/EtOAc (4M, 10mL,28.63 eq.) was stirred at 25℃for 0.5h. After completion, the mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg, crude material, HCl) as a solid.

Step 2: (S) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-pyrrolidin-3-yl]Methyl propionate (300 mg,1.35mmol,1 eq. HCl) and (3S) -2-tert-butoxycarbonyl-2-azaspiro [4.4]]To DMF (2 mL) and DCM (5 mL) of nonane-3-carboxylic acid (362.87 mg,1.35mmol,1 eq.) were added DMAP (329.19 mg,2.69mmol,2 eq.) and EDCI (516.56 mg,2.69mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether etoac=5:1 to 0:1) the residue was purified to give the product (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as an oil]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester (340 mg,777.09umol,57.68% yield).

Step 3: (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((S) -2-azaspiro [4.4] nonane-3-carboxamido) propionic acid methyl ester

The (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -2-azaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester (340 mg,777.09umol,1 eq.) in HCl/EtOAc (4M, 10mL,51.47 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (3S) -2-azaspiro [4.4] nonane-3-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (250 mg, crude material, HCl) as an oil.

Step 4: (S) -2- ((S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (3S) -2-azaspiro [4.4]]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,668.67 mol,1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (127.84 mg,668.67 mol,1 eq.) in DMF (2 mL) and DCM (6 mL) were added DMAP (163.38 mg,1.34mmol,2 eq.) and EDCI (256.37 mg,1.34mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ Petroleum ether etoac=0:1) the residue was purified to give the product (2S) -2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] as an oil]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (180 mg,352.54umol,52.72% yield). MS (ESI) m/z 511.2[ M+H ]] ⁺

Step 5: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamide

Ammonia (7M, 20mL,397.12 eq) containing methyl (2S) -2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (180 mg,352.54 mol,1 eq) was stirred at 80℃for 16H. After completion, the mixture was concentrated under reduced pressure to give the product (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamide (170 mg, crude material) as an oil.

Step 6: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamide

To (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]To DCM (3 mL) of nonane-3-carboxamide (170 mg,343.04umol,1 eq.) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (408.74 mg,1.72mmol,5 eq.). The mixture was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product (3S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]Nonane-3-carboxamide (25 mg,51.09umol,14.89% yield, 97.6% purity). MS (ESI) M/z478.2[ M+H] ⁺

¹ H NMR(400MHz,MMeOD-d ₄ )δ＝7.22-7.12(m,1H),7.11-6.98(m,2H),6.58-6.45(m,1H),5.11-4.95(m,1H),4.65-4.52(m,1H),3.94(s,3H),3.93-3.80(m,2H),3.28-3.18(m,1H),2.54-2.02(m,4H),2.01-1.48(m,12H)。

EXAMPLE 24 Synthesis of viral protease inhibitor Compound 225

Step 1: (2S) -2-amino-3- (3-methylimidazol-4-yl) propionic acid methyl ester

At 25 ℃, to (2S) -2- (tert-butyloxycarbonylamino) -3- (3-methyl)To a solution of methylimidazol-4-yl) propionic acid (300 mg,1.11mmol,1 eq.) in EtOAc (1.2 mL) was added HCl/EtOAc (4M, 2.79mL,10 eq.). The reaction mixture was stirred at 25℃for 1.5h. The resulting mixture was concentrated to give the product. Methyl (2S) -2-amino-3- (3-methylimidazol-4-yl) propionate (250 mg, crude material, HCl) was obtained as a solid and used directly in the next step. MS (ESI) m/z 183.2[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 8.94(s,1H),7.56(s,1H),4.51(t,J＝7.17Hz,1H),3.93(s,3H),3.87(s,3H),3.46-3.55(m,1H),3.32-3.42(m,1H)。

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl ] amino ] -3- (3-methylimidazol-4-yl) propanoic acid methyl ester

At 0℃under N ₂ To a mixture of methyl (2S) -2-amino-3- (3-methylimidazol-4-yl) propionate (250 mg,1.14mmol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoic acid (263.22 mg,1.14mmol,1 eq.) in THF (1 mL) and DCM (1 mL) and DIPEA (441.26 mg,3.41mmol, 594.69. Mu.L, 3 eq.) was added T3P (1.09 g,1.71mmol,1.02mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 10h. LCMS showed the reaction mixture was complete. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was washed with brine (3 ml x 3), dried over anhydrous sodium sulfate and concentrated to give the crude product. Obtaining (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl as an oil ]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (360 mg, crude) and used directly in the next step. MS (ESI) m/z 397.3[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- (3-methylimidazol-4-yl) propanoic acid methyl ester

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (360 mg, 907.99. Mu.L, 1 eq.) to a mixture of DCM (3.3 mL) was added TFA (1.04 g,9.08mmol, 672.27. Mu.L, 10 eq.). The mixture was stirred at 25℃for 1.5h. LCMS showed reactionThe mixture is completed. The reaction mixture was concentrated to give the product. Obtaining (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl group as an oil]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (370 mg, crude material, TFA) and used directly in the next step. MS (ESI) M/z297.2[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- (3-methylimidazol-4-yl) propionic acid methyl ester

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (370 mg,1.25mmol,1 eq. TFA) and 4-methoxy-1H-indole-2-carboxylic acid (238.69 mg,1.25mmol,1 eq.) were added in one portion EDCI (478.66 mg,2.50mmol,2 eq.) and DMAP (305.05 mg,2.50mmol,2 eq.) in a mixture of DMF (1.5 mL) and DCM (1.5 mL). The mixture was stirred at 25℃for 12h. To the resulting mixture was added water (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was washed with brine (3 ml x 3) and dried over anhydrous sodium sulfate and concentrated to give the crude product. By column chromatography (SiO ₂ Petroleum ether/etoac=2/1 to EtOAc/methanol=10/1). Obtaining (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as an oil]-4-methyl-pentanoyl]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (270 mg, crude material). MS (ESI) M/z469.5[ M+H ]] ⁺

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]Methyl 3- (3-methylimidazol-4-yl) propionate (235.00 mg,500.50umol,1 eq.) was added NH at one time ₃ MeOH (7M, 1.94mL,27.14 eq.). The mixture was stirred at 80 ℃ and for 12h. LCMS showed the reaction mixture was complete. The reaction mixture was cooled to 25 ℃ and concentrated to give the crude product. The residue was purified by preparative TLC. Obtaining N- [ (1S) -1- [ [ (1S) -2-amino-1- [ (3-methylimidazol-4-yl) methyl ] as a solid]-2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (170 mg, crude). MS (ESI) m/z 455.3[ M+H ]] ⁺

Step 6: n- [ (1S) -1- [ [ (1S) -1-cyano-2- (3-methylimidazol-4-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ (1S) -2-amino-1- [ (3-methylimidazol-4-yl) methyl]-2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (140 mg,308.02umol,1 eq.) in DCM (2 mL) was added the Buerger's reagent (293.61 mg,1.23mmol,4 eq.). The mixture was stirred at 25 ℃ for 12h and then concentrated to give the crude product. The crude product was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ (1S) -1-cyano-2- (3-methylimidazol-4-yl) ethyl in solid form]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (10.59 mg,23.82umol,7.73% yield, 98.2% purity). MS (ESI) m/z 437.2[ M+H ]] ⁺ 。

Preparative HPLC conditions:

column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-50％，6min

Column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：20％-45％，8min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.52-7.57(m,1H),7.28(s,1H),7.12-7.18(m,1H),7.03(d,J＝8.38Hz,1H),6.85-6.96(m,1H),6.52(d,J＝7.72Hz,1H),5.05-5.13(m,1H),4.55-4.62(m,1H),3.86-3.99(m,3H),3.68(s,3H),3.21(tt,J＝15.24,7.80Hz,2H),1.55-1.81(m,3H),0.86-1.07(m,6H)

EXAMPLE 25 Synthesis of viral protease inhibitor Compound 227

Step 1: (2S) -2-amino-3- (1-methylimidazol-4-yl) propionic acid methyl ester

At 25℃under N ₂ To a mixture of (2S) -2-amino-3- (1-methylimidazol-4-yl) propionic acid (0.5 g,2.96mmol,1 eq.) was added HCl/MeOH (4M, 7.39mL,10 eq.) in one portion. The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated to give the product. Methyl (2S) -2-amino-3- (1-methylimidazol-4-yl) propionate (0.6 g, crude material, HCl) was obtained as a solid and used directly in the next step. MS (ESI) m/z 184.1[ M+H ] ] ⁺

Step 2: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- (1-methylimidazol-4-yl) propionic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (498.76 mg,1.64mmol,1.2 eq) and methyl (2S) -2-amino-3- (1-methylimidazol-4-yl) propionate (0.3 g,1.37mmol,1 eq, HCl), DIPEA (882.53 mg,6.83mmol,1.19mL,5 eq) in THF (0.9 mL) and DCM (0.9 mL) was added T3P (1.30 g,2.05mmol,1.22mL,50% purity, 1.5 eq). The mixture was stirred at 25℃for 12h. The reaction mixture was added to saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was washed with brine (3 ml x 3) and dried over anhydrous sodium sulfate and concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]-4-methyl-pentanoyl]Amino group]Methyl 3- (1-methylimidazol-4-yl) propionate (100 mg,202.97umol,14.86% yield, 95.3% purity). MS (ESI) m/z 470.2[ M+H ]] ⁺

Preparative HPLC conditions:

column: kromasil C18 (250 x 50mm x 10 um); mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-50％，10min

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-1- [ (1-methylimidazol-4-yl) methyl ] -2-oxo-ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) ammoniaBase group]-4-methyl-pentanoyl]Amino group]Methyl-3- (1-methylimidazol-4-yl) propionate (100 mg, 212.98. Mu. Mol,1 eq.) was added NH at one time ₃ MeOH (7M, 10.00mL,328.67 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the product. Obtaining N- [ (1S) -1- [ [ (1S) -2-amino-1- [ (1-methylimidazol-4-yl) methyl ] as a solid]-2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (95.5 mg,190.57umol,89.48% yield, 90.7% purity) and used directly in the next step. MS (ESI) m/z 455.2[ M+H ]] ⁺

Step 4: n- [ (1S) -1- [ [ (1S) -1-cyano-2- (1-methylimidazol-4-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ (1S) -2-amino-1- [ (1-methylimidazol-4-yl) methyl]-2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (80.00 mg,176.01umol,1 eq.) in DCM (1 mL) was added the Bogis reagent (83.89 mg,352.02umol,2 eq.) in one portion. The mixture was stirred at 25℃for 12h. Water (0.3 mL) was added to the reaction mixture and stirred for 10min. The reaction mixture was then concentrated to give the crude product. The crude product was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ (1S) -1-cyano-2- (1-methylimidazol-4-yl) ethyl in solid form ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (26.39 mg,60.27umol,34.24% yield, 99.684% purity). MS (ESI) m/z 437.2[ M+H ]] ⁺

Preparative HPLC conditions:

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.35(s,1H),7.28(s,1H),7.12-7.20(m,1H),7.05(d,J＝8.38Hz,1H),6.91-6.98(m,1H),6.53(d,J＝7.72Hz,1H),5.01(t,J＝7.06Hz,1H),4.63(br dd,J＝9.59,4.96Hz,1H),3.94(s,3H),3.46-3.59(m,3H),3.00-3.13(m,2H),1.61-1.81(m,3H),0.89-1.07(m,6H)

EXAMPLE 26 Synthesis of viral protease inhibitor Compounds 231

Step 1: (S) -2-amino-3- (pyridin-3-yl) propionic acid methyl ester hydrochloride

At 25℃under N ₂ To a mixture of (2S) -2- (tert-butoxycarbonylamino) -3- (3-pyridyl) propionic acid (500 mg,1.88mmol,1 eq.) was added HCl/MeOH (4M, 20.80mL,44.31 eq.) in one portion. The mixture was stirred at 25 ℃ and for 12h. After completion, the reaction mixture was concentrated to give methyl (2S) -2-amino-3- (3-pyridinyl) propanoate (400 mg, crude material, HCl) as an oil and used directly in the next step. MS (ESI) M/z181.1[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (pyridin-3-yl) propionic acid methyl ester

At 0℃under N ₂ Downward methyl (2S) -2-amino-3- (3-pyridinyl) propanoate (0.3 g,1.66mmol,1 eq. HCl) and (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino)]To a mixture of 4-methyl-pentanoic acid (506.66 mg,1.66mmol,1 eq.) and DIPEA (1.08 g,8.32mmol,1.45mL,5 eq.) in THF (0.6 mL) and DCM (0.6 mL) was added T ₃ P (1.59 g,2.50mmol,1.49mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 12h. After completion, a saturated sodium bicarbonate solution (10 mL) was added to the reaction mixture and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by slurrying with petroleum ether (20 mL) and filtered to give a filter cake as product. Obtaining (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid ]-4-methyl-pentanoyl]Amino group]Methyl 3- (3-pyridyl) propionate (0.4 g, crude material) and used directly in the next step. MS (ESI) m/z 467.1[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- (pyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at a time to a mixture of methyl 3- (3-pyridyl) propionate (200.00 mg,428.70umol,1 eq.) ₃ MeOH (7M, 5mL,81.64 eq.). The mixture was stirred at 80℃for 4h. After completion, the reaction mixture was cooled to 25 ℃ and concentrated to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- (3-pyridylmethyl) ethyl as a solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (0.18 g,339.65umol,79.23% yield, 85.2% purity) and used directly in the next step. MS (ESI) m/z 452.2[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-cyano-2- (pyridin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- (3-pyridylmethyl) ethyl ]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (0.1 g,221.48umol,1 eq.) in DCM (1 mL) was added the Bogis reagent (105.56 mg,442.95umol,2 eq.) in one portion. The mixture was stirred at 25℃for 12h. To the above solution was added additional Prague reagent (105.56 mg,442.95umol,2 eq.) at 25℃and the reaction mixture was stirred for 1h at 25 ℃. After completion, water (0.5 mL) was added to the reaction mixture and stirred for 10min. The mixture was then concentrated to give the crude product. Purification of the crude product by preparative HPLC gave N- [ (1S) -1- [ [ (1S) -1-cyano-2- (3-pyridinyl) ethyl ] as a solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23.18 mg,52.94umol,23.90% yield, 99.009% purity). MS (ESI) m/z 434.2[ M+H ]] ⁺

Preparative HPLC conditions:

column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-60％，10min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 8.47-8.52(m,1H),8.34-8.45(m,1H),7.77-7.84(m,1H),7.28-7.38(m,1H),7.23-7.28(m,1H),7.12-7.19(m,1H),6.99-7.07(m,1H),6.52(d,J＝7.63Hz,1H),5.08-5.18(m,1H),4.48-4.61(m,1H),3.94(s,3H),3.12-3.29(m,2H),1.41-1.76(m,3H),0.87-1.03(m,6H)。

Step 5: (S) -2- (4-methoxy-1H-indole-2-carboxamide) -4-methylpentanoic acid tert-butyl ester

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (5 g,26.15mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (5.88 g,31.38mmol,1.2 eq., HCl), EDCI (6.52 g,34.00mmol,1.3 eq.), HOBt (4.59 g,34.00mmol,1.3 eq.) in DMF (30 mL) was added TEA (7.94 g,78.46mmol,10.92mL,3 eq.) in one portion. The mixture was stirred at 25 ℃ and stirred for 2h. After completion, water (90 mL) was added to the reaction mixture and extracted with ethyl acetate (25 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (25 mL) and 5% aqueous sodium bicarbonate (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Purification of the residue by column chromatography (SiO ₂ Petroleum ether: etoac=30:1 to 10:1), to give (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a solid]-4-methyl-pentanoic acid tert-butyl ester (5.93 g,16.45mmol,62.91% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

Step 6: (S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-valerate (0.5 g,1.39mmol,1 eq.) in DCM (0.33 mL) was added TFA (2.57 g,22.51mmol,1.67mL,16.23 eq.) and H in one portion ₂ O (166.71 mg,9.25mmol, 166.71. Mu.L, 6.67 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. After completion, the reaction mixture was concentrated to give (S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanoic acid as a solid(400 mg, crude, TFA) and used directly in the next step. MS (ESI) m/z 305.1[ M+H ]] ⁺

EXAMPLE 27 Synthesis of viral protease inhibitor Compound 599

Step 1: (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoic acid tert-butyl ester

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (5 g,26.15mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (5.88 g,31.38mmol,1.2 eq., HCl), EDCI (6.52 g,34.00mmol,1.3 eq.), HOBt (4.59 g,34.00mmol,1.3 eq.) in DMF (30 mL) was added TEA (7.94 g,78.46mmol,10.92mL,3 eq.) in one portion. The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (90 mL) and extracted with EtOAc (25 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (25 mL) and 5% aqueous sodium bicarbonate (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether etoac=30:1 to 10:1) purification residue. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoic acid tert-butyl ester (5.93 g,16.45mmol,62.91% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

Step 2: (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-valerate (2.00 g,5.55mmol,1 eq.) in DCM (8 mL) was added TFA (10.27 g,90.04mmol,6.67mL,16.23 eq.) and H in one portion ₂ O(66667mg,37.01mmol, 666.67. Mu.L, 6.67 eq.). The mixture was stirred at 25 ℃ and for 4h. The reaction mixture was concentrated to give a crude product. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoic acid (2.24 g,5.35mmol,96.50% yield, TFA) and used directly in the next step. MS (ESI) M/z305.1[ M+H] ⁺

Step 3:2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- (2-oxo-1H-quinolin-4-yl) propionic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (568.23 mg,1.36mmol,1.2 eq, TFA) and methyl 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionate (320 mg,1.13mmol,1 eq, HCl), DIPEA (731.40 mg,5.66mmol, 985.72. Mu.L, 5 eq) in THF (1 mL) and DCM (1 mL) was added T3P (1.08 g,1.70mmol,1.01mL,50% purity, 1.5 eq). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid ]-4-methyl-pentanoyl]Amino group]Methyl 3- (2-oxo-1H-quinolin-4-yl) propionate (0.2 g,375.53umol,33.18% yield). MS (ESI) m/z 533.2[ M+H ]] ⁺

Preparative HPLC conditions: column: kromasil C18 (250 x 50mm x 10 um); mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-60％，10min

Step 4: n- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-1H-quinolin-4-yl) methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at one time to a mixture of methyl-3- (2-oxo-1H-quinolin-4-yl) propionate (200.00 mg,375.53umol,1 eq) ₃ MeOH (7M, 10.00mL,186.41 eq.). The mixture was stirred at 80℃for 12h. Will be reversedThe mixture was cooled to 25 ℃ and concentrated to give the product. Obtaining N- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-1H-quinolin-4-yl) methyl in solid form]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (180 mg,326.21umol,86.87% yield, 93.8% purity) and used directly in the next step. MS (ESI) m/z 518.2[ M+H ]] ⁺

Step 5: n- [ (1S) -1- [ [ 1-cyano-2- (2-oxo-1H-quinolin-4-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-1H-quinolin-4-yl) methyl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (90 mg,173.89umol,1 eq.) in DCM (5 mL) was added the Bungeus reagent (207.19 mg,869.44umol,5 eq.) in one portion. The mixture was stirred at 25 ℃ for 12h and then concentrated to give the crude product.

The residue was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ 1-cyano-2- (2-oxo-1H-quinolin-4-yl) ethyl in solid form]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (20.74 mg,41.13umol,23.66% yield, 99.079% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺

Preparative HPLC conditions: column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-65％，10min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.93(br d,J＝8.16Hz,1H),7.50-7.58(m,1H),7.28-7.40(m,2H),7.26(dd,J＝11.47,0.66Hz,1H),7.11-7.19(m,1H),7.04(dd,J＝8.27,4.08Hz,1H),6.59-6.70(m,1H),6.46-6.56(m,1H),5.24-5.34(m,1H),4.53(td,J＝10.31,5.18Hz,1H),3.93(d,J＝4.41Hz,3H),3.40-3.59(m,3H),1.72(ddd,J＝15.16,9.87,5.18Hz,1H),1.53-1.66(m,2H),1.40-1.50(m,1H),0.87-1.01(m,5H)

Step 6: 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionic acid methyl ester

At 25℃under N ₂ Down 2-amino-3- (2-oxo-1H-quinolin-4-yl)) To propionic acid (400 mg,1.72mmol,1 eq.) was added HCl/MeOH (4M, 4.31mL,10 eq.) in one portion. The mixture was stirred at 25 ℃ and for 1h. The reaction mixture was concentrated to give the product. Methyl 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionate (370 mg, crude material, HCl) was obtained as a solid and was used directly in the next step.

EXAMPLE 28 Synthesis of viral protease inhibitor Compounds 249

Step 1: 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionic acid

At 25℃under N ₂ Down 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionic acid (200 mg,861.20umol,1 eq.) in H ₂ Pd/C (20 mg,861.20umol,10% purity) was added to a solution in O (1 mL). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (861.20 umol) (15 psi) was stirred at 70℃for 5h. The reaction mixture was cooled to 25 ℃ and filtered to give a filtrate. Concentrating the filtrate to obtain the product. 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionic acid (200 mg, crude material) was obtained as a solid and used directly in the next step. MS (ESI) m/z 235.0[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 1.92-2.03(m,1H)2.06-2.21(m,1H)2.45-2.62(m,1H)2.86(dd,J＝16.43,6.06Hz,1H)3.32-3.40(m,1H)3.83(br dd,J＝8.49,5.84Hz,1H)3.93(br t,J＝6.95Hz,1H)6.93(d,J＝7.72Hz,1H)7.01-7.10(m,1H)7.24(br t,J＝7.72Hz,1H)7.36(d,J＝7.06Hz,1H)

Step 2: 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionic acid methyl ester

At 25℃under N ₂ To 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionic acid (200 mg,853.79 mol,1 eq.) was added HCl/MeOH (4M, 9.91mL,46.45 eq.) in one portion. The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give a crude product. Obtain a yellow oily form Methyl 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionate (260 mg, crude material, HCl) and was used directly in the next step. MS (ESI) m/z 249.1[ M+H ]] ⁺

Step 3:2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionic acid methyl ester

At 0℃under N ₂ Downward methyl 2-amino-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionate (260 mg,913.12umol,1 eq. HCl) and (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a mixture of 4-methyl-pentanoic acid (277.90 mg,913.12umol,1 eq.) and DIPEA (590.07 mg,4.57mmol, 795.24. Mu.L, 5 eq.) in THF (0.6 mL) and DCM (0.6 mL) was added T ₃ P (871.61 mg,1.37mmol, 814.59. Mu.L, 50% purity, 1.5 eq.). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoyl]Amino group]-methyl 3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionate (85 mg,151.05umol,16.54% yield, 95% purity). MS (ESI) m/z 535.2[ M+H ] ] ⁺

^\ Preparative HPLC conditions: column: phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ Water (10 mM NH4HCO 3) -ACN]；B％：27％-47％，8min

Step 4: n- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at one time to a mixture of methyl-3- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) propionate (55 mg,102.88umol,1 eq) ₃ MeOH (7M, 1.83mL,124.74 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the product. Obtaining N-in solid form(1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) methyl]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (55 mg, crude) and used directly in the next step. MS (ESI) m/z 518.2[ M+H ]] ⁺

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) methyl ]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (75 mg,144.34umol,1 eq.) in DCM (0.1 mL) was added the Bogis reagent (103.19 mg,433.03umol,3 eq.) in one portion. The mixture was stirred at 25 ℃ and for 16h. Water (0.5 mL) was added to the reaction mixture and stirred for 10min. The mixture was then concentrated to give the crude product. The crude product was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ 1-cyano-2- (2-oxo-3, 4-dihydro-1H-quinolin-4-yl) ethyl in solid form]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (26.51 mg,52.85umol,36.62% yield, 100% purity). MS (ESI) M/z502.2[ M+H] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.51-11.61(m,1H),10.14-10.20(m,1H),8.84-9.01(m,1H),8.42-8.59(m,1H),7.32-7.42(m,1H),7.05-7.22(m,3H),6.81-7.04(m,3H),6.50(dd,J＝7.64,3.85Hz,1H),4.37-4.66(m,2H),3.83-3.95(m,3H),2.95-3.12(m,1H),2.63-2.82(m,1H),2.26-2.42(m,1H),1.88-2.08(m,2H),1.45-1.82(m,3H),0.81-1.02(m,6H)

Step 6: (S) -2- (4-methoxy-1H-indole-2-carboxamide) -4-methylpentanoic acid tert-butyl ester

At 25℃under N ₂ Downward 4-methoxy-1H-indole-2-carboxylic acid (15 g,78.46mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (21.07 g,94.15mmol, 1.2.)To a mixture of HCl in DMF (150 mL) were added EDCI (19.55 g,102.00mmol,1.3 eq.), HOBt (13.78 g,102.00mmol,1.3 eq.), TEA (23.82 g,235.38mmol,32.76mL,3 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (450 mL) and extracted with EtOAc (250 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (300 mL) and 5% aqueous sodium bicarbonate (300 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether etoac=30:1 to 10:1) purification residue. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoic acid tert-butyl ester (24 g,66.58mmol,84.87% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

Step 7: (S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-pentanoate (10 g,27.74mmol,1 eq.) in DCM (30 mL) was added TFA (61.60 g,540.26mmol,40mL,19.47 eq.) and H in one portion ₂ O (4.00 g,221.98mmol,4.00mL,8.00 eq.). The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated to give a crude product. The crude product was purified with petroleum ether: ethyl acetate=10:1 (20 mL) and filtered to give the product. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group as a solid]-4-methyl-pentanoic acid (6 g,19.22mmol,69.27% yield, 97.48% purity). MS (ESI) m/z 305.1[ M+H ]] ⁺

EXAMPLE 29 Synthesis of viral protease inhibitor Compound 600

Step 1: 2-amino-3- (2-oxo-1, 2-dihydropyridin-3-yl) propionic acid methyl ester

A mixture of 2-amino-3- (2-oxo-1H-pyridin-3-yl) propionic acid (500 mg,2.74mmol,1 eq.) and HCl/MeOH (4M, 30mL,43.72 eq.) was stirred at 25℃for 2H. Concentrating the reaction mixture under reduced pressure to give a yellow color The product methyl 2-amino-3- (2-oxo-1, 2-dihydropyridin-3-yl) propionate (650 mg, crude material, HCl) was used as an oil and directly in the next step. MS (ESI) m/z 197.0[ M+H ]] ⁺

Step 2: methyl-2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate

A mixture of methyl 2-amino-3- (2-oxo-1H-pyridin-3-yl) propionate (650 mg,2.79mmol,1 eq., HCl), (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoic acid (646.16 mg,2.79mmol,1 eq.), EDCI (1.07 g,5.59mmol,2 eq.), DMAP (682.62 mg,5.59mmol,2 eq.), DMF (2 mL), and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/EtOAc=0/1) the residue was purified to give the product methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate as a solid (900 mg,1.89mmol,67.68% yield, 86.02% purity). MS (ESI) m/z 410.1[ M+H ] ] ⁺

Step 3:2- ((S) -2-amino-4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoic acid methyl ester

A mixture of methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propionate (200 mg,488.43umol,1 eq) and HCl/EtOAc (4M, 30 mL) was stirred at 27℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give the product methyl 2- ((S) -2-amino-4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (170 mg, crude material, HCl) as a solid and used directly in the next step.

Step 4:2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoic acid methyl ester

Methyl 2- ((S) -2-amino-4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (1)A mixture of 70mg,491.58 mol,1 eq, HCl), 4-methoxy-1H-indole-2-carboxylic acid (93.98 mg,491.58 mol,1 eq), EDCI (188.47 mg,983.17 mol,2 eq), DMAP (120.11 mg,983.17 mol,2 eq), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and then extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/etoac=0/1) to give the compound methyl 2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (130 mg,269.41umol,54.81% yield) as a solid. MS (ESI) m/z 483.1[ M+H ]] ⁺

Step 5: n- ((2S) -1- ((1-amino-1-oxo-3- (2-oxo-1, 2-dihydropyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

Methyl 2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (190 mg,393.76umol,1 eq.) NH ₃ The mixture of MeOH (7M, 10 mL) was stirred at 80℃for 15h. The reaction mixture was concentrated under reduced pressure to give N- ((2S) -1- ((1-amino-1-oxo-3- (2-oxo-1, 2-dihydropyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (190 mg, crude material) as a solid. MS (ESI) m/z 468.2[ M+H ]] ⁺

Step 6: n- ((2S) -1- ((1-cyano-2- (2-oxo-1, 2-dihydropyridin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

A mixture of N- ((2S) -1- ((1-amino-1-oxo-3- (2-oxo-1, 2-dihydropyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (180 mg,385.01umol,1 eq), pragus reagent (917.53 mg,3.85mmol,10 eq) and DCM (30 mL) was stirred at 25℃for 8H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: PThe henomenex Gemini-NX C18 is 75 x 30mm x 3um; mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8 min) to give the product N- ((2S) -1- ((1-cyano-2- (2-oxo-1, 2-dihydropyridin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (24 mg,52.18umol,13.55% yield, 97.73% purity) as a solid. MS (ESI) m/z 450.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.90-11.40(m,2H),9.08-8.85(m,1H),8.55-8.35(m,1H),7.51-7.26(m,3H),7.16-7.05(m,1H),7.04-6.94(m,1H),6.51(d,J＝7.5Hz,1H),6.15(t,J＝6.6Hz,1H),5.19-5.01(m,1H),4.55-4.33(m,1H),3.89(s,3H),3.02-2.78(m,2H),1.75-1.33(m,3H),0.98-0.72(m,6H)

EXAMPLE 30 Synthesis of viral protease inhibitor Compounds 344C, 344D, 507 and 511

/>

Step of compound 344C: n- [ (1S) -1- [ [ (1S) -2-amino-2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (100 mg,180.79 mol,80% purity, 1 eq.) in DCM (10 mL) was added NH ₃ .H ₂ O (46.93 mg,361.58umol, 51.57. Mu.L, 27% purity, 2 eq.) and NH ₄ Cl (19.34 mg,361.58 mol,2 eq.). The mixture was stirred at 25℃for 30min, followed by the addition of KCN-containing H (94.18 mg,1.45mmol, 61.96. Mu.L) ₂ O (0.2 mL) and the mixture was stirred at 30deg.C for 16h. After the reaction was completed, H was then added at 0deg.C ₂ O (10 mL) to quench the reaction mixture, and then H was used ₂ O (10 mL) was diluted and extracted with EtOAc (30 mL x 2). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Drying and filteringAnd concentrated under reduced pressure to give a residue. NaOH was added to liquid water to adjust to ph=9, quenched with aqueous NaCl solution, and NaOH was then added to adjust to pH>14. Purification of the residue by HCl preparative HPLC gives the compound N- [ (1S) -1- [ [ (1S) -2-amino-2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,103.83 mol,57.43% yield, 97.3% purity). MS (ESI) m/z 469.2[ M+H ] ⁺

Preparative HPLC conditions:

column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ water (0.04% hcl) -ACN ]; b%:15% -40%,7min

¹ H NMR(400MHz,DMSO-d6)δ＝11.59(dd,J＝1.9,5.0Hz,1H),9.16-8.58(m,2H),8.54-8.26(m,2H),7.66(d,J＝9.0Hz,1H),7.37(dd,J＝2.0,4.2Hz,1H),7.14-7.06(m,1H),7.04-6.97(m,1H),6.51(d,J＝7.5Hz,1H),4.61-4.42(m,2H),4.39-4.21(m,1H),3.88(s,3H),3.20-2.98(m,2H),2.48-2.34(m,1H),2.14-1.88(m,2H),1.82-1.47(m,5H),0.92(dd,J＝6.0,14.8Hz,6H)

Step of Compound 511: n- [ (1S) -1- [ [ (1S) -2-cyano-2- (ethylamino) -1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (80 mg,108.47 mol,60% purity, 1 eq.) in DCM (5 mL) was added PdCl ₂ (3.85 mg,21.69umol,0.2 eq.) Na ₂ SO ₄ (53.93 mg, 379.66. Mu.L, 38.52. Mu.L, 3.5 eq.) and ethylamine (9.78 mg, 216.95. Mu.L, 14.19. Mu.L, 2 eq.). The resulting mixture was stirred at 25 ℃ for 30min, and then TMSCN (21.52 mg,216.95umol,27.14 μl,2 eq.) was added. The resulting mixture was stirred at 25℃for 1h. After the reaction was completed, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to give 70mg of a mixture. Purification of the mixture by SFC gives N- [ (1S) -1- [ [ (1S) -2-cyano-2 as an oil - (ethylamino) -1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (16 mg,28.20umol,26.00% yield, 87.525% purity) and N- [ (1S) -1- [ [ (1S) -2-cyano-2- (ethylamino) -1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (16 mg,31.44umol,28.98% yield, 97.569% purity). MS (ESI) m/z 497.3[ M+H ]] ⁺

Preparative HPLC conditions:

column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ water (0.04% hcl) -ACN ]; b%:25% -40%,7min

SFC conditions:

column: DAICEL CHIRALCEL OX (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-ETOH ]; b%:38% -38%,9min

Compound 511 isomer 1: ¹ H NMR(400MHz,DMSO-d6)δ＝11.56(br s,1H),8.37(br d,J＝7.7Hz,1H),8.29-8.20(m,1H),7.80-7.48(m,3H),7.35(br d,J＝2.0Hz,1H),7.17-6.96(m,2H),6.50(d,J＝7.7Hz,1H),4.53-4.40(m,1H),4.05(td,J＝3.9,7.7Hz,1H),3.88(s,3H),3.77(br dd,J＝4.9,10.1Hz,1H),3.18-2.97(m,2H),2.88-2.63(m,2H),2.40-2.24(m,1H),2.14-2.06(m,2H),1.82-1.31(m,5H),1.09-0.98(m,3H),0.91(br dd,J＝6.2,16.1Hz,6H)

compound 511 isomer 2: ¹ H NMR(400MHz,DMSO-d6)δ＝11.58(d,J＝1.5Hz,1H),8.41(br d,J＝7.9Hz,1H),8.17(br s,1H),7.63-7.50(m,1H),7.37(d,J＝1.8Hz,1H),7.14-7.05(m,1H),7.00(d,J＝8.2Hz,1H),6.50(d,J＝7.5Hz,1H),4.58-4.37(m,1H),4.25-3.99(m,1H),3.88(s,3H),3.81-3.51(m,1H),3.16-2.96(m,2H),2.89-2.54(m,2H),2.43-2.23(m,1H),2.20-1.99(m,1H),1.95-1.43(m,6H),1.10-0.98(m,3H),0.91(dd,J＝6.4,15.2Hz,6H)

step of compound 507: n- [ (1S) -1- [ [ (1S) -2- (benzylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl group-butyl group]To a mixture of 4-methoxy-1H-indole-2-carboxamide (150 mg,271.18umol,80% purity, 1 eq.) in DCM (15 mL) was added PdCl ₂ (9.62 mg,54.24umol,0.2 eq.) Na ₂ SO ₄ (134.82 mg,949.14umol, 96.30. Mu.L, 3.5 eq.) and BnNH ₂ (58.11 mg,542.36umol, 59.12. Mu.L, 2 eq.). The mixture was stirred at 25℃for 30min, followed by the addition of TMSCN (53.81 mg,542.36umol, 67.85. Mu.L, 2 eq.). The mixture was stirred at 25℃for 2h. After the reaction was completed, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by HCl preparative HPLC gave the compound N- [ (1S) -1- [ [ (1S) -2- (benzylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (30 mg,51.71umol,19.07% yield, 96.291% purity) and N- [ (1S) -1- [ [ (1S) -2- (benzylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl)]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (18 mg,31.04umol,11.44% yield, 96.329% purity). MS (ESI) m/z 559.3[ M+H ]] ⁺

Preparative HPLC conditions:

column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ water (0.04% hcl) -ACN ]; b%:38% -62%,7min

Compound 507 isomer 1: ¹ H NMR：(400MHz,DMSO-d6)δ＝11.58(d,J＝1.8Hz,1H),8.48-8.34(m,1H),8.23(br d,J＝9.5Hz,1H),7.69-7.53(m,1H),7.51-7.23(m,5H),7.14-7.05(m,1H),7.02-6.97(m,1H),6.50(d,J＝7.7Hz,1H),4.56-4.37(m,1H),4.23(br d,J＝9.3Hz,1H),4.13-3.91(m,2H),3.88(s,3H),3.84(br d,J＝13.2Hz,1H),3.17-2.95(m,2H),2.42-2.24(m,1H),2.16-1.98(m,1H),1.93-1.44(m,6H),0.90(dd,J＝6.3,16.2Hz,6H)

Compound 507 isomer 2: ¹ H NMR(400MHz,DMSO-d6)δ＝11.56(br d,J＝1.5Hz,1H),8.52-8.14(m,2H),7.69-7.55(m,1H),7.49-7.22(m,6H),7.13-7.05(m,1H),7.00(d,J＝8.4Hz,1H),6.50(d,J＝7.5Hz,1H),4.56-4.41(m,1H),4.21(br s,1H),4.06-3.94(m,2H),3.88(s,3H),3.83(br d,J＝12.8Hz,1H),3.17-2.97(m,2H),2.42-2.29(m,1H),2.17-2.00(m,2H),1.83-1.44(m,5H),0.90(dd,J＝6.3,17.8Hz,6H)

EXAMPLE 31 Synthesis of viral protease inhibitor Compound 129

Step 1.2- (trichloromethyl) -3H-imidazo [4,5-c ] pyridine

To a solution of pyridine-3, 4-diamine (2 g,18.33mmol,1 eq.) in AcOH (25 mL) was added methyl 2, 2-trichloroethyliminoate (3.88 g,21.99mmol,2.71mL,1.2 eq.). The solution was stirred at 100℃for 5h. Addition of H to the reactant ₂ O (90 mL) and extracted with ethyl acetate (70 mL. Times.3) and extracted with NaHCO ₃ (90 ml x 2) washing. The organic layer was concentrated carefully to give crude 2- (trichloromethyl) -3H-imidazo [4,5-c ] as a yellow solid]Pyridine (800 mg, crude material). The crude material was used directly in the next step. MS (ESI) M/z235.9[ M+H ]] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -3H-imidazo [4,5-c ] pyridine-2-carboxamide

To 2- (trichloromethyl) -3H-imidazo [4,5-c]Pyridine (150 mg,634.29umol,1 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (167.66 mg,634.29umol,1 eq.) in THF (5 mL) and H ₂ Na was added to the solution in O (2.5 mL) ₂ CO ₃ (201.68 mg,1.90mmol,3 eq.). The solution was stirred at 20℃for 1h. Adding H to the solution ₂ O (20 mL), extracted with ethyl acetate (40 mL x 3) and concentrated to give the crude material. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -23%,8 min) to give a 70% purity product and then continuing to pass through preparative HPLC (column: phenomenex Luna C1875×30mm×3um; mobile phase: [ Water (0.2% FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -30%,8 min) to give the product N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-3H-imidazo [4,5-c ]]Pyridine-2-carboxamide (3 mg,6.96umol,1.10% yield, 95% purity). MS (ESI) m/z 410.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.89-8.81(m,2H),8.77(d,J＝7.9Hz,1H),8.21(d,J＝5.4Hz,2H),7.54(s,1H),7.43(br d,J＝5.4Hz,1H),4.91-4.76(m,1H),4.44-4.32(m,1H),3.02-2.92(m,2H),2.25-2.16(m,1H),2.03-1.91(m,2H),1.78-1.38(m,4H),0.59(br s,1H),0.25(br d,J＝7.9Hz,2H),0.05-0.11(m,2H)。

EXAMPLE 32 Synthesis of viral protease inhibitor Compounds 389A and 389B

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propanamide

Tert-butyl N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamate (2 g,7.37mmol,1 eq.) in HCl/EtOAc (4M, 50mL,27.13 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionamide (1.2 g, crude material) as a solid.

Step 2: 2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

A solution of 2-tert-butoxycarbonyl-2-azaspiro [4.5] decane-3-carboxylic acid (3 g,10.59mmol,1 eq.) in HCl/MeOH (4M, 50mL,18.89 eq.) was stirred at 80℃for 2h. The mixture was concentrated under reduced pressure to give methyl 2-azaspiro [4.5] decane-3-carboxylate (2 g, crude material) as a yellow oil.

Step 3:2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

To 2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid methyl ester (2 g,10.14mmol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (2.33 g,12.17mmol,1.2 eq.) in DCM (30 mL) and DMF (5 mL) was added T3P (12.90 g,20.28mmol,12.06mL,50% purity, 2 eq.) and DIEA (3.93 g,30.41mmol,5.30mL,3 eq.). At 25 DEG CThe mixture was stirred for 2h. After completion, by adding H ₂ O (100 mL) to quench the reaction mixture and extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) to give the product 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a solid ]Decane-3-carboxylic acid methyl ester (3 g,8.10mmol,79.88% yield). MS (ESI) m/z 371.1[ M+H ]] ⁺

Step 4:2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxylic acid methyl ester (3 g,8.10mmol,1 eq.) in THF (45 mL) and H ₂ LiOH.H was added to the solution in O (15 mL) ₂ O (1.70 g,40.49mmol,5 eq.). The mixture was stirred at 25℃for 12h. After completion, by adding H ₂ O (50 mL) to quench the mixture, and then aqueous HCl (1M) was added to adjust to ph=3-4, and then extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (2.6 g, crude material). MS (ESI) m/z 357.1[ M+H ]] ⁺

Step 5: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 0deg.C]Decane-3-carboxylic acid (1 g,2.81mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a solution of propionamide (720.49 mg,4.21mmol,1.5 eq.) in DCM (30 mL) was added T3P (3.57 g,5.61mmol,3.34mL,50% purity, 2 eq.) and DIEA (1.09 g,8.42mmol,1.47mL,3 eq.). The mixture was stirred at 30℃for 1h. After completion, by adding H ₂ O (100 mL) to quench the mixture and then extract with DCM (50 mL x 3). Combined withThe organic layer was washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (700 mg,1.37mmol,48.96% yield). MS (ESI) m/z 510.3[ M+H ]] ⁺

Step 6: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxamide (700 mg,1.37mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (982.03 mg,4.12mmol,3 eq.). The mixture was stirred at 25℃for 2h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (500 mg,1.02mmol,74.05% yield). MS (ESI) m/z 492.3[ M+H ]] ⁺

Step 7: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,9 min) isolation of N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (500 mg,1.02 mmol) gives the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [ e.g.)4.5]Decane-3-carboxamide, isomer 1 (264 mg,537.04umol,52.80% yield). MS (ESI) m/z 492.3[ M+H ]] ⁺ ； ¹ H NMR (400 MHz, methanol-d) ₄ ) Delta = 7.28-6.76 (m, 3H), 6.60-6.38 (m, 1H), 5.05 (brdd, J = 5.2,10.2hz, 1H), 4.63-4.60 (m, 1H), 4.03-3.85 (m, 5H), 3.74-3.28 (m, 1H), 2.73 (brdd, J = 5.0,8.6hz, 1H), 2.51-2.28 (m, 2H), 2.27-2.08 (m, 1H), 1.96-1.72 (m, 2H), 1.69-1.38 (m, 11H), 1.37-1.09 (m, 1H); and

N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl in solid form]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5 ]]Decane-3-carboxamide, isomer 2 (140 mg,284.51umol,27.97% yield). MS (ESI) m/z 492.3[ M+H ]] ⁺ ； ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.30-6.81(m,3H),6.53(br d,J＝2.0Hz,1H),5.12-4.95(m,2H),4.70-4.55(m,2H),4.08-3.86(m,4H),3.84-3.72(m,1H),2.62-2.40(m,1H),2.36-2.18(m,2H),1.94-1.69(m,3H),1.68-1.34(m,11H)。

EXAMPLE 33 Synthesis of viral protease inhibitor Compounds 399

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester hydrochloride

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (130 mg,454.03umol,1 eq.) in HCl/dioxane (4M, 2.27mL,20 eq.) was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (173.4 mg,451.67umol,99.48% yield, HCl) as a yellow liquid product.

Step 2: (S) -7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

To (7S) -6-tert-Butoxycarbonyl-6-azaspiro [3.4]]Octane-7-carboxylic acid (105.34 mg,412.59umol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (158.4 mg, 41)To a solution of 2.59 mol,1 eq, HCl) in DCM (1.2 mL) and DMF (0.4 mL) was added DMAP (100.81 mg,825.19 mol,2 eq) and EDCI (158.19 mg,825.19 mol,2 eq). The reaction mixture was stirred at 25℃for 1h. Residue with H ₂ O (6 mL) was diluted and extracted with ethyl acetate (3 mL). The combined organic layers were washed with ethyl acetate (3 ml x 3), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the product (7S) -7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a liquid]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Tert-butyl octane-6-carboxylate (66.3 mg,156.55 mol,37.94% yield). MS (ESI) M/z424.0[ M+H ]] ⁺

Step 3: (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((S) -6-azaspiro [3.4] octane-7-carboxamido) propanoic acid methyl ester

A solution of (7S) -7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (66.3 mg,156.55 mol,1 eq) in HCl/MeOH (4M, 782.76. Mu.L, 20 eq) was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (7S) -6-azaspiro [3.4] octane-7-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (71.1 mg,156.09umol,99.71% yield, 79% purity, HCl) as a yellow liquid.

Step 4: (S) -2- ((S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (7S) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (62.8 mg,137.87umol,1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (26.36 mg,137.87umol,1 eq.) were added DMAP (33.69 mg,275.74umol,2 eq.) and EDCI (52.86 mg,275.74umol,2 eq.) in DCM (1.2 mL) and DMF (0.4 mL) for 1H at 25 ℃. The residue was diluted with brine (6 mL) and extracted with ethyl acetate (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the product (2S) -2- [ [ (7S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a white solid]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (33.2 mg,66.86umol,48.50% yield). MS (ESI) m/z 497.1[ M+H ]] ⁺

Step 5: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [ (7S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (23.0 mg,46.32umol,1 eq.) and ammonia (7M, 4mL,604.50 eq.) was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to give the product (7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carboxamide (15 mg, crude). MS (ESI) m/z 482.2[ M+H ]] ⁺

Step 6: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]A solution of octane-7-carboxamide (15 mg, 28.66. Mu. Mol,1 eq.) and the Buerger's reagent (13.66 mg, 57.32. Mu. Mol,2 eq.) was stirred at 25℃for 24h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -45%,8 min) to give the product (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide (3.01 mg,6.49umol,22.66% yield). M is MS(ESI)m/z 464.3[M+H] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δppm 6.95-7.24(m,3H)6.47-6.58(m,1H)5.01(br dd,J＝10.67,5.19Hz,1H)4.58(t,J＝7.09Hz,1H)3.82-4.19(m,5H)3.19(br t,J＝8.52Hz,1H)2.93-3.07(m,1H)2.28-2.56(m,3H)2.16-2.27(m,2H)1.94-2.14(m,6H)1.47-1.86(m,2H)。

EXAMPLE 34 Synthesis of viral protease inhibitor Compound 405

Step 1: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (225 mg,1.21mmol,1 eq.) in DMF (2 mL) and DCM (4 mL) was added TEA (733.62 mg,7.25mmol,1.01mL,6 eq.) and T ₃ P (1.15 g,3.62mmol,1.08mL,3 eq.) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (296.42 mg,1.21mmol,1 eq.). The solution was stirred at 25℃for 1h. Addition of H to the reactant ₂ O (40 mL) and extracted with ethyl acetate (50 mL. Times.3) and the organic layer was concentrated carefully to give crude compound (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (440 mg, crude) and was used directly in the next step. MS (ESI) m/z 414.1[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (440 mg,1.06mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 1h. TLC (DCM: meoh=10:1). The reaction was carefully concentrated to give the crude material. The solid compound (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (310 mg, crude)Substance) is used directly in the next step. MS (ESI) M/z314.3[ M+H] ⁺

Step 3: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (310 mg,989.18umol,1 eq.) in DMF (4 mL) and DCM (4 mL) were added EDCI (379.25 mg,1.98mmol,2 eq.) and DMAP (241.70 mg,1.98mmol,2 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (189.11 mg,989.18umol,1 eq.). The solution was stirred at 25℃for 3h. Addition of H to the reactant ₂ O (40 mL) and extracted with ethyl acetate (80 mL x 3) and the organic layer was carefully concentrated to give the crude material. By preparative TLC (SiO) ₂ The crude material was purified with ethyl acetate meoh=10:1 to give (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,411.05umol,41.55% yield). MS (ESI) m/z 487.2[ M-H] ⁺

Step 4: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (135 mg,277.46 mol,1 eq.) in NH ₃ A solution in MeOH (7M, 8mL,201.83 eq.) was stirred at 65℃for 16h. The reaction was carefully concentrated to give the crude material. The compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (130 mg, crude) was used directly in the next step. MS (ESI) m/z 472.3[ M+H ] ] ⁺ The method comprises the steps of carrying out a first treatment on the surface of the Preparative HPLC conditions: column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]；B％：35％-55％，8min

Step 5: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (130 mg,275.69umol,1 eq.) in DCM (7 mL) was added the Buerger's reagent (197.09 mg,827.06umol,3 eq.). The solution was stirred at 25℃for 1h. The reaction was carefully concentrated to give the crude material. Purification of the crude material by preparative HPLC (TFA) gives N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (36 mg,75.41umol,27.35% yield, 95% purity). MS (ESI) m/z 454.1[ M+H ]] ⁺ . Preparative HPLC conditions: column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ Water (0.04% HCl) -ACN]；B％：30％-55％，7min； ¹ H NMR (400 MHz, methanol-d) ₄ )δppm 1.02(s,9H)1.74-1.94(m,4H)2.21-2.37(m,2H)2.52-2.63(m,1H)3.16-3.26(m,2H)3.92(s,3H)4.63(dd,J＝8.49,4.30Hz,1H)4.98-5.06(m,1H)6.50(d,J＝7.72Hz,1H)7.02(d,J＝8.38Hz,1H)7.10-7.16(m,1H)7.23(d,J＝0.88Hz,1H)。

EXAMPLE 35 Synthesis of viral protease inhibitor Compounds 491 and 491A

Step 1: (2S) -2- [ [ 3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl propionate (1 g,4.22mmol,1 eq, HCl), 3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a mixture of propionic acid (1.5 g,5.06mmol,1.2 eq, HCl) and TEA (1.7 g,16.88mmol,2.35mL,4 eq) in DMF (5 mL) was added T ₃ P(5.3g，8.44mmol，5.02mL，50% purity, 2 equivalents). The mixture was stirred at 25℃for 16h. TLC (DCM: meoh=10:1/UV 254 nm) showed new spots were detected. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Purification of residue by flash silica gel chromatography12g/>Silica flash column, eluent: 100-25% ethyl acetate/MeOH at 30 mL/min). Obtaining the compound (2S) -2- [ [ 3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.9 g,3.84mmol,91.0% yield). By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,9.5 min) purification of (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (50 mg,0.10mmol,1 eq.). Obtaining the compound (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (50 mg,0.10mmol,1 eq.).

Step 2: n- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ 3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.00 g,1.73mmol,84% purity, 1 eq.) in NH ₃ The mixture in (7M, 24.77mL,100 eq.) 7M in MeOH was stirred at 80deg.C for 36h. Next, the reaction mixture was concentrated in vacuo. Obtaining the compound N- [2- [ [ (1S) -2-amino-2-oxo ] as a yellow solid1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (813 mg, crude material).

By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,7.8 min) purification of N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl)]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,0.10mmol,1 eq.). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (20.3 mg,42.5umol,39.9% yield, 98.4% purity).

Step 3: n- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

N- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A mixture of 4-methoxy-1H-indole-2-carboxamide (663.0 mg,1.41mmol,1 eq.) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (673.0 mg,2.82mmol,2 eq.) in DCM (8 mL) was stirred at 25℃for 16H. Next, methoxycarbonyl- (triethylammonium) sulfonyl-imide (336.5 mg,1.41mmol,1 eq.) was added to the mixture and the mixture was stirred at 25℃for 16hr. LC-MS showed detection of the desired compound. TLC (Petroleum ether: ethyl acetate=0:1/I) ₂ ) Indicating that a new blob was detected. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]；B％：23％-53％，9.5min) purifying the residue. Obtaining the compound N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a yellow solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (450 mg,0.98mmol,69.9% yield).

Step 4: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%) of purified N- [2- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (550.0 mg,1.22mmol,1 eq.). Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide, isomer 1 (147.1 mg,0.25mmol,22.1% yield). LCMS: rt=0.756 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,DMSO-d ₆ ) δ11.56 (br s, 1H), 8.90 (br d, j=8.0 hz, 1H), 8.49 (br d, j=7.4 hz, 1H), 7.52 (br s, 1H), 7.36 (s, 1H), 7.12-7.06 (m, 1H), 7.03-6.98 (m, 1H), 6.50 (d, j=7.6 hz, 1H), 5.17-4.96 (m, 1H), 4.56-4.33 (m, 1H), 3.88 (s, 3H), 3.09 (br s, 2H), 2.33-2.19 (m, 2H), 1.88-1.76 (m, 3H), 1.70 (br dd, j=3.8, 8.3hz, 1H), 1.57 (br s, 1H), 1.50-1.35 (m, 2H), 0.80 (br s, 3H), 0.03-0.25 hz, 1H); and is also provided with

Obtaining N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide, isomer 2 (113.1 mg,0.32mmol,28.8% yield, 100% purity). LCMS: rt=0.761 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.0[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,DMSO-d ₆ )δ11.57(s,1H),8.89(br d,J＝8.0Hz,1H),8.49(br d,J＝7.6Hz,1H),7.51(br s,1H),7.36(d,J＝1.6Hz,1H),7.13-7.06(m,1H),7.03-6.97(m,1H),6.50(d,J＝7.5Hz,1H),5.08-4.99(m,1H),4.52-4.42(m,1H),3.88(s,3H),3.08(br s,2H),2.23-2.13(m,2H),1.90-1.68(m,4H),1.64-1.36(m,3H),0.85-0.70(m,1H),0.45-0.33(m,2H),0.24-0.11(m,1H),0.13-0.03(m,1H)。

EXAMPLE 36 Synthesis of viral protease inhibitor Compound 531

Step 1: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (500 mg,1.68mmol,1 eq.) in DCM (10 mL) and DMF (2.5 mL) was added DMAP (616.30 mg,5.04mmol,3 eq.) in one portion. To the mixture were added 7-chloro-1H-indole-2-carboxylic acid (394.69 mg,2.02mmol,1.2 eq.) and EDCI (967.04 mg,5.04mmol,3 eq.). The resulting mixture was stirred at 25℃for 2h. The mixture was then concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (550 mg,1.16mmol,68.87% yield). MS (ESI) m/z 475.1[ M+H ]] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (500 mg,1.05mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,66.49 eq.) was stirred at 60℃for 16h. Concentrating the reaction mixture under reduced pressure to give a solidN- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as such]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (440 mg,956.68umol,90.87% yield). MS (ESI) m/z 460.3[ M+H ]] ⁺

Step 3: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 7-chloro-1H-indole-2-carboxamide (430 mg,934.94 mmol,1 eq.) in DCM (6 mL) was added the Bungeus reagent (445.61 mg,1.87mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃₎ -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to obtain 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-pyrrolidin-3-yl in solid form]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (180 mg,407.32umol,43.57% yield). MS (ESI) m/z 442.2[ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.71(br s,1H),9.01(d,J＝7.9Hz,1H),8.72(d,J＝7.5Hz,1H),7.71(s,1H),7.63(dd,J＝0.7,7.9Hz,1H),7.34-7.25(m,2H),7.07(t,J＝7.8Hz,1H),5.00(q,J＝7.9Hz,1H),4.58-4.49(m,1H),3.13(quin,J＝9.2Hz,2H),2.42-2.31(m,1H),2.22-2.05(m,2H),1.89-1.64(m,3H),1.57-1.46(m,1H),0.89-0.75(m,1H),0.50-0.37(m,2H),0.25-0.07(m,2H)。

EXAMPLE 37 Synthesis of viral protease inhibitor Compound 635

Step 1: (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -3-cyclopropyl-propionamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of benzyl carbamate (400 mg,0.92mmol,1 eq.) in MeOH (5 mL) was added Pd (200 mg,10% purity) and H ₂ (0.92 mmol). The mixture was stirred at 25℃for 1hr at 15 psi. The mixture was filtered to obtain a filtrate. Concentrating the mixture under reduced pressure to give the compound (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a solid]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (274 mg,0.92mmol,99.5% yield).

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (137 mg,0.46mmol,1 eq.) and 6-chloro-1H-indole-2-carboxylic acid (90.4 mg,0.46mmol,1 eq.) in DMF (2 mL) was added DIPEA (119.4 mg,0.92mmol,0.16mL,2 eq.) and HATU (210.9 mg,0.55mmol,1.2 eq.). The mixture was stirred at 25℃for 1hr. LCMS showed detection of one peak with the expected MS. The mixture was concentrated under reduced pressure. Through flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-1H-indole-2-carboxamide (200 mg,89.0% yield). LCMS: rt=0.780 min; c (C) ₂₃ H ₂₈ ClN ₅ O ₄ MS calculated: 473.18; MS experimental values: 474.1[ M+H ] ⁺ ]。

Step 3: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 6-chloro-1H-indole-2-carboxamide (47.5 mg,0.1mmol,1 eq.) in DCM (1 mL) was added the Buerger's reagent (71.6 mg,0.3mmol,3 eq.). The mixture was stirred at 25℃for 12hr. The mixture was concentrated under reduced pressure. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:31% -61%,7.8 min) to give the compound 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (64.33 mg,34.7% yield). LCMS: rt=0.832 min; c (C) ₂₃ H ₂₆ ClN ₅ O ₃ The method comprises the steps of carrying out a first treatment on the surface of the MS calculated: 455.17; MS experimental values: 456.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,DMSO-d ₆ )δ11.73(br s,1H),8.95(br d,J＝8.0Hz,1H),8.66(br d,J＝7.5Hz,1H),7.66(d,J＝8.5Hz,1H),7.53(br s,1H),7.44(s,1H),7.31(s,1H),7.05(dd,J＝1.8,8.5Hz,1H),5.11-4.96(m,1H),4.52-4.42(m,1H),3.09(br s,2H),2.34-2.21(m,2H),1.89-1.75(m,3H),1.74-1.65(m,1H),1.56(br s,1H),1.51-1.29(m,2H),0.79(br s,1H),0.42(br d,J＝7.0Hz,2H),0.23-0.01(m,2H)

EXAMPLE 38 Synthesis of viral protease inhibitor Compound 637

Step 1:4, 7-dichloro-2- (trichloromethyl) -1H-benzimidazole

To a solution of 3, 6-dichlorobenzene-1, 2-diamine (0.3 g,1.69mmol,1 eq.) in AcOH (12.57 g,209.2mmol,11.97mL,123.8 eq.) was added methyl 2, 2-trichloroethyliminoate (313.0 mg,1.77mmol,0.21mL,1.05 eq.) at 0deg.C). The mixture was stirred at 25℃for 16hr. The mixture obtained is treated with H ₂ O (40 mL) was diluted and filtered to give 4, 7-dichloro-2- (trichloromethyl) -1H-benzo [ d ] as a solid]Imidazole (300 mg, crude material).

Step 2:4, 7-dichloro-1H-benzimidazole-2-carboxylic acid

At 0deg.C, naOH (0.8 g,20.0mmol,20.2 eq.) was added to H ₂ To a solution of O (10 mL) was added 4, 7-dichloro-2- (trichloromethyl) -1H-benzo [ d ]]Imidazole (0.3 g,985.58umol,1 eq). The mixture was stirred at 25℃for 1hr. The pH of the mixture was adjusted to ph=2-3 with HCl (2M) and the mixture was then filtered to give 4, 7-dichloro-1H-benzo [ d ] as a solid]Imidazole-2-carboxylic acid (0.2 g, crude material).

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 6-dichloro-1H-benzimidazole-2-carboxamide

To (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropylpropionamide (130 mg,0.43mmol,1 eq.) and 4, 7-dichloro-1H-benzo [ d ]]To a solution of imidazole-2-carboxylic acid (101.3 mg,0.43mmol,1.0 eq.) in DMF (3 mL) was added HATU (250.1 mg,0.65mmol,1.5 eq.) and DIPEA (113.3 mg,0.87mmol,0.15mL,2.0 eq.). The mixture was stirred at 25℃for 1hr. TLC (dichloromethane: methanol=10/1) indicated 4, 7-dichloro-1H-benzo [ d ] ]Imidazole-2-carboxylic acid was completely depleted and a new spot formed. The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=100/1 to 10/1) to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 7-dichloro-1H-benzo [ d ] as a solid]Imidazole-2-carboxamide (0.2 g,0.39mmol,89% yield).

Step 4:4, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-benzoimidazole-2-carboxamide

To N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidine)3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 7-dichloro-1H-benzo [ d ]]To a solution of imidazole-2-carboxamide (100.00 mg,0.19mmol,1 eq.) in DCM (3.0 mL) was added the Buerger's reagent (140.3 mg,0.58mmol,3.0 eq.). The mixture was stirred at 25℃for 1hr. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05%NH3H2O+10mM NH4HCO3) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified for 7.8min, 20% -50% to give the product as a solid (22.11 mg,22% yield). LCMS: rt=0.824 min; c (C) ₂₂ H ₂₄ Cl ₂ N ₆ O ₃ MS calculated: 490.13; MS experimental values: 491.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.30(s,2H),5.22-5.09(m,1H),4.60(t,J＝7.1Hz,1H),3.27-3.19(m,2H),2.56-2.37(m,2H),2.06-1.88(m,3H),1.87-1.79(m,1H),1.73(td,J＝7.2,14.0Hz,2H),1.60-1.44(m,1H),0.96-0.75(m,1H),0.54(d,J＝6.9Hz,2H),0.21(dd,J＝4.8,10.4Hz,2H)。

Example 39 Synthesis of viral protease inhibitor Compounds 639 and 639A

/>

Step 1: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (1.07 g,4.65mmol,1.1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1 g,4.22mmol,1 eq., HCl) in DCM (10 mL) was added DMAP (1.55 g,12.67mmol,3 eq.) and EDCI (1.62 g,8.45mmol,2 eq.). The resulting mixture was stirred at 25℃for 1h. After completion, H is added to the solution ₂ O (30 mL) and then extracted with ethyl acetate (30 mL x 3). The combined organic phases were taken up in Na ₂ SO ₄ Dried, filtered and concentrated. By column chromatography (SiO ₂ The residue was purified with DCM/meoh=30/1 to 10/1 to give (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl as a yellow oilPropionyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.92mmol,68.97% yield, 100% purity). MS (ESI) m/z 412.3[ M+H ] ] ⁺ 。

Step 2: (2R) -N- (4- (tert-butyl) phenyl) -N- (2-oxo-1- (pyridin-3-yl) -2- ((pyridin-4-ylmethyl) amino) ethyl) pyrrolidine-2-carboxamide

Containing (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Ammonia (7M, 7.2mL,8.30 eq.) of methyl propionate (600 mg,1.46mmol,1 eq.) was stirred at 50℃for 14h. After completion, the solution was concentrated to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Tert-butyl carbamate (580 mg, crude). MS (ESI) m/z 397.3[ M+H ]] ⁺ 。

Step 3: (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -3-cyclopropyl-propionamide

Containing N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Tert-butyl carbamate (580 mg,1.46mmol,1 eq.) in HCl/MeOH (4M, 10.00mL,7.93 eq.) was stirred at 25℃for 1h. After completion, the solution was concentrated to give (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil ]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (380 mg, crude material). MS (ESI) m/z 297.2[ M+H ]] ⁺ 。

Step 4: (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -3-cyclopropyl-propionamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (380 mg,1.28mmol,1 eq.) in DC M (3 mL) was added 7-chloro-1H-indole-2-carboxylic acid (275.88 mg,1.41mmol,1.1 eq.) T ₃ P (1.22 g,1.93mmol,1.14mL,50% purity, 1.5 eq.) and DIEA (331.44 mg,2.56mmol, 446.68. Mu.L, 2 eq.). At 25 DEG CThe mixture was stirred for 2h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with DC M (30 mL. Times.3) and the combined organic phases were taken up in Na ₂ SO ₄ Dried, filtered and concentrated. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (350 mg,738.47 mol,57.59% yield, 100% purity). MS (ESI) m/z 474.3[ M+H ]] ⁺ 。

Step 5: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (350 mg,738.47 mmole, 1 eq.) in DCM (4 mL) was added the Buerger's reagent (527.94 mg,2.22mmol,3 eq.) and the solution stirred at 25℃for 6H. After completion, blow-drying was used to remove DCM. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mm X10 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH4HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give the product as a solid, which was purified by SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:33% -33%,8 min) to obtain:

7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl in solid form]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (250 mg,530.89umol,74.25% yield, 96.82% purity). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.58(d,J＝7.9Hz,1H),7.35-7.20(m,2H),7.06(t,J＝7.8Hz,1H),5.22-5.05(m,1H),4.57(t,J＝7.5Hz,1H),3.27-3.14(m,2H),2.61-2.34(m,2H),2.09-1.61(m,6H),1.59-1.43(m,1H),0.98-0.76(m,1H),0.55(dd,J＝1.3,8.2Hz,2H),0.31-0.09(m, 2H); and

7-chloro-N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl in solid form]Ethyl group]Amino group ]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (45 mg,98.70umol,13.37% yield, 100% purity). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.59(dd,J＝0.9,7.9Hz,1H),7.32-7.21(m,2H),7.07(t,J＝7.8Hz,1H),5.12-5.02(m,1H),4.59(dd,J＝6.4,7.9Hz,1H),3.21(dd,J＝4.6,7.7Hz,2H),2.44-2.23(m,2H),2.09-1.62(m,6H),1.60-1.47(m,1H),0.94-0.78(m,1H),0.62-0.43(m,2H),0.27-0.11(m,2H)。

EXAMPLE 40 Synthesis of viral protease inhibitor Compounds 643

Step 1: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (500 mg,2.11mmol,1 eq., HCl), (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (570.0 mg,2.32mmol,1.1 eq.) and TEA (855.0 mg,8.45mmol,1.18mL,4 eq.) in DMF (5 mL) was added T ₃ P (2.69 g,4.22mmol,2.51mL,50% purity, 2 eq.). The resulting mixture was stirred at 70℃for 16hr. TLC (petroleum ether: ethyl acetate=0:1/PMA) showed that new spots were detected. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography20g/>Silica flash column, eluent: gradient of 0-100% ethyl acetate/petroleum ether at 30mL/min ) The residue was purified. Obtaining the compound (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (436 mg,0.99mmol,47.2% yield, 97.9% purity).

Step 2: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

Methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (300 mg,0.70mmol,1 eq.) in HCl/dioxane (4M, 175.42uL,1 eq.) was stirred at 25℃for 2hr. The compound methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (250 mg, crude material, HCl) was obtained as a solid and used in the next step without further purification.

Step 3: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (310 mg,0.85mmol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (179.1 mg,0.93mmol,1.1 eq.), HATU (647.8 mg,1.70mmol,2 eq.) and DIPEA (440.4 mg,3.41mmol,0.60mL,4 eq.) in DCM (4 mL) was stirred at 25℃for 2hr. TLC (petroleum ether/ethyl acetate=0:1/UV 254 nm) showed that new spots were detected. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining an oily formThe compound (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (4571 mg,0.68mmol,80.1% yield) and confirmed by LC-MS.

Step 4: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (400 mg,0.79mmol,1 eq.) in MeOH ₃ (7M, 11.42mL,100 eq.). Next, the mixture was stirred at 80℃for 16hr. TLC (DCM: meoh=10:1/UV 254 nm) showed new spots were detected. The reaction mixture was concentrated in vacuo. Through flash silica gel chromatography 12g/>Silica flash column, eluent: 0-50% ethyl acetate/MeOH at 30 mL/min). Obtaining the compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (295 mg,0.60mmol,75.1% yield, 98.9% purity).

Step 5: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (290 mg,0.59mmol,1 eq.) in DCM (3 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (284.6 mg,1.19mmol,2 eq.). Next, the mixture was stirred at 25℃for 16hr. Next, methoxycarbonyl- (triethyl) was added to the mixtureAmmonium group) sulfonyl-imide (142.3 mg,0.59mmol,1 eq.) and the mixture was stirred at 25 ℃ for an additional 16hr. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: welch Xtimate C18X 25mm X5 um; mobile phase: [ water (0.05% ammonium hydroxide, v/v) -MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -85%,9.5 min) of the purified residue. Obtaining the compound N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (28.1 mg,59.3umol,9.9% yield, 98.7% purity). Rt=0.832 min; c (C) ₂₅ H ₃₃ N ₅ O ₄ MS calculated: 467.25, ms experimental value: 468.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.26-7.22(m,1H),7.18-7.12(m,1H),7.05-7.00(m,1H),6.51(d,J＝7.5Hz,1H),5.08(dd,J＝6.3,9.8Hz,1H),4.67-4.63(m,1H),3.93(s,3H),3.21-3.15(m,2H),2.47-2.38(m,2H),1.98-1.72(m,6H),1.70-1.58(m,1H),1.54-1.43(m,1H),1.02(s,8H),1.04-1.01(m,2H)。

EXAMPLE 41 Synthesis of viral protease inhibitor Compound 681

Step 1: (2S) -2- (2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,2.11mmol,1.1 eq, HCl) and 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid (684.45 mg,1.92mmol,1 eq.) in DMF (15 mL) was added N, N-Diisopropylethylamine (DIEA) (744.57 mg,5.76mmol,1.00mL,3 eq.) and hexafluorophosphoric acid 3-oxidation (1- [ bis (dimethylamino) methylene ]-1H-1,2, 3-triazolo [4,5-b]Pyridinium (HATU) (730.19 mg,1.92mmol,1 eq.). The mixture was stirred at 20℃for 1h. After the completion of this process,by adding H ₂ O (80 mL) to quench the two batches and extract with ethyl acetate (40 mL x 3). The combined organic layers were washed with 40mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.35 g, crude material). MS (ESI) m/z 539.3[ M+H ]] ⁺ 。

Step 2: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (650 mg,1.21mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 3.45mL,20 eq.) was stirred at 65℃for 17h. After completion, the two batches of the reaction mixture were concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a colorless oil ]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (1.22 g, crude). MS (ESI) m/z 524.3[ M+H ]] ⁺ 。

Step 3: n- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxamide (1.22 g,2.33mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (1.39 g,5.82mmol,2.5 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (3 mL) to quench the reaction mixture and then concentrate under reduced pressure to give a residue. By preparative HPLC (column: agela DuraShell C, 250X 70mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:43% -63%,20 min) to give the desired compound (490 mg) as a solid, which was purified by SFC (column: daicelCHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:58% -58%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid ]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5 ]]Decane-3-carboxamide, isomer 1 (201.77 mg,394.36umol,16.93% yield). MS (ESI) m/z506.3[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ ) Delta ppm 11.26 (br s, 1H) 8.50-8.85 (m, 1H) 7.23 (br s, 1H) 7.00-7.16 (m, 2H) 6.89 (br s, 1H) 6.52 (br d, j=7.46 hz, 1H) 4.86-5.06 (m, 1H) 4.48-4.79 (m, 1H) 3.80-3.98 (m, 4H) 3.59 (br d, j=4.65 hz, 1H) 3.09 (br s, 2H) 2.15-2.31 (m, 3H) 1.73-2.01 (m, 2H) 1.67 (br dd, j=12.17, 8.62hz, 2H) 1.33-1.61 (m, 12H); and is also provided with

Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5 ]]Decane-3-carboxamide, isomer 2 (200.95 mg,394.35umol,16.93% yield). MS (ESI) M/z506.3[ M+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.27(br s,1H)8.61(br d,J＝1.22Hz,1H)7.02-7.26(m,3H)6.91(br s,1H)6.53(d,J＝7.46Hz,1H)4.91-5.06(m,1H)4.62(br s,1H)3.82-3.98(m,4H)3.52-3.75(m,1H)3.09(br s,2H)2.09-2.28(m,3H)1.63-1.92(m,4H)1.33-1.62(m,12H)。

EXAMPLE 42 Synthesis of viral protease inhibitor Compounds 721

Step 1: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (2.49 g,10.14mmol,1.2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (2 g,8.45mmol,1 eq., HCl) in DCM (60 mL) was added DMAP (3.10 g,25.35mmol,3 eq.). Next, EDCI (3.24 g,16.90mmol,2 eq.) was added and the resulting mixture was stirred at 25℃for 1h. After the reaction is completed After this time, the mixture was quenched with water (400 mL), extracted with DCM (150 mL x 3) and then dried with saturated NaCl (50 mL). The resulting solution was concentrated in vacuo and passed through a column (SiO ₂ Petroleum ether ethyl acetate=2:1 to 0:1). The resulting residue was washed with HCl (1 m,150 ml), extracted with DCM (50 ml x 3), and then with saturated NaHCO ₃ (30 mL) the solution was adjusted to pH = -8. The resulting mixture was extracted with DCM (100 mL) and then concentrated in vacuo to give methyl (S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (3 g,6.32mmol,74.74% yield) as a solid. ¹ H NMR(400MHz,CDCl ₃ -d)δppm 7.61(d,J＝7.0Hz,1H),6.85-6.51(m,1H),6.22(s,1H),5.06-4.85(m,1H),4.63-4.47(m,1H),4.30-4.02(m,1H),3.79-3.66(m,3H),3.35-3.25(m,2H),2.42-2.24(m,1H),2.14-2.05(m,1H),1.96-1.66(m,4H),1.63-1.52(m,1H),1.43(s,9H),1.03-0.90(m,9H)。

Step 2: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.5 g,3.51mmol,1 eq.) in HCl/MeOH (4M, 20 mL) was stirred at 25℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.1 g, crude material, HCl) as a solid. ¹ H NMR(400MHz,D ₂ O)δppm 4.57(dd,J＝4.8,10.3Hz,1H),3.98(dd,J＝5.2,7.8Hz,1H),3.78-3.65(m,3H),3.29-3.14(m,2H),2.75-2.33(m,1H),2.24-1.47(m,8H),1.04-0.86(m,9H)。

Step 3: (S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (550 mg x 2, hydrochloride, 1.68mmol,1 eq.) and 7-chloro-1H-indole-2-carboxylic acid (394.29 mg,2.02mmol,1.2 eq.) in DCM (6 mL) was added DMAP (615.66 mg,5.04mmol,3 whenAmount). EDCI (644.05 mg,3.36mmol,2 eq.) was added to the mixture at 25℃and the mixture was stirred for 1h at 25 ℃. After the reaction was complete, the mixture was quenched with water (200 mL), extracted with DCM (70 ml×3), and then concentrated in vacuo. Through the column (SiO) ₂ Petroleum ether ethyl acetate=1:1 to 0:1) the resulting residue was purified, concentrated in vacuo, and then washed with 1M HCl (100 mL) and extracted with DCM (30 mL. Times.3). With saturated NaHCO ₃ (30 mL) the organic phase was adjusted to pH = -7 and then concentrated in vacuo to give methyl (S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamido) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (650 mg,1.16mmol,40% yield) as a solid. MS (ESI) m/z 505.2[ M+H ] ] ⁺ ； ¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.58(d,J＝7.8Hz,1H),7.32-7.17(m,2H),7.06(t,J＝7.8Hz,1H),4.73(dd,J＝3.8,8.6Hz,1H),4.55(dd,J＝4.0,11.7Hz,1H),3.71(s,3H),3.35(s,1H),3.24-3.01(m,2H),2.49-2.22(m,2H),2.02-1.40(m,8H),1.08-0.96(m,9H)。

Step 4: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (650 mg,1.29mmol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 10 mL) was stirred at 50deg.C for 16h. After the reaction was completed, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carboxamide (450 mg, crude material) as a pale yellow solid. MS (ESI) m/z 490.3[ M+H ]] ⁺

Step 5: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide

To N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carbonylTo a solution of amine (430 mg,877.56umol,1 eq.) in DCM (10 mL) was added the Buerger reagent (627.38 mg,2.63mmol,3 eq.). The reaction mixture was stirred at 25℃for 4h. After the reaction was completed, the mixture was quenched with water (10 mL), with N ₂ The stream was dried and purified by preparative HPLC (column: kromasil C18 (250 x 50mM x 10 um), mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide (205 mg,424.79umol,48.41% yield) as a white solid. MS (ESI) m/z 472.2[ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.70(s,1H),9.02(d,J＝8.0Hz,1H),8.71(d,J＝8.0Hz,1H),7.63(d,J＝8.0Hz,1H),7.52(s,1H),7.34-7.23(m,2H),7.07(t,J＝7.8Hz,1H),5.05(q,J＝8.2Hz,1H),4.63-4.54(m,1H),3.07(s,2H),2.30-2.18(m,2H),1.88-1.32(m,7H),0.95(s,9H)。

EXAMPLE 43 Synthesis of viral protease inhibitor Compound 133

Step 1: 7-chloro-1H-benzo [ d ] imidazole-2-carboxylic acid

To a solution of 3-chlorobenzene-1, 2-diamine (500 mg,3.51mmol,1 eq.) in AcOH (9 mL) was added dropwise methyl 2, 2-trichloroethyliminoate (619.29 mg,3.51mmol, 433.07. Mu.L, 1 eq.) and the mixture was stirred at 25℃for 2h. At 0deg.C with 10mL H ₂ The reaction mixture was quenched with O and the resulting precipitate was collected. H for solids ₂ O (2 x 10 ml) and dried in vacuo to give the product 7-chloro-1H-benzimidazole-2-carboxylic acid (500 mg, crude) as a solid. MS (ESI) m/z 195.1[ M-H ]] ⁺

Step 2: (S) -2- ((S) -2-amino-4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester hydrochloride

To a solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (300 mg,750.98umol,1 eq.) in Et OAc (2 mL) was added HCl/EtOAc (4 m,20mL,106.53 eq.) dropwise and the mixture stirred at 25 ℃ for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (250 mg, crude material, HCl) as a solid.

Step 3: (S) -2- ((S) -2- (7-chloro-1H-benzo [ d ] imidazole-2-carboxamido) -4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (250 mg,744.43 mol,1.0 eq, HCl) and 7-chloro-1H-benzimidazole-2-carboxylic acid (243.91 mg,744.43 mol,60% purity, 1 eq) in DMF (3 mL) was added EDCI (285.42 mg,1.49mmol,2.0 eq), DMAP (181.89 mg,1.49mmol,2.0 eq). After the addition of DCM (9 mL), the reaction was stirred at 25℃for 12h. By adding H at 0 ℃ ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 0/1) to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-benzimidazole-2-carbonyl) amino ] as a yellow solid]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (220 mg,327.28umol,43.96% yield, 71.1% purity). MS (ESI) m/z 478.0[ M+H ] ] ⁺

Step 4: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -7-chloro-1H-benzo [ d ] imidazole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-benzimidazole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (200 mg,418.46 mol,1 eq.) in ammonia (7M, 20mL,334.56 eq.) was stirred at 80℃for 12h. The reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino ] as a yellow solid-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-7-chloro-1H-benzimidazole-2-carboxamide (160 mg, crude). MS (ESI) m/z 463.2[ M+H ]] ⁺

Step 5: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -1H-benzo [ d ] imidazole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a solution of 7-chloro-1H-benzimidazole-2-carboxamide (80 mg,108.87umol,63% purity, 1 eq) in DCM (4 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (129.73 mg,544.36umol,5.0 eq) and the mixture was then stirred at 25 ℃ for 12H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,10 min) and preparative HPLC (column: phenomenex Luna C18, 75 x 30mm x 3um; mobile phase: [ Water (0.2% FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:10% -60%,8 min) to give 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-1H-benzimidazole-2-carboxamide (13.28 mg,29.85umol,27.42% yield, 100% purity). MS (ESI) m/z 445.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 13.64(br s,1H),8.76-9.00(m,2H),7.70(s,1H),7.51(br d,J＝6.2Hz,1H),7.25-7.42(m,2H),4.90-5.06(m,1H),4.55(br t,J＝7.4Hz,1H),3.05-3.18(m,2H),2.33-2.42(m,1H),2.05-2.23(m,2H),1.54-1.90(m,5H),0.92(br dd,J＝8.5,6.3Hz,6H)。

EXAMPLE 44 Synthesis of viral protease inhibitor Compound 145

/>

Step 1: 3-methoxybenzene-1, 2-diamine:

at 25 ℃, to 2-methoxy-6-nitro-aniline(1 g,5.95mmol,1.00mL,1 eq.) in EtOH (12 mL) and H ₂ NH was added at one time to the mixture in O (4 mL) ₄ Cl (1.59 g,29.74mmol,5 eq.) and then the reaction was heated to 80 ℃. Fe (1.66 g,29.74mmol,5 eq.) was added and stirred at 80℃for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, and then taken up with H ₂ O (10 mL) was diluted and extracted with 30mL (10 mL x 3) of ethyl acetate. The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 3-methoxybenzene-1, 2-diamine (770 mg,5.02mmol,84.34% yield, 90% purity) as a black oil. MS (ESI) m/z 139.1[ M+H ] ] ⁺

Step 2: 7-methoxy-1H-benzimidazole-2-carboxylic acid:

a mixture of 3-methoxybenzene-1, 2-diamine (750 mg,5.43mmol,1 eq.) and methyl 2, 2-trichloroethyliminoate (1.15 g,6.51mmol, 803.66. Mu.L, 1.2 eq.) in AcOH (8 mL) was added in one portion at 25 ℃. The mixture was stirred at 25℃for 2h. With Na ₂ CO ₃ The reaction mixture was brought to neutrality with the solution and then quenched with H ₂ Dilute with O (5 mL) and extract with ethyl acetate (5 mL x 3). The combined organic layers were washed with brine (10 ml x 1) and concentrated under reduced pressure to give the crude product. The crude material was purified by preparative HPLC (TFA conditions) to give 7-methoxy-1H-benzimidazole-2-carboxylic acid (300 mg,1.56mmol,28.76% yield) as a yellow solid. MS (ESI) m/z 193.1[ M+H ]] ⁺ . Column: phenomenex luna C18 100×40mm×5um; mobile phase: [ Water (0.1% TFA) -ACN]；B％：20％-55％，8min

Step 3: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-methoxy-1H-benzimidazole-2-carboxamide

To 7-methoxy-1H-benzimidazole-2-carboxylic acid (150 mg, 780.55. Mu. Mol,1 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl at 0 ℃ ]Ethyl group]To a mixture of 3-cyclopropyl-propionamide (711.44 mg,780.55umol,29% purity, 1 eq.) in DCM (3 mL) was added DIEA (302.64 mg,2.34mmol,407.88uL,3 eq.) and T ₃ P (745.07 mg,1.17mmol,696.33uL,50% purity, 1.5 eq.). The mixture was stirred at 0℃for 2h. H for the reaction mixture ₂ O (5 mL) was diluted and then extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (8 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. Purification of the crude material by preparative HPLC (neutral conditions) gave N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-methoxy-1H-benzimidazole-2-carboxamide (48 mg,109.47 mol,14.02% yield). MS (ESI) m/z 439.2[ M+H ]] ⁺ . Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：20％-40％，8min。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝13.29(br s,1H),9.09-8.90(m,1H),8.80-8.66(m,1H),7.79-7.67(m,1H),7.27-7.17(m,1H),7.09(d,J＝8.2Hz,1H),6.76(d,J＝7.9Hz,1H),5.06-4.83(m,1H),4.61-4.48(m,1H),3.98-3.88(m,3H),3.20-3.05(m,2H),2.44-2.30(m,1H),2.27-2.06(m,2H),1.96-1.84(m,1H),1.83-1.66(m,2H),1.65-1.55(m,1H),0.74(br s,1H),0.40(br d,J＝8.2Hz,2H),0.23--0.01(m,2H)

EXAMPLE 45 Synthesis of viral protease inhibitor Compound 163

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (500 mg,1.75mmol,1 eq.) in HCl/EtOAc (3 mL) was stirred at 25℃for 1h. After completion, the mixture was concentrated in vacuo to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (350 mg, crude, HCl) as a yellow gum.

Step 2: (2S, 3R) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -3-phenylpyrrolidine-1-carboxylic acid tert-butyl ester

To a solution of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (320 mg,1.44mmol,1 eq, HCl) and (2S, 3 r) -1-tert-butoxycarbonyl-3-phenyl-pyrrolidine-2-carboxylic acid (502.43 mg,1.72mmol,1.2 eq) in DCM (15 mL) was added DMAP (526.70 mg,4.31mmol,3 eq) and EDCI (1.38 g,7.19mmol,5 eq) and then the mixture was stirred at 25 ℃ for 1h. After completion, the mixture was poured into water (45 mL) and extracted with DCM (20 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ PE: ea=1:1 to 0:1 and then DCM: meoh=10:1 to 5:1) to give (2S, 3 r) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -3-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (500 mg,544.03umol,37.86% yield, 50% purity) as a white solid. MS (ESI) M/z460.3[ M+H ]] ⁺

Step 3: (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((2S, 3R) -3-phenylpyrrolidine-2-carboxamido) propionic acid methyl ester

A solution of (2S, 3R) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -3-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (500 mg,1.09mmol,1 eq.) in HCl/MeOH (4M, 5 mL) was stirred at 25℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((2S, 3 r) -3-phenylpyrrolidine-2-carboxamido) propanoate (340 mg, crude material, HCl) as a pale yellow solid.

Step 4: (S) -2- ((2S, 3R) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- ((2S, 3 r) -3-phenylpyrrolidine-2-carboxamido) propanoate (200 mg,278.23umol,50% purity, 1 eq) and 4-methoxy-1H-indole-2-carboxylic acid (63.83 mg,333.87umol,1.2 eq) in DCM (5 mL) was added DMAP (101.97 mg,834.68umol,3 eq) and EDCI (106.67 mg,556.45umol,2 eq) and then mixed with stirring at 25 ℃The reaction time was 1h. After the reaction was complete, the mixture was quenched with water (30 mL) and extracted with DCM (10 mL x 3). The resultant was concentrated in vacuo and purified by preparative TLC (SiO ₂ Ethyl acetate=1) to give methyl (S) -2- ((2S, 3 r) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (130 mg,216.51umol,77.82% yield, 88.7% purity) as a white solid. MS (ESI) m/z 533.3[ M+H ]] ⁺

Step 5: (2S, 3 r) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamide

Methyl (S) -2- ((2S, 3R) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (180 mg,337.97umol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 7.00 mL) was stirred at 25℃for 24h. After the reaction was completed, the mixture was concentrated in vacuo to give (2S, 3 r) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamide (160 mg, crude material) as a white solid. MS (ESI) m/z 518.3[ M+H ]] ⁺

Step 6: (2S, 3 r) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamide

To a solution of (2S, 3 r) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamide (160 mg,309.13umol,1 eq) in DCM (5 mL) was added the berges reagent (294.67 mg,1.24mmol,4 eq) and then the mixture was stirred at 45 ℃ for 4H. After the reaction was completed, the mixture was quenched with water (3 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge BEH C, 100 x 25mM x 5um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give (2S, 3 r) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H) as a white solid-indole-2-carbonyl) -3-phenylpyrrolidine-2-carboxamide (45 mg,89.18umol,28.85% yield, 99% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.69-11.50(m,1H),9.21-8.79(m,1H),7.76-7.49(m,1H),7.42-7.20(m,5H),7.17-6.72(m,3H),6.57-6.39(m,1H),5.00-4.76(m,1H),4.47(d,J＝6.8Hz,1H),4.17-3.72(m,5H),3.55-3.38(m,1H),3.17-2.77(m,2H),2.46-2.34(m,2H),2.30-2.01(m,3H),1.79-1.31(m,2H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δppm 11.33(s,1H),8.75(br s,1H),7.43-7.22(m,6H),7.17-7.03(m,2H),6.96(s,1H),6.52(d,J＝7.3Hz,1H),4.99-4.87(m,1H),4.63(s,1H),4.08(s,2H),3.90(s,3H),3.50(q,J＝6.8Hz,1H),3.17-3.06(m,2H),2.42(s,2H),2.25-2.03(m,3H),1.84-1.57(m,2H)。

EXAMPLE 46 Synthesis of viral protease inhibitor Compound 191

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.75mmol,1 eq.) in HCl/MeOH (4M, 7mL,16.03 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, and the residue was then dissolved with DCM (10 ml x 3). Concentrating the resulting product under reduced pressure to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a white oil]Methyl propionate (320 mg, crude material). MS (ESI) m/z 187.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3- (3-pyridinyl) propanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a mixture of methyl propionate (320 mg,1.44mmol,1.2 eq, HCl) in DCM (4 mL) and DMF (1 mL) was added (2S) -2- (tert-butoxycarbonylamino) -3- (3-pyri-dine)Bohdinyl) propionic acid (318.91 mg,1.20mmol,1 eq.), TEA (727.10 mg,7.19mmol,1.00mL,6 eq.) and T3P (1.14 g,1.80mmol,1.07mL,50% purity, 1.5 eq.) were stirred at 25℃for 1h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By chromatography on silica gel (SiO ₂ DCM: meoh=9:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1 to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3- (3-pyridinyl) propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (490 mg,1.13mmol,94.17% yield). MS (ESI) m/z 435.3[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [ (2S) -2-amino-3- (3-pyridinyl) propanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3- (3-pyridinyl) propanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (450 mg,1.04mmol,1 eq.) in HCl/MeOH (4M, 6mL,23.17 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-3- (3-pyridinyl) propionyl as a white oil ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (340 mg, crude material). MS (ESI) m/z 335.1[ M+H ]] ⁺ 。

Step 4: (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -3- (3-pyridinyl) propanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

Next, to (2S) -2- [ [ (2S) -2-amino-3- (3-pyridinyl) propanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (340 mg,916.86umol,1 eq, HCl) was added to a mixture of DCM (2 mL) and DMF (2 mL) 4-methoxy-1H-indole-2-carboxylic acid (210.35 mg,1.10mmol,1.2 eq), TEA (556.66 mg,5.50mmol, 765.70. Mu.L, 6 eq) and T3P (875.18 mg,1.38mmol, 817.93. Mu.L, 50% purity, 1.5 eq) and stirred at 25℃for 1.5H. After completion, reverseThe mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1) to give the product (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-3- (3-pyridyl) propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (180 mg,354.65umol,38.68% yield). MS (ESI) m/z 508.2[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -2-oxo-1- (3-pyridylmethyl) ethyl ] -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-3- (3-pyridyl) propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (165 mg,325.10umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 5mL,107.66 eq.) was stirred at 50deg.C for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]Amino group]-2-oxo-1- (3-pyridylmethyl) ethyl]-4-methoxy-1H-indole-2-carboxamide (150 mg, crude). MS (ESI) m/z 493.2[ M+H ]] ⁺ 。

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -2-oxo-1- (3-pyridylmethyl) ethyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ]Methyl group]Ethyl group]Amino group]-2-oxo-1- (3-pyridylmethyl) ethyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (126 mg,255.82umol,1 eq.) in DCM (3 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (243.86 mg,1.02mmol,4 eq.) and the reaction stirred at 40 ℃ for 2H. After completion, the mixture was quenched with water (1 mL) and with N ₂ And (5) blow-drying. By preparative HPLC (column: watersX bridge BEH C18X 25mm X5 um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-2-oxo-1- (3-pyridylmethyl) ethyl]-4-methoxy-1H-indole-2-carboxamide (30.52 mg,64.32umol,25.14% yield, 100% purity). MS (ESI) m/z 475.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝8.50(d,J＝1.5Hz,1H),8.41-8.34(m,1H),7.80(br d,J＝7.9Hz,1H),7.37(dd,J＝4.9,7.8Hz,1H),7.21(s,1H),7.13(d,J＝7.7Hz,1H),7.00(d,J＝8.2Hz,1H),6.50(d,J＝7.7Hz,1H),5.03(dd,J＝6.0,10.0Hz,1H),4.76(s,1H),3.92(s,3H),3.30-3.21(m,3H),3.17(dd,J＝8.8,13.9Hz,1H),2.56(dq,J＝5.5,9.3Hz,1H),2.36-2.21(m,2H),1.96-1.73(m,2H)。

EXAMPLE 47 Synthesis of viral protease inhibitor Compound 213

Step 1: methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (501 mg,1.75mmol,1 eq.) in HCl/EtOAc (4M, 10.02mL,22.91 eq.) was stirred at 25℃for 1h. After completion, the solution was concentrated to remove HCl/EA. The crude material was used directly in the next step without further purification. Methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg, crude material) was obtained as a yellow oil.

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (295.93 mg,1.59mmol,1.4 eq.) and (3S) -1-benzyloxycarbonyl hexahydropyridazine-3-carboxylic acid (300 mg,1.14mmol,1 eq.) in DCM (2 mL)/THF (2 mL) was cooled to 0deg.C and then T3P (1.08 g,1.70mmol,1.01mL,50% purity, 1.5 eq.) and DIEA were added(440.14 mg,3.41mmol, 593.18. Mu.L, 3 eq.). After stirring for 13H at 25℃the solution was treated with H ₂ O (20 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The crude material was used directly in the next step without further purification. Obtaining (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]Hexahydropyridazine-1-carboxylic acid benzyl ester (455 mg, crude material). MS (ESI) m/z 433.1[ M+H ]] ⁺ 。

To (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]To a solution of benzyl hexahydropyridazine-1-carboxylate (200 mg,462.46 mol,1 eq) in DCM (2 mL) was added DIEA (119.54 mg,924.92 mol,161.10 μl,2 eq), (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl chloride (121.56 mg,554.95 mol,1.2 eq) and the solution was then stirred at 25 ℃ for 1h. After completion, the solution is treated with H ₂ O (10 mL) was diluted, extracted with DCM (20 mL. Times.3) and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. Obtaining (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a yellow oil]-3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Hexahydropyridazine-1-carboxylic acid benzyl ester (160 mg,248.88umol,53.82% yield, 95.67% purity). MS (ESI) m/z 433.1[ M+H ]] ⁺ 。

Containing (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ]-3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Benzyl hexahydropyridazine-1-carboxylate (160 mg,260.14umol,1 eq.) of TFA (5 mL) was stirred at 75deg.C for 1h. After completion, the solution was concentrated to remove TFA, with NaHCO ₃ The solution was diluted and extracted with ethyl acetate (20 ml x 3). The combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The crude material was used directly in the next step without further purification. Obtaining (2S) -2- [ [ (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl ] as a yellow solid]Hexahydropyridazine-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (80 mg, crude material). MS (ESI) m/z 481.0[ M+H ]] ⁺ 。

Containing (2S) -2- [ [ (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Hexahydropyridazine-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Ammonia (7M, 4.00mL,168.32 eq.) of methyl propionate (80 mg,166.35umol,1 eq.) was stirred at 80℃for 17h. After completion, the solution was concentrated to remove MeOH. The crude material was used directly in the next step without further purification. Obtaining (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow oil ]Methyl group]Ethyl group]-2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]Hexahydropyridazine-3-carboxamide (75 mg, crude material). MS (ESI) M/z481.0[ M+H ]] ⁺ 。

To (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]To a solution of hexahydropyridazine-3-carboxamide (75 mg,160.98umol,1 eq.) in DCM (0.5 mL) was added the Bunges reagent (76.72 mg,321.95umol,2 eq.) and the solution was stirred at 25℃for 2h. After completion, the solution was concentrated to remove DCM. The residue was purified by preparative HPLC (neutral conditions). Column: phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ Water (10 m)M NH4HCO3)-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8min. Obtaining (3S) -2- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a white solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Hexahydropyridazine-3-carboxamide (20 mg,44.65umol,27.74% yield, 100% purity). ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.79-7.60(m,3H),7.32-7.22(m,2H),5.17(dd,J＝2.2,6.0Hz,1H),5.07(dd,J＝6.4,9.7Hz,1H),3.38-3.32(m,2H),3.12(br d,J＝13.7Hz,1H),2.90-2.74(m,1H),2.56(dq,J＝5.8,9.0Hz,1H),2.44-2.14(m,3H),2.08-1.79(m,3H),1.75-1.53(m,2H)。MS(ESI)m/z 448.2[M+H] ⁺ 。

Step 7: (E) -3- (4-chloro-2-fluorophenyl) acryloyl chloride

To a solution of (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid (120 mg, 598.22. Mu. Mol,1 eq.) in DCM (0.5 mL) was added DMF (437.26. Mu.g, 5.98. Mu. Mol, 0.46. Mu.L, 0.01 eq.) and the reaction was cooled to 0deg.C. Addition (COCl) ₂ (151.86 mg,1.20mmol,104.73uL,2 eq.) and the solution stirred at 25℃for 1h. After completion, the solution was concentrated to remove DCM and to give the crude material. The crude material was used directly in the next step without further purification. (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl chloride (125 mg, crude material) was obtained as a white solid.

EXAMPLE 48 Synthesis of viral protease inhibitor Compound 203

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (600 mg,2.10mmol,1 eq.) in HCl/EtOAc (20 mL) was stirred at 25℃for 1h. After completion, the mixture was concentrated in vacuo to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (530 mg, crude) as a yellow solid. MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4-fluoro-4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (530 mg,2.85mmol,1 eq.) in DMF (1 mL) and DCM (10 mL) was added (S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanoic acid (710.44 mg,2.85mmol,1 eq.), T3P (2.36 g,3.71mmol,2.20mL,50% purity, 1.3 eq.) and TEA (865.17 mg,8.55mmol,1.19mL,3 eq.) and the mixture was stirred at 25℃for 1h. After completion, by adding H ₂ O (50 mL) to quench the mixture and then extract with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue and purified by column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1) to give the crude product methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (730 mg,1.57mmol,55.19% yield, 89.95% purity) as a yellow oil. MS (ESI) M/z418.2[ M+H] ⁺

Step 3: (S) -2- ((S) -2-amino-4-fluoro-4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (530.00 mg,1.27mmol,1 eq.) in HC l/MeOH (20 mL) was stirred at 25℃for 1h. After completion, the reaction was concentrated in vacuo to give the crude product methyl (S) -2- ((S) -2-amino-4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (500 mg, crude) as a yellow solid. MS (ESI) m/z 318.2[ M+H ] ] ⁺

Step 4: (S) -2- ((S) -4-fluoro-2- (4-methoxy-1H-indole-2-carboxamide) -4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4-fluoro-4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (500.00 mg,1.58mmol,1 eq.) in ACN (20 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (301.21 mg,1.58mmol,1 equivalent), DMAP (384.96 mg,3.15mmol,2 equivalents), EDCI (604.06 mg,3.15mmol,2 equivalents) and the mixture was stirred at 25℃for 1h. After completion, the residue was poured into H ₂ O (50 mL) and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1) to yield the product methyl (S) -2- ((S) -4-fluoro-2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (340 mg,652.80umol,41.43% yield, 94.18% purity). MS (ESI) m/z 491.2[ M+H] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -4-fluoro-2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (330 mg,672.75umol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 10mL,104.05 eq.) was stirred at 25℃for 10h. After completion, the mixture was concentrated in vacuo to give the crude product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (280 mg, crude) as a yellow solid. MS (ESI) m/z 476.2[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (220 mg,462.66umol,1 eq) in DCM (10 mL) was added the bergs reagent (1.10 g,4.63mmol,10 eq) and the mixture stirred for 1H at 25 ℃. After completion, the mixture was concentrated in vacuo and purified by preparative HPLC (column Phenomenex Gemini-NX C18 75 x 30mm x 3 um (um); mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the product N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (10 mg,21.86umol,4.72% yield, 100% purity). MS (ESI) m/z 458.2[ M+H ]] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.22(s,1H),7.18-7.12(m,1H),7.03-7.02(m,1H),6.52-6.50(m,1H),5.06-5.03(m,1H),4.74-4.72(m,1H),3.93(s,3H),3.29-3.19(m,2H),2.32-2.31(m,1H),2.36-2.25(m,3H),2.24-2.14(m,1H),1.93-1.76(m,2H),1.48-1.46(m,3H),1.43-1.41(m,3H)

EXAMPLE 49 Synthesis of viral protease inhibitor Compound 223

Step 1: (2S) -2-amino-3- (1H-imidazol-5-yl) propionic acid methyl ester

To a solution of (2S) -2- (tert-butoxycarbonylamino) -3- (1H-imidazol-5-yl) propionic acid (0.5 g,1.96mmol,1 eq.) in MeOH (0.6 mL) was added HCl/MeOH (4M, 4.90mL,10 eq.) at 25 ℃. The reaction mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give methyl (2S) -2-amino-3- (1H-imidazol-5-yl) propionate (400 mg, crude material, HCl) as a white solid, which was used directly in the next step. MS (ESI) m/z 170.1[ M+H ]] ⁺

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (741.86 mg,1.77mmol,1 eq. TFA) and methyl (2S) -2-amino-3- (1H-imidazol-5-yl) propionate (0.3 g,1.77mmol,1 eq. HCl), DIPEA (1.15 g,8.87mmol,1.54mL,5 eq.) in THF (0.3 mL) and DCM (0.3 mL) was added T3P (1.69 g,2.66mmol,1.58mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 12h. Adding satiety to the reaction mixture And sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was washed with brine (3 ml x 3) and dried over anhydrous sodium sulfate and concentrated to give the crude product. Obtaining (2S) -3- (1H-imidazol-5-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]Methyl propionate (300 mg, crude) and was used directly in the next step. MS (ESI) m/z 456.2[ M+H ]] ⁺ .1H NMR (400 MHz, methanol-d) ₄ )δppm 7.48(s,1H),7.27(s,1H),7.11-7.18(m,1H),7.02(d,J＝8.16Hz,1H),6.85(s,1H),6.51(d,J＝7.72Hz,1H),4.60-4.71(m,2H),3.93(s,3H),3.68(s,3H),3.00-3.17(m,3H),1.62-1.78(m,3H),0.97(dd,J＝13.78,6.06Hz,6H)

At 25℃under N ₂ Downward (2S) -3- (1H-imidazol-5-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]-4-methyl-pentanoyl]Amino group]NH was added to methyl propionate (200 mg,439.07umol,1 eq.) at a time ₃ MeOH (7M, 11.76mL,187.56 eq.). The mixture was stirred at 80 ℃ and for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the crude product. Obtaining N- [ (1S) -1- [ [ (1S) -2-amino-1- (1H-imidazol-5-ylmethyl) -2-oxo-ethyl ] as a pale yellow solid]Carbamoyl group]-3-methyl-butyl ]-4-methoxy-1H-indole-2-carboxamide (170 mg,378.83 mol,86.28% yield, 98.16% purity) and used directly in the next step. MS (ESI) m/z 441.2[ M+H ]] ⁺

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ (1S) -2-amino-1- (1H-imidazol-5-ylmethyl) -2-oxo-ethyl]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (140 mg, 317.82. Mu.L, 1 eq.) in DCM (2 mL) was added TFAA (133.51 mg, 635.65. Mu.L, 88.41. Mu.L, 2 eq.). The mixture was stirred at 25℃for 2h. Concentrating the reaction mixture to obtain a crude productThe material, after 36H storage, was placed in N- [ (1S) -1- [ [ (1S) -1-cyano-2- (1H-imidazol-5-yl) ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide. Purification of the residue by preparative HPLC gave N- [ (1S) -1- [ [ (1S) -1-cyano-2- (1H-imidazol-5-yl) ethyl as a white solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23.89 mg,56.31umol,17.72% yield, 99.581% purity). MS (ESI) m/z 423.2[ M+H ] ] ⁺

Preparative HPLC conditions: column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-55％，10min； ¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.58(s,1H),7.30(s,1H),7.12-7.21(m,1H),6.99-7.09(m,2H),6.52(d,J＝7.72Hz,1H),5.05(t,J＝7.06Hz,1H),4.61(br dd,J＝9.70,4.85Hz,1H),3.94(s,3H),3.06-3.21(m,2H),1.60-1.83(m,3H),0.99(dd,J＝13.89,6.17Hz,6H)

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (5 g,26.15mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (5.88 g,31.38mmol,1.2 eq., HCl), EDCI (6.52 g,34.00mmol,1.3 eq.), HOBt (4.59 g,34.00mmol,1.3 eq.) in DMF (30 mL) was added TEA (7.94 g,78.46mmol,10.92mL,3 eq.) in one portion. The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (90 mL) and extracted with ethyl acetate (25 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (25 mL) and 5% aqueous sodium bicarbonate (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=30:1 to 10:1). Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]-4-methyl-pentanoic acid tert-butyl ester (5.93 g,16.45mmol,62.91% yield). MS (ESI) m/z 361.2[ M+H ] ] ⁺ 。 ¹ H NMR (400 MHz, chloroform-d) delta ppm 9.25 (br s, 1H), 7.10-7.16 (m, 1H), 6.93-7.00 (m, 2H), 6.56 (br d, J=8.31 Hz, 1H), 6.44 (d, J=7.70 Hz, 1H), 4.66 (td, J=3.1H)8.50,5.14Hz,1H),3.88(s,3H),1.62-1.75(m,2H),1.57-1.62(m,1H),1.42(s,9H),0.92(dd,J＝6.17,3.85Hz,6H)。

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-valerate (2.00 g,5.55mmol,1 eq.) in DCM (8 mL) was added TFA (10.27 g,90.04mmol,6.67mL,16.23 eq.) and H in one portion ₂ O (666.67 mg,37.01mmol, 666.67. Mu.L, 6.67 eq.). The mixture was stirred at 25 ℃ and for 4h. The reaction mixture was concentrated to give a crude product. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow solid]-4-methyl-pentanoic acid (2.24 g,5.35mmol,96.50% yield, TFA) and used directly in the next step. MS (ESI) m/z 305.1[ M+H ]] ⁺

EXAMPLE 50 Synthesis of viral protease inhibitor Compounds 237

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (5 g,26.15mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (5.88 g,31.38mmol,1.2 eq., HCl), EDCI (6.52 g,34.00mmol,1.3 eq.), HOBt (4.59 g,34.00mmol,1.3 eq.) in DMF (30 mL) was added TEA (7.94 g,78.46mmol,10.92mL,3 eq.) in one portion. The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (90 mL) and extracted with ethyl acetate (25 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (25 mL) and 5% aqueous sodium bicarbonate (25 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=30:1 to 10:1). Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]-4-methyl-pentanoic acid tert-butyl ester5.93g,16.45mmol,62.91% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-valerate (2.00 g,5.55mmol,1 eq.) in DCM (8 mL) was added TFA (10.27 g,90.04mmol,6.67mL,16.23 eq.) and H in one portion ₂ O (666.67 mg,37.01mmol, 666.67. Mu.L, 6.67 eq.). The mixture was stirred at 25 ℃ and for 4h. Concentrating the reaction mixture to give (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow solid]-4-methyl-pentanoic acid (2.24 g,5.35mmol,96.50% yield, TFA) which was used directly in the next step. MS (ESI) m/z 305.1[ M+H ] ] ⁺

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (568.23 mg,1.36mmol,1.2 eq, TFA) and methyl 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionate (320 mg,1.13mmol,1 eq, HCl), DIPEA (731.40 mg,5.66mmol, 985.72. Mu.L, 5 eq) in THF (1 mL) and DCM (1 mL) was added T3P (1.08 g,1.70mmol,1.01mL,50% purity, 1.5 eq). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]Methyl 3- (2-oxo-1H-quinolin-4-yl) propionate (0.2 g,375.53umol,33.18% yield). M is MS(ESI)m/z 533.2[M+H] ⁺

At 25℃under N ₂ Downward 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at one time to a mixture of methyl-3- (2-oxo-1H-quinolin-4-yl) propionate (200.00 mg,375.53umol,1 eq) ₃ MeOH (7M, 10.00mL,186.41 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give N- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-1H-quinolin-4-yl) methyl as a pale yellow solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (180 mg,326.21umol,86.87% yield, 93.8% purity) and used directly in the next step. MS (ESI) m/z 518.2[ M+H ]] ⁺

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ 2-amino-2-oxo-1- [ (2-oxo-1H-quinolin-4-yl) methyl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (90 mg,173.89umol,1 eq.) in DCM (5 mL) was added the Bungeus reagent (207.19 mg,869.44umol,5 eq.) in one portion. The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated and purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ 1-cyano-2- (2-oxo-1H-quinolin-4-yl) ethyl as a white solid ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (20.74 mg,41.13umol,23.66% yield, 99.079% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺

Step 6: 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionic acid methyl ester

At 25℃under N ₂ To 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionic acid (400 mg,1.72mmol,1 eq.) was added HCl/MeOH (4M, 4.31mL,10 eq.) in one portion. The mixture was stirred at 25 ℃ and for 1h. The reaction mixture was concentrated to give the product. Methyl 2-amino-3- (2-oxo-1H-quinolin-4-yl) propionate (370 mg, crude material, HCl) was obtained as a white solid and was used directly in the next step.

EXAMPLE 51 Synthesis of viral protease inhibitor Compound 241

Step 1: 2-amino-3- (1H-pyrazol-3-yl) propionic acid methyl ester

At 25℃under N ₂ To 2-amino-3- (1H-pyrazol-3-yl) propionic acid (0.5 g,2.19mmol,1 eq., 2 HCl) was added HCl/MeOH (4M, 17.01mL,31.03 eq.) in one portion. The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give a crude product. Methyl 2-amino-3- (1H-pyrazol-3-yl) propionate (530 mg, crude material, 2 HCl) was obtained as a yellow solid and was used directly in the next step. MS (ESI) m/z 170.1[ M+H ] ] ⁺

Step 2:2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- (1H-pyrazol-3-yl) propanoic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a mixture of 4-methyl-pentanoic acid (377.12 mg,1.24mmol,1 eq.) and methyl 2-amino-3- (1H-pyrazol-3-yl) propanoate (300 mg,1.24mmol,1 eq., 2 HCl), DIPEA (800.75 mg,6.20mmol,1.08mL,5 eq.) in THF (0.9 mL) and DCM (0.9 mL) was added T3P (1.18 g,1.86mmol,1.11mL,50% purity, 1.5 eq.). The mixture was stirred at 25 ℃ and for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]Methyl 3- (1H-pyrazol-3-yl) propionate (130 mg,285.40umol,23.03% yield). MS (ESI) m/z 456.2[ M+H ]] ⁺

Preparative HPLC conditions: column: phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-45％，8min

Step 3: n- [ (1S) -1- [ [ 2-amino-2-oxo-1- (1H-pyrazol-3-ylmethyl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at one time to a mixture of methyl 3- (1H-pyrazol-3-yl) propionate (100 mg,219.54 mol,1 eq.) ₃ MeOH (7M, 3.33mL,106.28 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the product. Obtaining N- [ (1S) -1- [ [ 2-amino-2-oxo-1- (1H-pyrazol-3-ylmethyl) ethyl ] as a pale yellow solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (95 mg, crude) and used directly in the next step. MS (ESI) m/z 441.2[ M+H ]] ⁺

Step 4: n- [ (1S) -1- [ [ 1-cyano-2- (1H-pyrazol-3-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ 2-amino-2-oxo-1- (1H-pyrazol-3-ylmethyl) ethyl]Carbamoyl group]-3-methyl-butyl]-4-To a mixture of methoxy-1H-indole-2-carboxamide (95 mg, 215.67. Mu.l, 1 eq.) and TEA (43.65 mg, 431.33. Mu.l, 2 eq.) in DCM (0.1 mL) was added TFAA (90.59 mg, 431.33. Mu.l, 60.00. Mu.l, 2 eq.). The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated to give a crude product. The crude product was purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ 1-cyano-2- (1H-pyrazol-3-yl) ethyl ] as a white solid ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (23.35 mg,54.93umol,25.47% yield, 99.384% purity). MS (ESI) m/z 423.2[ M+H ]] ⁺

Preparative HPLC conditions: column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:25% -55%,8min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.55(br d,J＝11.25Hz,1H),7.30(s,1H),7.13-7.22(m,1H),7.05(d,J＝7.95Hz,1H),6.54(d,J＝7.70Hz,1H),6.31(dd,J＝10.58,2.14Hz,1H),5.04-5.17(m,1H),4.56-4.64(m,1H),3.95(s,3H),3.13-3.30(m,2H),1.52-1.83(m,3H),0.90-1.08(m,6H)

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (15 g,78.46mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (21.07 g,94.15mmol,1.2 eq., HCl) in DMF (150 mL) was added EDCI (19.55 g,102.00mmol,1.3 eq.), HOBt (13.78 g,102.00mmol,1.3 eq.), TEA (23.82 g,235.38mmol,32.76mL,3 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (450 mL) and extracted with ethyl acetate (250 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (300 mL) and 5% aqueous sodium bicarbonate (300 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=30:1 to 10:1). Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid ]-4-methyl-pentanoic acid tert-butyl ester (24 g,66.58mmol,84.87% yield). MS (ESI) M/z361.2[ M+H] ⁺

Step 7: (S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-pentanoate (10 g,27.74mmol,1 eq.) in DCM (30 mL) was added TFA (61.60 g,540.26mmol,40mL,19.47 eq.) and H in one portion ₂ O (4.00 g,221.98mmol,4.00mL,8.00 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. The reaction mixture was concentrated to give a crude product. The crude product was purified by petroleum ether: ethyl acetate=10:1 (20 mL) and filtered to give the product. Obtaining (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]-4-methyl-pentanoic acid (6 g,19.22mmol,69.27% yield, 97.48% purity). MS (ESI) m/z 305.1[ M+H ]] ⁺

EXAMPLE 52 Synthesis of viral protease inhibitor Compound 245

Step 1: 2-amino-3- (1H-indazol-3-yl) propionic acid methyl ester

At 25℃under N ₂ To a mixture of 2-amino-3- (1H-indazol-3-yl) propionic acid (200 mg,827.56umol,1 eq., HCl) was added HCl/MeOH (4M, 2mL,9.67 eq.). The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give a crude product. Methyl 2-amino-3- (1H-indazol-3-yl) propionate (200 mg, crude material, HCl) was obtained as a pale yellow solid and used directly in the next step. MS (ESI) m/z 220.1[ M+H ] ] ⁺

Step 2:2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl ] amino ] -3- (1H-indazol-3-yl) propanoic acid methyl ester

At 0℃under N ₂ To a mixture of methyl 2-amino-3- (1H-indazol-3-yl) propionate (150 mg, 586.62. Mu. Mol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoic acid (203.52 mg, 879.94. Mu. Mol,1.5 eq.), DIPEA (379.09 mg,2.93mmol, 510.90. Mu.L, 5 eq.) in DCM (1.5 mL) and THF (1.5 mL) was added T3P (559.96 mg,879.94umol,523.33ul,50% purity, 1.5 eq). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2). The organic phase was concentrated to give 2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl as a pale yellow solid]Amino group]Methyl 3- (1H-indazol-3-yl) propionate (180 mg, crude). MS (ESI) m/z 433.2[ M+H ]] ⁺

Step 3:2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- (1H-indazol-3-yl) propanoic acid methyl ester

At 25℃under N ₂ Downward 2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl]Amino group]To a mixture of methyl 3- (1H-indazol-3-yl) propionate (180 mg, 416.17. Mu. Mol,1 eq.) was added HCl/MeOH (4M, 5.14mL,49.43 eq.) in one portion. The mixture was stirred at 25℃for 2h. Concentrating the reaction mixture to give 2- [ [ (2S) -2-amino-4-methyl-pentanoyl as a yellow oil ]Amino group]Methyl 3- (1H-indazol-3-yl) propionate (160 mg, crude, HCl) and used directly in the next step. MS (ESI) m/z 333.2[ M+H ]] ⁺

Step 4:3- (1H-indazol-3-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] propionic acid methyl ester

At 0℃under N ₂ Downward 2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]Methyl 3- (1H-indazol-3-yl) propionate (160 mg,433.77 mol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (99.52 mg,520.53 mol,1.2 eq), DIPEA (280.31 mg,2.17mmol, 377.78. Mu.L, 5 eq) were added in one portion to a mixture of DCM (1 mL) and THF (1 mL) T3P (414.05 mg,650.66 mol, 386.97. Mu.L, 50% purity, 1.5 eq). The mixture was stirred at 25℃for 4h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2). The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 3- (1H-indazol-3-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a pale yellow solid]-4-methyl-pentanoyl]Amino group]Methyl propionate (80 mg, crude material). MS (ESI) m/z 506.2[ M+H ]] ⁺

Preparative HPLC conditions: column: phenomenex Gemini-NX 8 0 x 40mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：35％-65％，8min

Step 5: n- [ (1S) -1- [ [1- (1H-indazol-3-ylmethyl) -2-nitroso-ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward 3- (1H-indazol-3-yl) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at one time to a mixture of methyl propionate (80 mg,158.24umol,1 eq) ₃ MeOH (7M, 1mL,44.24 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated. Obtaining N- [ (1S) -1- [ [1- (1H-indazol-3-ylmethyl) -2-nitroso-ethyl ] as a pale yellow solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (75 mg, crude) and used directly in the next step. MS (ESI) m/z 491.2[ M+H] ⁺

Step 6: n- [ (1S) -1- [ [ 1-cyano-2- (1H-indazol-3-yl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [1- (1H-indazol-3-ylmethyl) -2-nitroso-ethyl ]]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (75 mg,152.89 mol,1 eq.) in DCM (0.5 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (75.00 mg,314.72 mol,2.06 eq.). The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated and purified by preparative HPLC. Obtaining N- [ (1S) -1- [ [ 1-cyano-2- (1H-indazol-3-yl) ethyl ] as a white solid ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (12.0 mg,25.39umol,16.61% yield). MS (ESI) m/z 473.2[ M+H ]] ⁺

Preparative HPLC conditions: column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：28％-58％，10min

¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.95(br d,J＝8.82Hz,1H),11.59(br dd,J＝6.50,1.87Hz,1H),9.02(br dd,J＝14.11,7.94Hz,1H),8.39-8.51(m,1H),7.82(dd,J＝11.14,8.27Hz,1H),7.48-7.55(m,1H),7.31-7.41(m,1H),7.07-7.16(m,2H),6.99-7.05(m,1H),6.49-6.56(m,1H),5.24(quin,J＝7.77Hz,1H),4.39-4.57(m,1H),3.90(d,J＝3.97Hz,3H),3.37-3.62(m,2H),1.60-1.73(m,1H),1.43-1.53(m,1H),1.15-1.28(m,1H),0.84-0.98(m,3H),0.80(d,J＝6.39Hz,2H)

EXAMPLE 53 Synthesis of viral protease inhibitor Compound 1045

/>

Step 1: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (1.3 g,3.27mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 30min. The reaction mixture was concentrated under reduced pressure to give crude methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (900 mg,3.03mmol,92.54% yield) as a yellow oil.

Step 2: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-propoxy-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a mixture of methyl propionate (4478 mg,1.51mmol,1 eq.) and 4-propoxy-1H-indole-2-carboxylic acid (396.37 mg,1.81mmol,1.2 eq.) in DMF (2 mL) was added DCM (8 mL) and EDCI (866.48 mg,4.52mmol,3 eq.) in one portion. DMAP (552.19 mg,4.52mmol,3 eq.) was added to the mixture and stirred at 25℃for 2h. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) purification,to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-propoxy-1H-indole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (480 mg,962.75umol,63.90% yield). MS (ESI) m/z 499.2[ M+H ]] ⁺

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-propoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-propoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group ]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (480 mg,962.75umol,1 eq.) in NH ₃ The mixture in MeOH (7M) (3 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give crude N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-propoxy-1H-indole-2-carboxamide (380 mg,785.84umol,81.62% yield). MS (ESI) m/z 484.3[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-propoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-propoxy-1H-indole-2-carboxamide (380 mg,785.84umol,1 eq.) in DCM (7 mL) was added the Bogis reagent (1.12 g,4.72mmol,6 eq.) in one portion. The mixture was stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product was purified by preparative HPLC (neutral conditions) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-propoxy-1H-indole-2-carboxamide (120 mg,257.76umol,32.80% yield). MS (ESI) m/z 466.3[ M+H ]] ⁺

Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-60％，8min

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(br d,J＝1.7Hz,1H),9.07-8.85(m,1H),8.57(d,J＝7.6Hz,1H),7.83-7.61(m,1H),7.39(d,J＝1.6Hz,1H),7.14-6.90(m,2H),6.48(d,J＝7.6Hz,1H),5.09-4.86(m,1H),4.60-4.28(m,1H),4.04(t,J＝6.4Hz,2H),3.22-3.01(m,2H),2.45-2.03(m,3H),1.94-1.59(m,5H),1.58-1.34(m,1H),1.06(t,J＝7.4Hz,3H),0.95-0.69(m,1H),0.55-0.30(m,2H),0.28--0.02(m,2H)

EXAMPLE 54 Synthesis of viral protease inhibitor Compound 147

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (100 mg,349.26umol,1 eq.) in HCl/MeOH (4M, 2mL,22.91 eq.) was stirred at 25℃for 0.5h. The reaction mixture was concentrated to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (65 mg, crude) as a white solid. MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of (S) -2- ((tert-butoxycarbonyl) amino) -2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetic acid (100 mg,340.93umol,1 eq.) and methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (65 mg,349.07umol,1.02 eq.) in DMF (3 mL) was added TEA (206.99 mg,2.05mmol, 284.72. Mu.L, 6 eq.) and T ₃ P (325.43 mg, 511.40. Mu.mol, 304.14. Mu.L, 50% purity, 1.5 eq.). The reaction was stirred at 25℃for 1H and then with H ₂ O (20 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressureMethyl (S) -2- ((tert-butoxycarbonyl) amino) -2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (110 mg, crude) was obtained as a white solid. MS (ESI) m/z 462.2[ M+H ]] ⁺

Step 3: (S) -2- ((S) -2-amino-2- ((S) -2, 3-Dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (110 mg,238.35umol,1 eq) in HCl/dioxane (2 mL) was stirred at 25℃for 1h. The residue was concentrated in vacuo to give methyl (S) -2- ((S) -2-amino-2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (95 mg, crude material, HCl) as a white solid. MS (ESI) m/z 362.2[ M+H ] ] ⁺

Step 4: (S) -2- ((S) -2, 3-Dihydrobenzofuran-2-yl) -2- (4-methoxy-1H-indole-2-carboxamido) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-2- ((S) -2, 3-dihydrobenzofuran-2-yl) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (95 mg,238.78 mol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (45.65 mg,238.78 mol,1 eq) in DMF (3 mL) were added EDCI (91.55 mg,477.56 mol,2 eq) and DMAP (58.34 mg,477.56 mol,2 eq) followed by stirring at 25 ℃ for 1H. The residue was poured into water (20 mL). The aqueous phase was extracted with ethyl acetate (10 ml x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified for 20% -50%,10 min) to give methyl (S) -2- ((S) -2, 3-dihydrobenzofuran-2-yl) -2- (4-methoxy-1H-indole-2-carboxamido) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (21 mg,39.28umol,16.45% yield) as a pale yellow solid. MS (ESI) m /z 535.2[M+H] ⁺

Step 5: - ((S) -2- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1- ((S) -2, 3-dihydrobenzofuran-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2, 3-Dihydrobenzofuran-2-yl) -2- (4-methoxy-1H-indole-2-carboxamido) acetamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (19 mg, 35.54. Mu. Mol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 5mL,984.71 eq.) was stirred at 25℃for 12h. The reaction was concentrated to give N- ((S) -2- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1- ((S) -2, 3-dihydrobenzofuran-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide (19 mg, crude material) as a white solid. MS (ESI) m/z 520.1[ M+H ]] ⁺

Step 6: n- ((S) -2- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -1- ((S) -2, 3-dihydrobenzofuran-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide

A mixture of N- ((S) -2- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1- ((S) -2, 3-dihydrobenzofuran-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide (19 mg,36.57umol,1 eq), methoxycarbonyl- (triethylammonium) sulfonyl-imide (26.14 mg,109.71umol,3 eq) in DCM (2 mL) was stirred at 25℃for 3H. The reaction mixture was concentrated and purified by preparative HPLC (column Waters Xbridge Prep OBD C, 150 x 40mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to afford N- ((S) -2- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -1- ((S) -2, 3-dihydrobenzofuran-2-yl) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide (2.13 mg,3.74umol,10.22% yield, 88% purity) as a white solid. MS (ESI) m/z 502.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.29-7.32(m,1H)7.19-7.25(m,1H)7.17(d,J＝8.07Hz,1H)7.08-7.14(m,1H)7.05(d,J＝8.31Hz,1H)6.87(t,J＝7.40Hz,1H)6.74(d,J＝7.95Hz,1H)6.54(d,J＝7.70Hz,1H)5.04-5.24(m,2H)4.71-4.78(m,1H)4.63(s,1H)3.96(s,3H)3.35-3.51(m,2H)3.06-3.30(m,2H)2.68(ddt,J＝14.09,9.63,4.83,4.83Hz,0.4H)2.24-2.45(m,2H)2.13-2.22(m,0.6H)1.70-1.94(m,2H)

EXAMPLE 55 Synthesis of viral protease inhibitor Compound 491

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl propionate (240 mg,1.01mmol,1 eq., HCl), (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a mixture of propionic acid (412.2 mg,1.22mmol,1.2 eq, HCl) and TEA (410.4 mg,4.06mmol,0.56mL,4 eq) in DMF (3 mL) was added T ₃ P (1.2 g,2.03mmol,1.21mL,50% purity, 2 eq.). The mixture was stirred at 25℃for 16h. TLC (DCM: meoh=10:1/UV 254 nm) showed new spots were detected. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography12g/>Silica flash column, eluent: 100-25% ethyl acetate/MeOH at 30 mL/min). Obtaining the compound (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (256 mg,0.48mmol,48.2% yield, 92.5% purity).

In a sealed tube, (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (246.3 mg,0.47mmol,92.5% purity, 1 eq.) in NH ₃ The mixture in (7M, 6.72mL,100 eq.) 7M in MeOH was stirred at 80deg.C for 36h. LC-MS showed detection of the desired compound. The reaction mixture was concentrated in vacuo. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (220 mg, crude material) which was used in the next step without further purification.

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A mixture of 4-methoxy-1H-indole-2-carboxamide (250 mg,0.53mmol,1 eq.) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (444.0 mg,1.86mmol,3.5 eq.) in DCM (3 mL) was stirred at 25℃for 16H. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,9.5 min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (83 mg,0.18mmol,34.2% yield, 99.0% purity).

Isomer 1: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

Isomer 2: n- [ (1S) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

Isomer 3: n- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

Isomer 4: n- [ (1R) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10um; mobile phase: [0.1% NH.) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%, min) purification of N- [2- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ]]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,0.11mmol,1 eq.) gives three fragments.

Isomer 1: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide. Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (28.1 mg,62.2umol,56.2% yield, 100% purity). LCMS: rt=0.755 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,DMSO-d ₆ )δ11.57(s,1H),8.91(br d,J＝8.0Hz,1H),8.50(br d,J＝7.5Hz,1H),7.53(br s,1H),7.37(d,J＝1.4Hz,1H),7.15-7.06(m,1H),7.04-6.97(m,1H),6.51(d,J＝7.6Hz,1H),5.07(q,J＝8.2Hz,1H),4.49-4.40(m,1H),3.89(s,3H),3.15-3.01(m,2H),2.34-2.20(m,2H),1.91-1.76(m,3H),1.70(br dd,J＝4.4,8.7Hz,1H),1.64-1.53(m,1H),1.35(br s,1H),0.86-0.76(m,1H),0.48-0.35(m,2H),0.25-0.04(m,2H)。

Isomer 4: n- [ (1R) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide. Obtaining the compound N- [ (1R) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (6.1 mg,13.5umol,12.2% yield, 100% purity). LCMS: rt=0.752 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.27(s,1H),7.18-7.12(m,1H),7.03(d,J＝8.4Hz,1H),6.51(d,J＝7.6Hz,1H),5.12(dd,J＝6.4,7.7Hz,1H),4.85(br s,1H),3.93(s,3H),3.24-3.16(m,2H),2.50-2.32(m,2H),2.06-1.92(m,2H),1.92-1.82(m,2H),1.70(dt,J＝7.0,14.2Hz,2H),1.63-1.54(m,1H),1.31-1.31(m,1H),1.41-1.27(m,1H),0.91-0.80(m,1H),0.53(br d,J＝8.0Hz,2H),0.25-0.14(m,2H)。

By SFC (column: DAICEL CHIRALCEL OD-H (250 mm. Times.30 mm,5 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%, min) to purify a mixture of isomer 2 and isomer 3 (20.0 mg,44.3umol,1 eq) to give two fragments.

Isomer 3: n- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide. Obtaining the compound N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (5.1 mg,11.3umol,25.6% yield, 100% purity). LCMS: rt=0.754 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.28(s,1H),7.18-7.12(m,1H),7.03(d,J＝8.3Hz,1H),6.52(d,J＝7.5Hz,1H),5.06(dd,J＝6.5,9.8Hz,1H),4.81(br s,1H),3.93(s,3H),3.18(br s,2H),2.43-2.35(m,1H),2.45-2.27(m,1H),2.31(br s,1H),2.06-1.95(m,1H),1.94-1.78(m,3H),1.76-1.59(m,2H),1.58-1.45(m,1H),1.40(s,1H),1.29(s,1H),0.92-0.79(m,1H),0.58-0.44(m,2H),0.26-0.12(m,2H)。

Isomer 2: n- [ (1S) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide. Obtaining the compound N- [ (1S) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (6.3 mg,14.0umol,31.6% yield, 100% purity). LCMS: rt=0.754 min; c (C) ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.12(s,1H),7.01-6.96(m,1H),6.87(d,J＝8.3Hz,1H),6.35(d,J＝7.8Hz,1H),4.89(t,J＝7.2Hz,1H),4.43(dd,J＝6.3,8.3Hz,2H),3.77(s,3H),3.08-3.00(m,2H),2.32-2.22(m,1H),2.20-2.10(m,1H),2.27-2.07(m,1H),1.84-1.73(m,2H),1.72-1.62(m,2H),1.60-1.50(m,2H),1.43-1.34(m,1H),0.75-0.62(m,1H),0.40-0.27(m,2H),0.08 -0.04(m,2H)。

EXAMPLE 56 Synthesis of viral protease inhibitor Compounds 247

Step 1: (2S) -2- (Benzylmethoxycarbonylamino) -3- (1H-indol-3-yl) propionic acid

(2S) -2-amino-3- (1H-indol-3-yl) propionic acid (3 g,14.69mmol,1 eq.) is dissolved in NaOH (1M, 14.65 mL) and stirred at 0deg.C. CbzCl (2.51 g,14.73mmol,2.09mL,1 eq.) and NaOH (1M, 14.65 mL) were then added dropwise simultaneously. The mixture was stirred at 20℃for 17h. After completion, the solution was acidified to ph=1 with 6M HCl, followed by extraction of the product with EtOAc (80 ml x 3). The organic layers were combined, taken over Na ₂ SO ₄ Dried and evaporated. Crude productBy chromatography on silica gel (SiO ₂ DCM: meoh=7:1) and purified by preparative HPLC (HPLC: ET40319-84-P1D; column: xtime C1810u 250 mm.80mm; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:5% -35%,25 min) to give (2S) -2- (benzyloxycarbonylamino) -3- (1H-indol-3-yl) propionic acid (1.6 g,4.63mmol,31.55% yield, 98% purity) as a pale yellow solid. MS (ESI) m/z 339.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝10.79(br s,1H),7.57-7.44(m,1H),7.35-7.23(m,5H),7.21-7.13(m,1H),7.10(d,J＝2.1Hz,1H),7.07-7.01(m,1H),6.98-6.88(m,2H),6.98-6.88(m,1H),4.97(s,2H),4.09(dt,J＝4.6,7.7Hz,1H),3.22(dd,J＝4.3,14.4Hz,1H),3.00(dd,J＝8.0,14.5Hz,1H)。

Step 2: (2S) -2- (Benzyloxycarbonylamino) -3- (2-oxoindol-3-yl) propionic acid

To a mixture of (2S) -2- (benzyloxycarbonylamino) -3- (1H-indol-3-yl) propionic acid (1.5 g,4.00mmol,1 eq. HCl) in AcOH (60 mL) was added DMSO (469.01 mg,6.00mmol, 469.01. Mu.L, 1.5 eq.) and HCl (12M, 1.33mL,4 eq.) at 25℃and under N ₂ The mixture was stirred at 25℃for 3h. After completion, the mixture was quenched with water (100 mL), extracted with ethyl acetate (60 mL x 3), and the combined organic layers were washed with brine (200 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure at 40 ℃. The mixture was purified by preparative HPLC (column: welch Xtime C18250. Times.70 mm#10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:7% -37%,20 min) to give (2S) -2- (benzyloxycarbonylamino) -3- (2-oxoindol-3-yl) propionic acid (800 mg,2.03mmol,50.77% yield, 90% purity) as a white solid. MS (ESI) m/z 337.0[ M+H ]] ⁺

Step 3: n- [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl ] ethyl ] carbamic acid phenylmethyl ester

To a mixture of (2S) -2- (benzyloxycarbonylamino) -3- (2-oxoindolin-3-yl) propionic acid (600 mg,1.54mmol,1 eq. HCl) in DMF (20 mL) at 20deg.C1-hydroxybenzotriazole (207.45 mg,1.54mmol,1 eq.) and EDCI (323.74 mg,1.69mmol,1.1 eq.) were added. At N ₂ After stirring the mixture at 20℃for 2h, NH was added dropwise ₃ .H ₂ O (1.01 g,7.22mmol,1.11mL,25% purity, 4.7 eq.). The mixture was stirred at 20℃for 16h. After completion, the mixture was quenched with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure at 40 ℃ to give N- [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl as a white solid]Ethyl group]Benzyl carbamate (500 mg, crude material), which was used in the next step without further purification. MS (ESI) m/z 354.1[ M+H ]] ⁺

Step 4: (2S) -2-amino-3- (2-oxoindolin-3-yl) propanamide

At N ₂ Next, N- [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl was introduced into]Ethyl group]To a solution of benzyl carbamate (500 mg,1.41mmol,1 eq.) in i-PrOH (100 mL) was added Pd/C (339.59 mg, 282.99. Mu. Mol,10% purity, 0.2 eq.) and HCl (12M, 1.30mL,11 eq.). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (15 psi) and stirred at 50℃for 16h. After completion, the reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by preparative HPLC (HPLC: ET40319-96-P1C; column: waters Xbridge Prep OBD C18150. Times.40 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -20%,8 min) to give (2S) -2-amino-3- (2-oxoindolin-3-yl) propanamide (130 mg,549.67umol,38.85% yield, 92.7% purity) as a white solid. MS (ESI) m/z 220.1[ M+H ] ] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2-amino-3- (2-oxoindolin-3-yl) propanamide (60 mg,273.67umol,1 eq.) at 0deg.C, (2S) -3-cycloPropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionic acid (99.29 mg,328.41 mol,1.2 eq.) and Et ₃ N (166.16 mg,1.64mmol, 228.55. Mu.L, 6 eq.) to a mixture in DCM (10 mL) was added T3P (522.46 mg, 821.02. Mu.L, 488.28. Mu.L, 50% pure, 3 eq.) dropwise. At N ₂ The solution was stirred at 25℃for 1h. After completion, the mixture was quenched with water (20 mL) and extracted with DCM: meoh=7:1 (15 mL x 2). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure at 40 ℃. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl as a colorless oil]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (40 mg,55.61umol,20.32% yield, 70% purity). MS (ESI) m/z 504.3[ M+H ] ] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- (2-oxoindolin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

At 20℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (2-oxoindolin-3-yl) methyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (35 mg,69.51umol,1 eq.) in DCM (6 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (49.69 mg,208.52umol,3 eq.) in one portion and stirred at 20 ℃ for 2H. Methoxycarbonyl- (triethylammonium) sulfonyl-imide (82.82 mg,347.53 mol,5 eq.) was added at 20℃and stirred for 4h at 20 ℃. After completion, the crude material was purified with N ₂ Dried and purified by preparative HPLC (column Waters Xbridge Prep OBD C18 150 x 40mM x 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) and purified by preparative HPLC (column: phenomenex Luna C18, 75 x 30mm x 3um; mobile phase: [ Water (0.2% FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -70%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- (2-oxoindolin-3-yl) ethyl as a white solid ]Amino group]-1- (cyclopropyl) methylRadical) -2-oxo-ethyl radical]-4-methoxy-1H-indole-2-carboxamide (5 mg,10.23 mol,14.72% yield, 99.36% purity). MS (ESI) m/z 486.2[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.39-7.09(m,4H),7.07-6.95(m,2H),6.92-6.80(m,1H),6.51(dd,J＝3.1,7.5Hz,1H),5.37-5.14(m,1H),4.65-4.47(m,1H),3.93(dd,J＝1.4,3.4Hz,3H),3.70-3.52(m,1H),2.63-2.27(m,2H),1.92-1.60(m,2H),0.84(br s,1H),0.59-0.43(m,2H),0.27-0.10(m,2H)

EXAMPLE 57 Synthesis of viral protease inhibitor Compound 331

Step of isomers 1 and 2: n- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] -2-pyrrolidin-1-yl-ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (700 mg,1.27mmol,80% purity, 1 eq.) in EtOH (10 mL) was added pyrrolidine (180.01 mg,2.53mmol,211.28uL,2 eq.) and ZnCl2 (1M, 12.66uL,0.01 eq.). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (251.10 mg,2.53mmol,316.65ul,2 eq.) was added. The mixture was stirred at 25℃for 2h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave the compound N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (110 mg,199.95umol,15.80% yield, 95% purity) and N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl)]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (110 mg,199.95umol,15.80% yield, 95% purity). MS (ESI) m/z 523.4[ M+H ]] ⁺

Column: phenomenex luna CN 5u 100 x 30mm; mobile phase: [ hexane-IPA ]; b%:5% -40%,20min

Isomer 1: ¹ H NMR(400MHz,DMSO-d6)δ＝11.58(s,1H),8.43(d,J＝7.7Hz,1H),8.20(d,J＝9.4Hz,1H),7.68-7.49(m,1H),7.38(d,J＝1.2Hz,1H),7.18-6.93(m,2H),6.50(d,J＝7.6Hz,1H),4.57-3.99(m,3H),3.88(s,3H),3.19-2.95(m,2H),2.64-2.53(m,4H),2.38-2.27(m,1H),2.15-2.01(m,1H),1.85-1.44(m,10H),0.91(dd,J＝6.4,16.3Hz,6H)

isomer 2: ¹ H NMR(400MHz,DMSO-d6)δ＝11.59(br s,1H),8.39(br d,J＝7.6Hz,1H),8.01(br d,J＝9.1Hz,1H),7.69-7.49(m,1H),7.43-7.28(m,1H),7.16-6.86(m,2H),6.50(d,J＝7.6Hz,1H),4.59-4.24(m,3H),3.88(s,3H),3.19-2.94(m,2H),2.71-2.57(m,2H),2.49-2.32(m,3H),2.18-2.08(m,1H),2.06-1.93(m,1H),1.83-1.37(m,9H),0.90(dd,J＝6.5,15.2Hz,6H)

EXAMPLE 58 Synthesis of viral protease inhibitor Compound 389

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propanamide

Tert-butyl N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamate (2 g,7.37mmol,1 eq.) in HCl/EtOAc (4M, 50mL,27.13 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionamide (1.2 g, crude material) as a white solid.

Step 2: 2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

2-Boc-2-azaspiro [4.5] decane-3-carboxylic acid (3 g,10.59mmol,1 eq.) was added to HCl/MeOH (4M, 50mL,18.89 eq.). The mixture was stirred at 80℃for 2h. The mixture was concentrated under reduced pressure to give methyl 2-azaspiro [4.5] decane-3-carboxylate (2 g, crude material) as a yellow oil.

To 2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid methyl ester (2 g,10.14mmol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (2.33 g,12.17mmol,1.2 eq.) in DCM (30 mL) and DMF (5 mL) was added T3P (12.90 g,20.28mmol,12.06mL,50% purity, 2 eq.) and DIEA (3.93 g,30.41mmol,5.30mL,3 eq.). The mixture was stirred at 25℃for 2h. After completion, by adding H ₂ O (100 mL) to quench the reaction mixture and extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) purification of the residue to give the product 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid methyl ester (3 g,8.10mmol,79.88% yield). MS (ESI) m/z 371.1[ M+H ]] ⁺

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxylic acid methyl ester (3 g,8.10mmol,1 eq.) in THF (45 mL) and H ₂ LiOH.H was added to the solution in O (15 mL) ₂ O (1.70 g,40.49mmol,5 eq.). The mixture was stirred at 25℃for 12h. After completion, by adding H ₂ O (50 mL) to quench the mixture, and then aqueous HCl (1M) was added to adjust to ph=3-4 and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (2.6 g, crude material). MS (ESI) m/z 357.1[ M+H ]] ⁺

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 0deg.C]Decane-3-carboxylic acid (1 g,2.81mmol,1 equivalent) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of propionamide (720.49 mg,4.21mmol,1.5 eq.) in DCM (30 mL) was added T3P (3.57 g,5.61mmol,3.34mL,50% purity, 2 eq.) and DIEA (1.09 g,8.42mmol,1.47mL,3 eq.). The mixture was stirred at 30℃for 1h. After completion, by adding H ₂ O (100 mL) to quench the mixture and then extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (700 mg,1.37mmol,48.96% yield). MS (ESI) m/z 510.3[ M+H ]] ⁺

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.3)0mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,9 min) isolation of N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (500 mg,1.02 mmol) gave the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide isomer 1 (264 mg,537.04umol,52.80% yield, 100% purity). MS (ESI) m/z 492.3[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.28-6.76(m,3H),6.60-6.38(m,1H),5.05(br dd,J＝5.2,10.2Hz,1H),4.63-4.60(m,1H),4.03-3.85(m,5H),3.74-3.28(m,1H),2.73(br dd,J＝5.0,8.6Hz,1H),2.51-2.28(m,2H),2.27-2.08(m,1H),1.96-1.72(m,2H),1.69-1.38(m,11H),1.37-1.09(m,1H)。

N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] ]Decane-3-carboxamide isomer 2 (140 mg,284.51umol,27.97% yield, 99.9% purity). MS (ESI) m/z 492.3[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.30-6.81(m,3H),6.53(br d,J＝2.0Hz,1H),5.12-4.95(m,2H),4.70-4.55(m,2H),4.08-3.86(m,4H),3.84-3.72(m,1H),2.62-2.40(m,1H),2.36-2.18(m,2H),1.94-1.69(m,3H),1.68-1.34(m,11H)。

EXAMPLE 59 Synthesis of viral protease inhibitor Compound 513

Step 1: 4-Methoxyindoline-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester

To a mixture of 1-tert-butoxycarbonyl-4-hydroxy-indoline-2-carboxylic acid (300 mg,1.07mmol,1 eq.) in DMF (4 mL) was added K ₂ CO ₃ (445.37 mg,3.22mmol,3 eq.) and MeI (381.16 mg,2.69mmol,167.18uL,2.5 eq.) was added to the mixture at 0deg.C. At the position ofAfter stirring for 16H at 25℃the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 5/1) to give 4-methoxyindoline-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (220 mg,715.82umol,66.64% yield) as a yellow solid. MS (ESI) m/z 208.0[ M+H-Boc] ⁺

Step 2: 1-Boc-4-methoxy-indoline-2-carboxylic acid

To 4-methoxyindoline-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (200 mg,650.74umol,1 eq.) in THF (1 mL) and H at 25 ℃ ₂ To the mixture in O (1 mL) was added LiOH (46.75 mg,1.95mmol,3 eq.) in one portion. The mixture was stirred at 25℃for 16h. The reaction mixture was made acidic with HCl solution and extracted with ethyl acetate (2 ml x 3). The combined organic layers were washed with brine (5 ml 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 1-tert-butoxycarbonyl-4-methoxy-indoline-2-carboxylic acid (175 mg,596.63umol,45.84% yield) as a yellow oil. MS (ESI) m/z 237.9[ M+H-56 ]] ⁺

Step 3: 4-methoxyindoline-2-carboxylic acid

To a mixture of 1-tert-butoxycarbonyl-4-methoxy-indoline-2-carboxylic acid (150 mg,511.40 mol,1 eq.) was added HCl/dioxane (4 m,7.50ml,58.66 eq.). The reaction was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step to give the compound 4-methoxyindoline-2-carboxylic acid (110 mg,431.07umol,84.29% yield, 90% purity, HCl) as a yellow oil. MS (ESI) m/z 194.1[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-indoline-2-carboxamide

At 0deg.C, 4-methoxyindoline-2-carboxylic acid (110 mg,478.97umol,1 eq., HCl) and 2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a mixture of 3-cyclopropyl-propionamide (316.51 mg,478.97umol,40% purity, 1 eq.) in DCM (8 mL) was added DIEA (123.81 mg,957.94umol,166.86uL,2 eq.) and T ₃ P (457.20 mg,718.45umol,427.29uL,50% purity, 1.5 eq.). The mixture was stirred at 0℃for 1h. The mixture was stirred with EDTA (10 mL) at 25℃and the reaction mixture was quenched with H ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 2). The combined organic layers were concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-indoline-2-carboxamide (29 mg,63.81umol,13.32% yield, 96.7% purity) and N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl)]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-indoline-2-carboxamide (26 mg,55.61umol,11.61% yield, 94% purity). MS (ESI) m/z 440.2[ M+H ]] ⁺

Column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%: 20-50%, 10min

¹ H NMR(400MHz,DMSO-d6)δ＝8.99-8.83(m,1H),8.08-7.89(m,1H),7.71(s,1H),6.92(t,J＝8.0Hz,1H),6.25(dd,J＝4.4,7.9Hz,2H),5.91(d,J＝3.5Hz,1H),5.05-4.85(m,1H),4.42-4.14(m,2H),3.70(s,3H),3.28-2.97(m,3H),2.90-2.76(m,1H),2.43-2.26(m,1H),2.19-1.98(m,2H),1.87-1.54(m,3H),1.50-1.31(m,1H),0.79-0.54(m,1H),0.47-0.26(m,2H),0.20 -0.10(m,2H)

¹ H NMR(400MHz,DMSO-d6)δ＝8.88(d,J＝7.9Hz,1H),7.95(d,J＝8.0Hz,1H),7.70(s,1H),6.93(t,J＝8.0Hz,1H),6.26(dd,J＝4.5,7.9Hz,2H),5.92(d,J＝3.6Hz,1H),5.08-4.84(m,1H),4.50-4.17(m,2H),3.70(s,3H),3.27-2.99(m,3H),2.88-2.72(m,1H),2.40-2.25(m,1H),2.17-2.02(m,2H),1.87-1.57(m,3H),1.51-1.39(m,1H),0.70(br s,1H),0.49-0.26(m,2H),0.21 -0.14(m,2H)

EXAMPLE 60 Synthesis of viral protease inhibitor Compound 515

Step 1: 4-hydroxy-1H-indole-2-carboxylic acid

To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (500 mg,2.62mmol,1 eq.) in DCM (10 mL) at 0deg.C was added BBr ₃ (1.31 g,5.23mmol,2 eq.). The mixture was stirred at 25℃for 16h. H for mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 2). The combined organic layers were washed with 20mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 4-hydroxy-1H-indole-2-carboxylic acid (200 mg, crude material) as a red solid. MS (ESI) m/z 176.1[ M-H] ⁺

Step 2: 4-hydroxy-1H-indole-2-carboxylic acid methyl ester

4-hydroxy-1H-indole-2-carboxylic acid (200 mg,1.13mmol,1 eq.) was added to HCl/MeOH (4M, 10mL,35.43 eq.) and the mixture stirred at 70℃for 5H. The reaction mixture was concentrated under reduced pressure to give a crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=9/1 to 8/1) to give methyl 4-hydroxy-1H-indole-2-carboxylate (170 mg,800.28umol,70.89% yield, 90% purity) as a yellow solid. MS (ESI) m/z 190.1[ M-H] ⁺

Step 3:4- (2- (N-morpholinoethoxy) -1H-indole-2-carboxylic acid methyl ester

To a mixture of methyl 4-hydroxy-1H-indole-2-carboxylate (300 mg,1.57mmol,1 eq.) and 2- (N-morpholinoethanol (205.83 mg,1.57mmol,192.37uL,1 eq.) in THF (4 mL) was added PPh ₃ (452.73 mg,1.73mmol,1.1 eq.) at 0deg.C under N ₂ DIAD (317.30 mg,1.57mmol,305.10uL,1 eq.) was added. The mixture was stirred at 25℃for 60min. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 2). The combined organic layers were washed with brine (20 mL), filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (PE: ea=0:1) to give 4- (2-morpholinoethoxy) -1H-indole as a yellow solidMethyl indole-2-carboxylate (200 mg,591.44umol,37.69% yield, 90% purity). MS (ESI) M/z304.9[ M+H ]] ⁺

Step 4:4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid

To 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid methyl ester (200 mg,657.16umol,1 eq.) in THF (2 mL) and H at 25 ℃ ₂ LiOH.H was added to the mixture in O (1 mL) ₂ O (41.37 mg,985.74umol,1.5 eq.). The mixture was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to give a residue. The crude material was purified by HCl prep HPLC to give 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid (80 mg,261.79 mol,39.84% yield, 95% purity) as a white solid. MS (ESI) m/z 289.2[ M-H ] ] ⁺

Column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ water (0.04% hcl) -ACN ]; b%:1% -32%,6.5min

Step 5: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4- (2-morpholinoethoxy) -1H-indole-2-carboxamide

To 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid (70 mg,241.12umol,1 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl at 0deg.C]Ethyl group]To a mixture of 3-cyclopropyl-propionamide (159.33 mg,241.12umol,40% purity, 1 eq.) in DCM (2 mL) was added DIEA (93.49 mg,723.36umol,125.99uL,3 eq.) and T in one portion ₃ P (230.16 mg,361.68umol,215.10uL,50% purity, 1.5 eq.) and the mixture was stirred at 0deg.C for 2h. EDTA solution (2 mL) was added to the reaction mixture and stirred at 25℃for 10min, and then extracted with DCM (2 mL. Times.3). The combined organic layers were washed with brine (5 ml x 1) and concentrated under reduced pressure to give a residue. Purification of the residue by preparative HPLC (neutral conditions) gives N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4- (2-morpholinoethoxy) -1H-indole-2-carboxamide (13 mg,24.23 mol,10.05% yield). MS (ESI) m/z 537.3[ M+H ]] ⁺

Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：20％-50％，8min

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.57(s,1H),8.92(d,J＝7.9Hz,1H),8.60(br d,J＝7.5Hz,1H),7.79-7.68(m,1H),7.35(d,J＝1.5Hz,1H),7.14-6.93(m,2H),6.51(d,J＝7.5Hz,1H),4.98(q,J＝7.9Hz,1H),4.54-4.38(m,1H),4.21(br d,J＝3.5Hz,2H),3.59(t,J＝4.5Hz,4H),3.20-3.05(m,2H),2.78(t,J＝5.6Hz,2H),2.60-2.52(m,4H),2.43-2.28(m,1H),2.23-2.04(m,2H),1.92-1.60(m,3H),1.56-1.38(m,1H),0.80(br d,J＝5.3Hz,1H),0.51-0.30(m,2H),0.25-0.05(m,2H)

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.34-7.28(m,1H),7.18-7.11(m,1H),7.04(d,J＝8.4Hz,1H),6.53(d,J＝7.5Hz,1H),5.08(dd,J＝5.8,10.3Hz,1H),4.54(t,J＝7.4Hz,1H),4.30(t,J＝5.3Hz,2H),3.77-3.72(m,4H),3.30-3.27(m,2H),2.92(t,J＝5.3Hz,2H),2.75-2.59(m,5H),2.40-2.26(m,2H),1.99-1.79(m,3H),1.78-1.60(m,1H),0.93-0.76(m,1H),0.58-0.52(m,2H),0.20(br dd,J＝5.0,11.6Hz,2H)

EXAMPLE 61 Synthesis of viral protease inhibitor Compound 525

Step 1:4, 6-dichloro-2- (trichloromethyl) -1H-benzimidazole

A mixture of 3, 5-dichlorobenzene-1, 2-diamine (640.64 mg,3.62mmol,1 eq.) and methyl 2, 2-trichloroethyliminoate (766.16 mg,4.34mmol,535.78uL,1.2 eq.) in AcOH (5 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and filtered to give 4, 6-dichloro-2- (trichloromethyl) -1H-benzimidazole (860 mg,2.83mmol,78.07% yield) as a brown solid. MS (ESI) M/z304.5[ M+2H] ⁺

Step 2:4, 6-dichloro-1H-benzimidazole-2-carboxylic acid methyl ester

To 4, 6-dichloro-2- (trichloromethyl) -1H-benzimidazole (420 mg,1.38mmol,1 eq.) in MeOH (5 mL) was added Na in one portion ₂ CO ₃ (146.25 mg,1.38mmol,1 eq.). The mixture was heated to 70 ℃ and stirred for 14 hours. After completion, the reaction mixture was diluted with HCl (10 mL) and stirred at 25 ℃ for 1h and extracted with ethyl acetate (8 mL x 3). The combined organic layers were washed with brine (10 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give methyl 4, 6-dichloro-1H-benzimidazole-2-carboxylate (330 mg,1.35mmol,97.59% yield) as a brown solid. MS (ESI) m/z 245.0[ M+H ]] ⁺

Step 3:4, 6-dichloro-1H-benzimidazole-2-carboxylic acid

To 4, 6-dichloro-1H-benzimidazole-2-carboxylic acid methyl ester (330 mg,1.35mmol,1 eq.) in THF (2 mL) and H at 25 ℃ ₂ NaOH (161.58 mg,4.04mmol,3 eq.) was added in one portion to the mixture in O (2 mL). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was made acidic with 1M HCl solution (5 mL) and then extracted with ethyl acetate (5 mL x 3). The combined organic layers were washed with brine (10 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 4, 6-dichloro-1H-benzimidazole-2-carboxylic acid (200 mg,865.67umol,64.29% yield) as a brown solid. MS (ESI) m/z 229.0[ M-H] ⁺

Step 4: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-benzimidazole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (140 mg,470.83 mol,1 eq) and 4, 6-dichloro-1H-benzimidazole-2-carboxylic acid (197.78 mg,470.83 mol,55% purity, 1 eq) were added DMAP (172.56 mg,1.41mmol,3 eq) in a mixture of DCM (3 mL) and DMF (1 mL) at once. EDCI (270.78 mg,1.41mmol,3 eq.) was added to the mixture and stirred at 25℃for 2h. After completion, the reaction mixture was taken up with H ₂ Dilute with O (4 mL) and extract with ethyl acetate (5 mL x 3). The combined organic layers were washed with brine (8 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By preparative HPLC (alkaline conditions, column: phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-benzimidazole-2-carbonyl) amino) as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (70 mg,137.16umol,29.13% yield). MS (ESI) m/z 510.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 6-dichloro-1H-benzimidazole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-benzimidazole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (60 mg,117.56umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 8mL,476.34 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4, 6-dichloro-1H-benzimidazole-2-carboxamide (58 mg,117.09umol,99.60% yield). MS (ESI) m/z 495.2[ M+H ]] ⁺

Step 6:4, 6-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-benzoimidazole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4, 6-dichloro-1H-benzimidazole-2-carboxamide (50 mg,100.94umol,1 eq.) in DCM (1 mL) was added the bergs reagent (48.11 mg,201.87umol,2 eq.) in one portion. The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (basic conditions, column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,8 min) to give 4, 6-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-1H-benzimidazole-2-carboxamide (13 mg,27.23 mol,26.98% yield). MS (ESI) m/z 477.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝13.77(br s,1H),8.97-8.81(m,2H),7.71(s,1H),7.66-7.40(m,2H),5.05-4.91(m,1H),4.60-4.48(m,1H),3.21-3.03(m,2H),2.43-2.28(m,1H),2.22-2.06(m,2H),2.02-1.85(m,1H),1.84-1.54(m,3H),0.81-0.69(m,1H),0.48-0.34(m,2H),0.20-0.04(m,2H)

EXAMPLE 62 Synthesis of viral protease inhibitor Compound 529

Step 1: (2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To a mixture of 6-chloro-1H-indole-2-carboxylic acid (800 mg,4.08mmol,1 eq.) in DCM (6 mL) and DMF (3 mL) was added DMAP (1.50 g,12.24mmol,3 eq.) in one portion at 25 ℃. (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl was added in one portion to the mixture at 25 ℃]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.36 g,4.08mmol,1 eq., HCl) and EDCI (2.34 g,12.24mmol,3 eq.) and stirred for 2.5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. Obtaining the compound (2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (900 mg,1.89mmol,46.45% yield, 90% purity). MS (ESI) M/z475.1[ M+H ] ] ⁺ 。

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino ] at 25 ℃C]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (900 mg,1.89mmol,1 eq.) in NH ₃ Mixture in MeOH (7M, 10mL,94.99 eq.). The mixture was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-1H-indole-2-carboxamide (750 mg, crude) and used directly in the next step. MS (ESI) m/z 460.1[ M+H ]] ⁺ 。

Step 3: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 6-chloro-1H-indole-2-carboxamide (700 mg,1.52mmol,1 eq.) in DCM (7 mL) was added the Bungeus reagent (725.41 mg,3.04mmol,2.5 eq.) in one portion. The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Waters Xbridge Prep OBD C18150. 40 mM. 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) of the purified residue. Obtaining the compound 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (60 mg,133.90 mol,30.79% yield, 98.622% purity). MS (ESI) m/z 442.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.74(br s,1H),8.95(br d,J＝7.72Hz,1H),8.66(br d,J＝7.28Hz,1H),7.65-7.74(m,2H),7.43(s,1H),7.32(s,1H),7.05(dd,J＝8.49,1.87Hz,1H),4.95-5.03(m,1H),4.47(br dd,J＝13.67,7.94Hz,1H),3.07-3.18(m,2H),2.31-2.41(m,1H),2.07-2.18(m,2H),1.65-1.89(m,3H),1.42-1.54(m,1H),0.80(br s,1H),0.36-0.49(m,2H),0.07-0.24(m,2H),-0.69--0.69(m,1H)

EXAMPLE 63 Synthesis of viral protease inhibitor Compound 539

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (500 mg,1.75mmol,1 eq.) in HCl/MeOH (4M, 20mL,45.81 eq.) was stirred at 20deg.C for 1h and the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (350 mg, crude material, HCl) as a yellow solid.

Step 2: (2S, 4R) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-methylpyrrolidine-1-carboxylic acid tert-butyl ester

To (2S, 4R) -1-tert-butoxycarbonyl-4-methyl-pyrrolidine-2-carboxylic acid (250 mg,1.09mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (304.45 mg,1.64mmol,1.5 eq.) in DCM (10 mL) was added dropwise T3P (1.04 g,1.64mmol,972.75uL,50% purity, 1.5 eq.) and Et ₃ N (662.02 mg,6.54mmol,910.62uL,6 eq.) and the mixture stirred at 20℃for 2h. By adding H at 0 ℃ ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) the residue was purified to give (2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-pyrrolidine-3- ] as a colorless oilBase group]Methyl group]Ethyl group]Carbamoyl group]-4-methyl-pyrrolidine-1-carboxylic acid tert-butyl ester (320 mg,805.10umol,73.86% yield). MS (ESI) m/z 398.2[ M+H ]] ⁺ 。

Step 3: (S) -2- ((2S, 4R) -4-methylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

(2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A solution of tert-butyl 4-methyl-pyrrolidine-1-carboxylate (260 mg,654.15umol,1 eq.) in HCl/MeOH (4M, 8mL,48.92 eq.) was stirred at 20℃for 1h. The reaction mixture was concentrated under reduced pressure to give (2S) -2- [ [ (2S, 4R) -4-methylpyrrolidine-2-carbonyl ] as a colorless oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg, crude material, HCl). MS (ESI) m/z 298.2[ M+H ]] ⁺ 。

Step 4: (S) -2- ((2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4-methylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S, 4R) -4-methylpyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (200 mg,599.14umol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (229.09 mg,1.20mmol,2.0 eq) in DMF (2.0 mL) was added DMAP (219.59 mg,1.80mmol,3.0 eq) and EDCI (229.71 mg,1.20mmol,2 eq) and DCM (8.0 mL) and the mixture was stirred at 20 ℃ for 2H. By adding H at 0 ℃ ₂ O (50 mL) to quench the reaction mixture and then extract with DCM (40 mL x 3). The combined organic layers were washed with brine (60 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:1 to 0:1) to give (2S) -2- [ [ (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carbonyl as a yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,494.14umol,82.47% yield, 93% purity). MS (ESI) m/z 471.3[ M+H ]] ⁺ 。

Step 5: (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide

(2S) -2- [ [ (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (220 mg,434.84umol,93% purity, 1 eq.) in NH ₃ A solution in MeOH (7M, 20mL,321.96 eq.) was stirred at 60℃for 12h. The reaction mixture was concentrated under reduced pressure to give (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow solid]Methyl group]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (200 mg, crude). MS (ESI) m/z 456.2[ M+H ] ] ⁺ 。

Step 6: (2S, 4R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide

To (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (100 mg,219.54umol,1 eq.) in DCM (5 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (313.90 mg,1.32mmol,6 eq.) and stirred at 20 ℃ for 3H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -60%,8 min) to give (2S, 4 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (33 mg,75.43umol,34.36% yield, 100% purity). MS (ESI) m/z 438.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.73-11.47(m,1H),8.85(br d,J＝8.3Hz,1H),7.84-7.54(m,1H),7.24-6.84(m,3H),6.74-6.48(m,1H),5.10-4.47(m,2H),4.20-3.75(m,4H),3.47(t,J＝9.0Hz,1H),3.16(d,J＝7.9Hz,1H),2.61(s,1H),2.43-2.36(m,1H),2.27-1.43(m,7H),1.07(d,J＝6.4Hz,3H)。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.25-6.75(m,3H),6.59-6.40(m,1H),5.15-5.00(m,1H),4.84-4.61(m,1H),4.30-4.06(m,1H),3.98-3.84(m,3H),3.55(t,J＝8.9Hz,1H),3.30-3.24(m,1H),3.01-2.54(m,2H),2.46-2.09(m,4H),2.01-1.38(m,3H),1.15(br d,J＝6.6Hz,3H)。

EXAMPLE 64 Synthesis of viral protease inhibitor Compound 547

Step 1: (1S, 2S, 5R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-azabicyclo [3.2.0] heptane-3-carboxylic acid 9H-fluoren-9-ylmethyl ester

To (1S, 2S, 5R) -3- (9H-fluoren-9-ylmethoxycarbonyl) -3-azabicyclo [3.2.0]Heptane-2-carboxylic acid (150 mg,412.76umol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To DCM (2 mL) of propionitrile (75.87 mg,495.31umol,1.2 eq) were added T3P (394.00 mg,619.14umol,368.22uL,50% purity, 1.5 eq) and DIEA (160.04 mg,1.24mmol,215.69uL,3 eq) and the resulting mixture was stirred at 25℃for 2h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with ethyl acetate (20 mL. Times.3), and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give (1S, 2S,5 r) -2-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-3-azabicyclo [3.2.0]Heptane-3-carboxylic acid 9H-fluoren-9-ylmethyl ester (130 mg,260.74umol,63.17% yield, 100% purity). MS (ESI) m/z 499.3[ M+H ]] ⁺ 。

Step 2: (1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-azabicyclo [3.2.0] heptane-2-carboxamide

To (1S, 2S, 5R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group ]Carbamoyl group]-3-azabicyclo [3.2.0]To a solution of 9H-fluoren-9-ylmethyl heptane-3-carboxylate (250 mg,401.15umol,80% purity, 1 eq.) in DCM (2.5 mL) was added piperidine (68.31 mg,802.29 u)mol,79.23uL,2 eq) and the solution was stirred for 2h at 25 ℃. After completion, DCM was removed by blow-drying to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give (1S, 2S,5 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Ethyl group]-3-azabicyclo [3.2.0]Heptane-2-carboxamide (80 mg,289.51umol,72.17% yield, 100% purity). MS (ESI) m/z 277.2[ M+H ]] ⁺ 。

Step 3: (1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.2.0] heptane-2-carboxamide

To (1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-azabicyclo [3.2.0]To a solution of heptane-2-carboxamide (80 mg,289.51umol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (83.02 mg,434.26umol,1.5 eq) in DCM (1.5 mL) was added T3P (276.35 mg,434.26umol,258.27ul,50% purity, 1.5 eq) and DIEA (112.25 mg,868.52umol,151.28ul,3 eq). The resulting solution was stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with ethyl acetate (20 mL x 3), and the combined organic phases were over Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give (1S, 2S,5 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.2.0]Heptane-2-carboxamide (50 mg,111.23umol,38.42% yield, 100% purity). MS (ESI) m/z 449.9[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.57(br s,1H),9.23-8.65(m,1H),7.69(br s,1H),7.23-6.82(m,3H),6.52(br d,J＝7.4Hz,1H),5.08-4.84(m,1H),4.63(br d,J＝8.2Hz,1H),4.25(br s,1H),4.06(br s,1H),3.89(br s,3H),3.27-2.79(m,4H),2.28-1.53(m,9H)。

EXAMPLE 65 Synthesis of viral protease inhibitor Compounds 549

Step 1: (2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -4- (trifluoromethyl) pyrrolidine-1-carboxylic acid tert-butyl ester

At 0℃under N ₂ Downward (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (283.01 mg,1.27mmol,1.2 eq, HCl) and (2S, 4R) -1-tert-butoxycarbonyl-4- (trifluoromethyl) pyrrolidine-2-carboxylic acid (300 mg,1.06mmol,1 eq), DIEA (684.44 mg,5.30mmol,922.43uL,5 eq) in THF (3 mL) was added T3P (1.01 g,1.59mmol,944.87uL,50% purity, 1.5 eq). The mixture was stirred at 25℃for 1h. After completion, the residue was poured into saturated sodium bicarbonate solution (10 mL) and stirred for 1min. The aqueous phase was extracted with ethyl acetate (10 ml x 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give (2S, 4 r) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a pale yellow oil]Methyl group]Ethyl group]Carbamoyl group]-4- (trifluoromethyl) pyrrolidine-1-carboxylic acid tert-butyl ester (0.5 g, crude material) and used directly in the next step. MS (ESI) m/z 452.1[ M+H ]] ⁺ 。

Step 2: (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl ] -2- [ [ (2S, 4R) -4- (trifluoromethyl) pyrrolidine-2-carbonyl ] amino ] propionic acid methyl ester

At 25℃under N ₂ Downward (2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]To tert-butyl 4- (trifluoromethyl) pyrrolidine-1-carboxylate (0.5 g,1.11mmol,1 eq.) was added HCl/MeOH (4M, 3mL,10.83 eq.). The mixture was stirred at 25℃for 15min. After completion, the reaction mixture was concentrated to give the crude product (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl as a pale yellow oil]-2- [ [ (2 s,4 r) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]Methyl propionate (450 mg, crude material, HCl). MS (ESI) m/z 352.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [ (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 0℃under N ₂ Downward (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl]-2- [ [ (2 s,4 r) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]Methyl propionate (395.52 mg,1.02mmol,1.3 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (150 mg,784.59umol,1 eq) were added to a mixture of DIPEA (507.01 mg,3.92mmol,683.31ul,5 eq) in THF (3 mL) and DCM (3 mL) T3P (748.92 mg,1.18mmol,699.93ul,50% purity, 1.5 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was poured into saturated sodium bicarbonate solution (5 mL) and stirred for 2min. The aqueous phase was extracted with ethyl acetate (5 ml x 2). The combined organic phases were washed with brine (5 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was purified by preparative TLC (dichloromethane: methanol=10:1, r _f =0.43) to give (2S) -2- [ [ (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl as a pale yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg, crude material). MS (ESI) m/z 525.2[ M+H ]] ⁺ 。

Step 4: (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1- (nitrosomethyl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4- (trifluoromethyl) pyrrolidine-2-carboxamide

At 25℃under N ₂ Downward (2S) -2- [ [ (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added at one time to a mixture of methyl propionate (250 mg,476.65umol,1 eq) ₃ MeOH (7M, 3mL,44.06 eq.). The mixture was stirred at 80℃for 12h. After completion, the reaction mixture was cooled to 25 ℃ and concentrated to give the crude product. The crude product was purified by preparative TLC (dichloromethane: methanol=10:1, r _f =0.3) to give (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1- (nitrosomethyl) -2- ] as a pale yellow solid[ (3S) -2-oxo-pyrrolidin-3-yl]Ethyl group]-4- (trifluoromethyl) pyrrolidine-2-carboxamide (130 mg,247.51umol,51.93% yield, 97% purity). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

Step 5: (2S, 4R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide

At 25℃under N ₂ Downward (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1- (nitroso-methyl) -2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a mixture of 4- (trifluoromethyl) pyrrolidine-2-carboxamide (120 mg,235.54umol,1 eq.) in DCM (6 mL) was added at once the Bungeus reagent (112.26 mg,471.07umol,2 eq.). The mixture was stirred at 25℃for 4.5h. After completion, the residue was poured into water (0.5 mL) and stirred for 10min. The reaction mixture was then concentrated to give the crude product. The crude product was purified by preparative HPLC (column Phenomenex Gemini-NX 80X 40mM X3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8 min) to give (2S, 4 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide (22.56 mg,45.90umol,19.49% yield, 100% purity). MS (ESI) m/z 492.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.12-7.21(m,1H),6.84-7.10(m,2H),6.50(br s,1H),4.94-5.26(m,1H),4.75(br s,1H),4.07-4.47(m,2H),3.79-4.01(m,3H),3.45(br s,1H),2.16-2.98(m,6H),1.62-2.02(m,2H),1.39(br s,1H)。

EXAMPLE 66 Synthesis of viral protease inhibitor Compound 557

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (50)0mg,1.75mmol,1 eq.) in HCl/dioxane (4M, 8.73mL,20 eq.) and with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 20℃for 0.5h under an atmosphere. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl propionate (630 mg, crude material, HCl). MS (ESI) m/z 223.2[ M+H ]] ⁺ 。

Step 2:1- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) isoindoline-2-carboxylic acid tert-butyl ester

To 2-tert-Butoxycarbonyl isoindoline-1-carboxylic acid (436.93 mg,1.66mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a solution of methyl propionate (630 mg,1.74mmol,61.58% purity, 1.05 eq., HCl) in DCM (5 mL) and DMF (5 mL) was added T3P (1.58 g,2.49mmol,1.48mL,50% purity, 1.5 eq.) and TEA (1.01 g,9.96mmol,1.39mL,6 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (20 mL) to quench the reaction mixture and extract with ethyl acetate (10 mL x 3). The combined organic layers were washed with 15mL brine, over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives the product 1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Carbamoyl group]Isoindoline-2-carboxylic acid tert-butyl ester (720 mg, crude). MS (ESI) M/z432.2[ M+H] ⁺ 。

Step 3:

(2S) -2- (isoindoline-1-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A mixture of tert-butyl isoindoline-2-carboxylate (720 mg,1.67mmol,1 eq.) in HCl/dioxane (4M, 8.34mL,20 eq.) was degassed and purified with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 20℃for 0.5h under an atmosphere. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2- (isoindoline-1-carbonyl) as a brown oil Alkylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (770 mg, crude material, HCl). MS (ESI) m/z 332.3[ M+H ]] ⁺ 。

Step 4:

(2S) -2- (2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

4-methoxy-1H-indole-2-carboxylic acid (287.43 mg,1.50mmol,1 eq.) 2S-2- (isoindoline-1-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (770 mg,1.65mmol,79% purity, 1.1 eq., HCl), DMAP (367.34 mg,3.01mmol,2 eq.), EDCI (576.42 mg,3.01mmol,2 eq.) in DCM (8 mL) and DMF (2.7 mL) were degassed and purified with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred under an atmosphere at 20℃for 1h. After completion, by adding H ₂ O (25 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (15 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By neutral preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,10 min) to give (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl ] as a white solid ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (isomer 1:150mg,297.30umol,19.78% yield). MS (ESI) M/z505.3[ M+H ]] ⁺ The method comprises the steps of carrying out a first treatment on the surface of the And obtaining (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl ] as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (isomer 2:140mg,277.48umol,18.46% yield). MS (ESI) M/z505.3[ M+H ]] ⁺ 。

Step 5: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, 297.30. Mu. Mol,1 eq.) in MeOH/NH ₃ The solution in (7M, 849.44uL,20 eq.) was stirred at 45℃for 48h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (130 mg, crude). MS (ESI) m/z 490.3[ M+H ]] ⁺ 。

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (140 mg,277.48umol,1 eq.) in MeOH/NH ₃ The solution in (7M, 792.81uL,20 eq.) was stirred at 45℃for 24h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (110 mg, crude). MS (ESI) m/z 490.3[ M+H ]] ⁺ 。

Step 6: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (125 mg,255.35 mol,1 eq.) in DCM (8 mL) was added the berges reagent (273.84 mg,1.15mmol,4.5 eq.) and the resulting mixture was stirred at 30 ℃ for 20H. After completion, by adding H ₂ O (0.5 mL) to quench the reaction mixture, and then concentrated under reduced pressure to give a residue. By neutral prep HPLC (column: waters Xbridge BEH C18 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (31.50 mg, 66).81umol,26.16% yield, 100% purity). MS (ESI) m/z 472.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.53-11.83(m,1H)9.11-9.78(m,1H)7.31-7.78(m,5H)6.95-7.29(m,3H)6.42-6.63(m,1H)5.73(s,1H)5.27-5.41(m,1H)4.91-5.05(m,1H)3.76-3.99(m,3H)2.71-3.19(m,2H)2.00-2.30(m,3H)1.20-1.87(m,2H)。

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (105 mg,214.49umol,1 eq.) in DCM (6 mL) was added the berges reagent (204.47 mg,857.98umol,4 eq.). The mixture was stirred at 30℃for 7h. After completion, by adding H ₂ O (0.5 mL) to quench the reaction mixture, and then concentrated under reduced pressure to give a residue. By neutral preparative HPLC (column: waters Xbridge Prep OBD C18 150X 40mm X10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (34.83 mg,73.72umol,34.37% yield, 99.791% purity). MS (ESI) m/z 472.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.72(s,1H)9.19(d,J＝8.11Hz,1H)7.31-7.76(m,5H)6.92-7.29(m,3H)6.56(d,J＝7.75Hz,1H)5.74(s,1H)5.34(br d,J＝10.13Hz,1H)4.96(q,J＝8.23Hz,1H)3.86-3.89(m,1H)3.86-4.55(m,1H)3.84-4.01(m,3H)2.96-3.22(m,2H)2.25-2.41(m,1H)2.02-2.20(m,2H)1.47-1.87(m,2H)。

EXAMPLE 67 Synthesis of viral protease inhibitor Compounds 647

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (1.3 g,3.27mmol,1 eq.) in HCl/MeOH (15 mL) was stirred at 25℃for 30min. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (1.3 g, crude material) as a white solid.

Step 2: (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.68mmol,1 eq.) in DCM (6 mL) and DMF (2 mL) was added DMAP (616.30 mg,5.04mmol,3 eq.) at 25℃in one portion. To the mixture were added 4-chloro-1H-indole-2-carboxylic acid (394.69 mg,2.02mmol,1.2 eq.) and EDCI (967.04 mg,5.04mmol,3 eq.) and stirred at 25 ℃ for 2H. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By column chromatography (SiO ₂ The crude material was purified from petroleum ether/ethyl acetate=5/1 to ethyl acetate/methanol=10/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (760 mg,1.60mmol,95.16% yield). MS (ESI) M/z475.2[ M+H] ⁺ 。

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (700 mg,1.47mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 15mL,71.24 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Amino group]-1- (Ring)Propylmethyl) -2-oxo-ethyl]-4-chloro-1H-indole-2-carboxamide (660 mg,1.44mmol,97.36% yield). MS (ESI) m/z 460.2[ M+H ]] ⁺ 。

Step 4: 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-chloro-1H-indole-2-carboxamide (630 mg,1.37mmol,1 eq.) in DCM (10 mL) was added the Bungeus reagent (652.87 mg,2.74mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions, column: phenomenex Gemi ni-NX C18 75X 30mM X3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,8 min) to give 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (110 mg,248.92umol,18.17% yield). MS (ESI) m/z 442.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.96(br s,1H),8.93(d,J＝8.2Hz,1H),8.76(d,J＝7.7Hz,1H),7.78-7.67(m,1H),7.46-7.36(m,2H),7.21-7.09(m,2H),5.04-4.89(m,1H),4.55-4.43(m,1H),3.12(quin,J＝9.3Hz,2H),2.43-2.29(m,1H),2.19-2.07(m,2H),1.91-1.63(m,3H),1.54-1.41(m,1H),0.82(br dd,J＝5.6,7.4Hz,1H),0.50-0.34(m,2H),0.26-0.04(m,2H)。

EXAMPLE 68 Synthesis of viral protease inhibitor Compound 649

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (500 mg,968.64umol,77% purity, 1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 30min. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg, crude material) as a white solid.

Step 2: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propanoylamino ] propanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (288 mg,968.56umol,1 eq.) DMAP (354.98 mg,2.91mmol,3 eq.) was added to a mixture of DCM (5 mL) and DMF (2.5 mL) and 3- (3, 5-difluorophenyl) propionic acid (180.30 mg,968.56umol,1 eq.) and EDCI (928.37 mg,4.84mmol,5 eq.) were added to the mixture in one portion at 25 ℃. The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was taken up with H ₂ O (5 mL) was diluted and extracted with ethyl acetate (8 mL x 3). The combined organic layers were washed with brine (15 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiO ₂ The crude material was purified from petroleum ether/ethyl acetate=5/1 to ethyl acetate/methanol=5:1 to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (300 mg,547.81umol,56.56% yield, 85% purity). MS (ESI) m/z 466.2[ M+H ] ] ⁺ 。

Step 3: (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ] propionamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (300 mg,644.48umol,1 eq.) in NH ₃ In methanol (7M, 5.45mL,59.24 eq)The mixture was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]-3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ]]Propionamide (260 mg,577.16umol,89.55% yield). MS (ESI) m/z 451.2[ M+H ]] ⁺ 。

Step 4: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ] propanamide

To (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl at 25 ℃C]Methyl group]Ethyl group]-3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ]]POCl was added in one portion to a mixture of propionamide (70 mg,155.39umol,1 eq.) in ACN (1 mL) ₃ (47.65 mg,310.78umol,28.88uL,2 eq.). The mixture was stirred at 80℃for 0.5h. After completion, by addition of NaHCO at 25℃ ₃ (1 mL) the reaction mixture was quenched and then extracted with ethyl acetate (1 mL x 3). The combined organic layers were concentrated under reduced pressure to give the crude product. By preparative HPLC (neutral conditions, column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [3- (3, 5-difluorophenyl) propionylamino ]]Propionamide (7 mg,16.19umol,10.42% yield). MS (ESI) m/z 433.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.09-8.81(m,1H),8.15(br d,J＝7.5Hz,1H),7.83-7.70(m,1H),7.10-6.89(m,3H),4.99-4.83(m,1H),4.33-4.19(m,1H),3.19-3.04(m,2H),2.89-2.78(m,2H),2.46(br s,2H),2.39-2.03(m,3H),1.84-1.46(m,3H),1.40-1.19(m,1H),0.59(br s,1H),0.34(br s,2H),0.14 -0.05(m,2H)。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝6.84(br t,J＝5.7Hz,2H),6.74(tt,J＝2.2,9.3Hz,1H),5.06-4.92(m,1H),4.37-4.22(m,1H),3.38-3.32(m,2H),2.97-2.88(m,2H),2.71-2.57(m,2H),2.54-2.10(m,3H),2.01-1.77(m,2H),1.76-1.58(m,1H),1.55-1.36(m,1H),0.72-0.59(m,1H),0.53-0.36(m,2H),0.18-0.02(m,2H)。

EXAMPLE 69 Synthesis of viral protease inhibitor Compounds 653

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of (4-methoxy-1H-indole-2-carboxamide (1 g,1.81mmol,80% purity, 1 eq.) in EtOH (20 mL) was added 2-glycine (271.74 mg,3.62mmol,20.52uL,2 eq.), znCl ₂ (1M, 18.10uL,0.01 eq.). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (359.14 mg,3.62mmol,452.89ul,2 eq.) was added and the resulting mixture was stirred at 40 ℃ for 6h. After the reaction was completed, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex luna C, 18, 80X 40mm X3 um; mobile phase: [ water (0.04% HCl) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,7 min) to yield 400mg of the mixture product. The mixture was passed through SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,10 min) to give the compound 2- [ [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Amino group]Acetic acid (125 mg,235.87umol,13.03% yield, 99.363% purity) and 2- [ [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Amino group]Acetic acid (205 mg,373.82umol,20.65% yield, 96.023% purity). MS (ESI) m/z 527.3[ M+H ]] ⁺ 。

Isomer 1: ¹ H NMR(400MHz,DMSO-d6)δ＝11.56(d,J＝2.0Hz,1H),8.52-8.21(m,2H),7.58(s,1H),7.35(d,J＝1.7Hz,1H),7.14-7.05(m,1H),7.03-6.97(m,1H),6.50(d,J＝7.7Hz,1H),4.57-4.41(m,1H),4.14(tdd,J＝4.2,8.2,12.2Hz,1H),3.97-3.82(m,4H),3.52-3.36(m,2H),3.18-2.98(m,2H),2.41-2.27(m,1H),2.12-2.04(m,2H),1.82-1.36(m,5H),0.91(dd,J＝6.4,15.8Hz,6H)

isomer 2: ¹ H NMR(400MHz,DMSO-d6)δ＝11.57(d,J＝2.0Hz,1H),8.39(d,J＝7.8Hz,1H),8.20(d,J＝9.5Hz,1H),7.54(s,1H),7.37(d,J＝1.6Hz,1H),7.16-6.94(m,2H),6.50(d,J＝7.6Hz,1H),4.53-4.36(m,1H),4.18-4.01(m,1H),3.88(s,3H),3.77(d,J＝8.8Hz,1H),3.43-3.33(m,2H),3.15-2.96(m,2H),2.38-2.25(m,1H),2.08-2.01(m,1H),1.91-1.47(m,6H),0.91(dd,J＝6.4,14.8Hz,6H)

to N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (700 mg,1.27mmol,80% purity, 1 eq.) in EtOH (10 mL) was added pyrrolidine (180.01 mg,2.53mmol,211.28uL,2 eq.), znCl2 (1M, 12.66uL,0.01 eq.) and the resulting mixture was stirred at 25℃for 30min. After addition of TMSCN (251.10 mg,2.53mmol,316.65uL,2 eq.) the mixture was stirred at 25℃for 2h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (110 mg,199.95umol,15.80% yield, 95% purity) and N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl)]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (110 mg,199.95umol,15.80% yield, 95% purity). MS (ESI) m/z 523.4[ M+H ]] ⁺

example 70 Synthesis of viral protease inhibitor Compounds 655

Step 1: (2S, 4R) -4-hydroxypyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of (2 s,4 r) -1-tert-butoxycarbonyl-4-hydroxy-pyrrolidine-2-carboxylic acid (5 g,21.62mmol,1 eq.) in THF (75 mL) was added 2-tert-butyl-1, 3-diisopropyl-isourea (6.50 g,32.43mmol,1.5 eq.) at 25 ℃ and the resulting solution was then stirred at 60 ℃ for 2.5h. To the mixture was added 2-tert-butyl-1, 3-diisopropyl-isourea (6.50 g,32.43mmol,1.5 eq.) and then stirred at 60 ℃ for 14h. After completion, the reaction mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give di-tert-butyl (2 s,4 r) -4-hydroxypyrrolidine-1, 2-dicarboxylate (4.3 g,14.22mmol,65.75% yield, 95% purity) as a colorless oil. MS (ESI) m/z 288.2[ M+H ] ] ⁺

Step 2: (2S, 4S) -4-bromopyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

CBr is added to a solution of di-tert-butyl (2S, 4R) -4-hydroxypyrrolidine-1, 2-dicarboxylate (4 g,13.92mmol,1 eq.) in DCM (40 mL) at 25 ℃ ₄ (14.08 g,42.46mmol,3.05 eq.). The mixture was cooled to 0 ℃ and PPh was carefully added ₃ (11.32 g,43.15mmol,3.1 eq.). The reaction was stirred at 25℃for 15h. After completion, ethanol (4 mL) was added and the solution was stirred for 2h. MTBE (40 mL) was added dropwise to precipitate the phosphine oxide, which was filtered off, the filter cake was washed with DCM (30 mL x 2) and the filtrate was concentrated under reduced pressure to give a brown oil. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=100:0 to 10:1) to give (2 s,4 s) -4-bromopyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester (1.5 g,4.07mmol,29.23% yield, 95% purity) as a pale yellow oil.

Step 3: (2S, 4S) -4- (tert-butyl) pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

A mixture of phenylsulfanyl copper (1.58 g,9.14mmol,6.4 eq.) in anhydrous THF (30 mL) was cooled to-70℃and then treated by careful addition of t-BuLi (1.3M, 7.03mL,6.4 eq.). The resulting mixture was stirred for 30min, and a solution of pre-cooled (-20 ℃) di-tert-butyl (2S, 4S) -4-bromopyrrolidine-1, 2-carboxylate (500 mg,1.43mmol,1 eq.) in anhydrous THF (5 mL) was added. The solution was stirred at-70 ℃ for 5h and then at N ₂ The temperature is raised to 25 ℃ and maintained for 15 hours. After completion, by pouring saturated NH ₄ The reaction was quenched in aqueous Cl (30 mL). The aqueous mixture was vigorously stirred for 30min. The solid was filtered off and the phases were separated. The aqueous phase was extracted with MTBE (10 ml x 3) and the combined organic phases were extracted with saturated NaHCO ₃ Aqueous (10 mL) and brine (10 mL) washed with Na ₂ SO ₄ Drying and concentrating under reduced pressure to obtain crude material. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=100:0 to 10:1) to give di-tert-butyl (2 s,4 s) -4- (tert-butyl) pyrrolidine-1, 2-dicarboxylate (290 mg,797.05umol,55.83% yield, 90% purity) as an off-white solid.

Step 4: (2S, 4S) -4- (tert-butyl) pyrrolidine-2-carboxylic acid

A mixture of di-tert-butyl (2S, 4S) -4- (tert-butyl) pyrrolidine-1, 2-dicarboxylic acid (250 mg,763.46 mol,1 eq.) in HCl (6M, 2.5mL,19.65 eq.) was stirred at 100deg.C for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give (2 s,4 s) -4-tert-butylpyrrolidine-2-carboxylic acid (158 mg, crude, HCl) as a yellow solid.

Step 5: (2S, 4S) -1- (tert-Butoxycarbonyl) -4- (tert-butyl) pyrrolidine-2-carboxylic acid

To (2S, 4S) -4-tert-butylpyrrolidine-2-carboxylic acid (158 mg, 760.72. Mu. Mol,1 eq. HCl) in THF (1 mL) and H ₂ Mixing in O (1 mL)Adding K into the mixture ₂ CO ₃ (315.41 mg,2.28mmol,3 eq.) and Boc ₂ O (199.23 mg, 912.87. Mu.L, 209.72. Mu.L, 1.2 eq.). At N ₂ The reaction was stirred at 25℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give (2 s,4 s) -1- (tert-butoxycarbonyl) -4- (tert-butyl) pyrrolidine-2-carboxylic acid (650 mg, crude substance) as a yellow solid.

Step 6: (2S, 4S) -4- (tert-butyl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester

To a solution of (2S, 4S) -1- (tert-butoxycarbonyl) -4- (tert-butyl) pyrrolidine-2-carboxylic acid (630 mg,696.51umol,30% purity, 1 eq) in DCM (6 mL) and DMF (3 mL) was added TEA (422.88 mg,4.18mmol,581.68uL,6 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl)]Methyl propionate (186.11 mg,835.82umol,1.2 eq., HCl). After adding T3P (1.33 g,2.09mmol,1.24mL,50% purity, 3 eq.) at 0deg.C, the mixture was stirred at 25deg.C for 1h. After completion, the mixture was quenched with water (10.0 mL) and extracted with DCM (10.0 mL x 3). The organic layer was washed with brine (10.0 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) to give 4- (tert-butyl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl (2S, 4S) -tert-butyl ester (240 mg,546.02umol,78.39% yield) as a yellow solid. MS (ESI) m/z 440.3[ M+H ]] ⁺ 。

Step 7: (S) -2- ((2S, 4S) -4- (tert-butyl) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of tert-butyl (2S, 4S) -tert-butyl 4- (tert-butyl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylate (230 mg,523.27umol,1 eq) in HCl/MeOH (4M, 2.3mL,17.58 eq) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (S) as a pale yellow solid-methyl 2- ((2S, 4S) -4- (tert-butyl) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (196 mg, crude, HCl). MS (ESI) m/z 340.2[ M+H ]] ⁺ 。

Step 8: (S) -2- ((2S, 4S) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((2S, 4S) -4- (tert-butyl) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (196 mg,521.43umol,1 eq, HCl) in DCM (2 mL) and DMF (1 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (99.69 mg,521.43umol,1 eq), DMAP (127.41 mg,1.04mmol,2 eq) and then EDCI (199.92 mg,1.04mmol,2 eq) at 0 ℃. The mixture was then stirred at 25℃for 1h. After completion, the mixture was quenched with water (10.0 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Dichloromethane: methanol = 10:1 to 4:1) the residue was purified to give methyl (S) -2- ((2S, 4S) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (250 mg,414.56umol,79.50% yield, 85% purity) as a yellow solid. MS (ESI) m/z 513.3[ M+H ]] ⁺ 。

Step 9: (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

Methyl (S) -2- ((2S, 4S) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (235 mg,389.68umol,85% purity, 1 eq.) in NH ₃ A solution in methanol (7M, 5 mL) was stirred at 40℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (193 mg, crude) as a yellow solid. MS (ESI) m-z 498.3[M+H] ⁺ 。

Step 10: (2S, 4S) -4- (tert-butyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

To a solution of (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (193 mg,329.69umol,85% purity, 1 eq) in DCM (3 mL) was added the bergs reagent (235.71 mg,989.08umol,3 eq) and then stirred for 4H at 25 ℃. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified 30% -55%,10 min) to give (2S, 4S) -4- (tert-butyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (59.58 mg,124.24umol,37.68% yield, 100% purity) as a white solid. MS (ESI) m/z 480.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69-11.55(m,1H),9.17-8.75(m,1H),7.81-7.44(m,1H),7.16-7.07(m,1H),7.06-6.98(m,2H),6.55-6.46(m,1H),5.03-4.53(m,2H),4.04-3.74(m,4H),3.69-3.36(m,1H),3.22-2.55(m,2H),2.35-1.95(m,5H),1.83-1.51(m,3H),1.00-0.82(m,9H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.31(s,1H),8.68(s,1H),7.38(s,1H),7.18-7.02(m,2H),6.90(s,1H),6.60-6.47(m,1H),4.96(q,J＝7.6Hz,1H),4.72(s,1H),4.07-3.80(m,4H),3.66-3.50(m,1H),3.28-3.05(m,2H),2.32-1.97(m,5H),1.95-1.64(m,3H),0.95(s,9H)。

EXAMPLE 71 Synthesis of viral protease inhibitor Compounds 659

Step 1: (S) - (1-hydroxy-4, 4-dimethylpentan-2-yl) carbamic acid tert-butyl ester

To a solution of (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (5 g,20.38mmol,1 eq.) in THF (100 mL) was slowly added drop wise BH at 0deg.C ₃ -Me ₂ S (10M, 4.08mL,2.0 eq.) followed by stirring the mixture at 20deg.C for 15h. The reaction mixture was added to MeOH (40 mL) and stirred for 20min, then the mixture was concentrated. NaHCO for residue ₃ Aqueous solution (150 mL) was diluted and extracted with DCM (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 1:1) the residue was purified to give N- [ (1S) -1- (hydroxymethyl) -3, 3-dimethyl-butyl as a colorless oil ]Tert-butyl carbamate (2.5 g,10.81mmol,53.02% yield).

Step 2: (S) - (4, 4-dimethyl-1-oxopent-2-yl) carbamic acid tert-butyl ester

N- [ (1S) -1- (hydroxymethyl) -3, 3-dimethyl-butyl was prepared by the Dess-martin reaction at 0deg.C]To a solution of tert-butyl carbamate (2.4 g,10.37mmol,1 eq.) in DCM (40 mL) was added periodate (5.72 g,13.49mmol,4.18mL,1.3 eq.) and the reaction stirred for 1h. The mixture was warmed to 20 ℃ and stirred for 1h. By adding H at 0 ℃ ₂ O (60 mL) to quench the reaction mixture, and then NaHCO was added dropwise at 0deg.C ₃ The aqueous solution was brought to ph=8 and extracted with EtOAc (40 ml x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1 to 1:1) the residue was purified to give N- [ (1S) -1-formyl-3, 3-dimethyl-butyl as a colorless oil]Tert-butyl carbamate (1.6 g,6.98mmol,67.25% yield).

¹ H NMR(400MHz,DMSO-d ₆ )δppm 9.40(s,1H)7.30(br d,J＝8.00Hz,1H)3.91-3.82(m,1H)1.66(dd,J＝14.38,2.75Hz,1H)1.39(s,9H)1.32(br d,J＝9.26Hz,1H)0.90(s,9H)。

Step 3: (S) -2- (((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To N- [ (1S) -1-formyl-3, 3-dimethyl-butyl ]Carbamic acid tert-butyl ester (0.8 g,3.49mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (1.17 g,5.23mmol,1.5 eq. HCl) in DCE (20 mL) was added Et ₃ N (529.52 mg,5.23mmol,728.36uL,1.5 eq.) and NaBH (OAc) ₃ (2.22 g,10.47mmol,3 eq.) and the reaction stirred at 20℃for 2h.

By addition of NaHCO at 0deg.C ₃ The reaction mixture was quenched with aqueous solution (100 mL) and stirred for 0.5h, followed by extraction with DCM (60 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1 to 1:3) to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (450 mg,1.13mmol,32.29% yield). MS (ESI) m/z 400.3[ M+H ]] ⁺ 。

Step 4: (S) -2- (((S) -2-amino-4, 4-dimethylpentyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (200 mg,500.60 mol,1 eq.) in HCl/MeOH (4M, 4.00mL,31.96 eq.) was stirred at 20℃for 1h. The reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (168 mg, crude material, HCl) as a white solid.

Step 5: (S) -2- (((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (168 mg,500.20umol,1 eq, HCl) and 4-methoxy-1H-indole-2-carbonitrileTo a solution of acid (95.63 mg,500.20umol,1 eq.) in DMF (1 mL) were added DMAP (183.32 mg,1.50mmol,3.0 eq.) and EDCI (191.78 mg,1.00mmol,2 eq.) and DCM (3 mL) and the mixture stirred at 20deg.C for 2h. By adding 40mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (20 ml x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) to give (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow oil]-4, 4-dimethyl-pentyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg,301.54 mol,60.28% yield, 95% purity). MS (ESI) m/z 473.2[ M+H ]] ⁺ 。

Step 6: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4, 4-dimethylpentan-2-yl) -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (130 mg,275.09umol,1 eq.) in NH ₃ A solution in MeOH (7M, 15mL,381.70 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with ethyl acetate methanol=50:3 to give the product N- [ (1S) -1- [ [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid]Methyl group]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (60 mg,131.13umol,47.67% yield). MS (ESI) M/z458.3[ M+H] ⁺ 。

Step 7: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethylpentan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl [ (2-amino-3-carbonyl)]Methyl group]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,109.27 umol)1 equivalent) to a solution of T in EtOAc (2 mL) was added dropwise ₃ P (2.14 g,3.36mmol,2mL,50% purity, 30.77 eq.) and then stirring the mixture at 65℃for 12h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,8 min) and purified the residue by SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -25%,20 min) to give N- [ (1S) -1- [ [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (4.4 mg,9.92umol,29.07% yield, 99.1% purity). MS (ESI) m/z 440.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.22-6.99(m,3H)6.52(br d,J＝7.72Hz,1H)4.74-4.65(m,1H)4.61-4.48(m,1H)4.03-3.91(m,4H)3.62-3.51(m,1H)3.47-3.36(m,1H)3.27-3.19(m,1H)2.50-2.41(m,1H)2.29-2.18(m,1H)1.81(br s,1H)1.74-1.64(m,2H)1.60(br d,J＝10.14Hz,1H)1.34-1.28(m,1H)0.98(s,9H)。

Example 72 Synthesis of viral protease inhibitor Compounds 671

Step 1: chloroformic acid 2- (2-methoxyethoxy) ethyl ester

Triphosgene (4.93 g,16.61mmol,4.99e-1 eq.) Na ₂ CO ₃ A mixture of (3.53 g,33.29mmol,1 eq.) and DMF (95.00 mg,1.30mmol,0.1mL,3.90e-2 eq.) in toluene (50 mL) was cooled to 0deg.C and under N ₂ Stirring for 0.5h under an atmosphere. Next, a solution of 2- (2-methoxyethoxy) ethanol (4 g,33.29mmol,3.92mL,1 eq.) was added dropwise. The mixture was stirred at 0℃for 4h. After completion, the mixture was filtered and the filtrate was concentrated under reduced pressure to give 2- (2-methoxyethoxy) ethyl chloroformate (6 g) as a yellow oil.

Step 2: (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (600 mg,1.51mmol,1 eq.) in HCl/MeOH (4M, 12.00mL,31.80 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (450 mg) as a white solid.

Step 3: (11S, 14S) -11- (cyclopropylmethyl) -9, 12-dioxo-14- (((S) -2-oxopyrrolidin-3-yl) methyl) -2,5, 8-trioxa-10, 13-diazapenta-15-alkanoic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (450 mg,1.51mmol,1 eq.) in THF (10 mL) and H ₂ To a solution in O (1 mL) was added DIEA (391.19 mg,3.03mmol,527.20uL,2 eq.) and then 2- (2-methoxyethoxy) ethyl chloroformate (414.52 mg,2.27mmol,1.5 eq.) at 0deg.C. The mixture was stirred at 30℃for 2h. After completion, by adding H ₂ O (100 mL) to quench the reaction mixture and then extract with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -25%,10 min) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [2- (2-methoxyethoxy) ethoxycarbonylamino as a yellow oil]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg,901.94umol,59.60% yield). MS (ESI) m/z 444.2[ M+H ]] ⁺ 。

Step 4: ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid 2- (2-methoxyethoxy) ethyl ester

(2S) -2- [ [ (2S) -3 ]Cyclopropyl-2- [2- (2-methoxyethoxy) ethoxycarbonylamino]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg,901.94umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,77.61 eq.) was stirred at 70℃for 12h. After completion, the mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]2- (2-methoxyethoxy) ethyl carbamate (400 mg, crude). MS (ESI) m/z 429.2[ M+H ]] ⁺

Step 5: ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid 2- (2-methoxyethoxy) ethyl ester

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 2- (2-methoxyethoxy) ethyl carbamate (380 mg,886.86umol,1 eq.) in DCM (5 mL) was added the Buerger's reagent (422.69 mg,1.77mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, by adding H ₂ O (50 mL) to quench the reaction mixture and extract with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:10% -40%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]2- (2-methoxyethoxy) ethyl carbamate (150 mg, crude). MS (ESI) m/z 411.2[ M+H ]] ⁺

Step 6: ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid 2- (2-methoxyethoxy) ethyl ester

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase:[Neu-EtOH]the method comprises the steps of carrying out a first treatment on the surface of the B%:44% -44%,8 min) isolation of N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]2- (2-methoxyethoxy) ethyl carbamate (150 mg, crude material) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a colourless gum]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]2- (2-methoxyethoxy) ethyl carbamate (110 mg,262.36umol,71.79% yield, 97.9% purity). MS (ESI) m/z 411.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.81(br d,J＝7.8Hz,1H),7.72(s,1H),7.54(br d,J＝7.4Hz,1H),4.95(q,J＝8.2Hz,1H),4.08-3.86(m,3H),3.53(td,J＝4.6,15.2Hz,4H),3.47-3.39(m,2H),3.33(s,3H),3.19-3.05(m,2H),2.41-2.28(m,1H),2.19-2.03(m,2H),1.81-1.59(m,3H),1.28(td,J＝6.8,13.6Hz,1H),0.74(br d,J＝5.6Hz,1H),0.46-0.33(m,2H),0.18-0.01(m,2H)。

EXAMPLE 73 Synthesis of viral protease inhibitor Compound 691

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (13.00 g,45.40mmol,1 eq.) and HCl/MeOH (4M, 35mL,3.08 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl propionate (10 g, crude material, HCl). MS (ESI) m/z 223.1[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (9.71 g,43.62mmol,1 eq. HCl), (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (10 g,43.62 m)A solution of mol,1 eq.) and TEA (22.07 g,218.08mmol,30.35mL,5 eq.) in DCM (100 mL) was cooled to 0deg.C and then T3P (83.27 g,130.85mmol,77.82mL,50% purity, 3 eq.) was added to the solution. The mixture was stirred for 1h and gradually warmed to 20 ℃. After completion, H is added to the mixture ₂ O (100 mL) and then extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Then, the mixture was purified by column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1) the residue was purified to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (12 g,23.41mmol,53.67% yield, 77.53% purity). MS (ESI) m/z 398.2[ M+H ] ] ⁺ 。

Step 3: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (1.5 g,3.77mmol,1 eq.) in HCl/methanol (4M, 100mL,105.99 eq.) was stirred at 20deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.1 g, crude material, HCl). MS (ESI) m/z 298.2[ M+H ]] ⁺ 。

Step 4: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

4, 7-dichloro-1H-indole-2-carboxylic acid (650 mg,2.83mmol,1 eq.) was reacted with (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (943.18 mg,2.83mmol,1 eq., HCl), EDCI (1.08 g,5.65mmol,2 eq.) and DMAP (1.04 g,8.48mmol,3 eq.) in DCM (10 mL) was stirred at 20deg.C for 1h. After completion, H is added to the mixture ₂ O (50 mL) and connectExtraction was followed with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1) the residue was purified to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (550 mg,1.01mmol,35.92% yield, 93.99% purity). MS (ESI) m/z 509.1[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 7-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (550 mg,1.08mmol,1 eq.) in NH ₃ The mixture in methanol (7M, 154.25uL,1 eq.) was stirred at 60℃for 12h. After completion, the mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4, 7-dichloro-1H-indole-2-carboxamide (500 mg, crude). MS (ESI) m/z 494.1[ M+H ]] ⁺ 。

Step 6:4, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A mixture of 4, 7-dichloro-1H-indole-2-carboxamide (450 mg, 910.25. Mu. Mol,1 eq.) and Prague reagent (1.30 g,5.46mmol,6 eq.) in DCM (10 mL) was stirred at 20deg.C for 9H. After completion, the mixture was concentrated under reduced pressure to give a residue. The residue was then purified by preparative HPLC (column Phenomenex Gemini-NX 80X 40mM X3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-55％8 min) purification to give the product 4, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (300 mg,629.78 mol,69.19% yield, 100% purity). MS (ESI) M/z476.1[ M+H ] ] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.66-7.56(m,1H),7.52-7.45(m,1H),7.22-7.14(m,1H),5.16-5.05(m,1H),4.68-4.61(m,1H),3.36-3.32(m,2H),2.70-2.57(m,1H),2.40-2.27(m,2H),1.99-1.69(m,4H),0.91-0.79(m,1H),0.62-0.52(m,2H),0.27-0.15(m,2H)。

Example 74 Synthesis of viral protease inhibitor Compound 695

Step 1: methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate hydrochloride methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (2 g,6.99mmol,1 eq.) in HCl/EtOAc (4M, 40.00mL,22.91 eq.) the mixture was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (1.5 g, crude material, HCl) as a white solid.

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl at 0 ℃C]To a solution of methyl propionate (1.4 g,6.29mmol,1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (1.44 g,6.29mmol,1.00 eq) in DCM (30 mL) was added DIEA (3.25 g,25.15mmol,4.38mL,4 eq) and T was added dropwise ₃ P (12.00 g,18.86mmol,11.22mL,50% purity, 3 eq.) and then stirring the mixture at 20℃for 1h. After completion, by adding H at 0deg.C ₂ O (60 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). Combined withThe organic layer was washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.9 g,4.73mmol,75.27% yield, 99% purity). MS (ESI) m/z 398.4[ M+H ]] ⁺ 。

Step 3: (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester hydrochloride

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (0.8 g,2.01mmol,1 eq.) in HCl/MeOH (4M, 15mL,29.81 eq.) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (670 mg, crude material, HCl) as a white solid.

Step 4: (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (640 mg,2.01mmol,1.51 eq, HCl) and 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (300 mg,1.33mmol,1 eq) in DMF (5 mL) was added DMAP (487.32 mg,3.99mmol,3 eq), EDCI (509.78 mg,2.66mmol,2 eq) and DCM (15 mL), and the mixture was stirred at 20 ℃ for 2H. After completion, by adding H at 0deg.C ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give (2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole) as a yellow solid-2-carbonyl) amino]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (350 mg,658.47 mol,49.52% yield, 95% purity). MS (ESI) m/z 505.2[ M+H ]] ⁺ 。

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (320 mg,633.71umol,1 eq.) in NH ₃ A solution in MeOH (7M, 40mL,441.84 eq.) was stirred at 50deg.C for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-5-methoxy-1H-indole-2-carboxamide (290 mg, crude material). MS (ESI) m/z 490.2[ M+H ]] ⁺ 。

Step 6: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-5-methoxy-1H-indole-2-carboxamide (270 mg,551.08umol,1 eq.) in DCM (10 mL) was added the Bungeus reagent (393.97 mg,1.65mmol,3 eq.). After stirring at 20 ℃ for 7h, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the product 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-methoxy-1H-indole-2-carboxamide (139.27 mg,295.10umol,53.55% yield, 100% purity). MS (ESI) m/z 472.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.17(s,1H),7.07(d,J＝2.0Hz,1H),6.96(d,J＝2.1Hz,1H),5.08(dd,J＝6.0,10.3Hz,1H),4.55(t,J＝7.4Hz,1H),3.82(s,3H),3.30-3.27(m,2H),2.70-2.60(m,1H),2.40-2.28(m,2H),1.97-1.77(m,3H),1.72-1.60(m,1H),0.86(br s,1H),0.55(d,J＝8.0Hz,2H),0.20(dd,J＝4.8,9.4Hz,2H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.59(br s,1H),9.00(d,J＝7.9Hz,1H),8.66(d,J＝7.6Hz,1H),7.72(s,1H),7.17(s,1H),7.13(d,J＝2.2Hz,1H),7.00(d,J＝2.2Hz,1H),5.00(q,J＝7.9Hz,1H),4.60-4.45(m,1H),3.78(s,3H),3.18-3.05(m,2H),2.40-2.34(m,1H),2.21-2.06(m,2H),1.86-1.64(m,3H),1.50(ddd,J＝6.1,7.6,13.9Hz,1H),0.90-0.75(m,1H),0.50-0.37(m,2H),0.25-0.15(m,1H),0.13-0.04(m,1H)

EXAMPLE 75 Synthesis of viral protease inhibitor Compounds 711

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (900 mg,1.81mmol,80% purity, 1 eq.) in HCl/MeOH (4M, 12.00mL,26.50 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give (2S) -2-amino-3-cyclopropyl-propionyl as a white oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (600 mg, crude material, HCl). MS (ESI) m/z 298.1[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -3-cyclopropyl-2- (4, 5,6, 7-tetrahydro-1H-indole-2-carbonylamino) propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (600 mg,1.80mmol,1 eq, HCl) in DCM (7 mL) and DMF (0.5 mL) was added 4,5,6, 7-tetrahydro-1H-indole-2-carboxylic acid (415.68 mg,2.52mmol,1.4 eq), TEA (1.09 g,10.78mmol,1.50mL,6 eq) and T3P (1.72 g,2.70mmol,1.60mL,50% purity, 1.5 eq). After stirring for 3h at 25 ℃, the reaction mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1) to give the product (2S) -2- [ [ (2S) -3-cyclopropyl-2- (4, 5,6, 7-tetrahydro-1H-indole-2-carbonylamino) propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (350 mg,787.36umol,43.80% yield). MS (ESI) m/z 445.3[ M+H ]] ⁺ 。

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4,5,6, 7-tetrahydro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- (4, 5,6, 7-tetrahydro-1H-indole-2-carbonylamino) propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (350 mg,787.36umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,88.90 eq.) was stirred at 50deg.C for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4,5,6, 7-tetrahydro-1H-indole-2-carboxamide (300 mg, crude). MS (ESI) m/z 430.2[ M+H ]] ⁺ 。

Step 4: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4,5,6, 7-tetrahydro-1H-indole-2-carboxamide

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4,5,6,A mixture of 7-tetrahydro-1H-indole-2-carboxamide (290 mg,675.19umol,1 eq.) in T3P (3 mL,50% purity) and ethyl acetate (3 mL) was stirred at 40℃for 16H. After completion, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters X bridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4,5,6, 7-tetrahydro-1H-indole-2-carboxamide (61.92 mg,150.48umol,22.29% yield, 100% purity). MS (ESI) m/z 412.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝10.96(br s,1H),9.00-8.77(m,1H),7.89-7.66(m,2H),6.60(br s,1H),5.04-4.81(m,1H),4.48-4.28(m,1H),3.24-3.04(m,2H),2.47-1.96(m,7H),1.81-1.61(m,7H),1.40(br dd,J＝6.6,13.1Hz,1H),0.74(br s,1H),0.38(br s,2H),0.22-0.03(m,2H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝10.67(br s,1H),8.74-8.49(m,1H),7.53-7.28(m,2H),6.54(d,J＝2.2Hz,1H),5.05-4.84(m,1H),4.54-4.38(m,1H),3.17(br d,J＝7.2Hz,2H),2.54(br t,J＝6.1Hz,2H),2.43(br t,J＝5.6Hz,3H),2.28-2.08(m,2H),1.90-1.79(m,1H),1.77-1.65(m,6H),1.56(qd,J＝6.7,13.7Hz,1H),0.83-0.70(m,1H),0.42(br d,J＝7.8Hz,2H),0.20-0.04(m,2H)。

Example 76 Synthesis of viral protease inhibitor Compounds 719

Step 1:7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.08 g,4.57mmol,1 eq, HCl) and 6-tert-butoxycarbonyl-6-azaspiro [3.4]]To a solution of octane-7-carboxylic acid (1.4 g,5.48mmol,1.2 eq.) in DCM (15 mL) and DMF (1 mL) was added EDCI (1.75 g,9.14mmol,2 eq.) and DMAP (1.67 g,13.71mmol,3 eq.) and the mixture was stirred at 25℃for 1h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=2/1 to 0:1 to give 7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Tert-butyl octane-6-carboxylate (1.4 g,2.56mmol,56.02% yield, 80% purity). MS (ESI) m/z 438.3[ M+H ]] ⁺ 。

Step 2:7- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]A mixture of tert-butyl octane-6-carboxylate (0.7 g,1.60mmol,1 eq.) in HCl/MeOH (4M, 20mL,50.00 eq.) was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -2- (6-azaspiro [3.4] as a white solid]Octane-7-carbonylamino) -3- [ (3S) -2-oxo-3-piperidyl]Methyl propionate (0.6 g, crude material, HCl). MS (ESI) m/z 338.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- (6-azaspiro [3.4]]Octane-7-carbonylamino) -3- [ (3S) -2-oxo-3-piperidyl]To a mixture of methyl propionate (0.6 g,1.60mmol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (368.18 mg,1.93mmol,1.2 eq) in DCM (10 mL) and DMF (2 mL) was added EDCI (461.47 mg,2.41mmol,1.5 eq) and DMAP (588.18 mg,4.81mmol,3 eq). After stirring for 1h at 25℃the reaction mixture was diluted with water (50 mL) and treated with DCM20ml x 2) extraction. The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=2:1 to 0/1) to give (2S) -2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a yellow solid]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.65 g,1.15mmol,71.39% yield, 90% purity). MS (ESI) m/z 511.3[ M+H ]] ⁺ 。

Step 4: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.65 g,1.15mmol,90% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,61.10 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carboxamide (0.6 g, crude). MS (ESI) m/z 496.3[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]To a solution of octane-7-carboxamide (0.58 g,1.17mmol,1 eq.) in DCM (7 mL) was added the Buerger's reagent (1.39 g,5.85mmol,5 eq.) and the solution stirred at 25℃for 2h. After completion, the reaction mixture was diluted with water (30 mL) and extracted with DCM (20 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ Ethyl acetate meoh=20:1) the residue was separated to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxo1-3-piperidinyl]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 1 and N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 2.

By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) purification of N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl)]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 1, N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl, as a white solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 1 (92.10 mg,192.86umol,16.48% yield, 100% purity). MS (ESI) m/z 478.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.17-7.07(m,1H),7.03(d,J＝8.2Hz,1H),7.01-6.96(m,1H),6.55-6.44(m,1H),5.05-4.89(m,1H),4.43(t,J＝7.2Hz,1H),4.01-3.79(m,5H),3.13-2.76(m,2H),2.31-2.05(m,4H),2.03-1.73(m,8H),1.60-0.97(m,3H)；

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.49-11.19(m,1H),8.81-8.41(m,1H),7.31-7.20(m,1H),7.11(br d,J＝7.7Hz,1H),7.09-7.02(m,1H),7.02-6.81(m,1H),6.53(br d,J＝7.7Hz,1H),5.06-4.89(m,1H),4.53(br s,1H),4.07-3.79(m,5H),3.10-3.02(m,2H),2.19(br s,4H),2.06-1.31(m,11H)。

By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) purification of N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl)]Ethyl group ]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 2 gives N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 2 (30.29 mg,63.43umol,5.42% yield, 100% purity). MS (ESI) m/z 478.3[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, chloroform-d) δ=10.26-9.64 (m, 1H), 8.99-8.34 (m, 1H), 7.26-7.16 (m, 1H), 7.15-6.74 (m, 2H), 6.62-6.32 (m, 1H), 6.27-5.80 (m, 1H), 5.06-4.83 (m, 1H), 4.81-4.54 (m, 1H), 4.14-3.82 (m, 5H), 3.31-3.03 (m, 2H), 2.56-2.35 (m, 2H), 2.35-2.16 (m, 2H), 2.11-1.73 (m, 9H), 1.52-1.23 (m, 2H).

EXAMPLE 77 Synthesis of viral protease inhibitor Compounds 721

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (2.49 g,10.14mmol,1.2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (2 g,8.45mmol,1 eq., HCl) in DCM (60 mL) was added DMAP (3.10 g,25.35mmol,3 eq.). After EDCI (3.24 g,16.90mmol,2 eq.) was added, the mixture was stirred for 1h at 25 ℃. After the reaction was complete, the mixture was quenched with water (400 mL) and extracted with DCM (150 mL x 3). The organic layer was dried with saturated NaCl (50 mL), concentrated in vacuo and passed through a column (SiO ₂ Petroleum ether: ethyl acetate=2:1 to 0:1), washed with HCl (1 m,150 ml), extracted with DCM (50 ml×3), and then saturated NaHCO ₃ (30 mL) the pH was adjusted to-8. After extraction with DCM (100 mL), the residue was concentrated in vacuo to give methyl (S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (3 g,6.32mmol,74.74% yield, 90% purity) as a white solid.

¹ H NMR(400MHz,CDCl ₃ -d)δppm 7.61(d,J＝7.0Hz,1H),6.85-6.51(m,1H),6.22(s,1H),5.06-4.85(m,1H),4.63-4.47(m,1H),4.30-4.02(m,1H),3.79-3.66(m,3H),3.35-3.25(m,2H),2.42-2.24(m,1H),2.14-2.05(m,1H),1.96-1.66(m,4H),1.63-1.52(m,1H),1.43(s,9H),1.03-0.90(m,9H)。

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.5 g,3.51mmol,1 eq.) in HCl/MeOH (4M, 20 mL) was stirred at 25℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.1 g, crude material, HCl) as a white solid.

¹ H NMR(400MHz,D ₂ O)δppm 4.57(dd,J＝4.8,10.3Hz,1H),3.98(dd,J＝5.2,7.8Hz,1H),3.78-3.65(m,3H),3.29-3.14(m,2H),2.75-2.33(m,1H),2.24-1.47(m,8H),1.04-0.86(m,9H)。

Step 3:

(S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (550 mg x 2, hydrochloride, 1.68mmol,1 eq.) and 7-chloro-1H-indole-2-carboxylic acid (394.29 mg,2.02mmol,1.2 eq.) in DCM (6 mL) was added DMAP (615.66 mg,5.04mmol,3 eq.) and then EDCI (644.05 mg,3.36mmol,2 eq.) was added to the mixture at 25 ℃. After stirring for 1h at 25 ℃, the mixture was quenched with water (200 mL) and extracted with DCM (70 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ Petroleum ether ethyl acetate=1:1 to 0:1) purified and concentrated in vacuo, followed by washing with 1M HCl (100 mL) and extraction with DCM (30 mL x 3) and extraction with saturated NaHCO ₃ (30 mL) the pH of the organic phase was adjusted to pH 7. The residue was concentrated in vacuo to give methyl (S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (650 mg,1.16mmol,40% yield, 90% purity) as a pale yellow solid. MS (ESI) M/z505.2[ M+H] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.58(d,J＝7.8Hz,1H),7.32-7.17(m,2H),7.06(t,J＝7.8Hz,1H),4.73(dd,J＝3.8,8.6Hz,1H),4.55(dd,J＝4.0,11.7Hz,1H),3.71(s,3H),3.35(s,1H),3.24-3.01(m,2H),2.49-2.22(m,2H),2.02-1.40(m,8H),1.08-0.96(m,9H)。

Step 4:

n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (650 mg,1.29mmol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 10 mL) was stirred at 50deg.C for 16h. After the reaction was completed, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carboxamide (450 mg, crude material) as a pale yellow solid. MS (ESI) m/z 490.3[ M+H ] ] ⁺ 。

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-1H-indole-2-carboxamide (430 mg,877.56umol,1 eq) in DCM (10 mL) was added the bergs reagent (627.38 mg,2.63mmol,3 eq) and the reaction stirred at 25 ℃ for 4H. After the reaction was completed, the mixture was quenched with water (10 mL) and was purified by blowing N ₂ Dried and passed through preparative HPLC (column: kromasil C18 (250X 50mM X10 um)), mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide (205 mg,424.79umol,48.41% yield, 97.8% purity) as a white solid. MS (ESI) m/z 472.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.70(s,1H),9.02(d,J＝8.0Hz,1H),8.71(d,J＝8.0Hz,1H),7.63(d,J＝8.0Hz,1H),7.52(s,1H),7.34-7.23(m,2H),7.07(t,J＝7.8Hz,1H),5.05(q,J＝8.2Hz,1H),4.63-4.54(m,1H),3.07(s,2H),2.30-2.18(m,2H),1.88-1.32(m,7H),0.95(s,9H)。

EXAMPLE 78 Synthesis of viral protease inhibitor Compounds 723

Step 1:2, 2-difluoro-7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

(7S) -6-Boc-2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carboxylic acid (500 mg,1.72mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (406.29 mg,1.72mmol,1 eq., HCl), EDCI (987.17 mg,5.15mmol,3 eq.), DMAP (629.10 mg,5.15mmol,3 eq.) in DCM (5 mL) was degassed and taken up with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred for 2h at 20℃under an atmosphere. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (25 mL) and then extracted with DCM (25 mL x 3). The combined organic layers were washed with brine (25 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 1/1 to give (7S) -2, 2-difluoro-7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carboxylic acid tert-butyl ester (800 mg, crude material). MS (ESI) m/z 474.1[ M+H ]] ⁺ 。

Step 2: (2S) -2- (2, 2-difluoro-6-azaspiro [3.4] octane-7-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

(7S) -2, 2-difluoro-7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carboxylic acid tert-butyl ester (710 mg,1.50mmol,1 eq.) in HCl/MeOH (4M, 8mL,21.34 eq)) The solution in (2) was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2- [ [ (7S) -2, 2-difluoro-6-azaspiro [3.4] as a yellow oil]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (614 mg, crude, HCl). MS (ESI) m/z 374.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- (2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (7S) -2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (614 mg,1.50mmol,1 eq., HCl), 4-methoxy-1H-indole-2-carboxylic acid (286.41 mg,1.50mmol,1 eq.) and DMAP (549.06 mg,4.49mmol,3 eq.) in DCM (7 mL) was added ED CI (861.56 mg,4.49mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (25 mL) and then extracted with DCM (25 mL x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 1/1 to give (2S) -2- [ [ (7S) -2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a yellow solid]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (550 mg,1.01mmol,67.17% yield). MS (ESI) m/z 547.2[ M+H ]] ⁺ 。

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [ (7S) -2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (535 mg,978.85umol,1 eq.) in NH ₃ A solution in MeOH (7M, 10.70mL,76.52 eq.) was stirred at 30℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (7S) -N-)[ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (520 mg, crude). MS (ESI) m/z 532.2[ M+H ] ] ⁺ 。

Step 5: n- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

To (7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a solution of octane-7-carboxamide (515 mg,968.86umol,1 eq.) in EtOAc (2.5 mL) was added T3P (2.68 g,4.20mmol,2.5mL,50% purity, 4.34 eq.) and stirred at 20deg.C for 16h. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (25 mL) and then extracted with EtOAc (25 mL x 3). The combined organic layers were washed with brine (25 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,10 min) to give (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (188 mg,364.99umol,37.67% yield, 99.7% purity). MS (ESI) M/z514.3[ M+H ] ⁺ 。

Step 6: n- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

Isomer 1: by SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,10 min) isolation of (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (170 mg) to give (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]2, 2-difluoro-6- (4-)methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (42.5 mg,82.76umol,25.00% yield, 100% purity). MS (ESI) M/z514.3[ M+H] ⁺ 。

Isomer 1: ¹ H NMR(400MHz,MeOD-d ₄ ) Delta = 7.26-6.72 (m, 3H), 6.53 (d, J = 7.6hz, 1H), 5.03 (d, J = 5.7,10.5hz, 1H), 4.64 (d, J = 1.7hz, 1H), 4.25 (d, J = 10.1hz, 1H), 4.15-4.01 (m, 1H), 3.98-3.87 (m, 2H), 4.16-3.86 (m, 1H), 3.13 (s, 2H), 2.87-2.15 (m, 8H), 1.99-1.28 (m, 5H); and (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]-2, 2-difluoro-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]Octane-7-carboxamide (89.8 mg,173.47umol,52.40% yield, 99.2% purity). MS (ESI) m/z 514.3[ M+H ]] ⁺ 。

Isomer 2: ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.17-6.82(m,3H),6.56-6.44(m,1H),5.17-5.03(m,1H),4.61(t,J＝7.5Hz,1H),4.15(s,1H),4.01-3.78(m,4H),3.26-2.86(m,2H),2.75-2.14(m,8H),2.06-1.30(m,5H)。

example 79 Synthesis of viral protease inhibitor Compound 725

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (1.24 g,5.07mmol,1.2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1 g,4.22mmol,1 eq., HCl) in DCM (30 mL) was added DMAP (1.55 g,12.67mmol,3 eq.) and then EDCI (1.62 g,8.45mmol,2 eq.) was added. The mixture was stirred at 25℃for 1h. After completion, the mixture was quenched with water (400 mL) and extracted with DCM (150 mL x 3). After drying with saturated NaCl (50 mL), the reaction was concentrated in vacuo. The crude product was passed through a column (SiO ₂ Petroleum ether ethyl acetate=2:1 to 0:1) and washed with 1M HCl (100 mL), extracted with DCM (50 mL. Times.3), extracted with saturated NaHCO ₃ (50 mL) the pH was adjusted to pH 8, extracted with DCM (50 mL) and concentrated to give methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.4 g,2.95mmol,69.76% yield, 90% purity) as a white solid.

Step 2: ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamic acid tert-butyl ester

Methyl (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.4 g,3.27mmol,1 eq.) was reacted in NH ₃ A solution in MeOH (18 mL, 7M) was stirred at 50℃for 16h. After completion, the mixture was concentrated in vacuo to give tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate (1.1 g, crude material) as a white solid.

Step 3: (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide

A solution of tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate (1.5 g,3.64mmol,1 eq.) in HCl/MeOH (4M, 20 mL) was stirred at 25℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide (1.2 g, crude material) as a white solid.

Step 4: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -6, 7-dichloro-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl group]Ethyl group]To a mixture of 4, 4-dimethyl-pentanamide (900 mg,2.58mmol,1 eq, HCl) in DCM (8 mL) and DMF (3 mL) was added DMAP (945.50 mg,7.74mmol,3 eq) in one portion. Adding to the mixture6, 7-dichloro-1H-indole-2-carboxylic acid (593.47 mg,2.58mmol,1 eq.) and EDCI (1.48 g,7.74mmol,3 eq.) were added and the reaction stirred at 25℃for 2H. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1 to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-6, 7-dichloro-1H-indole-2-carboxamide (450 mg,858.06umol,33.26% yield). MS (ESI) m/z 524.2[ M+H ] ] ⁺ 。

Step 5:6, 7-dichloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a mixture of 6, 7-dichloro-1H-indole-2-carboxamide (400 mg,762.72 mol,1 eq.) in DCM (5 mL) was added the bergs reagent (363.53 mg,1.53mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 4h. After completion, by adding H ₂ O (3 mL) to quench the reaction mixture, and then the combined organic layers were concentrated under reduced pressure to give the crude product. By preparative HPLC (column: phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH3H2O +10mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give 6, 7-dichloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (165 mg,325.81 mol,42.72% yield). MS (ESI) m/z 506.1[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.54(d,J＝8.4Hz,1H),7.25-7.16(m,2H),5.13-5.05(m,1H),4.66(dd,J＝4.3,8.3Hz,1H),3.25-3.13(m,2H),2.50-2.35(m,2H),1.99-1.88(m,2H),1.87(d,J＝4.4Hz,1H),1.79(br dd,J＝8.4,14.6Hz,2H),1.71-1.56(m,1H),1.55-1.43(m,1H),1.03(s,9H)。

EXAMPLE 80 Synthesis of viral protease inhibitor Compounds 727

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (1.3 g,5.49mmol,1 eq, HCl) in DCM (12 mL) was added (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (1.51 g,6.59mmol,1.2 eq), TEA (3.33 g,32.95mmol,4.59mL,6 eq) and T3P (5.24 g,8.24mmol,4.90mL,50% purity, 1.5 eq). The reaction was stirred at 25℃for 2h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ DCM: meoh=10:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1 to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.99 g,4.11mmol,74.84% yield, 85% purity). MS (ESI) m/z 412.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1.20 g,2.48mmol,85% purity, 1 eq.) in HCl/MeOH (4M, 15mL,24.21 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give (2S) -2-amino-3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate(850 mg, crude material, HCl). MS (ESI) m/z 312.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (6, 7-dichloro-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (850 mg,2.44mmol,1 eq, HCl) in DCM (10 mL) and DMF (0.5 mL) was added 6, 7-dichloro-1H-indole-2-carboxylic acid (674.59 mg,2.93mmol,1.2 eq, 1.2), DMAP (746.35 mg,6.11mmol,2.5 eq) and EDCI (936.91 mg,4.89mmol,2 eq) and the resulting mixture was then stirred at 25 ℃ for 2H. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ DCM: meoh=10:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (6, 7-dichloro-1H-indole-2-carbonyl) amino ] as a yellow solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.24 g,1.50mmol,61.27% yield, 63% purity). MS (ESI) m/z 523.2[ M+H ]] ⁺ 。

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6, 7-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (6, 7-dichloro-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.38 g, parallel 3 batches, 726.01umol,1 eq.) in NH ₃ The mixture in MEOH (7M, 12.06mL,116.31 eq.) was stirred at 50deg.C for 48h. After completion, the mixture was concentrated under reduced pressure to give a residue, and then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6, 7-dichloro-1H-indole-2-carboxamide (1 g, crude). MS (ESI) m/z 508.2[M+H] ⁺ 。

Step 5:6, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A mixture of 6, 7-dichloro-1H-indole-2-carboxamide (1 g,1.97mmol,1 eq.) in T3P (5 mL,50% purity) and ethyl acetate (5 mL) was stirred at 40℃for 18H. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (20 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters X bridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product 6, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (288.22 mg,587.75umol,29.88% yield, 100% purity). MS (ESI) m/z 490.1[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.94(br s,1H),9.01(d,J＝7.9Hz,1H),8.76(br d,J＝7.5Hz,1H),7.66(d,J＝8.4Hz,1H),7.55(br s,1H),7.33-7.21(m,2H),5.21-4.90(m,1H),4.60-4.38(m,1H),3.16-3.01(m,2H),2.35-2.18(m,2H),1.90-1.65(m,4H),1.63-1.33(m,3H),0.80(br d,J＝5.5Hz,1H),0.49-0.35(m,2H),0.26-0.05(m,2H)。

EXAMPLE 81 Synthesis of viral protease inhibitor Compound 729

Step 1: (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (6-azaspiro [3.4] octane-7-carboxamide) propanoic acid methyl ester

7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoylRadical) -6-azaspiro [3.4]A solution of tert-butyl octane-6-carboxylate (1 g,2.29mmol,1 eq.) in HCl/MeOH (40 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove HCl/MeOH, and DCM (50 mL) was then added (three times). The reaction was concentrated under reduced pressure to give the crude product (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (6-azaspiro [ 3.4) as a yellow solid]Octane-7-carboxamido) propionic acid methyl ester (800 mg, crude material, HCl). MS (ESI) m/z 338.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- (6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (6-azaspiro [ 3.4)]To a solution of methyl octane-7-carboxamido) propionate (580 mg,1.72mmol,1 eq.) and 7-chloro-1H-indole-2-carboxylic acid (504.35 mg,2.58mmol,1.5 eq.) in DCM (10 mL) was added DMAP (420.00 mg,3.44mmol,2 eq.) and EDCI (494.29 mg,2.58mmol,1.5 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction was quenched with H ₂ O (100 mL) was quenched and then extracted with DCM (50 mL x 3). The combined organic phases were washed with brine (50 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1 to give (2S) -2- (6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4) as a yellow solid]Octane-7-formylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (680 mg,1.19mmol,69.13% yield, 90% purity). MS (ESI) m/z 515.2[ M+H ]] ⁺ 。

Step 3: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (6755 mg,1.31mmol,1 eq.) in NH ₃ (7M in MeOH, 29.53mL,157.72 eq). The mixture was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to giveResidue. DCM (50 mL) was added to the mixture (three times), and then the reaction was concentrated under reduced pressure to give a residue. The crude product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]-6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (700 mg, crude) was used in the next step and obtained as a yellow solid. MS (ESI) m/z 500.2[ M+H ]] ⁺ 。

Step 4:6- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -6-azaspiro [3.4] octane-7-carboxamide

At N ₂ Next, N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-6- (7-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a solution of octane-7-carboxamide (695 mg,1.39mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (1.66 g,6.95mmol,5 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure at 30 ℃ to give a residue. By preparative TLC (SiO) ₂ The residue was purified with ethyl acetate: meoh=20:1 to give the desired compound (450 mg,96% purity) as a yellow solid, which was purified by SFC (condition: column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%, min) to give 6- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide (145 mg,300.85umol,21.64% yield, 100% purity). MS (ESI) m/z 482.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.67-7.49(m,1H),7.31-7.23(m,1H),7.19-6.99(m,2H),5.14-4.95(m,1H),4.60-4.52(m,1H),4.07-3.77(m,2H),3.27-3.16(m,2H),2.56-1.50(m,15H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.27-11.09(m,1H),8.82-8.62(m,1H),7.72-7.53(m,1H),7.36-7.24(m,2H),7.19-7.02(m,2H),5.11-4.85(m,1H),4.67-4.42(m,1H),4.05-3.73(m,2H),3.10-3.06(m,2H),2.30-1.38(m,15H)。

6- (7-chloro-1H-indole) as a white solid-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide (170 mg,348.13umol,25.04% yield, 98.7% purity). MS (ESI) m/z 482.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.67-7.55(m,1H),7.31-7.25(m,1H),7.18-7.11(m,1H),7.10-7.04(m,1H),4.93(br s,1H),4.60-4.54(m,1H),4.13-3.79(m,2H),2.98(br s,2H),2.42-1.54(m,15H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.27-10.98(m,1H),8.88-8.54(m,1H),7.82-7.49(m,1H),7.34-6.98(m,4H),5.10-4.95(m,1H),4.69-4.39(m,1H),4.03-3.72(m,2H),3.10-3.05(m,2H),2.32-1.39(m,15H)。

EXAMPLE 82 Synthesis of viral protease inhibitor Compounds 731

Step 1:3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

At 0℃under N ₂ Downward (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,4.22mmol,1 eq. HCl) and 2-tert-butoxycarbonyl-2-azaspiro [4.5]]To a mixture of decane-3-carboxylic acid (1.26 g,4.44mmol,1.05 eq.) and DIPEA (2.73 g,21.12mmol,3.68mL,5 eq.) in THF (10 mL) was added T3P (4.03 g,6.34mmol,3.77mL,50% purity, 1.5 eq.). The mixture was stirred at 20℃for 2h. After completion, the residue was poured into saturated sodium bicarbonate solution (30 mL) and stirred for 2min. The aqueous phase was extracted with ethyl acetate (20 ml x 2). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a pale yellow oil]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (1.6 g, crude material))。MS(ESI)m/z 466.3[M+H] ⁺ 。

Step 2: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

At 0℃under N ₂ Downward 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]To a mixture of tert-butyl decane-2-carboxylate (1.6 g,3.44mmol,1 eq.) was added HCl/MeOH (4M, 16.00mL,18.62 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated to give (2S) -2- (2-azaspiro [4.5] as a pale yellow oil]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.5 g, crude material, HCl). MS (ESI) m/z 366.2[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

At 20℃under N ₂ Downward (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (1.5 g,3.73mmol,1 eq, HCl) and 7-chloro-1H-indole-2-carboxylic acid (729.99 mg,3.73mmol,1 eq), DIPEA (1.45 g,11.20mmol,1.95mL,3 eq) in DMF (10 mL) was added HATU (1.70 g,4.48mmol,1.2 eq). The mixture was stirred at 20℃for 2h. After completion, the residue was poured into ice water (10 mL) and stirred for 2min. The aqueous phase was extracted with ethyl acetate (10 ml x 3). The combined organic phases were washed with brine (5 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=2/1 to 0/1) to give (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a pale yellow oil]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.60 g, crude material). MS (ESI) m/z 543.2[ M+H ]] ⁺ 。

Step 4: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

At 20 DEG CUnder N ₂ Downward (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] ]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH was added to a mixture of methyl propionate (1.6 g,2.95mmol,1 eq.) in the form of a mixture ₃ MeOH (7M, 22.86mL,54.31 eq.). The mixture was stirred at 65℃for 12h. After completion, the mixture was cooled to 25 ℃ and concentrated under reduced pressure. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=5/1 to 0/1 to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a pale yellow solid]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.2 g, crude). MS (ESI) m/z 528.2[ M+H ]] ⁺ 。

Step 5:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

At 20℃under N ₂ Downward N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.2 g,2.27mmol,1 eq.) in DCM (5 mL) was added the Bungeus reagent (1.2 g,5.04mmol,2.22 eq.) in one portion. The mixture was stirred at 20℃for 2.5h. After completion, water (3 mL) was added to the mixture and stirred for 20min, followed by concentration to give a crude material. By preparative TLC (SiO) ₂ EtOAc: meoh=25:1) the residue was purified to give 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a pale yellow solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (0.75 g,1.45mmol,64.00% yield, 98.9% purity). MS (ESI) m/z 528.2[ M+H ]] ⁺ 。

Step 6:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

By chiral separation (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,6.7 min) to isolate 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (0.9 g,1.76mmol,1 eq.) gave 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide isomer 1 (298.31 mg,578.46umol,32.78% yield, 98.9% purity). MS (ESI) m/z 510.3[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.62(br d,J＝7.94Hz,1H),7.56-7.56(m,1H),7.22-7.30(m,1H),7.01-7.13(m,2H),5.11(br dd,J＝10.58,5.73Hz,1H),4.62(br dd,J＝9.81,7.83Hz,1H),3.83-3.96(m,1H),3.71(br d,J＝10.36Hz,1H),3.16-3.27(m,2H),2.40-2.62(m,2H),1.70-2.08(m,4H),1.29-1.65(m,12H)。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.15(br s,1H),8.71(br s,1H),7.62(br s,1H),7.18-7.35(m,2H),6.94-7.13(m,2H),4.96(br s,1H),4.62(br s,1H),3.51-3.87(m,2H),3.09-3.20(m,2H),2.09-2.36(m,3H),1.60-1.89(m,4H),1.19-1.55(m,12H)。

Obtaining 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide isomer 2 (252.99 mg,487.10umol,27.60% yield, 98.20% purity). MS (ESI) M/z510.3[ M+H] ⁺ ；

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.64(d,J＝7.72Hz,1H),7.21-7.33(m,1H),7.12(s,1H),7.04-7.10(m,1H),7.07(t,J＝7.83Hz,1H),5.02(dd,J＝10.25,6.06Hz,1H),4.62(dd,J＝9.70,7.72Hz,1H),3.95(br d,J＝10.14Hz,1H),3.77(br d,J＝10.58Hz,1H),3.01-3.22(m,2H),2.22-2.40(m,3H),1.86-2.04(m,2H),1.77-1.86(m,1H),1.72(br dd,J＝12.46,10.03Hz,1H),1.39-1.68(m,12H)。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.10(br s,1H),8.65(br d,J＝6.24Hz,1H),7.63(br d,J＝6.85Hz,1H),7.17-7.34(m,2H),7.08(br t,J＝7.70Hz,2H),4.99(br d,J＝7.46Hz,1H),4.61(br s,1H),3.56-3.89(m,2H),3.10(br s,2H),2.09-2.31(m,3H),1.64-1.95(m,4H),1.38-1.62(m,12H)。

EXAMPLE 83 Synthesis of viral protease inhibitor Compound 733

Step 1:3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

To a solution of methyl (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propanoate (500 mg,2.11mmol,1 eq. HCl) in DCM (4 mL) and DMF (2 mL) was added 2-t-butoxycarbonyl-2-azaspiro [4.5]]Decane-3-carboxylic acid (718.30 mg,2.53mmol,1.2 eq.) DMAP (774.22 mg,6.34mmol,3 eq.) and EDCI (809.90 mg,4.22mmol,2 eq.) were then added at 0deg.C. The mixture was then stirred at 25℃for 2h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1) to give 3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] as a yellow solid ]Decane-2-carboxylic acid tert-butyl ester (775 mg,1.50mmol,70.92% yield, 90% purity). MS (ESI) M/z466.3[ M+H] ⁺ 。

Step 2: (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

A mixture of tert-butyl 3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] decane-2-carboxylate (775 mg,1.50mmol,90% purity, 1 eq.) in HCl/MeOH (4M, 8mL,21.36 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [4.5] decane-3-carboxamido) propanoate (800 mg, crude material, HCl) as a yellow solid.

Step 3: (2S) -2- (2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [ 4.5)]To a solution of methyl decane-3-carboxamido) propionate (740 mg,1.38mmol,75% purity, 1 eq, HCl) in DCM (6 mL) and DMF (3 mL) was added 6-chloro-1H-indole-2-carboxylic acid (297.11 mg,1.52mmol,1.1 eq), DMAP (506.09 mg,4.14mmol,3 eq), followed by EDCI (529.42 mg,2.76mmol,2 eq) at 0 ℃ and the mixture was then stirred at 25 ℃ for 2H. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give (2S) -2- (2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [ 4.5) as a yellow solid]Decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (480 mg,1.35mmol,98.02% yield, 75% purity). MS (ESI) m/z 543.3[ M+H ]] ⁺ 。

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- (2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [ 4.5)]Decane-3-formylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (480 mg,1.35mmol,75% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 15mL,77.58 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (960 mg, crude). MS (ESI) m/z 528.2[ M+H ]] ⁺ 。

Step 5:2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (960 mg,1.36mmol, 75%)Purity, 1 eq.) to a solution of the primary Gibbs reagent (1.95 g,8.18mmol,6 eq.) in DCM (10 mL) and then stirred at 25℃for 4h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give 2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (280 mg,39.66% yield, 98.5% purity). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

Step 6:2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,8 min) purification of 2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ]Decane-3-carboxamide (280 mg,98.5% purity) gave 2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 1 (90.34 mg,175.89umol,12.90% yield, 99.3% purity). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.68(s,1H),9.10-8.79(m,1H),7.69(d,J＝8.4Hz,1H),7.58-7.47(m,1H),7.46-7.36(m,1H),7.14-6.62(m,2H),5.10-4.73(m,1H),4.51(t,J＝8.4Hz,1H),3.95-3.73(m,1H),3.65(d,J＝10.4Hz,1H),3.17-2.83(m,2H),2.35-2.07(m,3H),1.93-1.19(m,16H)。

¹ H NMR(400MHz,DMSO-d ₆ )(T＝273+80K)δ＝11.48(br s,1H),8.74(br s,1H),7.65(br s,1H),7.47(br s,1H),7.31(br s,1H),7.06(br d,J＝9.0Hz,2H),4.98(br s,1H),4.57(br s,1H),3.87(br d,J＝10.1Hz,1H),3.64(br s,1H),3.10-3.04(m,2H),2.39-2.11(m,3H),1.90-1.36(m,16H)。

Obtaining 2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 2 (143.12 mg,280.61umol,20.58% yield, 100% purity). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69(s,1H),9.12-8.72(m,1H),7.69(d,J＝8.6Hz,1H),7.57-7.40(m,2H),7.14-6.60(m,2H),5.08-4.78(m,1H),4.51(t,J＝8.4Hz,1H),3.92-3.78(m,1H),3.69(d,J＝10.4Hz,1H),3.13-2.92(m,2H),2.28-2.06(m,3H),1.87-1.29(m,16H)。

¹ H NMR(400MHz,DMSO-d ₆ )(T＝273+80K)δ＝11.49(br s,1H),8.69(br s,1H),7.79-7.57(m,1H),7.48(s,1H),7.27(br s,1H),7.06(br d,J＝8.4Hz,2H),4.97(br s,1H),4.57(br s,1H),3.88(d,J＝10.4Hz,1H),3.68(br s,1H),3.10-3.04(m,2H),2.20(br s,3H),1.91-1.31(m,16H)。

EXAMPLE 83a Synthesis of viral protease inhibitor Compound 743

(7S) -6-Boc-2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carboxylic acid (500 mg,1.72mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (406.29 mg,1.72mmol,1 eq., HCl), EDCI (987.17 mg,5.15mmol,3 eq.), DMAP (629.10 mg,5.15mmol,3 eq.) in DCM (5 mL) was degassed and taken up with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred for 2h at 20℃under an atmosphere. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (50 mL) and then extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (50 ml x 2), dried over Na ₂ SO ₄ Drying and filteringAnd concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 1/1 to give (7S) -2, 2-difluoro-7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carboxylic acid tert-butyl ester (800 mg, crude material). MS (ESI) m/z 474.1[ M+H ]] ⁺ 。

(7S) -2, 2-difluoro-7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]A solution of tert-butyl octane-6-carboxylate (800 mg,1.69mmol,1 eq.) in HCl/MeOH (4M, 8mL,18.94 eq.) was stirred at 20deg.C for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2- [ [ (7S) -2, 2-difluoro-6-azaspiro [3.4] as a yellow oil ]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (690 mg, crude material, HCl). MS (ESI) m/z 374.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- (6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3.4] octane-7-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (7S) -2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (69mg, 1.68mmol,1 eq., HCl), 7-chloro-1H-indole-2-carboxylic acid (329.30 mg,1.68mmol,1 eq.) and DMAP (617.03 mg,5.05mmol,3 eq.) in DCM (10 mL) was added EDCI (968.19 mg,5.05mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (35 mL), and then extracted (35 mL x 3). The combined organic layers were washed with brine (35 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 1/1 to give (2S) -2- [ [ (7S) -6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3 ] as a yellow solid.4]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]Methyl propionate (640 mg,1.16mmol,69.00% yield). MS (ESI) m/z 551.2[ M+H ]] ⁺ 。

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [ (7S) -6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (625 mg,1.13mmol,1 eq.) in NH ₃ Solution in MeOH (7M, 12mL,74.05 eq.). The mixture was stirred at 30℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (7S) -N- [ (1S) -2-amino-2-oxo-1-piperidinyl [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]-6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3.4]Octane-7-carboxamide (605 mg, crude). MS (ESI) M/z536.2[ M+H] ⁺ 。

Step 5:6- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2, 2-difluoro-6-azaspiro [3.4] octane-7-carboxamide

To (7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-6- (7-chloro-1H-indole-2-carbonyl) -2, 2-difluoro-6-azaspiro [3.4 ]To a solution of octane-7-carboxamide (585 mg,1.09mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (1.17 g,4.91mmol,4.5 eq.). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (30 mL) and then extracted with DCM (35 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give (7S) -6- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2, 2-difluoro-6-azaspiro [3.4]]Octane-7-carboxamide (135 mg,260.64umol, 23.8)8% yield). MS (ESI) m/z 518.2[ M+H ]] ⁺ 。

Step 6:6- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2, 2-difluoro-6-azaspiro [3.4] octane-7-carboxamide

Isomer 1: by SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,15 min) isolation of 6- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2, 2-difluoro-6-azaspiro [3.4] ]Octane-7-carboxamide (133 mg) gave (7S) -6- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2, 2-difluoro-6-azaspiro [3.4 ]]Octane-7-carboxamide (48.2 mg,93.06umol,36.24% yield). MS (ESI) m/z 518.2[ M+H ]] ⁺ 。

Isomer 1: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.34-11.12(m,1H),8.83-8.63(m,1H),7.71-7.55(m,1H),7.30(d,J＝7.1Hz,2H),7.13-7.04(m,1H),5.09-4.92(m,1H),4.71-4.52(m,1H),4.20-3.87(m,2H),3.09-3.05(m,1H),3.10-3.03(m,2H),2.91-2.52(m,4H),2.48-2.35(m,2H),2.29-2.08(m,2H),1.96-1.31(m,5H)。

isomer 2: to give (7S) -6- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2, 2-difluoro-6-azaspiro [3.4 ]]Octane-7-carboxamide (83.2 mg,160.63umol,62.56% yield). MS (ESI) m/z 518.2[ M+H ]] ⁺ 。

Isomer 2: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.32-11.13(m,1H),8.90-8.67(m,1H),7.69-7.48(m,1H),7.33-7.26(m,1H),7.19-6.89(m,2H),5.07-4.88(m,1H),4.74-4.51(m,1H),4.15-3.84(m,2H),3.11-3.06(m,2H),3.10-3.06(m,1H),2.82-2.55(m,4H),2.43(d,J＝3.2,5.2Hz,2H),2.32-2.07(m,2H),2.02-1.01(m,5H)。

example 84 Synthesis of viral protease inhibitor Compound 745

Step 1: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1 g,2.34mmol,1 eq.) in HCl/MeOH (4M) (10 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Obtaining the compound (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl as a white solid ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (900 mg, crude) and was used directly in the next step. MS (ESI) m/z 328.3[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

To a mixture of 6-chloro-1H-indole-2-carboxylic acid (400 mg,2.04mmol,1 eq.) in DCM (10 mL) and DMF (5 mL) was added DMAP (749.49 mg,6.13mmol,3 eq.) in one portion at 25 ℃. (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl was added to the mixture in one portion at 25 ℃C]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (892.94 mg,2.45mmol,1.20 eq. HCl) and EDCI (784.05 mg,4.09mmol,2 eq.) and the reaction stirred for 2.5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. Obtaining the compound (2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino ] as a yellow solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (800 mg,1.58mmol,77.47% yield). MS (ESI) m/z 505.2[ M+H ] ] ⁺ 。

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -6-chloro-1H-indole-2-carboxamide

One time at 25℃to (2S) -2- [ [ (2S) -2- [ (6-chloro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (605 mg, 1).20mmol,1 eq.) in NH ₃ In a mixture of MeOH (7M) (30.60 mg,1.80mmol,30.00uL,1.5 eq.). The mixture was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Obtaining the compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-6-chloro-1H-indole-2-carboxamide (600 mg, crude) and used directly in the next step. MS (ESI) m/z 490.1[ M+H ]] ⁺ 。

Step 4: 6-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl ]To a mixture of 6-chloro-1H-indole-2-carboxamide (500 mg,1.02mmol,1 eq.) in DCM (6 mL) was added the Buerger's reagent (607.94 mg,2.55mmol,2.5 eq.) and the mixture was stirred at 25℃for 3H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC { column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,8min } the residue was purified. The compound 6-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (202 mg,405.64umol,39.75% yield, 94.78% purity). MS (ESI) m/z 472.3[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.41-7.65(m,2H),7.01-7.22(m,2H),5.08(br s,1H),4.65(br s,1H),3.15-3.25(m,2H),2.43(br s,1H),1.46-2.05(m,8H),1.02(br s,9H)

EXAMPLE 85 Synthesis of viral protease inhibitor Compounds 791

Step 1:7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 25 ℃]To a mixture of methyl propionate (2.32 g,9.79mmol,1 eq., HCl) in DCM (30 mL) and DMF (10 mL) was added DMAP (3.59 g,29.38mmol,3 eq.) in one portion. Adding 6-t-Butoxycarbonyl-6-azaspiro [3.4] to the mixture ]Octane-7-carboxylic acid (3 g,11.75mmol,1.2 eq.) and EDCI (3.75 g,19.58mmol,2 eq.) were stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (40 mL) was diluted and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (80 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1 to give 7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carboxylic acid tert-butyl ester (5 g, crude material). MS (ESI) m/z 438.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- (6-azaspiro [3.4] octane-7-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

A mixture of 7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (1.6 g,3.66mmol,1 eq.) in HCl/MeOH (20 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- (6-azaspiro [3.4] octane-7-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (1.3 g, crude material) as a yellow solid.

Step 3: (2S) -2- [ [6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- (6-azaspiro [3.4]]Octane-7-carbonylamino) -3- [ (3S) -2-oxo-3-piperidyl]Methyl propionate (1.3 g,3.85mmol,1 eq.) and 6-chloro-1H-indole-2-carboxylic acid (904.35 mg,4.62mmol,1.2 eq.) in DCM (9 mL) and DMF (3)mL) was added DMAP (1.41 g,11.56mmol,3 eq.) and EDCI (1.48 g,7.71mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (25 mL x 3). The combined organic layers were washed with brine (20 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give (2S) -2- [ [6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a yellow oil]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.21mmol,57.45% yield, 95% purity). MS (ESI) M/z515.3[ M+H ]] ⁺

Step 4: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.33mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 15mL,45.06 eq.) was stirred at 50deg.C for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (480 mg,1.96mmol,84.12% yield). MS (ESI) m/z 500.2[ M+H ]] ⁺ 。

Step 5:6- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl group]Ethyl group]-6- (6-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a mixture of octane-7-carboxamide (480 mg,1.96mmol,1 eq.) in DCM (10 mL) was added the Bolus reagent (1.87 g,7.84mmol,4 eq.) in one portion. The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral Conditions; column: kromasil C18 (250 x 50mm x 10 um); mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,10 min) to give 6- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-6-azaspiro [3.4]]Octane-7-carboxamide (600 mg,1.24mmol,63.51% yield). MS (ESI) m/z 482.2[ M+H ]] ⁺

Step 6:6- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -6-azaspiro [3.4] octane-7-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%, min) to isolate the white solid to give 6- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-6-azaspiro [3.4]]Octane-7-carboxamide (140 mg,290.47 mol,23.33% yield) and 6- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-6-azaspiro [3.4]]Octane-7-carboxamide (110 mg,228.23umol,18.33% yield). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.41(br s,1H),8.60(br s,1H),7.65(br d,J＝8.3Hz,1H),7.49(s,1H),7.24-6.88(m,3H),5.10-4.82(m,1H),4.56(br s,1H),4.02-3.86(m,2H),3.09(br s,2H),2.36-2.26(m,1H),2.25-2.07(m,3H),2.07-1.77(m,8H),1.73-1.32(m,3H)。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.70-7.53(m,1H),7.46(s,1H),7.15-6.63(m,2H),5.02(dd,J＝6.0,10.6Hz,1H),4.65-4.52(m,1H),4.17-3.74(m,2H),3.25-2.90(m,2H),2.56-2.13(m,4H),2.11-1.74(m,8H),1.72-0.99(m,3H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.42(br s,1H),8.66(br s,1H),7.64(br s,1H),7.49(br s,1H),7.32-6.79(m,3H),4.97(br s,1H),4.57(br s,1H),3.93(br s,2H),3.11(br s,2H),2.38-2.11(m,4H),2.05-1.77(m,8H),1.73-1.34(m,3H)。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.64(d,J＝8.6Hz,1H),7.55-7.42(m,1H),7.13-6.99(m,2H),5.09(dd,J＝6.3,10.7Hz,1H),4.55(t,J＝7.5Hz,1H),4.12-3.95(m,2H),3.27-3.17(m,2H),2.63-2.36(m,3H),2.13-1.90(m,9H),1.80(br s,2H),1.51(br d,J＝9.3Hz,1H)。

EXAMPLE 86 Synthesis of viral protease inhibitor Compound 793

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (4.97 g,20.28mmol,1.2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (4 g,16.90mmol,1 eq., HCl) in DCM (60 mL) was added DMAP (6.19 g,50.70mmol,3 eq.) and then EDCI (6.48 g,33.80mmol,2 eq.) was added. The mixture was stirred at 20℃for 1h. After completion, the mixture was treated with H ₂ O (50 mL) was quenched, extracted with DCM (40 mL x 3), then washed with 1M HCl (40 mL) and extracted with DCM (80 mL x 3), and then dried with NaCl (100 mL), then concentrated in vacuo. The crude product was passed through a column (Plate 1, siO ₂ Petroleum ether ethyl acetate=2:1 to 0:1, i ₂ ，R _f =0.22), followed by concentration in vacuo, to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (5.75 g,11.43mmol,67.65% yield, 85% purity). MS (ESI) m/z 428.3[ M+H ]] ⁺

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (500 mg,1.17mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give a white solidIn the form of a body (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (425 mg, crude, HCl). MS (ESI) m/z 328.2[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (409.27 mg,1.12mmol,1 eq, HCl) and 4-chloro-1H-indole-2-carboxylic acid (220 mg,1.12mmol,1 eq) in DCM (15 mL) was added EDCI (646.84 mg,3.37mmol,3 eq) and then DMAP (412.22 mg,3.37mmol,3 eq). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (50 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with 1M HCl (20 ml x 2) and then dried over NaCl (10 ml x 2), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (321 mg,572.07umol,50.86% yield, 90% purity). MS (ESI) m/z 505.2[ M+H ]] ⁺ 。

Step 4: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (306 mg,605.93umol,1 eq.) in NH ₃ A solution in MeOH (7M, 3mL,34.66 eq.) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-chloro-1H-indole-2-carboxamide (250 mg, crude). MS (ESI) m/z 490.2[ M+H ]] ⁺ 。

Step 5: 4-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 4-chloro-1H-indole-2-carboxamide (230 mg,469.39 mmol,1 eq.) in DCM (3 mL) was added the Buerger's reagent (335.58 mg,1.41mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give 4-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (114 mg,241.54umol,51.46% yield, 100% purity). MS (ESI) m/z 472.2[ M+H ]] ⁺ 。

1H NMR(400MHz,MeOD-d ₄ )δ＝7.39(d,J＝8.4Hz,1H),7.28(s,1H),7.18(t,J＝7.8Hz,1H),7.12-7.04(m,1H),7.09(d,J＝7.4Hz,1H),5.16-5.02(m,1H),4.65(d,J＝4.4,8.4Hz,1H),3.24-3.16(m,2H),2.49-2.38(m,2H),2.00-1.73(m,5H),1.66(d,J＝8.4Hz,1H),1.55-1.45(m,1H),1.03(s,8H)

EXAMPLE 87 Synthesis of viral protease inhibitor Compounds 795

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (4.97 g,20.28mmol,1.2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (4 g,16.90mmol,1 eq., HCl) in DCM (120 mL) was added DMAP (6.19 g,50.70mmol,3 eq.) and then EDCI (6.48 g,33.80mmol,2 eq.) was added. The resulting mixture was stirred at 20℃for 1h. After completion, the mixture was treated with H ₂ O (500 mL) quenched and extracted with DCM (200 mL x 3), followed by washing with 1M HCl (200 mL) and extraction with DCM (80 mL x 3), and followed by drying with NaCl (100 mL), followed by concentration in vacuo. The crude product was passed through a column (Plate 1, siO ₂ Petroleum ether ethyl acetate=2:1 to 0:1, i ₂ ，R _f =0.22), followed by concentration in vacuo, gave methyl (S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (5.75 g,11.43mmol,67.65% yield, 85% purity) as a white solid. MS (ESI) m/z 428.3[ M+H ]] ⁺ 。

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (700 mg,1.64mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 1h. After completion, the reaction was concentrated in vacuo to give methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (630 mg, crude material, HCl) as a yellow solid. MS (ESI) m/z 328.2[ M+H ]] ⁺ 。

Step 3: (S) -2- ((S) -2- (4, 6-dichloro-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (630 mg,1.73mmol,1 eq, HCl) in DCM (20 mL) was added 4, 6-dichloro-1H-indole-2-carboxylic acid (438.12 mg,1.90mmol,1.1 eq), DMAP (634.54 mg,5.19mmol,3 eq) and EDCI (431.47 mg,2.25mmol,1.3 eq). The mixture was stirred at 25℃for 2h. After completion, by adding H ₂ O (100 mL) to quench the reaction and then extract with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure, and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=7/3 to 0/1) to give the product methyl (S) -2- ((S) -2- (4, 6-dichloro-1H-indole-2-carboxamido) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (400 mg,595.42umol,34.39% yield, 80.3% purity) as a yellow solid. MS (ESI) m/z 539.2[ M+H ]] ⁺ 。

Step 4: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (4, 6-dichloro-1H-indole-2-carboxamide) -4, 4-dimethylvalerylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (370 mg,685.88umol,1 eq.) was reacted in NH ₃ The solution in (7M, 20mL,204.12 eq.) was stirred at 30℃for 8h. After completion, the reaction was concentrated in vacuo to give the product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide (340 mg, crude material) as a yellow solid. MS (ESI) m/z 524.2[ M+H ]] ⁺ 。

Step 5:4, 6-dichloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide (320 mg,610.18umol,1 eq) in DCM (15 mL) was added the bergs reagent (1.02 g,4.27mmol,7 eq) and the mixture stirred at 40 ℃ for 3H. After completion, the reaction was concentrated in vacuo and purified by preparative HPLC (column Waters Xbridge Prep OBD C18 150 x 40mm x 10um; mobile phase: [ water (0.05% nh ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,8 min) to give 4, 6-dichloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide (110 mg,217.21umol,35.60% yield, 100% purity) as a white solid. MS (ESI) m/z 506.2[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.00(br s,1H),8.94-8.92(m,1H),8.81-8.80(m,1H),7.52(br s,1H),7.42(s,2H),7.24(s,1H),5.11-4.98(m,1H),4.54-4.49(m,1H),3.12-3.01(m,2H),2.34-2.19(m,2H),1.85-1.63(m,5H),1.58-1.45(m,1H),1.43-1.32(m,1H),0.94(s,9H)

EXAMPLE 88 Synthesis of viral protease inhibitor Compound 797

Step 1: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (800 mg,1.87mmol,1 eq.) and HCl/MeOH (4M, 25mL,53.44 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (650 mg, crude material, HCl). MS (ESI) m/z 328.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [ (2S) -2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (650 mg,1.79mmol,1.2 eq.) 4, 7-dichloro-1H-indole-2-carboxylic acid (342.45 mg,1.49mmol,1 eq.), EDCI (856.10 mg,4.47mmol,3 eq.) and DMAP (545.57 mg,4.47mmol,3 eq.) in DCM (10 mL) was stirred at 20deg.C for 1H. After completion, H is added to the mixture ₂ O (50 mL) and then extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1) the residue was purified to give (2S) -2- [ [ (2S) -2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino ] as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (650 mg,1.17mmol,78.79% yield, 97.34% purity). MS (ESI) m/z539.2[M+H] ⁺ 。

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4, 7-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4, 7-dichloro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (620 mg,1.15mmol,1 eq.) and NH ₃ The mixture of MeOH (7M, 20mL,121.81 eq.) was stirred at 65℃for 16h. After completion, the mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group ]-3, 3-dimethyl-butyl]-4, 7-dichloro-1H-indole-2-carboxamide (550 mg, crude). MS (ESI) m/z 524.2[ M+H ]] ⁺ 。

Step 4:4, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]A mixture of 4, 7-dichloro-1H-indole-2-carboxamide (530 mg,1.01mmol,1 eq.) and Prague reagent (722.50 mg,3.03mmol,3 eq.) in DCM (10 mL) was stirred at 20deg.C for 3.5H. After completion, the mixture was concentrated under reduced pressure to give a residue. The residue was then purified by preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,10 min) to give 4, 7-dichloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (170 mg,334.61umol,33.11% yield, 99.68% purity). MS (ESI) m/z 506.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.62-7.56(m,1H),7.52-7.46(m,1H),7.21-7.14(m,1H),5.14-5.07(m,1H),4.70-4.64(m,1H),3.25-3.17(m,2H),2.51-2.38(m,2H),2.02-1.85(m,3H),1.85-1.61(m,3H),1.58-1.44(m,1H),1.08-1.02(m,9H)

EXAMPLE 89 Synthesis of viral protease inhibitor Compound 799

Step 1: 7-chloro-4-methoxy-1H-indole-2-carboxylic acid

A mixture of methyl 7-chloro-4-methoxy-1H-indole-2-carboxylate (500 mg,2.09mmol,1 eq.) in NaOH (2M, 10.43mL,10 eq.) was then stirred at 100deg.C for 0.5H. After completion, the mixture was acidified with HCl (3M) to adjust the pH to about 3, and then the reaction was extracted with EtOAc (10 ml x 3). The organic layer was washed with water (10 mL), taken up in Na ₂ SO ₄ Dried, filtered, and concentrated under reduced pressure to give 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (400 mg, crude material) as a yellow solid.

Step 2: (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (800 mg,1.87mmol,1 eq.) in HCl/MeOH (4M, 8mL,17.10 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylvaleramide (810 mg, crude material, HCl) as a white solid.

Step 3: (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a solution of 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (440 mg,1.95mmol,1 eq.) in DCM (8 mL) and DMF (4 mL) was added (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide (851.53 mg,2.34mmol,1.2 eq.) DMAP (714.74 mg,5.85mmol,3 eq.) and then EDCI (747.67 mg,3.90mmol,2 eq.) was added at 0 ℃. Then atThe mixture was stirred at 25℃for 2h. After completion, the mixture was quenched with water (20 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10 mL), and dried over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gave a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=100:1 to 10:1 to give methyl (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propionate (1.3 g,1.82mmol,93.45% yield, 75% purity) as a yellow solid. MS (ESI) m/z 535.1[ M+H ]] ⁺ 。

Step 4: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.3 g,1.82mmol,75% purity, 1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 15mL,57.62 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide (1.25 g, crude material) as a yellow solid. MS (ESI) m/z 520.3[ M+H ]] ⁺ 。

Step 5: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide (1.21 g,1.75mmol,75% purity, 1 eq.) in EtOAc (6 mL) was added T3P (6.42 g,10.09mmol,6mL,50% purity, 5.78 eq.) and then the reaction stirred at 40 ℃ for 14H. After completion, the mixture was quenched with water (20 mL) and extracted with EtOAc (10 mL x 3). The organic layer was washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (496.09 mg,988.22umol,56.63% yield, 100% purity) as a white solid. MS (ESI) m/z 502.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.64(br s,1H),9.12-8.90(m,1H),8.72-8.54(m,1H),7.52(br s,1H),7.28(s,1H),7.20(d,J＝8.3Hz,1H),6.56(d,J＝8.3Hz,1H),5.05(q,J＝8.0Hz,1H),4.62-4.50(m,1H),3.89(s,3H),3.07(br s,2H),2.31-2.15(m,2H),1.88-1.63(m,5H),1.60-1.33(m,2H),1.06-0.85(m,9H)。

EXAMPLE 90 Synthesis of viral protease inhibitor Compound 801

Step 1: 4-chloro-1H-indole-2-carbonyl chloride

To a solution of 4-chloro-1H-indole-2-carboxylic acid (600 mg,3.07mmol,1 eq.) in DCM (9 mL) was added DMF (6.73 mg,92.02umol,7.08uL,0.03 eq.) and (COCl) ₂ (778.70 mg,6.13mmol,537.04uL,2 eq.) and stirred at 40℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give 4-chloro-1H-indole-2-carbonyl chloride (655 mg, crude) as a yellow oil.

Step 2:2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid

A mixture of 4-chloro-1H-indole-2-carbonyl chloride (650 mg,3.06mmol,1.1 eq.) in THF (6 mL) and DCM (6 mL) was poured into 2-azaspiro [ 4.5) ]Decane-3-carboxylic acid (611.20 mg,2.78mmol,1 eq. HCl), na ₂ CO ₃ (884.85 mg,2.78mmol,3 eq.) in DCM (6 mL) and H ₂ In a mixture of O (6 mL). At N ₂ The mixture was stirred at 15℃for 0.5h under an atmosphere. After completion ofThe reaction mixture was quenched by addition of HCl (1M) (15 mL) and extracted with DCM (10 mL x 4). The combined organic layers were concentrated under reduced pressure to give a residue. The crude product was wet-triturated with EtOAc (3 mL) for 15min at 20deg.C to give 2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (710 mg,1.97mmol,70.73% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺ 。

Step 3: (2S) -2- (2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (512.31 mg,2.16mmol,1.1 eq. HCl), 2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid (710 mg,1.97mmol,1 eq.) in DMF (15 mL) was added DIPEA (762.90 mg,5.90mmol,1.03mL,3 eq.) and HATU (748.17 mg,1.97mmol,1 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (40 mL) to quench the reaction mixture and then extract with ethyl acetate (20 mL x 4). The combined organic layers were washed with brine (20 ml x 1), dried over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives (2S) -2- [ [2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow oil]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,1.57mmol,79.55% yield).

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,1.57mmol,1 eq.) in MeOH/NH ₃ The solution in (7M, 11.05mL,49.42 eq.) was stirred at 65℃for 17h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a colorless oil]Methyl group]Ethyl group]-2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (820 mg, crude). MS (ESI) m/z 528.3[ M+H ]] ⁺ 。

Step 5:2- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (4-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (820 mg,1.55mmol,1 eq.) in DCM (15 mL) was added the Bungeus reagent (999.20 mg,4.19mmol,2.7 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (3 mL) was used to quench the reaction mixture and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,10 min) to give the desired compound as a white solid (450 mg), which was purified by SFC (column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%, min) to give 2- (4-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (168.83 mg,331.02umol,21.32% yield, 100% purity). MS (ESI) m/z 510.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.72(br s,1H)8.52-9.07(m,1H)6.72-7.49(m,5H)4.81-5.16(m,1H)4.43-4.78(m,1H)3.51-3.92(m,2H)2.10-2.39(m,3H)1.25-1.98(m,16H)。

Obtaining 2- (4-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (180.55 mg,354.00umol,22.80% yield, 100% purity). MS (ESI) m/z 510.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.56-11.85(m,1H)8.45-8.94(m,1H)7.43(br d,J＝8.16Hz,1H)7.04-7.35(m,3H)6.75-7.03(m,1H)4.42-5.12(m,2H)3.58-3.91(m,2H)2.06-2.30(m,3H)1.21-1.94(m,16H)。

EXAMPLE 91 Synthesis of viral protease inhibitor Compound 803

7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [ 3.4)]A solution of tert-butyl octane-6-carboxylate (1.2 g,2.47mmol,90% purity, 1 eq.) in HCl/MeOH (4M, 12mL,19.45 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (6-azaspiro [ 3.4) as a yellow solid]Octane-7-carboxamido) propionic acid methyl ester (1.3 g, crude material, HCl). MS (ESI) m/z 338.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- (6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (6-azaspiro [ 3.4)]To a solution of methyl octane-7-carboxamido) propionate (1.25 g,2.34mmol,70% purity, 1 eq, HCl) in DCM (8 mL) and DMF (4 mL) was added 4-chloro-1H-indole-2-carboxylic acid (457.78 mg,2.34mmol,1 eq), DMAP (857.77 mg,7.02mmol,3 eq) and then EDCI (897.29 mg,4.68mmol,2 eq). The mixture was then stirred at 25℃for 1h. After completion, the reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with dichloromethane: methanol=100:1 to 10:1 to give (2S) -2- (6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4) as a yellow solid]Octane-7-formylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (1.51 g,2.20mmol,93.96% yield, 75% purity). MS (ESI) M/z515.2[ M+H] ⁺ 。

Step 3: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- (6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4)]Octane-7-formylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (1.51 g,2.20mmol,75% purity, 1 eq.) in NH ₃ A solution in MeOH (7M, 15mL,47.75 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a yellow solid]Octane-7-carboxamide (1.5 g, crude). MS (ESI) m/z 500.3[ M+H ]] ⁺ 。

Step 4:6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4] octane-7-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -6- (4-chloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a solution of octane-7-carboxamide (1.5 g,2.10mmol,70% purity, 1 eq.) in EtOAc (8 mL) was added T3P (8.56 g,13.45mmol,8mL,50% purity, 6.41 eq.) and the reaction was then stirred at 40 ℃ for 14h. After completion, the mixture was quenched with water (25 mL) and extracted with EtOAc (15 mL x 3). The organic layer was washed with brine (15 mL), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,10 min) to give 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4] as a white solid]Octane-7-carboxamide (420 mg,865.32umol,41.20% yield, 99.3% purity). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

Step 5:6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4] octane-7-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O EtOH]；B％：60％-60％，min) isolation of 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4]Octane-7-carboxamide (420 mg,99.3% purity) to give 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4 ] as a white solid]Octane-7-carboxamide isomer 1 (8.72 mg,18.09umol,2.09% yield, 100% purity). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.49-7.34(m,1H),7.20(br t,J＝7.5Hz,1H),7.15-6.67(m,2H),5.15-5.00(m,1H),4.65-4.54(m,1H),4.15-3.78(m,2H),3.25-2.99(m,2H),2.58-1.25(m,15H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.96(br s,1H),9.06-8.67(m,1H),7.53(br d,J＝11.2Hz,1H),7.43(br d,J＝7.7Hz,1H),7.27-7.05(m,2H),7.04-6.54(m,1H),5.06-4.86(m,1H),4.57-4.36(m,1H),4.18-3.66(m,2H),3.08(br s,2H),2.37-2.11(m,4H),2.07-1.17(m,11H)。

To give 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4 ] as a white solid]Octane-7-carboxamide isomer 2 (197.12 mg,408.99umol,47.26% yield, 100% purity). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.46-7.34(m,1H),7.25-7.18(m,1H),7.17-6.65(m,2H),5.08-4.97(m,1H),4.58(t,J＝7.5Hz,1H),4.22-3.72(m,2H),3.24-2.87(m,2H),2.53-2.18(m,4H),2.13-1.75(m,8H),1.70-1.22(m,3H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.95(s,1H),9.30-8.55(m,1H),7.58-7.32(m,2H),7.31-7.07(m,2H),7.05-6.55(m,1H),5.16-4.85(m,1H),4.47(t,J＝7.2Hz,1H),4.13-3.68(m,2H),3.17-2.82(m,2H),2.34-2.10(m,4H),2.10-1.67(m,9H),1.63-1.01(m,2H)。

To give 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4 ] as a white solid]Octane-7-carboxamide isomer 3 (111.90 mg,232.17umol,26.83% yield, 100% purity). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.43-7.34(m,1H),7.22-7.16(m,1H),7.13-6.73(m,2H),5.10(dd,J＝5.7,10.3Hz,1H),4.57(t,J＝7.9Hz,1H),4.16-3.97(m,2H),3.27-3.19(m,2H),2.63-2.33(m,3H),2.30-2.19(m,1H),2.11-1.92(m,8H),1.85-1.68(m,2H),1.55-1.47(m,1H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.95(br s,1H),9.43-8.64(m,1H),7.63-7.33(m,2H),7.27-7.05(m,2H),7.04-6.56(m,1H),5.10-4.86(m,1H),4.46(br t,J＝7.4Hz,1H),4.08-3.60(m,2H),3.18-2.88(m,2H),2.36-2.09(m,4H),2.04-1.17(m,11H)。

To give 6- (4-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -6-azaspiro [3.4 ] as a white solid ]Octane-7-carboxamide isomer 4 (2.11 mg,4.24umol,0.49% yield, 96.8% purity). MS (ESI) m/z 482.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.44-7.36(m,1H),7.24-7.17(m,1H),7.16-6.69(mz,2H),5.21-5.01(m,1H),4.68-4.51(m,1H),4.12-3.81(m,2H),3.25-3.19(m,2H),2.56-2.15(m,3H),2.12-1.69(m,8H),1.64-1.26(m,4H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.96(br s,1H),9.02-8.65(m,1H),7.62-7.46(m,1H),7.45-7.34(m,1H),7.27-7.06(m,2H),7.04-6.57(m,1H),5.04-4.86(m,1H),4.57-4.37(m,1H),4.10-3.63(m,2H),3.17-2.83(m,2H),2.34-2.26(m,2H),2.23-2.10(m,2H),2.04-1.82(m,7H),1.80-1.37(m,2H),1.37-1.13(m,2H)。

EXAMPLE 92 Synthesis of viral protease inhibitor Compound 805

Step 1:2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxylic acid ethyl ester

To 8, 8-difluoro-2-azaspiro [4.5]]Dissolution of decane-3-carboxylic acid ethyl ester (1.5 g,6.07mmol,1 eq.) and 7-chloro-1H-indole-2-carboxylic acid (1.42 g,7.28mmol,1.2 eq.) in DCM (25 mL)DMAP (1.48 g,12.13mmol,2 eq.) and EDCI (2.33 g,12.13mmol,2 eq.) were added to the solution, followed by stirring the mixture at 20℃for 2h. After completion of the reaction, the mixture was quenched with water (20 mL) and extracted with DCM (10 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ Petroleum ether ethyl acetate=20:1 to 2.5:1) to give 2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] as a pink oil]Decane-3-carboxylic acid ethyl ester (1.6 g,3.58mmol,58.98% yield, 95% purity). MS (ESI) m/z 425.2[ M+H ]] ⁺ 。

Step 2:2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxylic acid

To 2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5 ]Decane-3-carboxylic acid ethyl ester (1.6 g,3.77mmol,1 eq.) in THF (12 mL) and H ₂ LiOH.H was added to the solution in O (6 mL) ₂ O (474.09 mg,11.30mmol,3 eq.) and then stirring the mixture at 20℃for 16h. After completion, the mixture was concentrated in vacuo and the pH was adjusted to pH = -1 with 1M HCl (30 mL). The reaction was wet-milled with DCM (30 mL) and then filtered and the filter cake dried in vacuo to give 2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (1.4 g,3.53mmol,93.69% yield).

Step 3:

(2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5]Decane-3-carboxylic acid (1.7 g,4.28mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.01 g,4.28mmol,1 eq., HCl) in DMF (30 mL) was added PyBOP (2.23 g,4.28mmol,1 eq.) and then TEA-containing DMF (5 mL) was added at-40 ℃. The mixture was stirred at-40℃for 2h. After completion of the reaction, the mixture was poured into water (100 mL) and extracted with DCM (40 mL x 3), followed by Na ₂ SO ₄ Dried and concentrated in vacuo and passed over a column (SiO ₂ Petroleum ether acetic acidEthyl ester=1:1 to DCM meoh=10:1) to give (2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [ 4.5) as a colorless solid]Decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (2.2 g,3.04mmol,70.95% yield, 80% purity). MS (ESI) m/z 579.3[ M+H ]] ⁺ 。

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [ 4.5)]Decane-3-formylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (400 mg,518.10umol,75% purity, 1 eq.) in NH ₃ A solution in MeOH (6 mL, 7M) was stirred at 30℃for 16h. After completion of the reaction, the mixture was concentrated in vacuo to give N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (1.4 g, batch 5, crude). MS (ESI) m/z 564.2[ M+H ]] ⁺

Step 5:2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -8, 8-difluoro-2-azaspiro [ 4.5)]To a solution of decane-3-carboxamide (1.4 g,2.48mmol,1 eq.) in DCM (30 mL) was added the Buerger's reagent (1.77 g,7.45mmol,3 eq.). The mixture was stirred at 30℃for 2h. After the reaction was completed, the reaction mixture was quenched with water (2 mL) and purified by blowing N ₂ To be dried. The concentrate was purified by preparative HPLC (column: welch Xtime C18X 70mM #10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,20 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (740 mg,1.33mmol,53.51% yield, 98% purity). MS (ESI) m/z 546.2[ M+H ]] ⁺

Step 6:2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [ Neu-ETOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,7 min) isolation of 2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5 ]Decane-3-carboxamide (740 mg,1.33mmol,53.51% yield, 98% purity) gave 2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide (isomer 1:340mg,622.70umol,45.95% yield, 100% purity). MS (ESI) m/z 546.1[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.67-7.47(m,1H),7.28(d,J＝7.6Hz,1H),7.18-6.82(m,2H),5.15-4.97(m,1H),4.82-4.58(m,1H),4.05-3.73(m,2H),3.27-2.92(m,2H),2.63-2.44(m,2H),2.39(dd,J＝7.7,12.5Hz,1H),2.07-1.72(m,11H),1.68-1.40(m,3H)。

2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -8, 8-difluoro-2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide (isomer 2:325mg,595.23umol,43.92% yield, 100% purity). MS (ESI) m/z 546.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.64(d,J＝8.0Hz,1H),7.28(d,J＝7.4Hz,1H),7.20-6.81(m,2H),5.02(dd,J＝6.2,10.1Hz,1H),4.66(dd,J＝7.9,9.4Hz,1H),4.08-3.81(m,2H),3.23-3.00(m,2H),2.55-2.23(m,3H),2.02-1.72(m,10H),1.71-1.59(m,3H),1.58-1.44(m,1H)。

EXAMPLE 93 Synthesis of viral protease inhibitor Compounds 806a

Step 1: (Z) -2-azido-3- (3-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester

A solution of NaOMe (1.90 g,35.17mmol,2 eq.) in MeOH (20 mL) was cooled to-10deg.C and a mixture of 3-chloro-2-methoxy-benzaldehyde (3 g,17.59mmol,1 eq.) and methyl azide acetate (4.12 g,35.17mmol,2 eq.) in MeOH (10 mL) was added dropwise. The mixture was stirred at 25℃for 3h. After completion, by adding H at 0deg.C ₂ O5 (0 mL) to quench the reaction mixture, and then H was used ₂ O (30 mL) was diluted and extracted with ethyl acetate (100 mL, extracted as 50mL x 2). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=20/1 to 10/1) to give (Z) -2-azido-3- (3-chloro-2-methoxy-phenyl) prop-2-enoic acid ester (2.1 g,7.45mmol,42.38% yield, 95% purity) as a yellow solid.

Step 2: 5-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

Dimethyl (20 mL) containing (Z) -2-azido-3- (3-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (2.1 g,7.85mmol,1 eq.) was stirred at 170 ℃ for 1.5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 5/1) the residue was purified to give the compound 5-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (1.7 g,6.38mmol,81.37% yield, 90% purity) as a white solid.

Step 3: 5-chloro-4-methoxy-1H-indole-2-carboxylic acid

To 5-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (1.2 g,5.01mmol,1 eq.) in THF (20 mL) and H ₂ LiOH.H was added to the mixture in O (10 mL) ₂ O (420.24 mg,10.01mmol,2 eq.). The mixture was stirred at 60℃for 2h. After completion, the pH of the reaction mixture was adjusted to ph=3 by addition of HCl, and then with H ₂ O (30 mL) dilution. The reaction was extracted with ethyl acetate (50 ml x 2). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 5-chloro-4-methoxy-propene as a white solid residue1H-indole-2-carboxylic acid (0.95 g,4.00mmol,79.88% yield, 95% purity).

Step 4: (2S) -2- [ [ (2S) -2- [ (5-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (4638 mg,1.29mmol,1 eq., HCl) and 5-chloro-4-methoxy-1H-indole-2-carboxylic acid (290.19 mg,1.29mmol,1 eq.) in DMF (10 mL) and DCM (20 mL) was added EDCI (493.11 mg,2.57mmol,2 eq.) and DMAP (314.25 mg,2.57mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (100 mL) was diluted and extracted with ethyl acetate (200 mL, extracted as 100mL x 2). The combined organic layers were washed with HCl (1M, 100 mL) and brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give the compound (2S) -2- [ [ (2S) -2- [ (5-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (600 mg,1.02mmol,79.35% yield, 91% purity). MS (ESI) m/z 535.2/537.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-methoxy-1H-benzimidazole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (5-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (450 mg,841.07umol,1 eq.) and NH ₃ The mixture of MeOH (7M, 15mL,124.84 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-5-chloro-4-methoxy-1H-indole-2-carboxamide (440 mg,719.20umol,85.51% Yield, 85% purity). MS (ESI) m/z 520.3[ M+H ]] ⁺

Step 6: 5-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a mixture of 5-chloro-4-methoxy-1H-indole-2-carboxamide (440 mg,846.12umol,1 eq.) in DCM (6 mL) was added the Bungeus reagent (604.92 mg,2.54mmol,3 eq.). The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and then extracted with DCM (20 mL x 2). The combined organic layers were concentrated and purified with N ₂ Blow-drying to obtain a residue. By neutral preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give 5-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (220 mg,430.07umol,50.83% yield, 98.134% purity). MS (ESI) m/z 502.2[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.85(br s,1H),8.96(d,J＝7.9Hz,1H),8.63(d,J＝8.1Hz,1H),7.67-7.38(m,2H),7.24-7.05(m,2H),5.16-4.92(m,1H),4.63-4.42(m,1H),4.11-4.02(m,3H),3.14-3.00(m,2H),2.36-2.17(m,2H),1.88-1.62(m,5H),1.59-1.29(m,2H),0.94(s,9H)

EXAMPLE 94 Synthesis of viral protease inhibitor Compound 808

Step 1: (Z) -2-azido-3- (2-chloro-3-methoxy-phenyl) prop-2-enoic acid methyl ester

A mixture of 2-chloro-3-methoxy-benzaldehyde (4 g,23.45mmol,1 eq.) and NaOMe (2.53 g,46.90mmol,2 eq.) with MeOH (20 mL) was cooled to-10deg.C, and then a mixture of methyl azide acetate (5.49 g,46.90mmol,2 eq.) in MeOH (50 mL) was added dropwise to the solution. The mixture was stirred at 25 ℃ for 16h and a white solid was observed. After completion, the reaction mixture was filtered to give the residual compound (Z) -2-azido-3- (2-chloro-3-methoxy-phenyl) prop-2-enoic acid methyl ester (3 g,10.09mmol,43.02% yield, 90% purity) as a white solid.

Step 2: 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (2-chloro-3-methoxy-phenyl) prop-2-enoic acid methyl ester (1 g,3.74mmol,1 eq.) in xylene (20 mL) was warmed to 170℃and stirred at 170℃for 1.5 h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was triturated with petroleum ether:ethyl acetate=5:1 at 25 ℃ to give 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester (450 mg,1.13mmol,30.16% yield, 60% purity) as a yellow solid.

Step 3: 4-chloro-5-methoxy-1H-indole-2-carboxylic acid

To 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester (450.00 mg,1.88mmol,1 eq.) in THF (10 mL) and H ₂ LiOH.H was added to the mixture in O (5 mL) ₂ O (157.59 mg,3.76mmol,2 eq.). The mixture was stirred at 60℃for 2h. After completion, the pH of the reaction mixture was adjusted to ph=3 by addition of HCl, and then with H ₂ O (30 mL) was diluted and extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the residual compound 4-chloro-5-methoxy-1H-indole-2-carboxylic acid (320 mg,992.78umol,52.87% yield, 70% purity) as a yellow solid.

Step 4: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (500 mg,1.17mmol,1 eq.) in HCl/MeOH (4M, 50.00mL,171.01 eq.) was stirred at 25℃for 1hr. After completion, the reaction mixture was concentrated under reduced pressure to give the residual compound methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (420 mg,1.15mmol,98.69% yield, HCl) as a white solid.

Step 5: (2S) -2- [ [ (2S) -2- [ (4-chloro-5-methoxy-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (420 mg,1.15mmol,1 eq, HCl) and 4-chloro-5-methoxy-1H-indole-2-carboxylic acid (260.43 mg,1.15mmol,1 eq) in DMF (10 mL) and DCM (20 mL) was added EDCI (442.53 mg,2.31mmol,2 eq) and DMAP (282.02 mg,2.31mmol,2 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was filtered and then taken up with H ₂ O (100 mL) was diluted and extracted with 300mL (150 mL. Times.2) ethyl acetate. The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-5-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (350 mg,588.75umol,51.01% yield, 90% purity). MS (ESI) m/z 535.3[ M+H ] ] ⁺

Step 6: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4-chloro-5-methoxy-1H-indole-2-carboxamide

In a sealed tube, (2S) -2- [ [ (2S) -2- [ (4-chloro-5-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (300.00 mg,560.72umol,1 eq.) and NH ₃ The mixture of MeOH (7M, 10mL,124 eq.) was stirred at 60℃for 16h. The reaction mixture was concentrated under reduced pressure to give the residual compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo- ] as a white solid1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-chloro-5-methoxy-1H-indole-2-carboxamide (290 mg,501.90umol,89.51% yield, 90% purity). MS (ESI) m/z 520.3[ M+H ]] ⁺

Step 7: 4-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-5-methoxy-1H-indole-2-carboxamide to N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a mixture of 4-chloro-5-methoxy-1H-indole-2-carboxamide (330 mg,634.59umol,1 eq.) in DCM (10 mL) was added the Bungeus reagent (453.69 mg,1.90mmol,3 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and extracted with DCM (40 mL, extracted as 20mL x 2). The combined organic layers were prepared by using N ₂ Blow-drying to concentrate to give a residue. By neutral preparative HPLC (column: waters Xbridge BEH C18 100. Times.30 mm. Times.10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the compound 4-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-5-methoxy-1H-indole-2-carboxamide (140 mg,276.09umol,43.51% yield, 99% purity). MS (ESI) m/z 502.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d6)δ＝11.74(s,1H),8.89(d,J＝8.1Hz,1H),8.68(d,J＝8.1Hz,1H),7.51(br s,1H),7.41-7.25(m,2H),7.13(d,J＝8.9Hz,1H),5.12-4.96(m,1H),4.52(dt,J＝3.8,8.4Hz,1H),3.91-3.76(m,3H),3.14-2.95(m,2H),2.37-2.13(m,2H),1.90-1.29(m,7H),1.01-0.81(m,9H)

EXAMPLE 95 Synthesis of viral protease inhibitor Compound 810

Step 1: 7-chloro-5-methoxy-1H-indole

at-40deg.C, to 2-chloro-4-methoxy-1-nitro-benzeneTo a solution of (4300 mg,22.92mmol,1 eq.) in THF (70 mL) was added magnesium bromide (1M, 80.23mL,3.5 eq.). The solution was stirred at-40℃for 2h. After completion, the solution was poured into NH ₄ Cl (200 mL) and concentrated, and extracted with ethyl acetate (80 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ Petroleum ether ethyl acetate=30:1 to 10:1) to afford the product 7-chloro-5-methoxy-1H-indole as a brown oil (1100 mg,11.56mmol,50.44% yield). MS (ESI) m/z 182.1[ M+H ] ] ⁺ 。

Step 2: 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole

To a solution of 7-chloro-5-methoxy-1H-indole (2100 mg,11.56mmol,1 eq.) in DMF (25 mL) was added NaH (739.94 mg,18.50mmol,60% purity, 1.6 eq.) at 0deg.C. The solution was stirred at 20℃for 0.5h. 4-Methylbenzenesulfonyl chloride (2.09 g,10.98mmol,0.95 eq.) was added and the solution stirred at 20℃for 1.5h. After completion, the solution is treated with H ₂ O (60 mL) was diluted and extracted with ethyl acetate (60 mL x 3) and then washed with brine (60 mL x 2) and concentrated to give the crude material. Through the column (SiO) ₂ Petroleum ether ethyl acetate=30:1 to 2:1) to afford 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole (280mg, 8.34mmol,72.11% yield) as a brown solid. MS (ESI) m/z 336.3[ M+H ]] ⁺

Step 3: 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole-2-carboxylic acid

To a solution of 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole (280mg, 8.34mmol,1 eq.) in THF (40 mL) at-70 ℃ was added LDA (1 m,16.68mL,2 eq.) and the solution stirred at-70 ℃ for 2.5h. After completion, the solution was poured quickly into dry ice and dried with H ₂ O (80 mL) was diluted and the solution was concentrated and extracted with ethyl acetate (80 mL) to recycle reactant 3. The aqueous layer was acidified with HCl (concentrate) to ph=5-6 and extracted with ethyl acetate (90 ml x 2) and Na ₂ SO ₄ Drying and concentration gave crude 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole-2-carboxylic acid (2300 mg, crude material) as a brown solid. The crude material was used directly in the next stepIs a kind of medium. MS (ESI) m/z 380.2[ M+H ]] ⁺

Step 4: 7-chloro-5-methoxy-1H-indole-2-carboxylic acid

A solution of 7-chloro-5-methoxy-1- (p-toluenesulfonyl) indole-2-carboxylic acid (210mg, 5.53mmol,1 eq.) and KOH (682.51 mg,12.16mmol,14.41uL,2.2 eq.) in MeOH (30 mL) was stirred at 70℃for 8h. After completion, the solution was concentrated and taken up with H ₂ O (40 mL) was diluted and acidified with HCl (1M) to ph=5-6 and the filter cake was filtered and collected to give 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (570 mg, crude material) as a brown solid. The crude material was used directly in the next step. MS (ESI) m/z 226.3[ M+H ]] ⁺

Step 5: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester; hydrochloride salt

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (500 mg,1.17mmol,1 eq.) in HCl/MeOH (20 mL) was stirred at 25℃for 1h. After completion, the solution was concentrated to give the crude product (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl as an off-white solid ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate; hydrochloride (420 mg, crude material). The crude material was used directly in the next step. MS (ESI) m/z 364.3[ M+H ]] ⁺

Step 6: (2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate; to a solution of hydrochloride (420 mg,1.15mmol,1 eq.) and DMAP (282.02 mg,2.31mmol,2 eq.) in DCM (20 mL) and DMF (10 mL) was added 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (299.49 mg,1.33mmol,1.15 eq.) and EDCI (442.54 mg,2.31mmol,2 eq.). The reaction was stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (40 mL) was diluted, extracted with ethyl acetate (50 mL x 3) and washed with brine (80 mL x 2) and concentrated to give the crude product. Through the column (SiO) ₂ The crude material was purified with ethyl acetate meoh=1:0 to 10:1 to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (370 mg,691.55umol,59.91% yield). MS (ESI) m/z 535.3[ M+H ] ] ⁺

Step 7: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -7-chloro-5-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (370 mg,691.55umol,1 eq.) in NH ₃ A solution in MeOH (7M, 16.44mL,166.45 eq.) was stirred at 60℃for 25h. After completion, the solution was concentrated to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as an off-white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-7-chloro-5-methoxy-1H-indole-2-carboxamide (350 mg, crude). The crude material was used directly in the next step. MS (ESI) M/z520.3[ M+H] ⁺

Step 8: 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -5-methoxy-1H-indole-2-carboxamide

N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]A solution of 7-chloro-5-methoxy-1H-indole-2-carboxamide (350 mg,673.05umol,1 eq.) and Prague reagent (641.57 mg,2.69mmol,4 eq.) in DCM (20 mL) was stirred at 25℃for 2H. After completion, the solution was washed with brine (30 ml x 2) and with N ₂ Blow-drying to obtain a crude product. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mm X10 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-5-methoxy-1H-indole-2-carboxamide (100 mg,199.20umol,29.60% yield). MS (ESI) m/z 502.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(br s,1H),9.11-8.94(m,1H),8.64(br d,J＝8.4Hz,1H),7.52(br s,1H),7.17-7.08(m,2H),6.98(d,J＝2.0Hz,1H),5.27-4.92(m,1H),4.69-4.37(m,1H),3.76(s,3H),3.05(br s,2H),2.30-2.16(m,2H),2.06(s,1H),1.83-1.66(m,4H),1.57-1.32(m,2H)

EXAMPLE 96 Synthesis of viral protease inhibitor Compound 812

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1 g,4.22mmol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (968.64 mg,4.22mmol,1 eq.) TEA (1.28 g,12.67mmol,1.76mL,3 eq.) in DCM (15 mL) was added T3P (8.07 g,12.67mmol,7.54mL,50% purity, 3 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H at 0deg.C ₂ O (20 mL) to quench the reaction mixture, the combined organic layers were washed with DCM (10 mL x 3), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=2/1 to 0/1 to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.08 g,2.05mmol,48.46% yield, 78% purity). MS (ESI) m/z 413.2[ M+H ]] ⁺ 。

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxoSubstituted-3-piperidyl]A solution of methyl propionate (1.04 g,2.53mmol,1 eq.) in HCl/MeOH (15 mL) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (879 mg, crude material, HCl). MS (ESI) m/z 313.2[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (879 mg,2.82mmol,1 eq.) and 4-chloro-1H-indole-2-carboxylic acid (552.18 mg,2.82mmol,1 eq.) TEA (856.96 mg,8.47mmol,1.18mL,3 eq.) in DCM (10 mL) was added T3P (5.39 g,8.47mmol,5.04mL,50% purity, 3 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H at 0deg.C ₂ O (20 mL) to quench the reaction mixture, the combined organic layers were washed with DCM (10 mL x 3), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=2/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (636 mg,1.04mmol,36.86% yield, 80% purity). MS (ESI) m/z 489.2[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-1H-indole-2-carbonyl) amino group ]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (621 mg,1.27mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 5mL,27.56 eq.) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino ] as a yellow solid-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-1H-indole-2-carboxamide (460 mg, crude). MS (ESI) m/z 474.2[ M+H ]] ⁺ 。

Step 5: 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 4-chloro-1H-indole-2-carboxamide (440 mg,928.37 mmole, 1 eq.) in DCM (8 mL) was added the Buerger's reagent (663.70 mg,2.79mmol,3 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give 4-chloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (105.2 mg,229.35umol,24.70% yield, 99.4% purity). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.95(s,1H),8.92(d,J＝8.4Hz,1H),8.81-8.70(m,1H),7.55-7.49(m,1H),7.44-7.37(m,2H),7.24-7.07(m,2H),5.14-5.00(m,1H),4.54-4.40(m,1H),3.18-2.99(m,2H),2.31-2.21(m,2H),1.93-1.66(m,4H),1.61-1.34(m,3H),0.89-0.76(m,1H),0.52-0.33(m,2H),0.24-0.04(m,2H)

Example 97 Synthesis of viral protease inhibitor Compound 814

Step 1: (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (6-azaspiro [3.4] octane-7-carboxamido) propionic acid methyl ester

A solution of 7- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (1.4 g,3.31mmol,1 eq.) in HCl/MeOH (4M, 14 mL) was stirred at 20deg.C for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (6-azaspiro [3.4] octane-7-carboxamido) propionate (1.29 g, crude material) as a pale yellow solid.

Step 2: (2S) -2- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (6-azaspiro [ 3.4)]To a solution of methyl octane-7-carboxamido) propionate (1.14 g,2.22mmol,70% purity, 1 eq, HCl) in DCM (25 mL) was added 6, 7-dichloro-1H-indole-2-carboxylic acid (612.19 mg,2.66mmol,1.2 eq) and DMAP (541.85 mg,4.44mmol,2 eq) and EDCI (850.23 mg,4.44mmol,2 eq). The mixture was stirred at 20℃for 2h. After completion of the reaction, the residue was poured into water (60 mL) and extracted with DCM (20 mL x 3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ ，DCM:MeOH＝10:1，R _f =0.35) to give (2S) -2- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4) as a pale yellow solid]Octane-7-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (800 mg,1.48mmol,66.70% yield, 99% purity). MS (ESI) m/z 535.2[ M+H ]] ⁺

Step 3: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4)]Octane-7-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (270 mg,504.28umol,1 eq.) in NH ₃ A solution in MeOH (6 mL, 7M) was stirred at 30℃for 20h. After completion of the reaction, the mixture was concentrated in vacuo to give N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a pale yellow solid]Octane-7-carboxamide (800 mg, crude). MS (ESI) m/z 520.2[ M+H ]] ⁺

Step 4: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a solution of octane-7-carboxamide (800 mg,1.54mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (1.10 g,4.61mmol,3 eq.) and the mixture stirred at 30℃for 4h. After the reaction was completed, the mixture was treated with H ₂ Quenched by O (2 mL) and blown N ₂ Dried and purified by preparative HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a white solid]Octane-7-carboxamide (380 mg,756.38umol,49.20% yield). MS (ESI) m/z 502.2[ M+H ]] ⁺ 。

Step 5: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

By SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O MEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%,15 min) isolation of N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4) ]Octane-7-carboxamide, N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [3.4 ] as a white solid]Octane-7-carboxamide (isomer 1:110mg,218.95umol,28.95% yield, 100% purity). MS (ESI) m/z 502.1[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.62(d,J＝8.6Hz,1H),7.23(d,J＝8.6Hz,1H),7.17(s,1H),5.01(dd,J＝5.8,10.3Hz,1H),4.58(t,J＝7.6Hz,1H),4.08-3.80(m,2H),3.15-2.58(m,1H),2.55-2.15(m,5H),2.11-1.74(m,9H)。

In the case of preparative HPLC (column Waters Xbridge BEH C18 100X 25mm 5um; mobile phase): [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,10 min) to give N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6- (6, 7-dichloro-1H-indole-2-carbonyl) -6-azaspiro [ 3.4) as a white solid]Octane-7-carboxamide (isomer 2:85mg,169.19umol,22.37% yield, 100% purity). MS (ESI) m/z 502.1[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δppm 7.61(d,J＝8.6Hz,1H),7.24-7.15(m,1H),7.13(s,1H),5.09-4.90(m,1H),4.78-4.51(m,1H),4.06-3.72(m,2H),2.83-2.63(m,1H),2.61-2.28(m,3H),2.22-1.76(m,10H),1.72-1.40(m,1H)。

EXAMPLE 98 Synthesis of viral protease inhibitor Compound 171

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (0.55 g,1.92mmol,1 eq.) and HCl/EtOAc (4M, 10mL,20.82 eq.) was stirred at 25℃for 0.5h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (0.35 g, crude material) as a yellow oil.

A mixture of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (0.15 g, 805.55. Mu. Mol,1 eq), (2S, 4S) -1-tert-butoxycarbonyl-4-phenyl-pyrrolidine-2-carboxylic acid (234.69 mg, 805.55. Mu. Mol,1 eq), DMAP (196.83 mg,1.61mmol,2 eq), EDCI (308.85 mg,1.61mmol,2 eq) in DMF (1 mL) and DCM (2 mL) was stirred at 25℃for 0.5h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtering andconcentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=2:1 to 0:1) to give (2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]Carbamoyl group]-4-phenyl-pyrrolidine-1-carboxylic acid tert-butyl ester (0.25 g,500.51umol,62.13% yield, 92% purity). MS (ESI) m/z 460.1[ M+H ]] ⁺ 。

(2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A mixture of tert-butyl 4-phenyl-pyrrolidine-1-carboxylate (0.25 g,544.03umol,1 eq.) and HCl/EtOAc (4M, 10mL,73.53 eq.) was stirred at 25℃for 0.5h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]Methyl propionate (0.2 g, crude material). MS (ESI) m/z 360.1[ M+H ]] ⁺ 。

(2S) -3- [ (3S) -2-oxopyrrolidin-3-yl]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]A mixture of methyl propionate (0.17 g,472.99 mol,1 eq), (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid (94.88 mg,472.99 mol,1 eq), T3P (451.48 mg,709.48 mol,421.95uL,50% purity, 1.5 eq), TEA (143.58 mg,1.42mmol,197.50uL,3 eq) in DMF (4 mL) was degassed and stirred at 25℃for 0.5h. After completion, the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=2:1 to 0:1) the residue was purified to give (2S) -2- [ [ (2S, 4S) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a white solid]-4-phenyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.11 g,162.36umol,34.33% yield, 80% purity). MS (ESI) m/z 542.1[ M+H ]] ⁺ 。

In a sealed tube, (2S) -2- [ [ (2S, 4S) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]-4-phenyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.1 g,184.50umol,1 eq.) in NH ₃ The mixture in MeOH (3 mL, 7M) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow oil]Methyl group]Ethyl group]-1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]-4-phenyl-pyrrolidine-2-carboxamide (0.09 g, crude material). MS (ESI) m/z 527.0[ M+H ] ] ⁺ 。

To (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl]To a solution of 4-phenyl-pyrrolidine-2-carboxamide (0.09 g,170.78umol,1 eq.) in DCM (1 mL) was added the berg reagent (203.50 mg,853.91umol,5 eq.) and the solution was then stirred at 25 ℃ for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give (2 s,4 s) -1- [ (E) -3- (4-chloro-2-fluoro-phenyl) prop-2-enoyl as a white solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-4-phenyl-pyrrolidine-2-carboxamide (29.73 mg,56.89umol,33.31% yield, 97.4% purity). MS (ESI) m/z 509.1[ M+H ]] ⁺ 。

EXAMPLE 99 Synthesis of viral protease inhibitor Compound 253

Step 1: 2-amino-3- (2-oxo-1, 2-dihydropyridin-3-yl) propionic acid methyl ester

A mixture of 2-amino-3- (2-oxo-1H-pyridin-3-yl) propionic acid (500 mg,2.74mmol,1 eq.) and HCl/MeOH (4M, 30mL,43.72 eq.) was stirred at 25℃for 2H. The reaction mixture was concentrated under reduced pressure to give methyl 2-amino-3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (650 mg, crude material, HCl) as a yellow oil and used directly in the next step. MS (ESI) m/z 197.0[ M+H ]] ⁺

A mixture of methyl 2-amino-3- (2-oxo-1H-pyridin-3-yl) propionate (650 mg,2.79mmol,1 eq., HCl), (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoic acid (646.16 mg,2.79mmol,1 eq.), EDCI (1.07 g,5.59mmol,2 eq.), DMAP (682.62 mg,5.59mmol,2 eq.), DMF (2 mL), and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the product methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (900 mg,1.89mmol,67.68% yield, 86.02% purity) as a white solid. MS (ESI) m/z 410.1[M+H] ⁺

A mixture of methyl-2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-methylpentanoylamino) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propionate (200 mg,488.43umol,1 eq) and HCl/EtOAc (4M, 30 mL) was stirred at 27℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give methyl 2- ((S) -2-amino-4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (170 mg, crude material, HCl) as a white solid and used directly in the next step.

A mixture of methyl 2- ((S) -2-amino-4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (170 mg,491.58 mol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (93.98 mg,491.58 mol,1 eq.), EDCI (188.47 mg,983.17 mol,2 eq.), DMAP (120.11 mg,983.17 mol,2 eq.), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give methyl 2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (130 mg,269.41umol,54.81% yield) as a white solid. MS (ESI) m/z 483.1[ M+H ]] ⁺

Methyl 2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- (2-oxo-1, 2-dihydropyridin-3-yl) propanoate (190 mg,393.76umol,1 eq.) NH ₃ /MeThe mixture of OH (7M, 10 mL) was stirred at 80℃for 15h. The reaction mixture was concentrated under reduced pressure to give the residue N- ((2S) -1- ((1-amino-1-oxo-3- (2-oxo-1, 2-dihydropyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (190 mg, crude material) as a yellow solid. MS (ESI) m/z 468.2[ M+H ]] ⁺

A mixture of N- ((2S) -1- ((1-amino-1-oxo-3- (2-oxo-1, 2-dihydropyridin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (180 mg,385.01umol,1 eq), pragus reagent (917.53 mg,3.85mmol,10 eq) and DCM (30 mL) was stirred at 25℃for 8H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8 min) to give the product N- ((2S) -1- ((1-cyano-2- (2-oxo-1, 2-dihydropyridin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (24 mg,52.18umol,13.55% yield, 97.73% purity) as a yellow solid. MS (ESI) m/z 450.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.90-11.40(m,2H),9.08-8.85(m,1H),8.55-8.35(m,1H),7.51-7.26(m,3H),7.16-7.05(m,1H),7.04-6.94(m,1H),6.51(d,J＝7.5Hz,1H),6.15(t,J＝6.6Hz,1H),5.19-5.01(m,1H),4.55-4.33(m,1H),3.89(s,3H),3.02-2.78(m,2H),1.75-1.33(m,3H),0.98-0.72(m,6H)。

Example 100 Synthesis of viral protease inhibitor Compounds 267 and 267A

Step 1:3- [2- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -1H-pyrrole-2-carboxylic acid tert-butyl ester

To tert-butyl-but-3-ynyloxy-dimethyl-silane (5.00 g,27.10mmol,1.5 eq.) and Ag ₂ CO ₃ To a solution of (498 mg,1.81mmol,0.1 eq.) in dioxane (8 mL) was added tert-butyl 2-isocyanoacetate (2.55 g,18.06mmol,2.63mL,1 eq.). Next, the mixture was stirred at 80℃for 1hr. TLC (petroleum ether/ethyl acetate=10/1, uv) showed complete exhaustion of starting material and formation of new spots. The reaction mixture was filtered and the filtrate concentrated in vacuo. Through flash silica gel chromatography 25g/>Silica flash column, eluent: 0-10% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining 3- [2- [ tert-butyl (dimethyl) silyl ] as a white solid]Oxyethyl radical]-1H-pyrrole-2-carboxylic acid tert-butyl ester (2.5 g,42.5% yield).

Step 2:3- (2-hydroxyethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester

At 0℃to 3- [2- [ tert-butyl (dimethyl) silyl ]]Oxyethyl radical]To a solution of tert-butyl-1H-pyrrole-2-carboxylate (2.5 g,7.68mmol,1 eq.) in THF (20 mL) was added TBAF (1M, 15.3mL,2 eq.) and the mixture was then stirred at 25℃for 16hr. TLC (petroleum ether/ethyl acetate=5/1, uv) showed complete exhaustion of starting material and formation of new spots. The reaction mixture was concentrated in vacuo. Through flash silica gel chromatography40g/>Silica flash column, eluent: 0-10% DCM/MeOH at 30 mL/min). 3- (2-hydroxyethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester (1.3 g,80.1% yield) was obtained as a colorless oil.

Step 3:3- (2-Oxoethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester

To a solution of 3- (2-hydroxyethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester (1.15 g,5.44mmol,1 eq.) in DCM (20 mL) was added DMP (3.23 g,7.62mmol,1.4 eq.) and the mixture stirred at 25℃for 1hr. LCMS showed starting material remained and-60% of the desired product was detected. TLC (petroleum ether/ethyl acetate=5/1, uv) showed complete exhaustion of starting material and formation of new spots. The reaction mixture was filtered and the filtrate concentrated in vacuo. The residue was diluted with ethyl acetate (50 mL) and taken up in H ₂ O (10 mL), brine (10 mL) and washed with Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography25g />Silica flash column, eluent: 0-20% ethyl acetate/petroleum ether gradient at 30 mL/min). 3- (2-oxoethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester (1.5 g,65.8% yield) was obtained as a colorless oil.

¹ H NMR(400MHz,CDCl ₃ )δ9.80-9.60(m,1H),9.48(br s,1H),6.91(t,J＝2.76Hz,1H),6.16(t,J＝2.51Hz,1H),3.82(d,J＝1.76Hz,2H,),1.56(s,9H)。

Step 4:3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] amino ] ethyl ] -1H-pyrrole-2-carboxylic acid tert-butyl ester

A solution of 3- (2-oxoethyl) -1H-pyrrole-2-carboxylic acid tert-butyl ester (1.5 g,7.17mmol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionic acid methyl ester (1.29 g,7.17mmol,1 eq., HCl) in MeOH (20 mL) was stirred at 25℃for 0.5hr. Next, naBH is added to the mixture ₃ CN (900.9 mg,14.34mmol,2 eq.) and the resulting solution was stirred at 25℃for 16hr. LCMS showed complete depletion of starting material and 40% of the desired product was detected. TLC (petroleum ether/ethyl acetate=5/1, uv) showed complete exhaustion of starting material and formation of new spots. H for the reaction mixture ₂ O (10 mL) was quenched and extracted with ethyl acetate (15 mL. Times.3). The combined organic phasesBy H ₂ O (10 mL) and brine (10 mL. Times.2), washed with Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography25g/>Silica flash column, eluent: 0-20% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining 3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] as a colourless oil]Amino group]Ethyl group]-1H-pyrrole-2-carboxylic acid tert-butyl ester (0.6 g,24.8% yield).

¹ H NMR(400MHz,CDCl ₃ )δ8.98(br s,1H),6.91-6.65(m,1H),6.15(t,J＝2.56Hz,1H),3.71(s,3H),3.40(t,J＝6.69Hz,1H),2.99-2.92(m,2H),2.82-2.90(m,1H),2.78-2.69(m,1H),1.68-1.63(m,1H),1.57(s,9H),1.50-1.42(m,1H),0.76-0.66(m,1H),0.48-0.36(m,2H),0.11-0.01(m,2H)。

Step 5:3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] amino ] ethyl ] -1H-pyrrole-2-carboxylic acid

To a solution of 3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] amino ] ethyl ] -1H-pyrrole-2-carboxylic acid tert-butyl ester (0.2 g,0.59mmol,1 eq.) in dioxane (1 mL) was added HCl/dioxane (4 m,1.49mL,10 eq.) and the mixture stirred at 25 ℃ for 16hr. LCMS showed complete depletion of starting material and 88% of the desired product was detected. The reaction mixture was concentrated in vacuo. The crude product was used directly in the next step. 3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] amino ] ethyl ] -1H-pyrrole-2-carboxylic acid (0.15 g,90% yield) was obtained as a dark brown oil.

Step 6: (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2,3-c ] pyridin-6-yl) propionic acid methyl ester

To 3- [2- [ [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ]]Amino group]Ethyl group]To a solution of 1H-pyrrole-2-carboxylic acid (150 mg,0.53mmol,1 eq.) in DMF (1 mL) was added HOBt (108.4 mg,0.802mmol,1.5 gAmount), DIEA (207.4 mg,1.61mmol,0.28ml,3 eq.) and EDCI (153.8 mg,0.80mmol,1.5 eq.). The mixture was stirred at 25℃for 16hr. LCMS showed complete depletion of starting material and 45% of the desired product was detected. TLC (petroleum ether/ethyl acetate=2/1, uv) showed complete exhaustion of starting material and formation of new spots. H for the reaction mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (20 mL x 3). Combined organic phases with H ₂ O (10 mL), brine (10 mL) and washed with Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography24gSilica flash column, eluent: 0-50% ethyl acetate/petroleum ether gradient at 30 mL/min). (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c) was obtained as a colourless oil]Methyl pyridin-6-yl) propionate (85 mg,58.1% yield).

LCMS：Rt＝0.773min；C ₁₄ H ₁₈ N ₂ O ₃ MS calculated: 262.13; MS experimental values: 263.0[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ6.97-6.85(m,1H),6.04(d,J＝2.26Hz,1H,),5.09(dd,J＝10.26,5.27Hz,1H),3.71(s,3H),3.67-3.58(m,2H),2.93-2.74(m,2H),2.02-1.87(m,1H),1.81-1.70(m,1H),0.83-0.68(m,1H),0.56-0.39(m,2H),0.22-0.07(m,2H)。

Step 7: (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2,3-c ] pyridin-6-yl) propionic acid

To (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c)]To a solution of methyl pyridin-6-yl) propionate (60 mg,0.228mmol,1 eq.) in MeOH (2 mL) was added K-containing ₂ CO ₃ (94.8 mg,0.686mmol,3 eq.) H ₂ O (1 mL) and the mixture was stirred at 25℃for 16hr. LCMS showed complete depletion of starting material and detection of 100% of the desired product. H for the reaction mixture ₂ O (5 mL) dilution with 0.5M aqueous HClAdjust to ph=3 and extract with ethyl acetate (15 ml x 3). Combined organic phases with H ₂ O (5 mL), brine (5 mL) and washed with Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was used directly in the next step. Obtaining (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c) as a white solid]Pyridin-6-yl) propionic acid.

LCMS：Rt＝0.706min；C ₁₃ H ₁₆ N ₂ O ₃ MS calculated: 248.12; MS experimental values: 248.9[ M+H ] ⁺ ]。

267A: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2,3-c ] pyridin-6-yl) propanamide

267: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2,3-c ] pyridin-6-yl) propanamide

To (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2,3-c ] at 25 ℃ ]Pyridin-6-yl) propionic acid (40 mg,0.16mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of propionitrile (30.5 mg,0.16mmol,1 eq, HCl) in DMF (1 mL) was added TEA (32.6 mg,0.32mmol,44uL,2 eq) and T ₃ P (153.7 mg,0.241mmol,0.14mL,50% purity, 1.5 eq.) and the mixture was stirred at 25℃for 1hr. LCMS showed complete depletion of starting material and 86% of the desired product was detected. The reaction mixture was concentrated in vacuo. The residue was checked by HPLC and purified by preparative HPLC (column Phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ water (0.05% NH ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,7.8 min). Obtaining (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c)]Pyridin-6-yl) propionamide (22 mg,35.6% yield).

By chiral SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um): mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -30%, min) purifying the crude productThe product is obtained. Obtaining (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c) ]Pyridin-6-yl) propionamide (2.0 mg,8.7% yield) and (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl were obtained as a white solid]Ethyl group]-3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c)]Pyridin-6-yl) propionamide (15.1 mg,68.2% yield).

267A：

LCMS：Rt＝0.746min；C ₂₀ H ₂₅ N ₅ O ₃ MS calculated: 383.20; MS experimental values: 384.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ6.91(d,J＝2.50Hz,1H)。6.04(d,J＝2.38Hz,1H),5.15(dd,J＝8.44,6.94Hz,1H),5.04(br d,J＝6.75Hz,1H),3.66-3.55(m,2H),3.33(br s,2H),2.88-2.76(m,2H),2.55-2.42(m,1H),2.39-2.23(m,2H),1.96-1.83(m,2H),1.82-1.74(m,2H),0.70(br s,1H),0.46(t,J＝7.88Hz,2H),0.15(d,J＝4.38Hz,2H)。

267：

LCMS：Rt＝0.751min；C ₂₀ H ₂₅ N ₅ O ₃ MS calculated: 383.20; MS experimental values: 384.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ6.90(d,J＝2.38Hz,1H),6.03(d,J＝2.25Hz,1H),5.02(dd,J＝10.13,6.63Hz,2H),3.66(tq,J＝13.12,6.34Hz,2H),3.30-3.18(m,2H),2.80(br t,J＝6.19Hz,2H),2.59-2.44(m,1H),2.37-2.21(m,2H),1.97-1.69(m,4H),0.78-0.67(m,1H),0.60-0.42(m,2H),0.17(d,J＝4.50Hz,2H)。

EXAMPLE 101 Synthesis of viral protease inhibitor Compounds 481 and 269A

Step 1: 4-chloro-1H-imidazo [4,5-c ] pyridine

2-chloropyridine-3, 4-diamine (3 g,20.90mmol,1 eq.) and HCl (2.06 g,20.90mmol,2.0mL,37% purity, 1 eq.) were combined in diethoxyA mixture in methoxyethane (30.9 g,208.95mmol,34.7mL,10 eq.) was degassed and treated with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 25℃for 12hr under an atmosphere. The precipitate formed was filtered off and washed with PE. No purification was performed. Obtaining the compound 4-chloro-1H-imidazo [4,5-c ] as a white solid]Pyridine (3 g,93.4% yield).

Step 2:1, 5-Dihydroimidazo [4,5-c ] pyridin-4-one

A solution of 4-chloro-1H-imidazo [4,5-c ] pyridine (3 g,19.54mmol,1 eq.) and HCl (1.9 g,19.54mmol,1.8mL,37% purity, 1 eq.) in MeOH (10 mL). The mixture was stirred at 50deg.C for 30hr. The reaction mixture was concentrated under reduced pressure to remove HCl/MeOH. The crude product was wet-milled with PE at 25℃for 150min. The compound 1, 5-dihydroimidazo [4,5-c ] pyridin-4-one (2.5 g, crude material) was obtained as a yellow solid.

Step 3:3- (2-trimethylsilylethoxymethyl) -5H-imidazo [4,5-c ] pyridin-4-one

To 1, 5-dihydroimidazo [4,5-c ]]To a solution of pyridin-4-one (2.5 g,18.50mmol,1 eq.) and SEM-Cl (3.0 g,18.50mmol,3.2mL,1 eq.) in THF (1 mL) was added NaH (2.2 g,55.50mmol,60% purity, 3 eq.). The mixture was stirred at 25℃for 2hr. TLC (petroleum ether/ethyl acetate=0:1, uv 254) indicated starting material remained and a new spot formed. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography25g/>Silica flash column, eluent: 0-100% petroleum ether/ethyl acetate at 35 mL/min). Obtaining the compound 3- (2-trimethylsilylethoxymethyl) -5H-imidazo [4,5-c ] as a yellow solid]Pyridin-4-one (1.8 g,32.2% yield, 88% purity).

Step 4: 3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ] pyridin-5-yl ] propionic acid methyl ester

3- (2-trimethylsilylethoxymethyl) -5H-imidazo [4,5-c ]To a solution of pyridin-4-one (1.5 g,5.65mmol,1 eq.) and methyl (2R) -2-bromo-3-cyclopropyl-propionate (1.1 g,5.65mmol,1 eq.) in DMF (4 mL) was added K ₂ CO ₃ (1.5 g,11.30mmol,2 eq.). The mixture was stirred at 25℃for 16hr. TLC (petroleum ether/ethyl acetate=3:1, uv 254) indicated starting material remained and a new spot formed. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-40% petroleum ether/ethyl acetate at 35 mL/min). Obtaining the compound 3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ] as a white solid]Pyridin-5-yl]Methyl propionate (865 mg,36.7% yield, 94% purity).

Step 5: 3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ] pyridin-5-yl ] propionic acid

3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c]Pyridin-5-yl]Methyl propionate (865 mg,2.21mmol,1 eq.) in H ₂ LiOH.H was added to a solution of O (1 mL) and THF (1 mL) ₂ O (185.4 mg,4.42mmol,2 eq.). The mixture was stirred at 25℃for 16hr. The reaction mixture was concentrated under reduced pressure to remove THF. Residue with H ₂ O (2 mL) was diluted and HCl (2 mL, 2N) was added. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2)By anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The crude product was wet-milled with PE at 25℃for 60min. Obtaining the compound 3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ] as a white solid]Pyridin-5-yl]Propionic acid (746 mg,86.7% yield, 97% purity).

481 (N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl)]Ethyl group]-3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ]]Pyridin-5-yl]Acrylamide): (2S) -3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c]Pyridin-5-yl]Propionic acid (600 mg,1.59mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of propionitrile (301.4 mg,1.59mmol,1 eq., HCl), HATU (604.3 mg,1.59mmol,1 eq.), DIPEA (410.8 mg,3.18mmol,0.55mL,2 eq.) in DCM (1 mL) was degassed and taken up with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 25℃for 1hr under an atmosphere. TLC (petroleum ether/ethyl acetate=3:1, uv 254) indicated starting material remained and a new spot formed. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-45% petroleum ether/ethyl acetate at 35 mL/min). Obtaining the compound N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [ 4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c ]]Pyridin-5-yl]Propionamide (640 mg,66.4% yield, 84% purity). />

269A: to N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-2- [4-oxo-3- (2-trimethylsilylethoxymethyl) imidazo [4,5-c]Pyridin-5-yl]To a solution of propionamide (600 mg,1.17mmol,1 eq.) in THF (1 mL) was added TBAF (1 m,2.3mL,2 eq.). The mixture was stirred at 60℃for 2hr. The reaction mixture was concentrated under reduced pressure to remove THF. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:5% -35%,9.5 min) of the purified residue. Obtaining compound C as a white solid ₁₉ H ₂₂ N ₆ O ₃ (34 mg,7.6% yield, 100% purity).

Purification of (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl by SFC]Ethyl group]-3-cyclopropyl-2- (4-oxo-3H-imidazo [4, 5-c)]Pyridin-5-yl) propionamide (34 mg,88.9umol,1 eq). By preparative HPLC (column (s, s) WHELK-O1 (250 mm. Times.30 mm,5 um); mobile phase: [0.1% NH) ₃ H ₂ O EtOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%, min) purification residue. Obtaining the compound (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (4-oxo-3H-imidazo [4, 5-c)]Pyridin-5-yl) propionamide (18.56 mg,54.5% yield, 100% purity).

LCMS：Rt＝0.627min；C ₁₉ H ₂₂ N ₆ O ₃ MS calculated: 382.42; MS experimental values: 383.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ9.28(br s,1H),7.94-7.79(m,1H),6.86(br d,J＝7.3Hz,1H),5.74-5.50(m,2H),4.62-4.18(m,2H),3.50-3.32(m,1H),3.14(br s,1H),2.66-2.37(m,1H),2.28(br s,1H),2.16-1.95(m,3H),1.92-1.72(m,2H),0.62(br s,1H),0.41(br d,J＝3.8Hz,2H),0.18(br s,1H),0.03(br d,J＝4.5Hz,1H)。

EXAMPLE 102 Synthesis of viral protease inhibitor Compound 269

Purification of (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl by SFC]Ethyl group]-3-cyclopropyl-2- (4-oxo-3H-imidazo [4, 5-c)]Pyridin-5-yl) propionamide (28 mg,73.2umol,1 eq). By preparative HPLC (column (s, s) WHELK-O1 (250)mm 30mm,5 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%, min) purification residue. Obtaining the compound (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-3-cyclopropyl-2- (4-oxo-3H-imidazo [4, 5-c)]Pyridin-5-yl) propionamide (15.52 mg,55.4% yield, 100% purity).

LCMS：Rt＝0.647min；C ₁₉ H ₂₂ N ₆ O ₃ MS calculated: 382.42; MS experimental values: 383.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ9.29(br s,1H),7.88(br d,J＝6.3Hz,1H),6.87(br d,J＝6.5Hz,1H),5.89-5.41(m,1H),4.74-4.29(m,1H),3.48(br s,1H),3.30-3.09(m,1H),2.67-2.42(m,1H),2.39-2.21(m,1H),2.21-1.99(m,3H),1.94-1.55(m,1H),0.63(br s,1H),0.42(br s,2H),0.27-0.08(m,2H)。

Example 103 Synthesis of viral protease inhibitor Compounds 271 and 271A

271A isomer 1: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5R) -1-methyl-6-oxo-1, 7-diazaspiro [4.4] non-7-yl ] propionamide

271A isomer 2: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5S) -1-methyl-6-oxo-1, 7-diazaspiro [4.4] non-7-yl ] propionamide

271 isomer 3: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5R) -1-methyl-6-oxo-1, 7-diazaspiro [4.4] non-7-yl ] propionamide

271 isomer 4: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5S) -1-methyl-6-oxo-1, 7-diazaspiro [4.4] non-7-yl ] propionamide

By preparative SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH3H2O ETOH)]；B％：20％-20%, min) purification 270 to give 271A (30 mg) and 271 (20 mg). By preparative SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -25%, min) to purify 271A isomers 1 and 2, to give 271A isomer 1 (2.65 mg,2% yield) and 271A isomer 2 (2.76 mg,2% yield) as two white solids. By preparative SFC (column: DAICEL CHIRALPAK IG (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O MEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%, min) to purify 271 isomer 1 and 271 isomer 2 to give 271 isomer 1 (15.96 mg,15% yield) and 271 isomer 2 (13.71 mg,13% yield) as two white solids.

271A isomer 1: LCMS: rt= 1.208min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]； ¹ H NMR(400MHz,CD ₃ OD)δ4.96(dd,J＝6.8,9.3Hz,1H),4.66-4.61(m,1H),3.50(dd,J＝5.6,8.1Hz,2H),3.37-3.31(m,2H),3.11-3.02(m,1H),2.91-2.81(m,1H),2.53-2.42(m,1H),2.40-2.31(m,4H),2.30-2.09(m,3H),2.02-1.81(m,7H),1.61(td,J＝7.2,14.1Hz,1H),0.69-0.60(m,1H),0.55-0.40(m,2H),0.20-0.13(m,2H)。

271A isomer 2: LCMS: rt=1.180 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]； ¹ H NMR(400MHz,CD ₃ OD)δ5.01(dd,J＝6.3,9.9Hz,1H),4.57(t,J＝7.8Hz,1H),3.55-3.47(m,2H),3.37-3.31(m,2H),3.11-2.99(m,1H),2.90-2.80(m,1H),2.60-2.46(m,1H),2.37-2.14(m,6H),2.09-1.72(m,8H),1.63-1.50(m,1H),0.74-0.63(m,1H),0.58-0.44(m,2H),0.24-0.15(m,2H)。

271 isomer 1: LCMS: rt=1.217 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]； ¹ H NMR(400MHz,CD ₃ OD)δ5.04-4.92(m,1H),4.67-4.60(m,1H),3.73-3.39(m,2H),3.37-3.32(m,2H),3.15-3.00(m,1H),2.88(d,J＝6.5Hz,1H),2.62-2.42(m,1H),2.40-2.15(m,6H),2.11-1.76(m,8H),1.68-1.51(m,1H),0.75-0.57(m,1H),0.57-0.39(m,2H),0.23-0.11(m,2H)。

271 isomer 2: LCMS: rt=1.222 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]； ¹ H NMR(400MHz,CD ₃ OD)δ5.01(dd,J＝6.1,9.9Hz,1H),4.56(t,J＝7.8Hz,1H),3.70-3.61(m,1H),3.49-3.40(m,1H),3.37-3.32(m,1H),3.30-3.23(m,1H),3.06-2.98(m,1H),2.87-2.77(m,1H),2.53(dq,J＝5.5,9.3Hz,1H),2.37-2.16(m,6H),2.10-1.75(m,8H),1.65-1.54(m,1H),0.72-0.61(m,1H),0.57-0.46(m,2H),0.21-0.11(m,2H)。

Example 104 Synthesis of viral protease inhibitor Compounds 273A, 273B and 273C

Step 1: 3-cyclopropyl-2- (1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) propionic acid

A solution of 1 (0.7 g,1.91mmol,1 eq.) in HCl/dioxane (4M, 10mL,20.9 eq.) was stirred at 25℃for 0.5hr. LC-MS showed complete depletion of 1 and 45% of the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product was used in the next step without further purification. 3-cyclopropyl-2- (1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) propionic acid (500 mg, crude material) was obtained as a colourless oil.

Step 2: (2S) -3-cyclopropyl-2- (6-methyl-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) propionic acid

3-cyclopropyl-2- (1-oxo-2, 6-diazaspiro [4.5]]To a solution of dec-2-yl) propionic acid (0.5 g,1.88mmol,1 eq.) in MeOH (4 mL) was added Pd/C (50 mg,0.37mmol,10% purity) and formaldehyde (1.52 g,18.7mmol,1.4mL,37% purity, 10 eq.) and degassed in vacuo and with H ₂ Purging several times. The mixture is put in H ₂ (15 psi) at 25℃for 0.5 hour. By TLC (dichloromethane: methanol=5/1, kmno) ₄ ) A spot is detected. LC-MS showed complete exhaustion of 2 and 71% of the desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. General purpose medicineColumn chromatography (SiO) ₂ The residue was purified with methylene chloride methanol=100/1 to 5/1 to give (2S) -3-cyclopropyl-2- (6-methyl-1-oxo-2, 6-diazaspiro [4.5] as a white solid]Dec-2-yl) propionic acid (0.4 g,76% yield).

Step 3: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (6-methyl-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) propanamide

To (2S) -3-cyclopropyl-2- (6-methyl-1-oxo-2, 6-diazaspiro [4.5] ]To a solution of dec-2-yl) propionic acid (0.3 g,1.0mmol,1 eq.) in DCM (6 mL) was added HATU (610.3 mg,1.61mmol,1.5 eq.), DIPEA (276.5 mg,2.14mmol,0.37mL,2.0 eq.) and 3a (243.51 mg,1.28mmol,1.2 eq., HCl). The mixture was stirred at 25℃for 1hr. LC-MS showed 3 was completely depleted and 15% of the desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:17% -47%,9.5 min) to give (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (6-methyl-1-oxo-2, 6-diazaspiro [4.5]]Dec-2-yl) propanamide (60 mg,13% yield).

273A: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5R) -6-methyl-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl ] propionamide

273B: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5S) -6-methyl-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl ] propionamide

273C: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5R) -6-methyl-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl ] propionamide

By preparative SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -20%, min) purification of 4 to give 273A and 273B (20 mg) toAnd 273C (2.79 mg,6.5umol,4% yield, 97% purity). By preparative SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%: from 40% to 40%, min) purified 273A and 273B (20 mg) to give 273A (2.50 mg,4.1% yield) and 273B (2.59 mg,4% yield).

273A：LCMS：Rt＝1.362min；C ₂₂ H ₃₃ N ₅ O ₃ MS calculated: 415.26; MS experimental values: 416.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ4.79(s,1H),4.52-4.47(m,1H),3.34-3.24(m,2H),3.20-3.15(m,2H),2.60-2.44(m,1H),2.35-2.24(m,1H),2.22-2.00(m,4H),1.94(s,3H),1.83-1.64(m,4H),1.60-1.31(m,7H),0.55-0.38(m,1H),0.37-0.20(m,2H),0.06-0.12(m,2H)。

273B：LCMS：Rt＝1.353min；C ₂₂ H ₃₃ N ₅ O ₃ MS calculated: 415.26; MS experimental values: 416.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ4.83(dd,J＝6.8,9.3Hz,1H),4.52-4.49(m,1H),3.47-3.38(m,1H),3.34-3.25(m,1H),3.23-3.17(m,2H),2.60-2.50(m,1H),2.41-2.27(m,1H),2.25-2.03(m,4H),1.97(s,3H),1.82-1.31(m,11H),0.50-0.40(m,1H),0.37-0.23(m,2H),0.05 -0.06(m,2H)。

273C：LCMS：Rt＝1.363min；C ₂₂ H ₃₃ N ₅ O ₃ MS calculated: 415.26; MS experimental values: 416.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ4.82(dd,J＝6.0,10.0Hz,1H),4.42-4.38(m,1H),3.41-3.23(m,2H),3.18-3.14(m,1H),3.12-3.06(m,1H),2.57-2.46(m,1H),2.45-2.29(m,1H),2.17-1.95(m,4H),1.93(s,3H),1.80-1.58(m,4H),1.57-1.26(m,7H),0.57-0.41(m,1H),0.40-0.23(m,2H),0.06-0.07(m,2H)。

EXAMPLE 105 Synthesis of viral protease inhibitor Compounds 278

7-amino-5, 6,7, 8-tetrahydroquinoline-7-carbonitrile

To 6, 8-dihydro-5H-quinolin-7-one (350 mg,1.91 mmol)To a solution of 1 eq, HCl) in DCM (7 mL) was added NH ₃ (7M, 2.72mL,10 eq.) and Ti (i-PrO) ₄ (650.0 mg,2.29mmol,0.67mL,1.2 eq.) and stirred at 25℃for 2hr. TMSCN (283.6 mg,2.86mmol,0.35mL,1.5 eq.) was added and the solution stirred at 25℃for 16hr. LC-MS showed complete depletion of starting material and detected a major peak with the desired MS. Ethyl acetate (50 mL) and H were added ₂ O (2.0 mL), the reaction mixture was filtered, and the filtrate was concentrated to reduced pressure. The compound 7-amino-6, 8-dihydro-5H-quinoline-7-carbonitrile (260 mg, crude material) was obtained as a yellow solid.

278: n- (1- ((7-cyano-5, 6,7, 8-tetrahydroquinolin-7-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

7-amino-6, 8-dihydro-5H-quinoline-7-carbonitrile (80 mg,0.46mmol,1 eq.) and (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]A solution of propionic acid (153.5 mg,0.50mmol,1.1 eq.) and pyridine (365.3 mg,4.62mmol,0.37mL,10 eq.) in THF (2 mL) was stirred at 25℃for 15min. Drop wise addition of POCl at 0deg.C ₃ After (177.0 mg,1.15mmol,0.10mL,2.5 eq.) the reaction mixture was stirred at 25℃for 16 hours. LC-MS showed starting material remaining and one peak with the expected MS was detected. The reaction mixture was saturated with NaHCO ₃ Alkalization to ph=8 and extraction with ethyl acetate (30 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,9.5 min) to give the title compound as a pale yellow solid. The compound N- [2- [ (7-cyano-6, 8-dihydro-5H-quinolin-7-yl) amino was obtained as a pale yellow solid ]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (2.32 mg,1.08% yield, 98.6% purity).

LCMS：Rt＝0.754min；C ₂₆ H ₂₇ N ₅ O ₃ MS calculated: 457.21; MS experimental values: 458.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.37-8.24(m,1H),7.65-7.56(m,1H),7.29-7.12(m,3H),7.03(d,J＝8.3Hz,1H),6.52(d,J＝7.8Hz,1H),4.64-4.60(m,1H),3.93(s,3H),3.76-3.57(m,1H),3.45-3.33(m,1H),3.17-2.94(m,2H),2.60-2.36(m,2H),1.88-1.78(m,1H),1.75-1.60(m,1H),0.89-0.72(m,1H),0.56-0.41(m,2H),0.24-0.12(m,2H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ11.56(s,1H),8.79(d,J＝15.8Hz,1H),8.53-8.43(m,1H),8.36(dd,J＝4.6,11.4Hz,1H),7.56(d,J＝7.5Hz,1H),7.35(d,J＝14.3Hz,1H),7.25-7.15(m,1H),7.14-7.06(m,1H),7.05-6.98(m,1H),6.51(d,J＝7.5Hz,1H),4.61-4.45(m,1H),3.89(s,3H),3.59(d,J＝16.8Hz,1H),3.23(d,J＝16.8Hz,1H),2.98-2.83(m,2H),2.42(dd,J＝6.1,12.9Hz,1H),2.36-2.18(m,1H),1.87-1.68(m,1H),1.57-1.34(m,1H),0.88-0.64(m,1H),0.46-0.25(m,2H),0.23-0.01(m,2H)

EXAMPLE 106 Synthesis of viral protease inhibitor Compound 323

323 isomers 1 and 2 steps: n- [ (1S) -1- [ [ (1S) -2- (tert-butylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (100 mg,180.79umol,80% purity, 1 eq.) in DCM (5 mL) was added PdCl ₂ (6.41 mg,36.16 mol,0.2 eq.) Na ₂ SO ₄ (89.88 mg, 632.76. Mu.L, 64.20. Mu.L, 3.5 eq.) and 2-methylpropan-2-amine (26.44 mg, 361.58. Mu.L, 37.99. Mu.L, 2 eq.). The mixture was stirred at 25℃for 30min, followed by the addition of TMSCN (35.87 mg, 361.58. Mu.L, 45.23. Mu.L, 2 eq.) and the mixture was stirred at 25℃for 2h. After the reaction was completed. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by hexane-IPA preparative HPLC gave the formation of a white solid The compound N- [ (1S) -1- [ [ (1S) -2- (tert-butylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (16.10 mg,25.59umol,14.16% yield, 83.4% purity) and N- [ (1S) -1- [ [ (1S) -2- (tert-butylamino) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl)]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (7.92 mg,12.27umol,6.79% yield, 81.3% purity). MS (ESI) m/z 524.8[ M+H ]] ⁺

Column: phenomenex luna CN 5u 100 x 30mm; mobile phase: [ hexane-IPA ]; b%:5% -60%,10min

1H NMR(400MHz,DMSO-d6)δ＝11.57(s,1H),8.45-8.35(m,1H),7.98(d,J＝9.3Hz,1H),7.56(s,1H),7.42-7.30(m,1H),7.15-7.05(m,1H),7.03-6.96(m,1H),6.50(d,J＝7.6Hz,1H),4.51-4.41(m,1H),3.99-3.91(m,1H),3.88(s,3H),3.63(dd,J＝7.7,10.2Hz,1H),3.16-2.99(m,2H),2.37-2.21(m,1H),2.16-2.03(m,1H),1.90-1.45(m,6H),1.05-1.00(m,9H),0.97-0.84(m,6H)

1H NMR(400MHz,DMSO-d6)δ＝11.81-11.42(m,1H),8.62-7.84(m,2H),7.69-7.47(m,1H),7.42-7.28(m,1H),7.20-6.94(m,2H),6.50(d,J＝7.6Hz,1H),4.61-4.40(m,1H),4.13-3.58(m,5H),3.23-2.91(m,2H),2.38-1.98(m,3H),1.91-1.37(m,5H),1.12-1.00(m,9H),0.97-0.79(m,6H)

Example 107 Synthesis of viral protease inhibitor Compounds 325

N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] -2- (2, 2-trifluoroethylamino) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (180 mg,325.42umol,80% purity, 1 eq.) in EtOH (2 mL) was added 2, 2-trifluoroethylamine (64.47 mg,650.84umol,51.17uL,2 eq.) and ZnCl ₂ (8.87mg65.08 mol,3.05uL,0.2 eq). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (64.57 mg,650.84umol,81.42ul,2 eq.) was added. The mixture was stirred at 25℃for 2h. Purification of the residue by HCl preparative HPLC gave the compound N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]-2- (2, 2-trifluoroethylamino) ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (120 mg,215.78 mol,66.31% yield, 99% purity). MS (ESI) m/z 551.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d6)δ＝11.56(dd,J＝2.0,5.5Hz,1H),8.39(br t,J＝8.6Hz,1H),8.32-8.16(m,1H),7.59(br d,J＝17.6Hz,1H),7.37(dd,J＝1.5,5.7Hz,1H),7.15-6.92(m,2H),6.50(d,J＝7.7Hz,1H),4.57-4.36(m,1H),4.25-4.02(m,1H),4.00-3.81(m,4H),3.78-3.40(m,2H),3.19-2.94(m,2H),2.42-1.94(m,3H),1.88-1.36(m,5H),0.91(dd,J＝6.3,15.1Hz,6H)。

EXAMPLE 108 Synthesis of viral protease inhibitor Compounds 327

/>

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (180 mg,325.42umol,80% purity, 1 eq.) in EtOH (4 mL) was added ZnCl ₂ (8.87 mg, 65.08. Mu.L, 3.05. Mu.L, 0.2 eq.) and aniline (60.61 mg, 650.84. Mu.L, 59.42. Mu.L, 2 eq.) and stirring the mixture at 25℃for 30min. After the addition of TMSCN (64.57 mg,650.84umol,81.42uL,2 eq.) the mixture was stirred at 25℃for 2h. After the reaction was completed. The reaction mixture was filtered to give the product. The reaction mixture was purified by preparative HPLC to give the product N- [ (1S) -1- [ [ (1S) -2-anilino-2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid ]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (70 mg,122.10umol,37.52% yield, 95% purity). MS (ESI) m/z 545.3[ M+H ]] ⁺

1H NMR(400MHz,DMSO-d6)δ＝11.59(br d,J＝2.0Hz,1H),8.44(br d,J＝7.7Hz,1H),8.26(d,J＝9.5Hz,1H),7.63-7.51(m,1H),7.38(d,J＝1.8Hz,1H),7.26-6.94(m,4H),6.80-6.65(m,3H),6.51(d,J＝7.5Hz,1H),6.34(d,J＝9.9Hz,1H),4.59-4.20(m,3H),3.89(s,3H),3.18-2.95(m,2H),2.44-2.30(m,1H),2.24-2.00(m,1H),1.97-1.43(m,6H),0.99-0.82(m,6H)

EXAMPLE 109 Synthesis of viral protease inhibitor Compounds 329

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of (4-methoxy-1H-indole-2-carboxamide (100 mg, 180.79. Mu. Mol,80% purity, 1 eq.) in EtOH (4 mL) was added (1S) -1-phenylethanamine (43.82 mg, 361.58. Mu. Mol, 46.02. Mu.L, 2 eq.), znCl ₂ (4.93 mg, 36.16. Mu. Mol, 1.69. Mu.L, 0.2 eq.). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (35.87 mg,361.58umol,45.24ul,2 eq.) was added. After stirring the mixture at 25 ℃ for 2h, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by HCl preparative HPLC gave the compound N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]-2- [ [ (1S) -1-phenylethyl]Amino group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (90 mg,154.01umol,85.19% yield, 98% purity). MS (ESI) m/z 573.2[ M+H ] ] ⁺

Column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ water (0.04% hcl) -ACN ]; b%:40% -70%,7min

1H NMR(400MHz,DMSO-d6)δ＝11.39(br s,1H),8.46-7.80(m,2H),7.52-6.89(m,9H),6.51(br d,J＝7.5Hz,1H),4.64-4.35(m,1H),4.26-4.03(m,1H),3.96-3.83(m,4H),3.36-3.03(m,3H),2.37-1.51(m,8H),1.39-1.21(m,3H),0.90(br dd,J＝5.7,14.6Hz,6H)

EXAMPLE 110 Synthesis of viral protease inhibitor Compound 331

/>

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of (4-methoxy-1H-indole-2-carboxamide (100 mg, 180.79. Mu. Mol,80% purity, 1 eq.) in EtOH (4 mL) was added pyrrolidine (25.72 mg, 361.58. Mu. Mol, 30.18. Mu.L, 2 eq.) ZnCl ₂ (1M, 1.81uL,0.01 eq.). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (35.87 mg,361.58umol,45.24ul,2 eq.) was added. The mixture was stirred at 25 ℃ for 2h, and then the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by hexane-IPA preparative HPLC gave the compound N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (19.34 mg,33.19umol,18.36% yield, 89.7% purity) and N- [ (1S) -1- [ [ (1S) -2-cyano-1- [ [ (3S) -2-oxopyrrolidin-3-yl) ]Methyl group]-2-pyrrolidin-1-yl-ethyl]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (10.41 mg,13.70umol,7.58% yield, 68.8% purity). MS (ESI) m/z 523.4[ M+H ]] ⁺

Column: phenomenex luna CN 5 μ 100×30mm; mobile phase: [ hexane-IPA ]; b%:5% -60%,10min

1H NMR(400MHz,DMSO-d6)δ＝11.58(s,1H),8.43(d,J＝7.7Hz,1H),8.19(d,J＝9.4Hz,1H),7.61-7.50(m,1H),7.38(d,J＝1.8Hz,1H),7.14-6.95(m,2H),6.50(d,J＝7.6Hz,1H),4.54-4.35(m,1H),4.17-4.00(m,1H),3.99-3.92(m,1H),3.88(s,3H),3.14-2.94(m,2H),2.64-2.53(m,4H),2.39-2.27(m,1H),2.17-2.02(m,1H),1.88-1.66(m,7H),1.63-1.44(m,3H),0.91(dd,J＝6.3,16.2Hz,6H)

1H NMR(400MHz,DMSO-d6)δ＝11.56(br d,J＝1.8Hz,1H),8.43-8.30(m,1H),8.00(d,J＝9.2Hz,1H),7.60(s,1H),7.35(d,J＝1.8Hz,1H),7.16-6.94(m,2H),6.50(d,J＝7.6Hz,1H),4.55-4.35(m,1H),4.13-4.03(m,1H),4.02-3.94(m,1H),3.88(s,3H),3.13-3.01(m,2H),2.70-2.57(m,2H),2.43-2.29(m,1H),2.17-1.94(m,2H),1.88-1.34(m,9H),0.90(dd,J＝6.5,15.2Hz,6H)

EXAMPLE 111 Synthesis of viral protease inhibitor Compound 345

/>

Step 1: (S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamic acid tert-butyl ester

At 25℃under N ₂ Downward (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (2.00 g,6.99mmol,1 eq.) in THF (20 mL) was added LiBH4 (2M, 6.99mL,2 eq.). The mixture was then stirred at 25℃for 1h. NH for mixture ₄ Aqueous Cl (20.0 mL) was quenched and extracted with EtOAc (20.0 mL x 5). The organic layer was washed with brine (20.0 mL), and dried over Na ₂ SO ₄ Dried, filtered, and concentrated under reduced pressure to give a residue. The residue was wet-milled with ethyl acetate: petroleum ether=1:2 for 1h at 25 ℃ to give tert-butyl ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamate (1.57 g, crude material) as a white solid. MS (ESI) m/z 259.2[ M+H ] ] ⁺ 。

Step 2: (S) -3- ((S) -2-amino-3-hydroxypropyl) pyrrolidin-2-one

A solution of tert-butyl ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamate (2.39 g,9.25mmol,1 eq.) in HCl/MeOH (4M, 23.9mL,10.33 eq.) was stirred at 25℃for 1h. The mixture was concentrated under reduced pressure to give (S) -3- ((S) -2-amino-3-hydroxypropyl) pyrrolidin-2-one (1.8 g, crude material, HCl) as a yellow oil.

Step 3: (2S, 4S) -2- (((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester

To (S) -3- ((S) -2-amino-3-hydroxypropyl) pyrrolidin-2-one (1.8 g,9.25mmol,1 eq. HCl) in DMFTo a solution of TEA (5.61 g,55.48mmol,7.72mL,6 eq), (2S, 4S) -1-tert-butoxycarbonyl-4-phenyl-pyrrolidine-2-carboxylic acid (2.69 g,9.25mmol,1 eq.) in DCM (6 mL) was added followed by T3P (17.65 g,27.74mmol,16.5mL,50% purity, 3 eq.) at 0deg.C. After stirring the mixture at 0 ℃ for 1h, the mixture was quenched with water (40 mL) and extracted with EtOAc (20 mL x 5). The organic layer was washed with brine (20.0 mL), and dried over Na ₂ SO ₄ Drying, filtering, concentrating under reduced pressure, and purifying by column chromatography (SiO ₂ DCM: meoh=10:1 to 1:1) to give (2S, 4S) -2- (((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (3.04 g,6.69mmol,72.38% yield, 95% purity) as a yellow solid. MS (ESI) m/z 432.2[ M+H ]] ⁺ 。

Step 4: (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide

A mixture of (2S, 4S) -2- (((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (3.04 g,7.04mmol,1 eq.) in HCl/EtOAc (4M, 30mL,17.03 eq.) was stirred at 25℃for 1h. The mixture was concentrated under reduced pressure to give (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide (2.59 g, crude material, HCl) as a white solid. MS (ESI) m/z 332.2[ M+H ]] ⁺ 。

Step 5: (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide

To a solution of (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide (2.58 g,7.01mmol,1 eq, HCl) in DMF (16 mL) and DCM (8 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (1.34 g,7.01mmol,1 eq), DMAP (1.71 g,14.03mmol,2 eq) and EDCI (2.69 g,14.03mmol,2 eq). The mixture was stirred at 0℃for 1h. The mixture was quenched with water (50.0 mL) and extracted with EtOAc (20.0 mL x 4). The organic layer was washed with brine (20.0 mL), and dried over Na ₂ SO ₄ Dried, filtered, concentrated under reduced pressure and purified by column chromatography (SiO ₂ DCM: meoh=10:1 to 3:1) to give (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide (2.1 g,3.79mmol,54.00% yield, 91% purity) as a pale yellow solid. MS (ESI) m/z 505.1[ M+H ]] ⁺ 。

Step 6: (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide

To a mixture of (2S, 4S) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide (200 mg,376.55umol,95% purity, 1 eq.) in DMSO (2 mL) was added TFA (64.40 mg,564.83umol,41.82ul,1.5 eq.) and IBX (332.97 mg,1.13mmol,95% purity, 3 eq.) and the mixture stirred at 25 ℃ for 14H. NaHCO is used for the mixture ₃ Aqueous (15.0 mL) was quenched and extracted with EtOAc (10.0 mL x 3). The organic layer was washed with brine (10.0 mL), and dried over Na ₂ SO ₄ Dried, filtered, concentrated under reduced pressure and purified by preparative HPLC to give (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide as a white solid (58 mg,111.95umol,29.73% yield, 97.0% purity). MS (ESI) m/z 503.2[ M+H ] ] ⁺ 。

Preparative HPLC conditions: column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:20% -50%,10min.

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.66-11.41(m,1H),9.51-7.92(m,1H),7.82-7.47(m,1H),7.43-7.20(m,5H),7.17-6.98(m,2H),6.98-6.78(m,1H),6.55-6.39(m,1H),5.83-5.67(m,1H),4.84-4.59(m,1H),4.48-4.35(m,1H),4.32-4.12(m,1H),3.94-3.66(m,4H),3.64-3.39(m,1H),3.20-3.03(m,1H),2.98-2.54(m,1H),2.47-2.12(m,3H),2.03-1.78(m,1H),1.72-1.22(m,2H)

Step 7: (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide

To a mixture of (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-phenylpyrrolidine-2-carboxamide (90 mg,179.08umol,1 eq.) in DCM (1 mL) was added NaHSO ₃ (74.54 mg, 716.33. Mu. Mol, 50.37. Mu.L, 4 eq.) in H2O (0.5 mL) and then KCN (46.64 mg, 716.33. Mu. Mol, 30.69. Mu. L,4 eq.) was added at 0deg.C. The mixture was stirred at 25℃for 14h. The mixture was quenched with water (15.0 mL) and extracted with EtOAc (10.0 mL x 3). The organic layer was washed with brine (10.0 mL), and dried over Na ₂ SO ₄ Dried, filtered, concentrated under reduced pressure and purified by preparative HPLC to give (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide as a white solid (29 mg,54.76umol,30.58% yield, 100% purity). MS (ESI) m/z 530.2[ M+H ] ] ⁺ 。

Preparative HPLC conditions: column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ Water (0.04% HCl) -ACN ]; b%:32% -48%,7min.

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.66-11.42(m,1H),8.63-8.22(m,1H),7.62(d,J＝5.6Hz,0.5H),7.44(s,0.5H),7.41-7.29(m,4H),7.29-7.21(m,1H),7.15-7.07(m,1H),7.06-6.99(m,1H),6.98-6.92(m,0.5H),6.86-6.78(m,0.5H),6.70(s,1H),6.53-6.41(m,1H),5.32-5.18(m,0.5H),4.86-4.66(m,0.5H),4.66-4.54(m,0.5H),4.45-4.34(m,1H),4.24-4.15(m,0.5H),4.12-3.96(m,1H),3.95-3.87(m,0.5H),3.87-3.76(m,3H),3.76-3.66(m,0.5H),3.64-3.56(m,0.5H),3.52-3.42(m,1H),3.20-3.09(m,1H),2.85-2.74(0.5,1H),2.62-2.54(m,0.5H),2.46-2.35(m,1.5H),2.30-1.98(m,2H),1.92-1.80(m,0.5H),1.68-1.19(m,2.5H)。

Step 8: (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide

Purification of (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3-)((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide (27 mg,50.98umol,1 eq) gave (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide (13.03 mg,22.71umol,44.54% yield, 92.3% purity) as a white solid. MS (ESI) m/z 530.2[ M+H ]] ⁺ (2S, 4S) -N- ((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamide (12.12 mg,19.86umol,38.96% yield, 86.8% purity) as a white solid. MS (ESI) m/z 530.2[ M+H ]] ⁺ 。

Isomer 1: ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.64-11.47(m,1H),8.62-8.30(m,1H),7.64(s,0.5H),7.45(s,0.5H),7.42-7.30(m,4H),7.30-7.21(m,1H),7.16-7.06(m,1H),7.05-6.99(m,1H),6.97-6.92(m,0.5H),6.86-6.80(m,0.5H),6.75-6.65(m,1H),6.53-6.41(m,1H),5.32-5.23(m,0.5H),4.85-4.78(m,0.5H),4.66-4.54(m,1H),4.45-4.35(m,0.5H),4.24-4.14(m,0.5H),4.13-3.98(m,1H),3.95-3.87(m,0.5H),3.87-3.75(m,3H),3.75-3.66(m,0.5H),3.64-3.56(m,0.5H),3.53-3.41(m,0.5H),3.22-3.11(m,1H),2.80(t,J＝8.8Hz,0.5H),2.62-2.53(m,0.5H),2.46-2.36(m,2H),2.29-2.15(m,1.5H),2.14-1.97(m,1H),1.69-1.54(m,0.5H),1.51-1.12(m,2.5H)

isomer 2: ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.64-11.506(m,1H),8.48(d,J＝9.6Hz,0.5H),8.29(d,J＝9.6Hz,0.5H),7.62(s,0.5H),7.44(s,0.5H),7.40-7.31(m,4H),7.29-7.23(m,1H),7.11(q,J＝8.4Hz,1H),7.05-6.99(m,1H),6.98-6.94(s,0.5H),6.84-6.78(m,0.5H),6.77-6.69(m,1H),6.52-6.42(m,1H),5.23(d,J＝7.2Hz,0.5H),4.70(d,J＝6.8Hz,0.5H),4.48-4.32(m,1H),4.26-4.15(m,0.5H),4.13-3.95(m,1H),3.94-3.88(m,0.5H),3.86-3.76(m,3H),3.76-3.69(m,0.5H),3.66-3.53(m,0.5H),3.51-3.40(m,0.5H),3.20-3.09(m,1H),2.84-2.74(m,1H),2.45-2.35(m,2H),2.31-2.11(m,2H),1.92-1.80(m,1H),1.60-1.21(m,3H)

EXAMPLE 112 Synthesis of viral protease inhibitor Compounds 355

Step 1: n- ((2S) -1- (((2S) -1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (150 mg, 338.98. Mu. Mol,1 eq.) in DCM (2 mL) was added saturated NaHSO ₃ (35.27 mg, 338.98. Mu. Mol, 23.83. Mu.L, 1 eq.) and the mixture was stirred at 25℃for 30min. KCN (100 mg,1.54mmol,65.79uL,4.53 eq.) was added to H ₂ A solution in O (0.5 mL) and the mixture was stirred at 25℃for 2h. After completion, the organic phase was collected and the aqueous layer was extracted with DCM (30 ml×3). The combined organic phases were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Drying and concentration gave the crude material. NaOH was added to liquid water to ph=9 followed by quenching with aqueous NaClO followed by NaOH to pH>14. The crude material was used directly in the next step without further purification. Obtaining N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl ]-4-methoxy-1H-indole-2-carboxamide (140 mg, crude). MS (ESI) m/z 470.1[ M+H ]] ⁺ 。

Step 2: propylcarbamic acid (2S) -1-cyano-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propyl ester

N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] was added dropwise to a solution of 1-isocyanatopropane (27.19 mg,319.47 mol,30.21uL,5 eq.) in anhydrous toluene (0.1 mL) at 0deg.C]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]A solution of 4-methoxy-1H-indole-2-carboxamide (30 mg, 63.89. Mu.L, 1 eq.) in dry toluene (0.5 mL) was followed by the addition of TEA (64.65. Mu.g, 6.39 e-1. Mu.L, 8.89 e-2. Mu.L, 0.01 eq.) and N ₂ The solution was stirred at 25℃for 17h. At the position ofAfter completion, the solution was concentrated to give a crude material. The residue was purified by preparative HPLC (FA conditions), column: phenomenex Luna C18 200 x 40mm x 10um; mobile phase: [ Water (0.2% FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -80%,8min. Obtaining N-propylcarbamic acid [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Propyl group]The ester (8 mg,14.15umol,22.15% yield, 98.124% purity).

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.27(s,1H),7.19-7.10(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.7Hz,1H),5.48-5.40(m,1H),4.64-4.53(m,1H),4.46-4.34(m,1H),3.93(s,3H),3.29-3.19(m,2H),3.15-3.03(m,2H),2.72-2.57(m,1H),2.34-2.11(m,2H),1.89-1.44(m,7H),1.07-0.88(m,9H)。MS(ESI)m/z 555.3[M+H] ⁺ 。

Step 3: n- ((2S) -1- (((1S) -1- (4-imino-2-oxo-3-propyloxazolidin-5-yl) -2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

N-propylcarbamic acid [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Esters (50 mg,90.15umol,1 eq.) in NH ₄ HCO ₃ The mixture of (0.01M, 45.07mL,5 eq.) and ACN (5 mL) was stirred at 25℃for 17h. After completion, the solution was extracted with EA (40 ml x 3) and the combined organic phases were extracted with Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The residue was purified by preparative HPLC, column: waters Xb ridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8min. Obtaining N- [ (1S) -1- [ [ (1S) -1- (4-imino-2-oxo-3-propyl-oxazolidin-5-yl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (20 mg,34.21umol,16.06% yield, 94.871% purity). ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.31-7.21(m,1H),7.19-7.10(m,1H),7.06-6.98(m,1H),6.56-6.46(m,1H),5.16-5.03(m,1H),4.79-4.37(m,2H),3.96-3.88(m,3H),3.58-3.40(m,2H),3.28-3.13(m,2H),2.65-2.51(m,1H),2.41-2.05(m,2H),1.90-1.40(m,7H),1.07-0.87(m,9H)。

EXAMPLE 113 Synthesis of viral protease inhibitor Compound 357

Step 1: (2S) -1-cyano-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propyl isopropyl carbamate

To a solution of 2-isocyanatopropane (10.88 mg,127.79umol,12.53ul,3 eq.) in anhydrous toluene (0.1 mL) was added dropwise a solution of N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide (20 mg,42.60umol,1 eq.) in anhydrous toluene (0.5 mL) at 0 ℃. After the addition of TEA (4.31 mg, 42.60. Mu. Mol, 5.93. Mu.L, 1 eq.) the solution was stirred under dry argon at 25℃for 16h. After completion, the solution was concentrated to remove toluene. The residue was purified by preparative HPLC (FA conditions), column: phenomenex Luna C18, 75 x 30mm x 3um; mobile phase: [ water (0.2% fa) -ACN ]; b%:30% -70%,8min. N-isopropylcarbamic acid [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propyl ] ester (8 mg,14.30umol,33.57% yield, 99.129% purity) was obtained as a white solid.

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.27(s,1H),7.19-7.10(m,1H),7.02(d,J＝8.4Hz,1H),6.51(d,J＝7.7Hz,1H),5.48-5.39(m,1H),4.64-4.53(m,1H),4.44-4.33(m,1H),3.93(s,3H),3.80-3.64(m,1H),3.28-3.17(m,2H),2.72-2.58(m,1H),2.34-2.10(m,2H),1.88-1.58(m,5H),1.23-1.09(m,6H),1.01(td,J＝5.7,11.5Hz,6H)。MS(ESI)m/z555.3[M+H] ⁺ 。

Step 2: n- ((2S) -1- (((1S) -1- (4-imino-3-isopropyl-2-oxooxazolidin-5-yl) -2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

Will contain N-isopropyl carbamic acid [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]NH of ester (110 mg,198.33umol,1 eq) ₄ HCO ₃ (0.01M, 30mL,1.51 eq.) ACN (5 mL) was stirred at 25℃for 17h. After completion, the solution was extracted with ethyl acetate (30 ml x 3) and the combined organic phases were taken up over Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The residue was purified by preparative HPLC (neutral conditions), column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH4HCO 3) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8min. Obtaining N- [ (1S) -1- [ [ (1S) -1- (4-imino-3-isopropyl-2-oxo-oxazolidin-5-yl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (27 mg,48.55 mol,24.48% yield, 99.738% purity). ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.33-7.21(m,1H),7.19-7.10(m,1H),7.07-6.97(m,1H),6.57-6.46(m,1H),5.06-4.96(m,1H),4.73(br d,J＝11.2Hz,1H),4.64-4.53(m,1H),4.44-4.28(m,1H),3.96-3.89(m,3H),3.27-3.15(m,2H),2.66-2.49(m,1H),2.43-2.15(m,2H),1.85-1.24(m,4H),1.07-0.91(m,6H)

EXAMPLE 114 Synthesis of viral protease inhibitor Compound 359

Step 1: (2S) -1-cyano-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propyl phenylcarbamate

N- [ (1S) -1- [ [ (1S) -2-cyano-2-hydroxy-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] was added dropwise to a solution of isocyanatobenzene (127 mg,1.07mmol,115.32uL,5 eq.) in anhydrous toluene (0.2 mL) at 0deg.C]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indoleA solution of the indole-2-carboxamide (100 mg, 212.98. Mu. Mol,1 eq.) in dry toluene (1 mL) was followed by the addition of TEA (215.51 ug, 2.13. Mu. Mol,2.96 e-1. Mu.L, 0.01 eq.). After stirring the solution at 25℃for 16H under a dry argon atmosphere, the solution was treated with H ₂ O (10 mL) was quenched, extracted with ethyl acetate (20 mL. Times.3), and the combined organic phases were taken up over Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The crude material was used directly in the next step without further purification. (2S) -1-cyano-2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propyl phenylcarbamate (50 mg,84.94umol,1 eq.) was obtained as a white solid. MS (ESI) m/z 589.2[ M+H ] ] ⁺ 。

Step 2: n- [ (1S) -1- [ [ (1S) -1- (4-imino-2-oxo-3-phenyl-oxazolidin-5-yl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

The N-phenylcarbamic acid [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]NH of ester (50 mg,84.94umol,1 eq) ₄ HCO ₃ A solution of (0.01M, 42.47mL,5 eq.) and ACN (3 mL) was stirred at 25℃for 17h. After completion, the solution was extracted with ethyl acetate (40 ml x 3) and the combined organic phases were taken up over Na ₂ SO ₄ Drying, filtration and concentration gave a crude material, the residue was purified by preparative HPLC (neutral conditions), column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -50%,8min. Obtaining N- [ (1S) -1- [ [ (1S) -1- (4-imino-2-oxo-3-phenyl-oxazolidin-5-yl) -2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (6 mg,9.70umol,11.42% yield, 95.16% purity). ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.60-7.38(m,4H),7.38-7.20(m,2H),7.18-7.10(m,1H),7.07-6.98(m,1H),6.55-6.46(m,1H),5.29-5.15(m,1H),4.85-4.74(m,1H),4.61-4.47(m,1H),3.98-3.87(m,3H),3.29-3.18(m,2H),2.72-2.56(m,1H),2.48-2.20(m,2H),1.91-1.42(m,5H),1.08-0.85(m,6H)MS(ESI)m/z 589.3[M+H] ⁺ 。

EXAMPLE 115 Synthesis of viral protease inhibitor Compound 361

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.25mmol,1 eq.) in HCl/EtOAc (20 mL) was stirred at 25deg.C for 1h. TLC showed the reaction was complete. The reaction was concentrated to give crude (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl as a colorless oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (360 mg, crude material). The crude product was used without further purification. MS (ESI) m/z 299.2[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -4-methyl-2- [ [4- (trifluoromethoxy) -1H-indole-2-carbonyl ] amino ] pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To a mixture of methyl (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg,901.91 mol,90% purity, 1 eq.) and 4- (trifluoromethoxy) -1H-indole-2-carboxylic acid (221.11 mg,901.91 mol,1 eq.) in DCM (12 mL) and DMF (4 mL) was added EDCI (691.58 mg,3.61mmol,4 eq.) and DMAP (440.74 mg,3.61mmol,4 eq.). The mixture was stirred at 25 ℃ and for 3 hours.

LCMS showed the reaction was complete. The residue was concentrated in vacuo. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -70%,10 min) to give (2S) -2- [ [ (2S) -4-methyl-2- [ [4- (trifluoromethoxy) -1H-indole-2-carbonyl ] as a white solid]Amino group]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,427.35 um)ol,47.38% yield, 90% purity). MS (ESI) m/z 526.2[ M+H ]] ⁺

Step 3: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4- (trifluoromethoxy) -1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -4-methyl-2- [ [4- (trifluoromethoxy) -1H-indole-2-carbonyl ] carbonyl]Amino group]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (250 mg,474.83umol,1 eq.) in ammonia (29.14 g,1.71mol,28.57mL,3603.85 eq.) was stirred at 80℃and stirred for 16 hours. LCMS showed the reaction was complete. Concentrating the reaction to give crude N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl ]-4- (trifluoromethoxy) -1H-indole-2-carboxamide (200 mg, crude) (white solid). The crude product was used without further purification. MS (ESI) m/z 511.2[ M+H ]] ⁺

Step 4: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4- (trifluoromethoxy) -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4- (trifluoromethoxy) -1H-indole-2-carboxamide (35 mg,68.43umol,1 eq.) in DCM (2 mL) was added the berg reagent (65.23 mg,273.71umol,4 eq.). The mixture was stirred at 25 ℃ and for 3 hours. LCMS and HPLC showed the reaction was complete. The reaction was concentrated and purified by preparative HPLC (column Phenomenex Gemini-NX 18C 75 x 30mM x 3um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -55%,8 min) to give N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4- (trifluoromethoxy) -1H-indole-2-carboxamide (10.02 mg,20.30umol,29.67% yield, 100% purity) (white solid). MS (ESI) m/z 493.2[ M+H ] ] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 12.04(s,1H),8.94-9.08(m,1H),8.72-8.74(d,J＝7.60Hz,1H),7.71-7.76(m,1H),7.44-7.46(d,J＝7.60Hz,2H),7.21 -7.23(m,1H),7.02-7.05(d,J＝7.60Hz,1H),4.97-5.01(m,1H),4.47-4.50(m,1H),3.10-3.14(m,2H),2.14-2.15(m,1H),2.01-2.10(m,2H),1.67-1.70(m,1H)1.67-1.70(m,1H),1.69-1.72(m,4H)，0.92-0.95(m,3H)，1.69-1.72(m,3H)

EXAMPLE 116 Synthesis of viral protease inhibitor Compounds 363

Step 1:2- (trichloromethyl) -7- (trifluoromethoxy) -1H-benzo [ d ] imidazole

To a solution of 3- (trifluoromethoxy) benzene-1, 2-diamine (500 mg,2.60mmol,1 eq.) in AcOH (15 mL) was added dropwise methyl 2, 2-trichloroethyliminoate (459.12 mg,2.60mmol,321.06uL,1.00 eq.) and the reaction was then stirred at 25℃for 12h. By adding H at 0 ℃ ₂ O (50 mL) was used to quench the reaction mixture and then extracted with EtOAc (25 mL x 3). The combined organic layers were washed with brine (30 mL), filtered and concentrated under reduced pressure to give the product 2- (trichloromethyl) -7- (trifluoromethoxy) -1H-benzimidazole (720 mg, crude) as a yellow solid. MS (ESI) m/z 320.8[ M+H ]] ⁺

Step 2:7- (trifluoromethoxy) -1H-benzo [ d ] imidazole-2-carboxylic acid methyl ester

To a solution of 2- (trichloromethyl) -7- (trifluoromethoxy) -1H-benzimidazole (720 mg,2.25mmol,1 eq.) in MeOH (10 mL) was added Na ₂ CO ₃ (238.85 mg,2.25mmol,1 eq.) and the mixture was stirred at 70℃for 14h. 1N HCl was added to the solution and the reaction stirred for 0.5h. The mixture was extracted with EtOAc (30 mL x 3) and the combined organic layers were washed with brine (30 mL), over Na ₂ SO ₄ Dried, filtered and the filtrate concentrated under reduced pressure to give the product 7- (trifluoromethoxy) -1H-benzimidazole-2-carboxylic acid methyl ester (520 mg, crude material) as a yellow solid. MS (ESI) m/z 260.8[ M+H ]] ⁺

Step 3:7- (trifluoromethoxy) -1H-benzo [ d ] imidazole-2-carboxylic acid

To 7- (trifluoromethoxy) -1H-benzimidazole-2-carboxylic acid methyl ester (300 mg,1.15mmol,1 eq.) in THF (6 mL) and H ₂ To a solution in O (2 mL) was added LiOH (165.69 mg,6.92mmol,6 eq.) and the mixture was stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:2% -40%,9 min) to give the product 7- (trifluoromethoxy) -1H-benzimidazole-2-carboxylic acid (150 mg,591.12umol,51.26% yield, 97% purity) as a white solid. MS (ESI) m/z 245.1[ M-H] ⁺

¹ H NMR(DMSO-d ₆ ,400MHz)：δppm 7.46(d,J＝8.2Hz,1H),7.22(t,J＝7.9Hz,1H),7.10-7.14(m,1H)

Step 4: (S) -2- ((S) -2-amino-4-methylpentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester hydrochloride

To a solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (250 mg,625.81umol,1 eq.) in EtOAc (0.5 mL) was added HCl/EtOAc (4 m,10mL,63.92 eq.) dropwise and the mixture was stirred at 25 ℃ for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (200 mg, crude material, HCl) as a white solid.

Step 5: (S) -2- ((S) -4-methyl-2- (4- (trifluoromethoxy) -1H-benzo [ d ] imidazole-2-carboxamide) pentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg, 595.55. Mu. Mol,1.22 eq, HCl) and 7- (trifluoromethoxy) -1H-benzimidazole-2-carboxylic acid (120 mg, 487.52. Mu. Mol,1 eq.) were added Et dropwise to a solution of DMF (1 mL) and DCM (6 mL) ₃ N (295.99 mg,2.93mmol,407.14uL,6.0 eq.) and T ₃ P (930.72 mg,1.46mmol,869.83uL,50% purity, 3.0 eq.). The mixture was stirred at 25℃for 2h.By adding H at 0 ℃ ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=10/1 to 0/1 to give the product (2S) -2- [ [ (2S) -4-methyl-2- [ [7- (trifluoromethoxy) -1H-benzimidazole-2-carbonyl ] as a colorless oil]Amino group]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (180 mg,307.11umol,62.99% yield, 90% purity). MS (ESI) m/z 528.2[ M+H ] ] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )：δppm 7.58(br d,J＝7.9Hz,1H),7.40(br t,J＝8.0Hz,1H),7.21-7.33(m,1H),4.64(br t,J＝6.9Hz,1H),4.55-4.59(m,1H),3.72(s,3H),3.22-3.30(m,2H),2.60(br d,J＝9.0Hz,1H),2.27-2.37(m,1H),2.15-2.24(m,1H),1.72-1.92(m,5H),1.02(br dd,J＝12.8,6.1Hz,6H)

Step 6: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -4-methyl-1-oxopent-2-yl) -4- (trifluoromethoxy) -1H-benzo [ d ] imidazole-2-carboxamide

A solution of methyl (2S) -2- [ [ (2S) -4-methyl-2- [ [7- (trifluoromethoxy) -1H-benzimidazole-2-carbonyl ] amino ] pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (170 mg,322.28umol,1 eq) in ammonia (7M, 17mL,369.24 eq) was stirred at 80℃for 12H.

The reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-7- (trifluoromethoxy) -1H-benzimidazole-2-carboxamide (150 mg, crude). MS (ESI) m/z 513.2[ M+H ]] ⁺ 。

Step 7: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -4-methyl-1-oxopent-2-yl) -4- (trifluoromethoxy) -1H-benzo [ d ] imidazole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidine-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a solution of 7- (trifluoromethoxy) -1H-benzimidazole-2-carboxamide (100 mg,195.13umol,1 eq.) in DCM (6 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (232.51 mg,975.65umol,5.0 eq.). The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -70%,8 min) to give the product N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-7- (trifluoromethoxy) -1H-benzimidazole-2-carboxamide (35.5 mg,70.86umol,36.32% yield, 98.7% purity). MS (ESI) m/z 495.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 13.73(s,1H),8.86-9.04(m,2H),7.71(s,1H),7.56(d,J＝7.6Hz,1H),7.37-7.44(m,1H),7.26-7.37(m,1H),4.98(dd,J＝6.9,1.2Hz,1H),4.54(br s,1H),3.07-3.19(m,2H),2.33-2.43(m,1H),2.14(br dd,J＝8.8,4.9Hz,2H),1.55-1.90(m,5H),0.92(dd,J＝8.8,6.2Hz,6H)

EXAMPLE 117 Synthesis of viral protease inhibitor Compounds 365

Step 1: (2S) -2- [ [ (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ] amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate; EDCI (359.62 mg,1.88mmol,2 eq.) and DMAP (229.19 mg,1.88mmol,2 eq.) were added to a mixture of HCl (315.00 mg,937.99umol,1 eq.) and 3- (4-chlorophenyl) -3-hydroxy-butyric acid (201.33 mg,937.99umol,1 eq.) in DCM (3 mL) and DMF (6 mL). The mixture was stirred at 25℃for 2h. After the reaction was completed. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification by neutral prep HPLCThe residue gives the compound (2S) -2- [ [ (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ] as a white solid ]Amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (90 mg,172.38umol,18.38% yield, 95% purity) and (2S) -2- [ [ (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl)]Amino group]-4-methyl-pentanyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (120 mg,229.84umol,24.50% yield, 95% purity). MS (ESI) m/z 496.3[ M+H ]] ⁺

Column: kromasil C18 (250 x 50mm x 10 um); mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:25% -55%,10min.

Step 2:3- (4-chlorophenyl) -3-hydroxy-butyric acid

To 3- (4-chlorophenyl) -3-hydroxy-butanoic acid ethyl ester (500 mg,2.06mmol,1 eq.) in H ₂ LiOH.H was added to a mixture of O (3 mL) and THF (6 mL) ₂ O (172.90 mg,4.12mmol,2 eq.). The mixture was stirred at 25℃for 1h. After the reaction was completed. H for the reaction mixture ₂ O (20 mL) was diluted and extracted with 60mL ethyl acetate (30 mL x 2). The combined organic layers were washed with brine (930 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the residual compound 3- (4-chlorophenyl) -3-hydroxy-butyric acid (400 mg,1.68mmol,81.41% yield, 90% purity) as a white solid.

Step 3: (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- [3- (4-chlorophenyl) -3-hydroxy-butyryl ] amino ] -4-methyl-pentanamide

To (2S) -2- [ [ (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ]]Amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to a mixture of methyl propionate (80 mg,153.23umol,95% purity, 1 eq) ₃ MeOH (7M) (7M, 9.50mL,434.00 eq.). The mixture was stirred at 80 ℃ for 16h and then the reaction mixture was concentrated under reduced pressure to give the residual compound (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow oil]Methyl group]Ethyl group]-2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ]]Amino group]4-methyl-pentanamide (70 mg,130.98umol,85.48% yield, 90% purity). MS (ESI) m/z 481.2[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ] amino ] -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -4-methyl-pentanamide

To (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ]]Amino group]To a mixture of 4-methyl-pentanamide (70 mg,145.54umol,1 eq.) in DCM (4 mL) was added the Buerger reagent (69.36 mg,291.07umol,2 eq.). After stirring the mixture at 25℃for 60min, the reaction mixture was taken up in H ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 2). The combined organic layers were concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave the compound (2S) -2- [ [3- (4-chlorophenyl) -3-hydroxy-butyryl ] as a white solid]Amino group]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-4-methyl-pentanamide (11 mg,23.76umol,16.33% yield, 100% purity). MS (ESI) m/z 463.2[ M+H ]] ⁺

Isomer 1:1H NMR (400 MHz, methanol-d 4) delta=7.51-7.42 (m, 2H), 7.36-7.26 (m, 2H), 4.96 (dd, J=6.0, 10.1Hz, 1H), 4.27-4.17 (m, 1H), 3.30-3.23 (m, 2H), 2.83-2.63 (m, 2H), 2.51 (dq, J=5.3, 9.3Hz, 1H), 2.34-2.17 (m, 2H), 1.94-1.72 (m, 2H), 1.57 (s, 3H), 1.54-1.26 (m, 3H), 0.93-0.77 (m, 6H)

Isomer 2:1H NMR (400 MHz, methanol-d 4) delta=7.44 (d, J=8.6 Hz, 2H), 7.34-7.24 (m, 2H), 5.06-4.93 (m, 1H), 4.26-4.13 (m, 1H), 3.38-3.32 (m, 1H), 3.29-3.24 (m, 1H), 2.85-2.62 (m, 2H), 2.53 (dq, J=5.5, 9.3Hz, 1H), 2.42-2.17 (m, 2H), 1.98-1.74 (m, 2H), 1.52 (s, 3H), 1.49-1.36 (m, 2H), 1.31-1.18 (m, 1H), 0.90-0.79 (m, 3H), 0.72 (d, J=6.5 Hz, 3H)

EXAMPLE 118 Synthesis of viral protease inhibitor Compound 265

Step 1: (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2,3-c ] pyridin-6-yl) propionic acid methyl ester

To (2S) -3-cyclopropyl-2- (7-oxo-4, 5-dihydro-1H-pyrrolo [2, 3-c)]To a solution of methyl pyridin-6-yl) propionate (20 mg,76.2umol,1 eq) in dioxane (2 mL) was added DDQ (51.9 mg,0.22mmol,3 eq) and the mixture stirred under microwaves at 100 ℃ for 1hr. The reaction mixture was concentrated in vacuo. The residue was diluted with ethyl acetate (30 mL), washed with 10% aqueous NaOH (10 mL), brine (10 mL) and over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was purified by preparative TLC (petroleum ether/ethyl acetate=1/1). (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2, 3-c) was obtained as a yellow oil]Methyl pyridin-6-yl) propionate (10 mg,38umol,50% yield, 100% purity).

LCMS：Rt＝0.746min；C ₁₄ H ₁₆ N ₂ O ₃ MS calculated: 260.12; MS experimental values: 260.9[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CDCl ₃ )δ11.17(br s,1H),7.29(t,J＝2.76Hz,1H),6.98(d,J＝7.03Hz,1H),6.64(d,J＝7.03Hz,1H),6.39(t,J＝2.38Hz,1H),5.61(dd,J＝9.79,5.52Hz,1H),3.74(s,3H),2.17-2.07(m,1H),2.06 -1.99(m,1H),0.71-0.55(m,1H),0.49-0.32(m,2H),0.18-0.10(m,1H),0.05-0.00(m,1H)。

Step 2: (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2,3-c ] pyridin-6-yl) propionic acid

To (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2, 3-c)]To a solution of methyl pyridin-6-yl) propionate (10 mg, 38.4. Mu. Mol,1 eq.) in MeOH (0.5 mL) was added K-containing ₂ CO ₃ (15.9 mg,0.115mmol,3 eq.) H ₂ O (0.2 mL) and the mixture was stirred at 25℃for 16hr. After concentrating the reaction mixture in vacuo, the residue was taken up with H ₂ O (5 mL) was diluted, pH was adjusted to about 4 with 1M aqueous HCl and extracted with ethyl acetate (5 mL x 3). The combined organic phases were washed with brine (5 mL) and dried over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was used directly in the next step. (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2, 3-c) as a white solid was obtained]Pyridin-6-yl) propionic acid (8 mg,32.1umol,83.7% yield, 99% purity)。

LCMS：Rt＝0.701min；C ₁₃ H ₁₄ N ₂ O ₃ MS calculated: 246.10; MS experimental values: 246.9[ M+H ] ⁺ ]。

265: n- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2,3-c ] pyridin-6-yl) propanamide

To (2S) -3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2, 3-c)]Pyridin-6-yl) propionic acid (8 mg,32.4umol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of propionitrile (4.9 mg, 26.2. Mu. Mol, HCl) in DMF (1 mL) were added TEA (3.2 mg, 32.4. Mu. Mol, 4. Mu.L, 1 eq.) and T ₃ P (31.0 mg,48.7umol,28uL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 1hr. The reaction mixture was concentrated in vacuo. The crude product was checked by HPLC and purified by preparative HPLC (column Phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ water (0.05% nh) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:14% -44%,9.5 min). Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (7-oxo-1H-pyrrolo [2, 3-c)]Pyridin-6-yl) propionamide (2.9 mg,23.5% yield).

LCMS：Rt＝0.702min；C ₂₀ H ₂₃ N ₅ O ₃ MS calculated: 381.18; MS experimental values: 382.0[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.21-7.15(m,1H),7.11-7.02(m,1H),6.60-6.49(m,1H),6.28-6.19(m,1H),5.55-5.35(m,1H),4.95-4.78(m,1H),3.12-2.94(m,2H),2.39-2.27(m,1H),2.16 -1.99(m,2H),1.90-1.82(m,2H),1.76-1.61(m,2H),0.56-0.43(m,1H),0.31-0.17(m,2H),0.04-0.02(m,1H),0.02-0.00(m,1H)。

Example 118a Synthesis of viral protease inhibitor Compound 369

Step 1: (2S) -2- ((2S) -4-methyl-2- (4, 4-trifluoro-3-hydroxy-3-phenylbutyrylamino) pentanoylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (140 mg,467.66 mol,1 eq), 4-trifluoro-3-hydroxy-3-phenyl-butanoic acid (164.27 mg,701.48 mol,1.5 eq) in DCM (1.5 mL) and THF (1.5 mL) was added T ₃ P (446.40 mg,701.48umol,417.19uL,50% purity, 1.5 eq.) and DIEA (181.32 mg,1.40mmol,244.37uL,3 eq.). The mixture was stirred at 25℃for 2h. LCMS showed the reaction was complete and the desired MS was observed. The reaction mixture was concentrated under reduced pressure to remove the solvent. Purification of the residue by neutral prep HPLC gave the product (2S) -2- [ [ (2S) -4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino ] as a white solid ]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (120 mg,232.77umol,49.77% yield). MS (ESI) m/z 516.2[ M+H ]] ⁺ 。

Step 2: (2S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-methyl-2- (4, 4-trifluoro-3-hydroxy-3-phenylbutyrylamino) pentanamide

(2S) -2- [ [ (2S) -4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino]Pentanoyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (120 mg, 232.77. Mu. Mol,1 eq.) in MeOH/NH ₃ (7M, 5mL,150.36 eq.) in solution. The mixture was stirred at 80℃for 15h. LCMS showed the reaction was complete and the desired MS was observed. The reaction mixture was filtered and concentrated under reduced pressure to give the residue as a colorless oily product, (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3R) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino group]Pentanamide (120 mg, crude material). MS (ESI) m/z 501.2[ M+H ]] ⁺ 。

Step 3:

(2S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -4-methyl-2- (4, 4-trifluoro-3-hydroxy-3-phenylbutyrylamino) pentanamide

To (2S) -N- [ (1S) -2-amino-2-oxo 1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino group]To a solution of valeramide (120 mg,239.76umol,1 eq.) in DCM (3 mL) was added the Buerger's reagent (114.27 mg,479.51umol,2 eq.). The mixture was stirred at 25℃for 1h. LCMS showed the reaction was complete and the desired MS was observed. By adding 5mL of H ₂ O quench the reaction mixture and then extract with DCM (2.5 ml x 3). The combined organic layers were washed with brine (3 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave the product (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino group]Pentanamide (20.18 mg,38.77umol,16.17% yield, 92.698% purity). MS (ESI) m/z 483.3[ M+H ]] ⁺ 。

Preparative HPLC conditions:

column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-55％，8min

Column: phenomenex Luna C18, 75 x 30mm x 3um; mobile phase: [ water (0.2% fa) -ACN ]; b%:40% -65%,8min

¹ H NMR(400MHz,DMSO-d6)δppm 8.83(d,J＝7.72Hz,1H)8.37(d,J＝8.16Hz,1H)7.72(s,1H)7.53(br d,J＝7.06Hz,2H)7.29-7.42(m,3H)7.16(s,1H)4.77-4.99(m,1H)4.15-4.28(m,1H)3.03-3.20(m,4H)2.16-2.27(m,1H)1.95-2.09(m,2H)1.57-1.78(m,2H)1.29-1.44(m,3H)0.69-0.88(m,6H)。

Obtaining (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-4-methyl-2- [ (4, 4-trifluoro-3-hydroxy-3-phenyl-butyryl) amino group]Pentanamide (13.28 mg,27.20umol,11.34% yield, 98.809% purity). MS (ESI) M/z483.3[ M+H] ⁺ 。

¹ H NMR(400MHz,DMSO-d6)δppm 8.86(d,J＝7.94Hz,1H)8.58(d,J＝8.16Hz,1H)7.73(s,1H)7.51-7.62(m,2H)7.31-7.42(m,3H)6.92(s,1H)4.86-4.96(m,1H)4.11(ddd,J＝9.65,8.32,5.18Hz,1H)3.29(br d,J＝14.55Hz,1H)3.06-3.20(m,2H)2.89(d,J＝14.55Hz,1H)2.21-2.36(m,1H)2.02-2.17(m,2H)1.62-1.82(m,2H)1.20-1.38(m,2H)1.02-1.14(m,1H)0.73(d,J＝6.62Hz,3H)0.49(d,J＝6.39Hz,3H)。

EXAMPLE 119 Synthesis of viral protease inhibitor Compound 375

A mixture of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (1.2 g,4.19mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 30min. The reaction mixture was concentrated under reduced pressure to give crude methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (770 mg,4.14mmol,98.67% yield) as a yellow oil.

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl at 0 ℃C]To a mixture of methyl propionate (750 mg,4.03mmol,1 eq.) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (923.45 mg,4.03mmol,1 eq.) in DCM (3 mL) was added TEA (2.04 g,20.14mmol,2.80mL,5 eq.) in one portion. T is added to the mixture at 0 DEG C ₃ P (3.84 g,12.08mmol,3.59mL,3 eq.) and stirring at 25℃for 2h. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic layers were washed with 10mL brine (10 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give crude (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.5 g,3.77mmol,93.70% yield). MS (ESI) m/z 398.3[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-cyclopropyl-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (448 mg,1.51mmol,1 eq.) and 4-cyclopropyl-1H-indole-2-carboxylic acid (364.61 mg,1.81mmol,1.2 eq.) in DCM (8 mL) was added DMF (2 mL) and EDCI (868.40 mg,4.53mmol,3 eq.) in one portion. DMAP (553.43 mg,4.53mmol,3 eq.) was added to the mixture and the reaction stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product was purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2-cyclopropyl-2- [ (4-cyclopropyl-1H-indole-2-carbonyl) amino as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (500 mg,1.04mmol,68.90% yield). MS (ESI) m/z 481.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-cyclopropyl-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-cyclopropyl-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (500 mg,1.04mmol,1 eq.) in NH ₃ The mixture in MeOH (7M) (7 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-cyclopropyl-1H-indole-2-carboxamide (390 mg,837.73umol,80.52% yield). MS (ESI) m/z 466.3[ M+H ]] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-cyclopropyl-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-cyclopropyl-1H-indole-2-carboxamide (390 mg,837.73umol,1 eq.) in DCM (7 mL) was added the bergs reagent (1.20 g,5.03mmol,6 eq.) in one portion. The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure to give a residue. Purification of the residue by preparative HPLC (neutral conditions) gives N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-cyclopropyl-1H-indole-2-carboxamide (68 mg,151.95umol,18.14% yield). MS (ESI) m/z 448.3[ M+H ]] ⁺

Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-55％，8min

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(s,1H),9.12-8.84(m,1H),8.59(d,J＝7.6Hz,1H),7.84-7.65(m,1H),7.48(d,J＝1.3Hz,1H),7.21(d,J＝8.2Hz,1H),7.06(t,J＝7.7Hz,1H),6.64(d,J＝7.2Hz,1H),5.09-4.87(m,1H),4.69-4.36(m,1H),3.20-3.06(m,2H),2.46-2.07(m,4H),1.95-1.39(m,4H),1.01(br dd,J＝2.2,8.3Hz,2H),0.92-0.74(m,3H),0.55-0.34(m,2H),0.28-0.00(m,2H)

Example 120 Synthesis of viral protease inhibitor Compounds 377

Step 1: (2S) -2- [ [ (2S) -2- [ (4-ethoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate; EDCI (257.46 mg,1.34mmol,2 eq.) and DMAP (164.08 mg,1.34mmol,2 eq.) are added to a mixture of hydrochloride (250.00 mg,671.53umol,1 eq.) and 4-ethoxy-1H-indole-2-carboxylic acid (165.36 mg,805.83umol,1.2 eq.) in DCM (10 mL) and DMF (5 mL). After stirring the mixture at 25℃for 2H, the reaction mixture was taken up in H ₂ O (20 mL) was diluted and extracted with ethyl acetate (60 mL, extracted as 30mL x 2). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ Petroleum ether ethyl acetate=0:1) the residue was purified to give (2S) -2- [ [ (2S) -2- [ (4-ethoxy-1H-indole-2-carbonyl) amino as a yellow oil]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,369.94umol,55.09% yield, 90% purity).

Step 2: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-ethoxy-1H-indole-2-carboxamide

To (2S) -2- [ [ (2S) -2- [ (4-ethoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to a mixture of methyl propionate (120 mg,246.63umol,1 eq) ₃ MeOH (7M) (4.20 mg,246.63umol,20mL,1 eq.) and stirred at 80℃for 16h. After completion, the mixture was concentrated under reduced pressure to give a residue which was used directly in the next step. The compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl was obtained as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-ethoxy-1H-indole-2-carboxamide (112 mg,213.76umol,86.67% yield, 90% purity).

Step 3: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4-ethoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-ethoxy-1H-indole-2-carboxamide (111.11 mg,212.07umol,90% purity, 1 eq.) in DCM (2 mL) was added the berg reagent (151.61 mg,636.20umol,3 eq.). After stirring the mixture at 25 ℃ for 3h, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Purification of the residue by HCl preparative HPLC gave the compound N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4-ethoxy-1H-indole-2-carboxamide (38 mg,81.87umol,38.61% yield, 97.716% purity). MS (ESI) m/z 454.2[ M+H ]] ⁺

Column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:30% -60%,10min

H NMR(400MHz,DMSO-d6)δ＝11.56(br s,1H),8.90(d,J＝8.1Hz,1H),8.52(br d,J＝7.6Hz,1H),7.72(s,1H),7.39(s,1H),7.11-7.03(m,1H),7.01-6.96(m,1H),6.48(d,J＝7.6Hz,1H),5.04-4.92(m,1H),4.57-4.37(m,1H),4.14(q,J＝7.0Hz,2H),3.21-3.03(m,2H),2.43-2.28(m,1H),2.21-2.04(m,2H),1.82-1.46(m,5H),1.41(t,J＝7.0Hz,3H),0.91(dd,J＝6.4,19.5Hz,6H)

EXAMPLE 121 Synthesis of viral protease inhibitor Compound 379

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (1.3 g,3.27mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 30min. The reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (900 mg,3.03mmol,92.54% yield) as a yellow oil.

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (4478 mg,1.51mmol,1 eq.) and 4-propoxy-1H-indole-2-carboxylic acid (396.37 mg,1.81mmol,1.2 eq.) in DMF (2 mL) was added DCM (8 mL) and EDCI (866.48 mg,4.52mmol,3 eq.) in one portion. DMAP (552.19 mg,4.52mmol,3 eq.) was added to the mixture and the reaction stirred at 25℃for 2h. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (30 mL, added extraction as 10mL x 3). The combined organic layers were washed with brine (10 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2-cyclopropyl-2- [ (4-propoxy-1H-indole-2-carbonyl) amino as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (480 mg,962.75umol,63.90% yield). MS (ESI) M/z499.2[ M+H ] ⁺

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-propoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (480 mg,962.75umol,1 eq.) in NH ₃ The mixture in MeOH (7M) (3 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give crude N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-propoxy-1H-indole-2-carboxamide (380 mg,785.84umol,81.62% yield). MS (ESI) m/z 484.3[ M+H ]] ⁺

Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-60％，8min

EXAMPLE 122 Synthesis of viral protease inhibitor Compound 383

Step 1: (2S) -2- [ [ (2S) -2- [ (4, 4-difluorocyclohexyl) methoxycarbonylamino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To a mixture of bis (trichloromethyl) carbonate (940 mg,3.17mmol,1.36 eq.) in THF (2 mL) at 25deg.C DIEA (602.47 mg,4.66mmol,811.95uL,2 eq.) was added, and then THF (2 mL) containing (4, 4-difluorocyclohexyl) methanol (350 mg,2.33mmol,1 eq.) was added at 0deg.C. After stirring the mixture at 0 ℃ for 15min, the reaction was heated to 25 ℃ and stirred for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (4, 4-difluorocyclohexyl) methyl chloroformate (400 mg,1.51mmol,64.57% yield, 80% purity) as a yellow oil residue.

At 0 ℃, to (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate; to a mixture of hydrochloride (300 mg,893.32umol,1 eq.) in THF (3 mL) was added THF (2 mL) containing DIEA (346.37 mg,2.68mmol,466.80uL,3 eq.) and methyl chloroformate (4, 4-difluorocyclohexyl) (356.14 mg,1.34mmol,80% purity, 1.5 eq.). The mixture was stirred at 0℃for 10min and at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=8/1 to 0/1 to give (2S) -2- [ [ (2S) -2- [ (4, 4-difluorocyclohexyl) methoxycarbonylamino as a yellow oil]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (320 mg,605.65umol,67.80% yield, 90% purity). MS (ESI) m/z 476[ M+H ]] ⁺

Step 2: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] carbamic acid (4, 4-difluorocyclohexyl) methyl ester

To (2S) -2- [ [ (2S) -2- [ (4, 4-difluorocyclohexyl) methoxycarbonylamino]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added to a mixture of methyl propionate (300 mg,630.88umol,1 eq) ₃ MeOH (7M) (10.74 mg,630.89umol,1 eq.). After stirring the mixture at 80 ℃ for 16h, the reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step. The compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl was obtained as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl](4, 4-difluorocyclohexyl) methyl carbamate (280 mg,364.81umol,57.83% yield, 60% purity). MS (ESI) m/z 461.3[ M+H ]] ⁺

Step 3: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] carbamic acid (4, 4-difluorocyclohexyl) methyl ester

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of (4, 4-difluorocyclohexyl) methyl carbamate (230 mg,299.67umol,60% purity, 1 eq.) in DCM (6 mL) was added the berg reagent (142.83 mg,599.33umol,2 eq.). The mixture was stirred at 25℃for 60min. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with DCM (20 mL). The combined organic layers were concentrated under reduced pressure to give a residue. Purification of the residue by neutral prep HPLC gave the compound N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]Carbamoyl group]-3-methyl-butyl](4, 4-difluorocyclohexyl) methyl carbamate (48 mg,100.77umol,33.63% yield, 92.9% purity). MS (ESI) m/z 443.3[ M+H ]]+

Column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:25% -45%,8min

H NMR(400MHz,DMSO-d6)δ＝8.81(d,J＝8.0Hz,1H),7.81-7.66(m,1H),7.40(br d,J＝7.8Hz,1H),5.01-4.81(m,1H),4.03-3.88(m,1H),3.83(br d,J＝6.1Hz,2H),3.21-3.03(m,2H),2.40-2.22(m,1H),2.18-1.94(m,4H),1.90-1.54(m,8H),1.53-1.30(m,2H),1.29-1.10(m,2H),0.87(dd,J＝6.5,12.8Hz,6H)

EXAMPLE 123 Synthesis of viral protease inhibitor Compound 385

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (350 mg,1.22mmol,1 eq.) in HCl/EtOAc (4M, 5mL,16.36 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (272 mg, crude, HCl) as a yellow oil.

Step 2:

(2S) -4- (bicyclo [1.1.1] pent-1-yl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (272 mg,1.22mmol,1 eq., HCl) in DCM (6 mL) was added DMAP (298.47 mg,2.44mmol,2 eq.) and EDCI (468.34 mg,2.44mmol,2 eq.) and then (2S) -4- (1-bicyclo [1.1.1 eq.) was added ]Pentanyl) -1-tert-butoxycarbonyl-pyrrolidine-2-carboxylic acid (343.68 mg,1.22mmol,1 eq). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was poured into H at 25 ℃ ₂ O (20 mL) and then extracted with DCM (25 mL x 3). The combined organic layers were washed with brine (25 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 0/1) to give (2S) -4- (1-bicyclo [ 1.1.1) as a yellow solid]Pentanyl) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]Pyrrolidine-1-carboxylic acid tert-butyl ester (450 mg,1.00mmol,81.95% yield). MS (ESI) m/z 450.1[ M+H ]] ⁺

Step 3:

(2S) -2- ((2S) -4- (bicyclo [1.1.1] pent-1-yl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of (2S) -4- (1-bicyclo [1.1.1] pentanyl) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] pyrrolidine-1-carboxylic acid tert-butyl ester (350 mg,778.58 mol,1 eq) in HCl/MeOH (5 mL) was stirred at 25℃for 0.5h. After completion, the reaction mixture was concentrated to give methyl (2S) -2- [ [ (2S) -4- (1-bicyclo [1.1.1] pentanyl) pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (300 mg, crude, HCl) as a yellow solid.

Step 4:

(2S) -2- ((2S) -4- (bicyclo [1.1.1] pent-1-yl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (222.95 mg,1.17mmol,1.5 eq.) 2S-2- [ [ (2S) -4- (1-bicyclo [ 1.1.1)]Pentanyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (300 mg,777.43umol,1 eq, HCl) in DCM (6 mL) was added DMAP (284.94 mg,2.33mmol,3 eq) and CDI (378.18 mg,2.33mmol,3 eq). The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was poured into H at 25 ℃ ₂ O (20 mL) and then extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (25 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM HCOONH) ₄ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -55%,10 min) to give (2S) -2- [ [ (2S) -4- (1-bicyclo [ 1.1.1) as a yellow solid]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (200 mg,373.91umol,48.09% yield, 97.70% purity) and (2S) -2- [ [ (2S) -4- (1-bicyclo [ 1.1.1)]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (70 mg,127.20umol,16.36% yield, 94.96% purity). MS (ESI) m/z 523.2[ M+H ]] ⁺

Step 5: (2S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (bicyclo [1.1.1] pent-1-yl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

Isomer 1: (2S) -2- [ [ (2S) -4- (1-bicyclo [ 1.1.1)]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,382.71umol,1 eq.) in NH ₃ In MeOH (7M, 8mL,146.33 eq.)Is stirred at 80℃for 24h. After completion, the reaction mixture was concentrated to give (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-4- (1-bicyclo [ 1.1.1)]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (150 mg, crude). MS (ESI) M/z508.2[ M+H] ⁺

Isomer 2: (2S) -2- [ [ (2S) -4- (1-bicyclo [ 1.1.1) ]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (70 mg,133.95umol,1 eq.) in NH ₃ A solution in MeOH (7M, 4mL,209.04 eq.) was stirred at 80℃for 24hr. After completion, the reaction mixture was concentrated to give (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-4- (1-bicyclo [ 1.1.1)]Pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (50 mg, crude). MS (ESI) M/z508.2[ M+H] ⁺

Step 6: (2S) -4- (bicyclo [1.1.1] pent-1-yl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

Isomer 1: to (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4- (1-bicyclo [ 1.1.1)]To a solution of pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (145 mg,285.67umol,1 eq.) in DCM (4 mL) was added the berges reagent (680.76 mg,2.86mmol,10 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was poured into H at 25 ℃ ₂ O (20 mL) and then extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,8 min) to give (2S) -4- (1-bicyclo [ 1.1.1) as a white solid]Pentanyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-Formamide (46 mg,93.11umol,32.59% yield, 99.09% purity). MS (ESI) m/z 490.2[ M+H ]] ⁺ ； ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.24-6.79(m,3H),6.56-6.41(m,1H),5.05(s,1H),4.63(d,J＝4.4Hz,1H),4.34-3.36(m,6H),3.03-1.50(m,15H),1.37(d,J＝8.4Hz,1H)。

Isomer 2: to (2S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4- (1-bicyclo [ 1.1.1)]To a solution of pentanyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (50 mg,98.51umol,1 eq.) in DCM (2 mL) was added a berges reagent (70.42 mg,295.52umol,3 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was poured into H at 25 ℃ ₂ O (20 mL) and then extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,8 min) to give (2S) -4- (1-bicyclo [ 1.1.1) as a white solid]Pentanyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (10 mg,20.14umol,20.45% yield, 98.61% purity). MS (ESI) m/z 490.2[ M+H ]] ⁺ ； ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.16(d,J＝7.9Hz,1H),7.10-7.00(m,2H),6.53(d,J＝7.7Hz,1H),5.01(s,1H),4.66(d,J＝5.4,8.3Hz,1H),4.18-4.05(m,1H),3.95(s,3H),3.89(s,1H),3.79(d,J＝6.4,9.9Hz,1H),2.70-2.60(m,1H),2.51(d,J＝17.0Hz,2H),2.43(d,J＝4.3,8.5,12.5Hz,1H),2.30(d,J＝6.6,13.7Hz,1H),2.20-2.11(m,1H),2.04-1.83(m,3H),1.81-1.69(m,7H)。

Example 124 Synthesis of viral protease inhibitor Compound 387

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (0.3 g,1.05mmol,1 eq.) in HCl/EtOAc (4M, 5mL,19.09 eq.) was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (0.2 g, crude material) as a yellow gum.

Step 2:3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -8-oxa-2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (0.18 g,808.38umol,1 eq, HCl) in DMF (1 mL) and DCM (2 mL) was added DMAP (197.52 mg,1.62mmol,2 eq), 2-tert-butoxycarbonyl-8-oxa-2-azaspiro [4.5]]Decane-3-carboxylic acid (230.66 mg,808.38umol,1 eq.) and EDCI (309.93 mg,1.62mmol,2 eq.) and then stirring the resulting solution at 25℃for 2h. After completion, the reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:1 to 0:1) to give 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]Carbamoyl group]-8-oxa-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (0.3 g,562.26umol,69.55% yield, 85% purity). MS (ESI) m/z 454.2[ M+H ]] ⁺ 。

Step 3: (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (8-oxa-2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

A mixture of 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -8-oxa-2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester (0.25 g,551.23umol,1 eq.) in HCl/EtOAc (5 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- (8-oxa-2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (0.2 g, crude material, HCl) as a yellow oil.

Step 4: (2S) -2- (2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- (8-oxa-2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (0.2 g,512.99umol,1 eq, HCl) in DMF (1 mL) and DCM (2 mL) was added DMAP (125.34 mg,1.03mmol,2 eq) and then 4-methoxy-1H-indole-2-carboxylic acid (107.88 mg,564.29umol,1.1 eq) and EDCI (196.68 mg,1.03mmol,2 eq). The solution was stirred at 25℃for 1h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether ethyl acetate=0:1 to DCM meoh=10:1 to give (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.13 g,232.06umol,45.24% yield, 94% purity). MS (ESI) m/z 527.2[ M+H ]] ⁺ 。

Step 5: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.13 g,246.88umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 3mL,85.06 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5]Decane-3-carboxamide (0.12 g, crude). MS (ESI) m/z 512.2[ M+H ]] ⁺ 。

Step 6: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5]To a solution of decane-3-carboxamide (0.12 g,234.57 mol,1 eq.) in DCM (2 mL) was added the Bungeus reagent (167.70 mg,703.72 mol,3 eq.) and the solution was then stirred for 1h at 25 ℃. After completion, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,10 min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5]Decane-3-carboxamide (44.25 mg,88.76umol,37.84% yield, 99% purity). MS (ESI) m/z 494.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.23-7.08(m,2H),7.08-6.98(m,1H),6.53(br d,J＝7.6Hz,1H),5.02(br dd,J＝5.7,10.1Hz,1H),4.72-4.62(m,2H),4.19-4.03(m,1H),3.98-3.81(m,4H),3.77-3.62(m,4H),3.29-3.17(m,1H),2.52-2.20(m,3H),2.02-1.42(m,8H)。

EXAMPLE 125 Synthesis of viral protease inhibitor Compound 389

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propanamide

A solution of tert-butyl N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamate (2 g,7.37mmol,1 eq.) in HCl/EtOAc (4M, 50mL,27.13 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionamide (1.2 g, crude material) as a white solid.

Step 2: 2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

A solution of 2-tert-butoxycarbonyl-2-azaspiro [4.5] decane-3-carboxylic acid (3 g,10.59mmol,1 eq.) was added to HCl/MeOH (4M, 50mL,18.89 eq.) and stirred at 80℃for 2h. The mixture was concentrated under reduced pressure to give methyl 2-azaspiro [4.5] decane-3-carboxylate (2 g, crude material) as a yellow oil.

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxylic acid methyl ester (3 g,8.10mmol,1 eq.) in THF (45 mL) and H ₂ LiOH.H was added to the solution in O (15 mL) ₂ O (1.70 g,40.49mmol,5 eq.). The mixture was stirred at 25℃for 12h. After completion, by adding H ₂ O (50 mL) to quench the mixture, and then aqueous HCl (1M) was added to adjust the pH to about 3-4, and then extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (2.6 g, crude material). MS (ESI) m/z 357.1[ M+H ]] ⁺

To 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 0deg.C]Decane-3-carboxylic acid (1 g,2.81mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of propionamide (720.49 mg,4.21mmol,1.5 eq.) in DCM (30 mL) was added T ₃ P (3.57 g,5.61mmol,3.34mL,50% purity, 2 eq.) and DIEA (1.09 g,8.42mmol,1.47mL,3 eq.). The mixture was stirred at 30℃for 1h. After completion, by adding H ₂ O (100 mL) to quench the mixture and then extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=1:0 to 10:1 to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (700 mg,1.37mmol,48.96% yield). MS (ESI) m/z 510.3[ M+H ]] ⁺

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,9 min) isolation of N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (500 mg,1.02 mmol) gave the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide isomer 1 (264 mg,537.04umol,52.80% yield, 100% purity). MS (ESI) m/z 492.3[ M+H ]] ⁺

EXAMPLE 126 Synthesis of viral protease inhibitor Compounds 391

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (350 mg,1.22mmol,1 eq.) was added HCl/EtOAc (12 mL) and the mixture was stirred at 25℃for 1h. After completion, the mixture was concentrated in vacuo to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (330 mg, crude) as a yellow oil. MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (2S, 3S) -3-ethyl-2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) azetidine-1-carboxylic acid tert-butyl ester

To a solution of methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate (330 mg,1.77mmol,1 eq), (2S, 3S) -1- (tert-butoxycarbonyl) -3-ethylazetidine-2-carboxylic acid (406.32 mg,1.77mmol,1 eq.) in DMF (2 mL) and DCM (10 mL) were added EDCI (679.47 mg,3.54mmol,2 eq.) and DMAP (433.02 mg,3.54mmol,2 eq.). After stirring the mixture at 25℃for 1H, the mixture was prepared by adding H ₂ O (50 mL) to quench the mixture and then extract with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue and purified by preparative TLC (SiO ₂ Petroleum ether ethyl acetate=0:1) to give the crude (2S, 3S) -3-ethyl-2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) azetidine-1-carboxylic acid tert-butyl ester (270 mg,679.31umol,38.33% yield) as a yellow oil. MS (ESI) m/z 398.2[ M+H ]] ⁺

Step 3: (S) -2- ((2S, 3S) -3-Ethylazetidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S, 3S) -3-ethyl-2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxo-pyrrolidin-3-yl)To a solution of tert-butyl propan-2-yl) azetidine-1-carboxylate (240 mg,603.83 mol,1 eq.) in DCM (1 mL) was added TFA (4.13 g,36.23mmol,2.68mL,60 eq.) and the resulting mixture was stirred at 25℃for 1h. After completion, the residue was poured into NaHCO ₃ (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give the crude product methyl (S) -2- ((2S, 3S) -3-ethylazetidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (200 mg, crude) as a white solid. MS (ESI) m/z 298.2[ M+H ] ] ⁺

Step 4: (S) -2- ((2S, 3S) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((2S, 3S) -3-ethylazetidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (200 mg,672.61 mol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (128.59 mg,672.61 mol,1 eq) in DCM (1 mL) were added EDCI (257.88 mg,1.35mmol,2 eq) and DMAP (164.34 mg,1.35mmol,2 eq) and the mixture was stirred at 25 ℃ for 1H. After completion, the residue was poured into H ₂ O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ Petroleum ether ethyl acetate=0:1) to give the product methyl (S) -2- ((2S, 3S) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (90 mg,191.28umol,28.44% yield) as a white solid. MS (ESI) m/z 471.2[ M+H ]] ⁺

Step 5: (2S, 3S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamide

(S) -2- ((2S, 3S) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamido) -3- ((S) -2-oxoPyrrolidin-3-yl) propionic acid methyl ester (80 mg,170.03umol,1 eq.) in NH ₃ A solution in MeOH (7M, 16.00mL,658.72 eq.) was stirred at 80℃for 16h. After completion, the mixture was concentrated in vacuo to give the product (2S, 3S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamide (66 mg, crude material) as a white solid. MS (ESI) m/z 456.2[ M+H ]] ⁺

Step 6: (2S, 3S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamide

To a solution of (2S, 3S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamide (66 mg,144.89umol,1 eq) in DCM (3 mL) was added the beggar reagent (414.35 mg,1.74mmol,12 eq) and the mixture was then stirred at 25 ℃ for 3H. After completion, the mixture was concentrated in vacuo and purified by preparative HPLC (column Phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ water (0.05% nh ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,8 min) to give (2S, 3S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -3-ethyl-1- (4-methoxy-1H-indole-2-carbonyl) azetidine-2-carboxamide (5 mg,11.43umol,7.89% yield). MS (ESI) m/z 438.2[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.24-7.11(m,1H),7.09-6.61(m,2H),6.52-6.51(m,1H),5.08-4.87(m,0.5H),4.75-4.73(m,1.5H),4.56-4.43(m,1H),4.42-4.00(m,1H),3.93(s,3H),3.22-2.90(m,1H),2.65-2.63(m,2H),2.42-2.07(m,2H),2.04-1.49(m,5H),1.01-0.99(m,3H)

EXAMPLE 127 Synthesis of viral protease inhibitor Compound 395

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg,1.05mmol,1 eq.) was added HCl/EtOAc (4M, 30 mL) at 25deg.C and the mixture was stirred at 25deg.C for 1h. Concentrating the reaction mixture under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate as a yellow oil; hydrochloride (230 mg, crude) and used directly in the next step.

Step 2:

(S) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4, 4-dimethylpyrrolidine-1-carboxylic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (230 mg,1.03mmol,1 eq., HCl), (2S) -1-tert-butoxycarbonyl-4, 4-dimethyl-pyrrolidine-2-carboxylic acid (251.31 mg,1.03mmol,1 eq.), DMAP (252.38 mg,2.07mmol,2 eq.), EDCI (396.02 mg,2.07mmol,2 eq.), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give (S) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4, 4-dimethylpyrrolidine-1-carboxylic acid tert-butyl ester (200 mg,486.04umol,47.05% yield) as a yellow oil. MS (ESI) m/z 412.2[ M+H ]] ⁺ 。

Step 3: (S) -methyl-2- ((S) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate

A mixture of (S) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4, 4-dimethylpyrrolidine-1-carboxylic acid tert-butyl ester (200 mg,486.04umol,1 eq) and HCl/EtOAc (4M, 20 mL) was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product (S) -methyl-2- ((S) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (170 mg, crude material, HCl) as a yellow oil and used directly in the next step.

Step 4: (S) -2- ((S) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of (S) -methyl-2- ((S) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (170 mg, 488.74. Mu. Mol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (93.44 mg, 488.74. Mu. Mol,1 eq.), DMAP (119.42 mg, 977.47. Mu. Mol,2 eq.), EDCI (187.38 mg, 977.47. Mu. Mol,2 eq.), DMF (2 mL) and DCM (4 mL) was stirred at 25℃for 1H. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the compound (S) -methyl-2- ((S) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (180 mg,371.48umol,76.01% yield) as a yellow solid. MS (ESI) m/z 485.2[ M+H ]] ⁺ 。

Step 5: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide

(S) -methyl-2- ((S) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (180 mg,371.48umol,1 eq.) and NH ₃ The mixture of MeOH (7M, 7 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give the product (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide (170 mg, crude material) as a yellow solid. MS (ESI) m/z 470.2[ M+H ]] ⁺ 。

Step 6: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide

A mixture of (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide (160 mg,340.76umol,1 eq.) and Buerger' S reagent (649.66 mg,2.73mmol,8 eq.) and DCM (25 mL) was stirred at 25℃for 1H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -40%,8 min) to give the product (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide isomer 1 (27 mg,58.95umol,17.30% yield, 98.58% purity) as a white solid. MS (ESI) m/z 452.3[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.76-11.39(m,1H),9.18-8.79(m,1H),7.85-7.46(m,1H),7.21-6.67(m,3H),6.58-6.35(m,1H),5.13-4.81(m,1H),4.74-4.31(m,1H),3.97-3.55(m,5H),3.31-3.05(m,2H),2.47-1.96(m,4H),1.85-1.27(m,3H),1.25-0.80(m,6H)。

(S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) -4, 4-dimethylpyrrolidine-2-carboxamide isomer 2 (3 mg,6.41umol,1.88% yield, 96.44% purity) as a white solid. MS (ESI) m/z 452.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.78-11.34(m,1H),9.33-8.76(m,1H),7.91-7.53(m,1H),7.23-6.67(m,3H),6.61-6.31(m,1H),5.09-4.80(m,1H),4.61-4.43(m,1H),4.01-3.67(m,5H),3.20-2.99(m,2H),2.43-1.91(m,4H),1.86-1.55(m,3H),1.33-0.83(m,6H)。

Example 128 Synthesis of viral protease inhibitor Compound 397

Step 1: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamic acid tert-butyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1 g,3.49mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 15 mL) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure. Obtaining N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Methyl group]Ethyl group]Tert-butyl carbamate (900 mg, crude material). MS (ESI) m/z 272.2[ M+H ]] ⁺ 。

Step 2: (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propanamide

N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]A solution of tert-butyl carbamate (900 mg,3.32mmol,1 eq.) in HCl/EA (4M, 15 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Propionamide (650 mg, crude, HCl). MS (ESI) m/z 172.1[ M+H ]] ⁺ 。

Step 3: (6S) -6- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -5-azaspiro [2.4] heptane-5-carboxylic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionamide (400 mg,1.93mmol,1 eq. HCl), (6S) -5-tert-butoxycarbonyl-5-azaspiro [ 2.4)]A solution of heptane-6-carboxylic acid (464.77 mg,1.93mmol,1 eq.) and TEA (974.58 mg,9.63mmol,1.34mL,5 eq.) was dissolved in DCM (8 mL) and DMF (3 mL) and the solution was then cooled to 0deg.C. Adding T to the solution ₃ After P (3.68 g,5.78mmol,3.44mL,50% purity, 3 eq.) the mixture was stirred for 1h and gradually warmed to 25 ℃. After completion, H is added to the mixture ₂ O (50 mL) and then extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to obtain the final productThe product (6S) -6- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-5-azaspiro [2.4]]Heptane-5-carboxylic acid tert-butyl ester (200 mg, crude material). MS (ESI) m/z 395.2[ M+H ] ] ⁺ 。

Step 4: (6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -5-azaspiro [2.4] heptane-6-carboxamide

(6S) -6- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-5-azaspiro [2.4]]A solution of tert-butyl heptane-5-carboxylate (200 mg,464.12umol,1 eq. HCl) in HCl/EtOAc (4M, 15 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]-5-azaspiro [2.4]]Heptane-6-carboxamide (140 mg, crude, HCl). MS (ESI) m/z 295.2[ M+H ]] ⁺ 。

Step 5: (6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -5- (4-methoxy-1H-indole-2-carbonyl) -5-azaspiro [2.4] heptane-6-carboxamide

To (6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-5-azaspiro [2.4]]To a solution of heptane-6-carboxamide (140 mg,423.20umol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (80.91 mg,423.20umol,1 eq), EDCI (202.82 mg,1.06mmol,2.5 eq) was added DCM (3 mL) containing DMAP (155.11 mg,1.27mmol,3 eq) and the reaction was then stirred at 25 ℃ for 1H. After completion, H is added to the mixture ₂ O (30 mL) and then extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-5- (4-methoxy-1H-indole-2-carbonyl) -5-azaspiro [2.4]Heptane-6-carboxamide (80 mg,117.37umol,27.73% yield, 68.59% purity). MS (ESI) m/z 468.2[ M+H ]] ⁺ 。

Step 6: (6S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -5- (4-methoxy-1H-indole-2-carbonyl) -5-azaspiro [2.4] heptane-6-carboxamide

(6S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-5- (4-methoxy-1H-indole-2-carbonyl) -5-azaspiro [2.4]A solution of heptane-6-carboxamide (80 mg,171.12umol,1 eq) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (163.11 mg,684.47umol,4 eq) in DCM (5 mL) was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated and concentrated under reduced pressure to give (6S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-5- (4-methoxy-1H-indole-2-carbonyl) -5-azaspiro [2.4 ]Heptane-6-carboxamide (15.5 mg,34.44umol,20.13% yield, 99.88% purity). MS (ESI) m/z 450.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.23-7.12(m,1H),6.87-7.10(m,2H),6.59-6.39(m,1H),5.35-5.07(m,2H),4.85-4.69(m,1H),4.10-3.61(m,5H),3.03-2.17(m,4H),2.13-1.62(m,3H),1.62-1.22(m,1H),0.87-0.57(m,4H)。

EXAMPLE 129 Synthesis of viral protease inhibitor Compounds 399

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (130 mg,454.03umol,1 eq.) in HCl/dioxane (4M, 2.27mL,20 eq.) was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (173.4 mg,451.67umol,99.48% yield, 58% purity, HCl) as a yellow liquid.

To (7S) -6-tert-Butoxycarbonyl-6-azaspiro [3.4]]Octane-7-carboxylic acid (105.34 mg,412.59umol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (158.4 mg,412.59umol,58% purity, 1 eq., HCl) in DCM (1.2 mL) and DMF (0.4 mL) was added DMAP (100.81 mg,825.19umol,2 eq.) and EDCI (158.19 mg,825.19umol,2 eq.). After stirring the mixture at 25℃for 1H, the residue was taken up in H ₂ O (6 mL) was diluted and extracted with ethyl acetate (3 mL). The combined organic layers were washed with ethyl acetate (3 ml x 3), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give (7S) -7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow liquid]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Tert-butyl octane-6-carboxylate (66.3 mg,156.55 mol,37.94% yield). MS (ESI) m/z 424.0[ M+H ]] ⁺

A solution of (7S) -7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (66.3 mg,156.55 mol,1 eq) in HCl/MeOH (4M, 782.76ul,20 eq) was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (7S) -6-azaspiro [3.4] octane-7-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (71.1 mg,156.09umol,99.71% yield, 79% purity, HCl) as a yellow liquid.

To (2S) -2- [ [ (7S) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (62.8 mg,137.87umol,79% purity, 1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (26.36 mg,137.87umol,1 eq.) inDMAP (33.69 mg, 275.74. Mu. Mol,2 eq.) and EDCI (52.86 mg, 275.74. Mu. Mol,2 eq.) were added to a solution of DCM (1.2 mL) and DMF (0.4 mL) and maintained at 25℃for 1h. The residue was diluted with brine (6 mL) and extracted with EtOAc (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the product (2S) -2- [ [ (7S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a white solid]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (33.2 mg,66.86umol,48.50% yield). MS (ESI) m/z 497.1[ M+H ]] ⁺

(2S) -2- [ [ (7S) -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (23.0 mg,46.32umol,1 eq.) and ammonia (7M, 4mL,604.50 eq.) was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to give (7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carboxamide (15 mg, crude). MS (ESI) m/z 482.2[ M+H ]] ⁺

(7S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]A solution of octane-7-carboxamide (15 mg,28.66umol,92% purity, 1 eq.) and the Buerger's reagent (13.66 mg,57.32umol,2 eq.) was stirred at 25℃for 24h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]；B％:20% -45%,8 min) to give (7S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide (3.01 mg,6.49umol,22.66% yield, 100% purity). MS (ESI) m/z 464.3[ M+H ]] ⁺

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 6.95-7.24(m,3H)6.47-6.58(m,1H)5.01(br dd,J＝10.67,5.19Hz,1H)4.58(t,J＝7.09Hz,1H)3.82-4.19(m,5H)3.19(br t,J＝8.52Hz,1H)2.93-3.07(m,1H)2.28-2.56(m,3H)2.16-2.27(m,2H)1.94-2.14(m,6H)1.47-1.86(m,2H)。

EXAMPLE 130 Synthesis of viral protease inhibitor Compound 401

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

Methyl (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (400 mg,1.40mmol,1 eq.) in HCl/EtOAc (4M, 10mL,28.63 eq.) was stirred at 25℃for 0.5h. After completion, the mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (300 mg, crude material, HCl) as a yellow solid.

To (2S) -2-amino-3- [ (3S) -2-oxo-pyrrolidin-3-yl]Methyl propionate (300 mg,1.35mmol,1 eq. HCl) and (3S) -2-tert-butoxycarbonyl-2-azaspiro [4.4] ]To DMF (2 mL) and DCM (5 mL) of nonane-3-carboxylic acid (362.87 mg,1.35mmol,1 eq.) were added DMAP (329.19 mg,2.69mmol,2 eq.) and EDCI (516.56 mg,2.69mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester (340 mg,777.09umol,57.68% yield).

The (3S) -3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -2-azaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester (340 mg,777.09umol,1 eq.) in HCl/EtOAc (4M, 10mL,51.47 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (3S) -2-azaspiro [4.4] nonane-3-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (250 mg, crude material, HCl) as a yellow oil.

To (2S) -2- [ [ (3S) -2-azaspiro [4.4]]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg,668.67 mol,1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (127.84 mg,668.67 mol,1 eq.) in DMF (2 mL) and DCM (6 mL) were added DMAP (163.38 mg,1.34mmol,2 eq.) and EDCI (256.37 mg,1.34mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion, by adding H ₂ O (10 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with PE: ea=0:1) to give the product (2S) -2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] as a yellow oil]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (180 mg, 35)2.54umol,52.72% yield). MS (ESI) M/z511.2[ M+H ]] ⁺

Ammonia (7M, 20mL,397.12 eq) containing methyl (2S) -2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (180 mg,352.54 mol,1 eq) was stirred at 80℃for 16H. After completion, the mixture was concentrated under reduced pressure to give the product (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamide (170 mg, crude material) as a yellow oil.

To (3S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]To DCM (3 mL) of nonane-3-carboxamide (170 mg,343.04umol,1 eq.) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (408.74 mg,1.72mmol,5 eq.). The mixture was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product (3S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]Nonane-3-carboxamide (25 mg,51.09umol,14.89% yield, 97.6% purity). MS (ESI) m/z478.2[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.22-7.12(m,1H),7.11-6.98(m,2H),6.58-6.45(m,1H),5.11-4.95(m,1H),4.65-4.52(m,1H),3.94(s,3H),3.93-3.80(m,2H),3.28-3.18(m,1H),2.54-2.02(m,4H),2.01-1.48(m,12H)。

Step 7: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4] nonane-3-carboxamide

Isomer 1: by preparative SFC (column: DAICEL CHIRALPAK IC (250 mm. Times.30 mm,5 um); mobile phase: [ Neu-ETOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%,15 min) isolation of (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [ 4.4)]Nonane-3-carboxamide (30 mg, 62.82. Mu. Mol) gives the product (3S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]Nonane-3-carboxamide (12.11 mg,24.62umol,39.20% yield, 97.1% purity). MS (ESI) M/z478.2[ M+H] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.20-7.11(m,1H),7.08-6.85(m,2H),6.59-6.42(m,1H),5.05(br dd,J＝5.6,10.4Hz,1H),4.58(br dd,J＝7.4,9.6Hz,1H),3.97-3.92(m,3H),3.88-3.52(m,2H),3.28(br s,1H),2.87-2.65(m,1H),2.47-2.29(m,2H),2.25-2.16(m,1H),2.03-1.53(m,11H),1.34-1.20(m,1H)。

Isomer 2: by preparative SFC (column: DAICEL CHIRALPAK IC (250 mm. Times.30 mm,5 um); mobile phase: [ Neu-ETOH) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%,15 min) isolation of (S) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [ 4.4)]Nonane-3-carboxamide (30 mg, 62.82. Mu. Mol) gives the product (3S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.4]Nonane-3-carboxamide (16.81 mg,34.46umol,54.86% yield, 97.9% purity). MS (ESI) M/z478.2[ M+H] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23-7.13(m,1H),7.10-6.84(m,2H),6.52(d,J＝7.7Hz,1H),5.03(br dd,J＝5.7,10.4Hz,1H),4.67-4.54(m,1H),4.00-3.57(m,5H),3.27-3.16(m,1H),2.55-2.39(m,1H),2.37-2.04(m,3H),2.02-1.44(m,11H),1.43-1.16(m,1H)。

EXAMPLE 131 Synthesis of viral protease inhibitor Compound 405

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (225 mg,1.21mmol,1 eq.) in DMF (2 mL) and DCM (4 mL) was added TEA (733.62 mg,7.25mmol,1.01mL,6 eq.) and T ₃ P (1.15 g,3.62mmol,1.08mL,3 eq.) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (296.42 mg,1.21mmol,1 eq.). The solution was stirred at 25℃for 1h. Reactant H ₂ O (40 mL) was diluted and extracted with ethyl acetate (50 mL. 3) and the organic layer was carefully concentrated to give crude compound (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a yellow solid ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (440 mg, crude) and was used directly in the next step. MS (ESI) m/z 414.1[ M+H ]] ⁺

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (440 mg,1.06mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 1h. TLC (DCM: meoh=10:1) showed the desired product and the reaction was carefully concentrated to give the crude material. The compound (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] as a yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (310 mg, crude) was used directly in the next step. MS (ESI) m/z 314.3[ M+H ]] ⁺

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (310 mg,989.18umol,1 eq.) in DMF (4 mL) and DCM (4 mL) was added EDCI (379.25 mg,1.98mmol,2 eq.) and DMAP (241.70 mg,1.98mmol,2 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (189.11 mg,989.18umol,1 eq.). The solution was stirred at 25℃for 3H and then the reaction was taken up with H ₂ O (40 mL) was diluted and extracted with ethyl acetate (80 mL x 3) and the organic layer was carefully concentrated to give the crude material. By preparative TLC (SiO) ₂ EA: meoh=10:1) to give the product. Obtaining the compound (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,411.05umol,41.55% yield). MS (ESI) m/z 487.2[ M-H] ⁺

(2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (135 mg,277.46 mol,1 eq.) in NH ₃ A solution in MeOH (7M, 8mL,201.83 eq.) was stirred at 65℃for 16h. HPLC showed the desired product. The reaction was carefully concentrated to give the crude material. Obtaining the compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (130 mg, crude) and used directly in the next step. MS (ESI) m/z 472.3[ M+H ] ] ⁺

Preparative HPLC conditions: column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]；B％：35％-55％，8min

To N- [ (1S) -1- [ [ (1S) -2-ammonia1- [ [ (3S) -2-oxo-pyrrolidin-3-yl-2-oxo-1- [ (3S) -yl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (130 mg,275.69umol,1 eq.) in DCM (7 mL) was added the Buerger's reagent (197.09 mg,827.06umol,3 eq.). The solution was stirred at 25℃for 1h. The reaction was carefully concentrated to give the crude material, and the crude material was purified by preparative HPLC (TFA) to give N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (36 mg,75.41umol,27.35% yield, 95% purity). MS (ESI) M/z454.1[ M+H] ⁺ 。

Preparative HPLC conditions: column: phenomenex luna C18, 80 x 40mm x 3um; mobile phase: [ Water (0.04% HCl) -ACN ]; b%:30% -55%,7min

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 1.02(s,9H)1.74-1.94(m,4H)2.21-2.37(m,2H)2.52-2.63(m,1H)3.16-3.26(m,2H)3.92(s,3H)4.63(dd,J＝8.49,4.30Hz,1H)4.98-5.06(m,1H)6.50(d,J＝7.72Hz,1H)7.02(d,J＝8.38Hz,1H)7.10-7.16(m,1H)7.23(d,J＝0.88Hz,1H)。

EXAMPLE 132 Synthesis of viral protease inhibitor Compound 409

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propanamide

A mixture of tert-butyl N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamate (300 mg,1.11mmol,1 eq.) in HCl/EtOAc (4M, 10mL,36.17 eq.) was stirred at 25℃for 0.5h. After completion, the mixture was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionamide (200 mg, crude material, HCl) as a white solid.

Step 2: (2S, 4R) -2- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -4-ethoxypyrrolidine-1-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl at 0 ℃C]To propionamide (200 mg,963.12umol,1 eq, HCl) and DMF (4 mL) and DCM (8 mL) of (2 s,4 r) -1-tert-butoxycarbonyl-4-ethoxy-pyrrolidine-2-carboxylic acid (249.74 mg,963.12umol,1 eq) were added TEA (487.29 mg,4.82mmol,670.27ul,5 eq) and T3P (1.84 g,2.89mmol,1.72mL,50% purity, 3 eq). The mixture was stirred at 25℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give the product (2S, 4 r) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-4-ethoxypyrrolidine-1-carboxylic acid tert-butyl ester (140 mg,339.41umol,35.24% yield). MS (ESI) m/z 413.1[ M+H ]] ⁺

Step 3: (2S, 4R) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-ethoxypyrrolidine-2-carboxamide

A mixture of (2S, 4R) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -4-ethoxypyrrolidine-1-carboxylic acid tert-butyl ester (100 mg,242.44umol,1 eq) in HCl/EtOAc (4M, 10mL,164.99 eq) was stirred at 25℃for 0.5h. After completion, the mixture was concentrated under reduced pressure to give the product (2S, 4 r) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -4-ethoxy-pyrrolidine-2-carboxamide (80 mg, crude, HCl) as a white solid.

Step 4: (2S, 4R) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4-ethoxy-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

To (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-4-ethoxy-pyrrolidine-2-carboxamide (80 mg,229.34umol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid(65.77 mg, 344.01. Mu. Mol,1.5 eq.) DMAP (56.04 mg, 458.68. Mu. Mol,2 eq.) and EDCI (87.93 mg, 458.68. Mu. Mol,2 eq.) are added to a mixture of DCM (3 mL) and DMF (1 mL). The mixture was stirred at 25℃for 1h. After completion, by adding H ₂ O (30 mL) to quench the mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give the product (2S, 4 r) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow oil]Methyl group]Ethyl group]-4-ethoxy-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (100 mg, crude). MS (ESI) m/z 486.2[ M+H ]] ⁺

Step 5: (2S, 4R) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -4-ethoxy-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

To (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ]Methyl group]Ethyl group]To a mixture of 4-ethoxy-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (80 mg,164.77umol,1 eq.) in DCM (3 mL) was added a bergiut reagent (196.33 mg,823.84umol,5 eq.). The mixture was stirred at 25℃for 2h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -45%,10 min) to give the product (2S, 4 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]-4-ethoxy-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (28 mg,58.81umol,35.69% yield, 98.2% purity). MS (ESI) m/z 468.2[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.19-7.13(m,1H),7.09-6.86(m,2H),6.57-6.42(m,1H),5.17-5.01(m,1H),4.69-4.58(m,1H),4.36-4.18(m,1H),4.16-3.97(m,2H),3.96-3.85(m,3H),3.68-3.44(m,2H),3.00-2.54(m,2H),2.50-2.31(m,2H),2.25-2.02(m,2H),2.01-1.72(m,2H),1.69-1.26(m,1H),1.25-1.13(m,3H)。

EXAMPLE 133 Synthesis of viral protease inhibitor Compound 433

Step 1: 2-amino-2- (3-pyridinyl) acetic acid methyl ester

At 25℃under N ₂ To 2- (t-butoxycarbonylamino) -2- (3-pyridyl) acetic acid (0.5 g,1.98mmol,1 eq.) was added HCl/MeOH (4M, 20mL,40.36 eq.) in one portion. The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give a crude product. The crude product was used in the next step without purification. Methyl 2-amino-2- (3-pyridinyl) acetate (400 mg, crude material, HCl) was obtained as a yellow oil and used directly in the next step. MS (ESI) m/z 167.1[ M+H ] ] ⁺

Step 2:2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -2- (3-pyridinyl) acetic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of 4-methyl-pentanoic acid (600.76 mg,1.97mmol,1 eq.) and methyl 2-amino-2- (3-pyridinyl) acetate (400 mg,1.97mmol,1 eq., HCl), DIPEA (1.28 g,9.87mmol,1.72mL,5 eq.) in THF (1.2 mL) and DCM (1.2 mL) was added T ₃ P (1.88 g,2.96mmol,1.76mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by preparative HPLC. Obtaining 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]Methyl 2- (3-pyridyl) acetate (0.3 g, crude material). MS (ESI) m/z 453.2[ M+H ]] ⁺

Preparative HPLC conditions: column: kromasil C18 (250 x 50mm x 10 um); mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-50％，10min。

Step 3: n- [ (1S) -1- [ [ 2-amino-2-oxo-1- (3-pyridinyl) ethyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward 2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4-methyl-pentanoyl]Amino group]NH was added at a time to a mixture of methyl-2- (3-pyridyl) acetate (0.2 g,441.99umol,1 eq.) ₃ MeOH (7M, 6mL,95.03 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooled to 25 ℃ and concentrated to give the product. By preparative TLC (SiO) ₂ ，DCM:MeOH＝10:1，R _f =0.22) purification residue. Obtaining N- [ (1S) -1- [ [ 2-amino-2-oxo-1- (3-pyridinyl) ethyl ] as a pale yellow solid]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (70 mg, crude). MS (ESI) m/z 438.2[ M+H ]] ⁺

Step 4: n- [ (1S) -1- [ [ cyano (3-pyridinyl) methyl ] carbamoyl ] -3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- [ [ 2-amino-2-oxo-1- (3-pyridinyl) ethyl]Carbamoyl group]-3-methyl-butyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (60 mg,137.15umol,1 eq.) in DCM (0.2 mL) was added the Bogis reagent (65.37 mg,274.29umol,2 eq.) in one portion. The mixture was stirred at 25℃for 16h. The reaction mixture was concentrated to give a crude product. The crude product was purified twice by preparative HPLC. Obtaining N- [ (1S) -1- [ [ cyano (3-pyridinyl) methyl ] as a white solid ]Carbamoyl group]-3-methyl-butyl]-4-methoxy-1H-indole-2-carboxamide (12.78 mg,29.52umol,21.52% yield, 96.878% purity). MS (ESI) m/z 423.2[ M+H ]] ⁺

Preparative HPLC conditions: column: waters Xbridge BEH C18 100×25mm×5um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-55％，10min。

Column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-55％，8min。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.61(dd,J＝7.03,1.77Hz,1H),9.49(dd,J＝17.24,7.83Hz,1H),8.59-8.71(m,2H),8.53(d,J＝7.82Hz,1H),7.85-7.93(m,1H),7.47-7.55(m,1H),7.38(t,J＝2.51Hz,1H),7.06-7.14(m,1H),7.01-7.01(m,1H),7.01(dd,J＝8.25,3.24Hz,1H),6.51(dd,J＝7.70,1.34Hz,1H),6.32(dd,J＝12.41,7.76Hz,1H),4.44-4.61(m,1H),3.89(d,J＝1.10Hz,3H),1.62-1.81(m,2H),1.46-1.60(m,1H),0.81-1.03(m,7H)。

At 25℃under N ₂ To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (15 g,78.46mmol,1 eq.) and (2S) -2-amino-4-methyl-pentanoic acid tert-butyl ester (21.07 g,94.15mmol,1.2 eq., HCl) in DMF (150 mL) was added EDCI (19.55 g,102.00mmol,1.3 eq.), HOBt (13.78 g,102.00mmol,1.3 eq.), TEA (23.82 g,235.38mmol,32.76mL,3 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. To the reaction mixture was added water (450 mL) and extracted with ethyl acetate (250 mL x 3) to give an organic phase. The organic phase was washed with 5% citric acid (300 mL) and 5% aqueous sodium bicarbonate (300 mL) and dried over anhydrous sodium sulfate, filtered and concentrated to give the product. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=30:1 to 10:1) to give (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid ]-4-methyl-pentanoic acid tert-butyl ester (24 g,66.58mmol,84.87% yield). MS (ESI) m/z 361.2[ M+H ]] ⁺

Step 7: (S) -2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanoic acid

At 0℃under N ₂ Downward (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of tert-butyl-4-methyl-pentanoate (10 g,27.74mmol,1 eq.) in DCM (30 mL) was added TFA (61.60 g,540.26mmol,40mL,19.47 eq.) and H in one portion ₂ O (4.00 g,221.98mmol,4.00mL,8.00 eq.). The mixture was stirred at 25 ℃ and stirred for 2h. The reaction mixture was concentrated to give a crude product. The crude product was purified by slurrying with petroleum ether: ethyl acetate=10:1 (20 mL) and filtered to give the product. (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) was obtained as a pale yellow solidRadical) amino group]-4-methyl-pentanoic acid (6 g,19.22mmol,69.27% yield, 97.48% purity). MS (ESI) M/z305.1[ M+H] ⁺

EXAMPLE 134 Synthesis of viral protease inhibitor Compound 439

Step 1: (2S) -2-amino-3- (2-pyridinyl) propanoic acid methyl ester

At 25℃under N ₂ To a mixture of (2S) -2- (tert-butoxycarbonylamino) -3- (2-pyridinyl) propionic acid (1 g,3.76mmol,1 eq.) was added HCl/MeOH (4M, 10mL,10.65 eq.) in one portion. The mixture was stirred at 25℃for 1h. The reaction mixture was concentrated to give the product. Methyl (2S) -2-amino-3- (2-pyridinyl) propanoate (900 mg,3.48mmol,92.79% yield, 98% purity, 2 HCl) was obtained as a white solid and used directly in the next step. MS (ESI) m/z 181.1[ M+H ] ] ⁺

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (2-pyridinyl) propanoic acid methyl ester

At 0℃under N ₂ To a mixture of methyl (2S) -2-amino-3- (2-pyridinyl) propanoate (0.9 g,3.56mmol,1 eq., 2 HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propanoate (978.23 mg,4.27mmol,1.2 eq.) and DIPEA (2.30 g,17.78mmol,3.10mL,5 eq.) in DCM (6 mL) and THF (6 mL) was added T ₃ P (3.39 g,5.33mmol,3.17mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 2h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. Obtaining (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a pale yellow solid]Amino group]Methyl 3- (2-pyridinyl) propionate (1.1 g,2.81mmol,79.03% yield) and used directly in the next step. MS (ESI) m/z 392.2[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (2-pyridinyl) propanoic acid methyl ester

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]To a mixture of methyl 3- (2-pyridyl) propionate (1.1 g,2.81mmol,1 eq.) was added HCl/MeOH (4M, 11mL,15.66 eq.) in one portion. The mixture was stirred at 25℃for 1h. The reaction mixture was concentrated to give the product. Obtaining (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl group as brown solid ]Amino group]Methyl 3- (2-pyridinyl) propionate (1 g, crude material, HCl) and used directly in the next step. MS (ESI) m/z 292.2[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- (2-pyridinyl) propanoic acid methyl ester

At 0℃under N ₂ Downward (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]Methyl 3- (2-pyridinyl) propionate (0.8 g,2.20mmol,1 eq, 2 HCl) and 4-methoxy-1H-indole-2-carboxylic acid (461.86 mg,2.42mmol,1.1 eq) and DIPEA (1.42 g,10.98mmol,1.91mL,5 eq) in a mixture of DCM (0.5 mL) and THF (0.5 mL) was added T ₃ P (2.10 g,3.29mmol,1.96mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 12h. To the reaction mixture was added saturated sodium bicarbonate solution (10 mL) and extracted with DCM (10 mL x 2) to give an organic phase. The organic phase was concentrated to give the crude product. The residue was purified by flash chromatography on silica gel. Obtaining (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a pale yellow solid]Propionyl radical]Amino group]Methyl 3- (2-pyridinyl) propanoate (0.8 g,1.50mmol,68.38% yield, 87.2% purity). MS (ESI) m/z 465.2[ M+H ] ] ⁺

Step 5: n- [ (1S) -1- (cyclopropylmethyl) -2- [ [ (1S) -1- (nitrosomethyl) -2- (2-pyridinyl) ethyl ] amino ] -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]NH was added at a time to a mixture of methyl 3- (2-pyridyl) propionate (0.2 g,430.56umol,1 eq.) ₃ MeOH (7M, 4mL,65.03 eq.). The mixture was stirred at 80℃for 12h. The reaction mixture was cooledBut to 25 ℃ and concentrated to give the crude product. Obtaining N- [ (1S) -1- (cyclopropylmethyl) -2- [ [ (1S) -1- (nitrosomethyl) -2- (2-pyridinyl) ethyl ] as a pale yellow solid]Amino group]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (200 mg, crude) and used directly in the next step. MS (ESI) m/z 450.2[ M+H ]] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- (2-pyridinyl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -1- (cyclopropylmethyl) -2- [ [ (1S) -1- (nitrosomethyl) -2- (2-pyridinyl) ethyl]Amino group]-2-oxo-ethyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (0.1 g,222.47umol,1 eq.) in DCM (1 mL) was added the Bogis reagent (212.06 mg,889.88umol,4 eq.) in one portion. The mixture was stirred at 25℃for 12h. The reaction mixture was concentrated to give a crude product. The crude product was purified by preparative HPLC. Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- (2-pyridinyl) ethyl ] as a white solid ]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (25.44 mg,58.27umol,26.19% yield, 98.833% purity). MS (ESI) M/z432.2[ M+H] ⁺

Preparative HPLC conditions: column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：30％-50％，8min

¹ H NMR(400MHz,MeOD-d ₄ )δppm 8.27-8.39(m,1H),7.64-7.73(m,1H),7.31-7.39(m,1H),7.23-7.30(m,1H),7.12-7.23(m,2H),7.00-7.07(m,1H),6.52(d,J＝7.50Hz,1H),5.28(t,J＝7.17Hz,1H),4.51-4.63(m,1H),3.87-3.98(m,3H),3.30-3.31(m,2H),1.57-1.83(m,2H),0.62-0.85(m,1H),0.34-0.54(m,2H),0.05-0.22(m,2H)。

¹ H NMR (400 MHz, chloroform-d) delta ppm 9.35 (br s, 1H), 8.50-8.68 (m, 1H), 8.04-8.26 (m, 1H), 7.51 (td, J=7.69, 1.75Hz, 1H), 6.93-7.11 (m, 4H), 6.77-6.90 (m, 2H), 6.34-6.42 (m, 1H), 5.11-5.23 (m, 1H), 4.61-4.71 (m, 1H), 3.76-3.87 (m, 3H), 3.07-3.25 (m, 2H), 1.55-1.69 (m, 2H), 0.48-0.67 (m, 1H), 0.28-0.40 (m, 2H), 0.09-0.08 (m, 2H).

EXAMPLE 135 Synthesis of viral protease inhibitor Compound 448

Step 1: (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid methyl ester

To a solution of 4-methyl-3-nitro-1H-pyridin-2-one (500 mg,3.24mmol,1 eq.) in DMF (10 mL) was added NaH (181.6 mg,4.54mmol,60% purity, 1.4 eq.) at 0deg.C and the reaction mixture was stirred at 25deg.C for 0.5hr. Next, (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester (671.7 mg,3.24mmol,1 eq.) was added at 0deg.C. At N ₂ The mixture was stirred at 25℃for 16h. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (50 mL) was quenched and extracted with ethyl acetate (150 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography24g />Silica flash column, eluent: 0-50% ethyl acetate/petroleum ether gradient, at 35 mL/min) to give methyl (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanoate (457 mg,45.1% yield) as a yellow solid.

LCMS：Rt＝0.780min；C ₁₃ H ₁₆ N ₂ O ₅ MS calculated: 280.11; MS experimental values: 281.0[ M+H ] ⁺ ]。

¹ H NMR(400MHz,DMSO-d ₆ )δ7.93(d,J＝7.03Hz,1H),6.43(d,J＝7.03Hz,1H),5.30(t,J＝7.65Hz,1H),3.65(s,3H),2.23(s,3H),1.99(t,J＝7.40Hz,2H),0.56-0.45(m,1H),0.38-0.25(m,2H),0.15-0.13(m,2H)。

Step 2: (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid

Methyl (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanoate (255 mg,0.90mmol,1 eq.) LiOH. H ₂ O (151.5 mg,3.61mmol,4 eq.) in THF (2.1 mL), meOH (0.7 mL), H ₂ The mixture in O (0.7 mL) was degassed and replaced with N ₂ Purge 3 times and then at N ₂ The mixture was stirred at 25℃for 1hr under an atmosphere. LCMS showed detection of one peak with the expected MS. Adding H to the mixture ₂ O (5 mL), followed by addition of 2M HCl (2 mL) to the mixture to adjust the pH to about 6-7. Adding H to the mixture ₂ O (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid (207 mg,77.9% yield) as a yellow solid.

LCMS：Rt＝0.732min；C ₁₂ H ₁₄ N ₂ O ₅ MS calculated: 266.09; MS experimental values: 267.0[ M+H ] ⁺ ]。

Step 3: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide

To a solution of (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid (207 mg,0.77mmol,1 eq.) in DMF (2 mL) was added T ₃ P (989.5 mg,1.55mmol,0.92mL,50% purity, 2 eq), TEA (314.6 mg,3.11mmol,0.43mL,4 eq) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (147.4 mg,0.77mmol,1 eq., HCl). The mixture was stirred at 25℃for 4h. LCMS showed detection of a peak with the expected MS. H for mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By flash silica gel chromatography12gSilica flash column, eluent: purification of the residue with 0-10% DCM/MeOH at 30mL/min gave the compound (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid ]Ethyl group]-3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (60 mg,17.8% yield).

LCMS：Rt＝1.336min；C ₁₉ H ₂₃ N ₅ O ₅ MS calculated: 401.17; MS experimental values: 402.1[ M+H ] ⁺ ]。

(2S) -2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-propionamide

To (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (60 mg,0.14mmol,1 eq.) in THF (2 mL) was added Pd/C (70 mg,65.7umol,10% purity, 0.44 eq.). At H ₂ The mixture was stirred at 25℃for 15min. LCMS showed detection of one peak with the expected MS. The reaction mixture was filtered and the filtrate was concentrated. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:13% -43%,9.5 min) to give (2S) -2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a brown solid]Ethyl group]-3-cyclopropyl-propionamide (7.45 mg,19.7umol,13.2% yield, 98.4% purity).

LCMS：Rt＝0.698min；C ₁₉ H ₂₅ N ₅ O ₃ MS calculated: 371.20; MS experimental values: 372.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ6.94-6.82(m,1H),6.11-6.01(m,1H),5.40-5.23(m,1H),4.86(br dd,J＝6.0,9.8Hz,1H),3.14-3.08(m,2H),2.47-2.27(m,1H),2.23-2.03(m,2H),1.99-1.91(m,3H),1.83-1.57(m,4H),0.48(br d,J＝7.3Hz,1H),0.34-0.19(m,2H),0.02-0.16(m,2H)。

EXAMPLE 136 Synthesis of viral protease inhibitor Compound 449

To (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (345.0 mg,0.85mmol,1 eq.) in THF (5 mL) was added Pd/C (233.1 mg,0.21mmol,10% purity). At H ₂ The mixture was stirred at 25℃for 25min. LCMS showed detection of one peak with the expected MS. The reaction mixture was filtered and the filtrate was taken up with H ₂ O (20 mL) was quenched and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give the product (203 mg). By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%, min) to isolate 70mg of the product to give 2- [ (1S) -3-amino-4-methyl-2-oxo-1-pyridinyl) as a white solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (20.08 mg,6.2% yield) and 2- [ (1R) -3-amino-4-methyl-2-oxo-1-pyridinyl ]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (23.04 mg,7.0% yield).

Isomer 1: LCMS: rt= 0.659min; c (C) ₁₉ H ₂₅ N ₅ O ₃ MS calculated: 371.20; MS experimental values: 394.1[ M+Na ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.02(d,J＝7.0Hz,1H),6.22(d,J＝7.1Hz,1H),5.50(t,J＝7.8Hz,1H),5.04-4.98(m,1H),3.37-3.32(m,2H),2.52-2.46(m,1H),2.38-2.24(m,2H),2.11(s,3H),1.94-1.81(m,4H),0.61-0.56(m,1H),0.42-0.38(m,2H),0.13-0.02(m,2H)。

Isomer 2: LCMS: rt=0.704 min; c (C) ₁₉ H ₂₅ N ₅ O ₃ MS calculated: 371.20; MS experimental values: 372.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.03(d,J＝7.1Hz,1H),6.20(d,J＝7.0Hz,1H),5.41(dd,J＝7.1,8.4Hz,1H),5.00(br dd,J＝6.1,10.0Hz,1H),3.29-3.24(m,2H),2.49(dq,J＝5.4,9.3Hz,1H),2.31-2.21(m,2H),2.09(s,3H),1.98-1.76(m,4H),0.69-0.57(m,1H),0.50-0.41(m,2H),0.17-0.04(m,2H)。

Example 137 Synthesis of viral protease inhibitor Compounds 450

To a solution of 4-methyl-3-nitro-1H-pyridin-2-one (1 g,6.49mmol,1 eq.) in DMF (15 mL) was added NaH (363.3 mg,9.08mmol,60% purity, 1.4 eq.) at 0deg.C and the reaction mixture stirred at 25deg.C for 0.5hr. Next, (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester (1.34 g,6.49mmol,1 eq.) was added at 0deg.C. At N ₂ The mixture was stirred at 25℃for 16h. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography24g/>Silica flash column, eluent: 0-50% ethyl acetate/petroleum ether gradient, at 35 mL/min) to give (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propan as a yellow solid Methyl acid ester (867 mg,47.4% yield).

LCMS：Rt＝0.785min；C ₁₃ H ₁₆ N ₂ O ₅ MS calculated: 280.11; MS experimental values: 281.1[ M+H ] ⁺ ]。

Methyl (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanoate (867 mg,3.09mmol,1 eq.) LiOH. H ₂ O (519.2 mg,12.37mmol,4 eq.) in THF (6 mL), meOH (2 mL), H ₂ The mixture in O (2 mL) was degassed and replaced with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred under an atmosphere at 25℃for 1h. LCMS showed detection of one peak with the expected MS. Adding H to the mixture ₂ O (5 mL), followed by addition of 2M HCl (4 mL) to the mixture to adjust the pH to about 6-7. The mixture was extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product. The compound (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid was obtained as a yellow solid (791 mg,94.8% yield).

LCMS：Rt＝0.735min；C ₁₂ H ₁₄ N ₂ O ₅ MS calculated: 266.09; MS experimental values: 267.0[ M+H ] ⁺ ]。

Step 3: n- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide

To a solution of (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid (791 mg,2.97mmol,1 eq.) in DCM (10 mL) was added HATU (1.36 g,3.57mmol,1.2 eq.), DIPEA (1.15 g,8.91mmol,1.55mL,3 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (676.0 mg,3.57mmol,1.2 eq. HCl). The mixture was stirred at 25℃for 2h. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (20 mL) was quenched and extracted with DCM (40 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography24gSilica flash column, eluent: gradient of 0-10% DCM/MeOH ether at 35 mL/min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Ethyl group]-3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (838 mg,64.5% yield). LCMS: rt=0.741 min; c (C) ₁₉ H ₂₃ N ₅ O ₅ MS calculated: 401.17; MS experimental values: 402.1[ M+H ] ⁺ ]。

Step 4:2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-propionamide

To N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (838 mg,2.09mmol,1 eq.) in THF (10 mL) was added Pd/C (566.5 mg,0.53mmol,10% purity). At H ₂ The mixture was stirred at 25℃for 1h. LCMS showed detection of one peak with the expected MS. The mixture was filtered and concentrated under reduced pressure to give 2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-propionamide (616 mg,68.7% yield).

LCMS：Rt＝0.703min；C ₁₉ H ₂₅ N ₅ O ₃ MS calculated: 371.20; MS experimental values: 372.1[ M+H ] ⁺ ]。

N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ 4-methyl-2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl ] propanamide

To 2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (100 mg,0.26mmol,1 eq.) inNa was added to the solution in DMA (5 mL) ₂ CO ₃ (730.5 mg,6.89mmol,25.60 eq.) and 2, 2-trifluoroethyl triflate (1.6 g,6.89mmol,25.6 eq.). The mixture was stirred at 40℃for 16h. The mixture was filtered and then the filtrate was taken up with H ₂ O (20 mL) was quenched and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,7.8 min). Obtaining the compound N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [ 4-methyl-2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl]Propionamide (71.7 mg,57.9% yield). LCMS: rt=0.794 min; c (C) ₂₁ H ₂₆ F ₃ N ₅ O ₃ MS calculated: 453.20; MS experimental values: 454.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.24(dd,J＝3.9,7.2Hz,1H),6.22(dd,J＝5.5,7.0Hz,1H),5.52-5.32(m,1H),5.01(dd,J＝6.1,9.9Hz,1H),4.03-3.73(m,2H),3.36-3.32(m,1H),3.29-3.21(m,1H),2.56-2.45(m,1H),2.41-2.22(m,2H),2.21(d,J＝5.3Hz,3H),2.04-1.91(m,2H),1.91-1.71(m,2H),0.67-0.55(m,1H),0.48-0.35(m,2H),0.18 -0.02(m,2H)。

EXAMPLE 138 Synthesis of viral protease inhibitor Compound 451

N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ 4-methyl-2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl ] propionamide (69 mg,0.15mmol,1 eq.) was isolated by SFC (conditions: column DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH3H2O ETOH ];:. B%:45% -45%, min) as a white solid (17.12 mg,24.8% yield) of (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ 4-methyl-2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl ] propionamide.

Isomer 1: LCMS: rt=0.799 min; c (C) ₂₁ H ₂₆ F ₃ N ₅ O ₃ MS calculated: 453.20; MS experimental values: 454.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.24(d,J＝7.0Hz,1H),6.23(d,J＝7.3Hz,1H),5.45(t,J＝7.8Hz,1H),5.01(dd,J＝6.7,9.4Hz,1H),4.03-3.70(m,2H),3.36-3.32(m,2H),2.56-2.46(m,1H),2.41-2.24(m,2H),2.21(s,3H),1.98-1.93(m,2H),1.93-1.76(m,2H),0.64-0.50(m,1H),0.44-0.33(m,2H),0.17-0.04(m,2H)。

Isomer 2: LCMS: rt=0.800 min; c (C) ₂₁ H ₂₆ F ₃ N ₅ O ₃ MS calculated: 453.20; MS experimental values: 454.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.25(d,J＝7.3Hz,1H),6.22(d,J＝7.0Hz,1H),5.38(dd,J＝7.0,8.5Hz,1H),5.01(dd,J＝6.0,10.0Hz,1H),3.89(q,J＝9.5Hz,2H),3.30-3.21(m,2H),2.50(dq,J＝5.3,9.3Hz,1H),2.32-2.22(m,2H),2.20(s,3H),2.06-1.90(m,2H),1.89-1.68(m,2H),0.69-0.57(m,1H),0.50-0.36(m,2H),0.22-0.04(m,2H)。

EXAMPLE 139 Synthesis of viral protease inhibitor Compounds 455

To (3R, 6S) -6-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]To a solution of pyridine-3-carboxamide (40.0 mg,0.11mmol,1 eq.) in DMF (0.5 mL) was added Na ₂ CO ₃ (24.1 mg,0.22mmol,2 eq.) and 2, 2-trifluoroethyl triflate (26.4 mg,0.11mmol,1 eq.). The mixture was stirred at 25℃for 1h. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: pheNOMenex Gemini-NX 80.times.30 mm.times.3 um; mobile phase: [ Water (0.05% NH) ₃ H ₂ O+10mM NH4HCO3)-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:8% -38%,9.5 min). Obtaining (3R, 6S,8 aS) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-5-oxo-6- (2, 2-trifluoroethylamino) -2,3,6,7,8 a-hexahydrothiazolo [3,2-a ] ]Pyridine-3-carboxamide (8.02 mg,18.5umol,16.2% yield, 100% purity).

LCMS：Rt＝0.686min；C ₁₇ H ₂₂ F ₃ N ₅ O ₃ S MS calculated: 433.45; MS experimental values: 434.0[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ5.02(dd,J＝10.79,5.27Hz,1H),4.90-4.98(m,2H),4.77-4.83(m,1H),3.33-3.49(m,4H),3.20-3.29(m,1H),3.11-3.20(m,1H),2.67(qd,J＝9.29,5.27Hz,1H),2.17-2.45(m,4H),1.72-1.99(m,4H)。

EXAMPLE 140 Synthesis of viral protease inhibitor Compound 457

(3R, 6S,8 aS) -6-amino-N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -5-oxohexahydro-2H-thiazolo [3,2-a ] pyridine-3-carboxamide

To a solution of 9H-fluoren-9-ylmethyl N- [ (3 r, 6S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ] pyridin-6-yl ] carbamate (50 mg,87.1umol,1 eq) in DCM (0.2 mL) was added piperidine (14.8 mg,0.17mmol,17ul,2 eq). The mixture was stirred at 25℃for 0.5hr. The compound (3 r, 6S) -6-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ] pyridine-3-carboxamide (30 mg, crude material) was obtained as a yellow oil.

Benzyl (((3R, 6S,8 aS) -3- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) carbamoyl) -5-oxohexahydro-2H-thiazolo [3,2-a ] pyridin-6-yl) carbamate

To (3R, 6S) -6-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]To a solution of pyridine-3-carboxamide (30 mg,85.3umol,1 eq.) in DCM (1 mL) was added benzyl chloroformate (29.1 mg,0.17mmol,24uL,2 eq.) and TEA (25.9 mg,0.25mmol,35uL,3 eq.). The mixture was stirred at 25℃for 2h. LCMS detected the desired compound. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini: 25 mm. Times.10 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH4HCO3)-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,9.5 min). The residue was then purified by prep HPLC (column: waters Xbridge 150 x 25mm x 5um; mobile phase: [ water (0.04% NH) ₃ H ₂ O+10mM NH4HCO3)-MeOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -80%,9.5 min). The compound N- [ (3R, 6S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid]Ethyl group]Carbamoyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridin-6-yl]Benzyl carbamate (1.41 mg,2.8umol,3.3% yield, 99% purity). LCMS: rt=0.751 min; c (C) ₂₃ H ₂₇ N ₅ O ₅ S MS calculated: 485.56; MS experimental values: 486.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CDCl ₃ )δ8.32(br s,1H),7.37(br s,5H),6.07(br s,1H),5.67(br s,1H),5.38(br s,1H),5.17(br d,J＝10.26Hz,2H),4.90(br s,1H),4.80(br s,1H),3.97(br s,1H),3.52(br s,1H),3.25(br s,1H),3.33(br s,3H),2.44(br s,1H),2.33(br d,J＝15.38Hz,1H),1.97-2.13(m,2H),1.85(br s,3H)。

EXAMPLE 141 Synthesis of viral protease inhibitor Compound 459

N- ((3R, 6S,8 aS) -3- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) carbamoyl) -5-oxohexahydro-2H-thiazolo [3,2-a ] pyridin-6-yl) -5-methylisoxazole-3-carboxamide

A mixture of 5-methylisoxazole-3-carboxylic acid (36.1 mg,0.28mmol,2 eq), HATU (108.2 mg,0.28mmol,2 eq) and DIEA (73.5 mg,0.56mmol,99uL,4 eq) in DMF (1 mL) was stirred at 25℃for 0.5h and then (3R, 6S) -6-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was added to the reaction mass]Ethyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridine-3-carboxamide (50.0 mg,0.14mmol,1 eq.). The resulting mixture was stirred at 25℃for 2h. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:7% -37%,9.5 min) of the purified residue. The compound N- [ (3R, 6S,8 aS) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid ]Ethyl group]Carbamoyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridin-6-yl]-5-methyl-isoxazole-3-carboxamide (15.28 mg,33.0umol,23.2% yield, 99.7% purity). LCMS: rt=0.698 min; c (C) ₂₀ H ₂₄ N ₆ O ₅ S MS calculated: 460.51; MS experimental values: 461.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δppm 6.52(d,J＝0.75Hz,1H),4.98-5.07(m,3H),4.44(dd,J＝11.17,6.90Hz,1H),3.41(dd,J＝11.67,7.65Hz,1H),3.23-3.29(m,3H),2.58-2.69(m,1H),2.48(s,3H),2.27-2.44(m,4H),2.08-2.21(m,1H),1.79-2.01(m,3H)。

EXAMPLE 142 Synthesis of viral protease inhibitor Compound 465

(2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propionic acid methyl ester

To a solution of 3-nitro-1H-pyridin-2-one (1 g,7.14mmol,1 eq.) in DMF (10 mL) was added NaH (428.2 mg,10.71mmol,60% purity, 1.5 eq.) at 0deg.C for 15min. Next, (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester (1.6 g,7.85mmol,1.1 eq.) was added to the mixture and the mixture was stirred at 25℃for 2hr. TLC (petroleum ether: ethyl acetate=1:1) showed that a new spot was detected. The reaction mixture was quenched by the addition of H2O (10 mL) at 0 ℃ and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography25g />Silica flash column, eluent: the residue was purified by a gradient of 0-50% petroleum ether/ethyl acetate at 30mL/min to give methyl (2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propanoate (552 mg,28.7% yield, 98.9% purity) as a yellow solid.

(2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propionic acid

To a solution of methyl (2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propanoate (230 mg,0.86mmol,1 eq.) in THF (1 mL) and MeOH (0.2 mL) was added LiOH. H ₂ H of O (108.7 mg,2.59mmol,3 eq) ₂ O (0.2 mL). The mixture was stirred at 0deg.C for 10min. LC-MS showed detection of the desired compound. The reaction was adjusted to ph=4 with 4m hcl. H for the reaction mixture ₂ Dilute with O (5 mL) and extract with ethyl acetate (5 mL x 3). The combined organic phases were washed with brine (5 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was used in the next step without further purification. The compound (2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propionic acid was obtained as a yellow solid (210 mg,96.3% yield).

(2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propanamide

To a solution of (2S) -3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propionic acid (260 mg,1.03mmol,1 eq.) in DCM (3 mL) was added HATU (470.3 mg,1.24mmol,1.2 eq.), DIPEA (266.4 mg,2.06mmol,0.35mL,2 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]Propionitrile (234.5 mg,1.24mmol,1.2 eq. HCl). The mixture was stirred at 25℃for 16h. TLC (DCM/meoh=10:1) showed that a new spot was detected. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography4g />Silica flash column, eluent: gradient of 0-10% DCM/MeOH ether at 20 mL/min) to give (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Ethyl group]-3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propanamide (225 mg,54.0% yield, 96% purity).

(2S) -2- (3-amino-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-propionamide

At N ₂ Next, (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-2- (3-nitro-2-oxo-1-pyridinyl) propanamide (200 mg,0.51mmol,1 eq.) in THF (0.5 mL) was added Pd/C (200 mg,0.18mmol,10% purity, 3.64e-1 eq.). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (15 psi) at 25℃for 10min. LC-MS showed detection of the desired compound. TLC (DCM/meoh=10:1) showed that a new spot was detected. The resulting product was dissolved in MeOH (5 mL) and filtered to removeRemoving insoluble substances. The filtrate was concentrated in vacuo. Through flash silica gel chromatography4g/>Silica flash column, eluent: 0-5% Petroleum ether/ethyl acetate ether gradient, at 20 mL/min) to give (2S) -2- (3-amino-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a brown solid]Ethyl group]-3-cyclopropyl-propionamide (119 mg,64.3% yield, 99.7% purity).

LCMS：Rt＝0.669min；C ₁₈ H ₂₃ N ₅ O ₃ MS calculated: 357.18; MS experimental values: 358.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.06-7.01(m,1H),6.70-6.64(m,1H),6.24(s,1H),5.56-5.41(m,1H),5.06-4.97(m,1H),3.30-3.24(m,2H),2.57-2.43(m,1H),2.38-2.18(m,2H),2.04-1.85(m,3H),1.85-1.69(m,1H),0.70-0.54(m,1H),0.50-0.36(m,2H),0.21-0.12(m,1H),0.10-0.02(m,1H)。

EXAMPLE 143 Synthesis of viral protease inhibitor Compound 465

The residue was purified by SFC. LC-MS showed detection of the desired compound. By SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O EtOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -30%, min) purification residue.

Isomer 1: 2- [ (1S) -3-amino-2-oxo-1-pyridinyl as brown solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (2.84 mg,6.3% yield, 98.9% purity). LCMS: rt=0.660 min; c (C) ₁₈ H ₂₃ N ₅ O ₃ MS calculated: 357.41; MS experimental values: 358.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.03(dd,J＝1.4,7.0Hz,1H),6.68(dd,J＝1.4,7.2Hz,1H),6.26(t,J＝7.1Hz,1H),5.53(t,J＝7.7Hz,1H),5.02(dd,J＝6.8,9.3Hz,1H),3.38-3.32(m,2H),2.56-2.46(m,1H),2.36(m,1H),2.32-2.23(m,1H),1.97-1.87(m,3H),1.87-1.79(m,1H),0.67-0.54(m,1H),0.45-0.34(m,2H),0.19-0.10(m,1H),0.07-0.02(m,1H)。

Isomer 2: obtaining the compound 2- [ (1R) -3-amino-2-oxo-1-pyridinyl) as a brown solid]-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (21.3 mg,46.5% yield). LCMS: rt=0.671 min; c (C) ₁₈ H ₂₃ N ₅ O ₃ MS calculated: 357.41; MS experimental values: 358.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.04(dd,J＝1.5,7.0Hz,1H),6.67(dd,J＝1.5,7.3Hz,1H),6.24(t,J＝7.1Hz,1H),5.44(t,J＝7.7Hz,1H),5.01(dd,J＝6.1,10.1Hz,1H),3.30-3.24(m,2H),2.49(dq,J＝5.4,9.3Hz,1H),2.32-2.20(m,2H),2.01-1.83(m,3H),1.83-1.70(m,1H),0.70-0.59(m,1H),0.51-0.38(m,2H),0.20-0.12(m,1H),0.10-0.00(m,1H)。

EXAMPLE 144 Synthesis of viral protease inhibitor Compound 466

N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ 2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl ] propanamide

To 2- (3-amino-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-propionamide (110 mg,0.30mmol,1 eq.) in DMA (1 mL) was added Na ₂ CO ₃ (326.2 mg,3.08mmol,10 eq.) and 2, 2-trifluoroethyl triflate (2.1 g,9.23mmol,30 eq.). The mixture was stirred at 40℃for 16h. TLC (DCM: meoh=10:1) showed new spots were detected. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. General purpose medicine Supermaking TLC (SiO) ₂ DCM: meoh=10:1) the residue was purified to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [ 2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl]Propionamide (23 mg,16.8% yield, 98.8% purity).

LCMS：Rt＝0.797min；C ₂₀ H ₂₄ F ₃ N ₅ O ₃ MS calculated: 439.18; MS experimental values: 440.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.06(dt,J＝1.5,6.8Hz,1H),6.65-6.55(m,1H),6.37-6.27(m,1H),5.56-5.40(m,1H),5.05-4.98(m,1H),3.88(dq,J＝6.0,9.2Hz,2H),3.34(br d,J＝3.0Hz,1H),3.30-3.24(m,1H),2.57-2.42(m,1H),2.39-2.20(m,2H),2.08-1.88(m,3H),1.86-1.74(m,1H),0.71-0.52(m,1H),0.50-0.36(m,2H),0.22-0.11(m,1H),0.10-0.03(m,1H)。

EXAMPLE 145 Synthesis of viral protease inhibitor Compound 467

The residue was further separated by SFC. By SFC (column: DAICEL CHIRALCEL OD-H (250 mm. Times.30 mm,5 um); mobile phase: [0.1% NH) ₃ H ₂ OETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -20%, min) to further separate the residue.

Isomer 1: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [ 2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl]Propionamide (2.56 mg,12.3% yield). LCMS: rt=0.837 min; c (C) ₂₃ H ₃₁ N ₅ O ₅ MS calculated: 457.23; MS experimental values: 458.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.06(dt,J＝1.5,6.8Hz,1H),6.65-6.55(m,1H),6.37-6.27(m,1H),5.56-5.40(m,1H),5.05-4.98(m,1H),3.88(dq,J＝6.0,9.2Hz,2H),3.34(br d,J＝3.0Hz,1H),3.30-3.24(m,1H),2.57-2.42(m,1H),2.39-2.20(m,2H),2.08-1.88(m,3H),1.86-1.74(m,1H),0.71-0.52(m,1H),0.50-0.36(m,2H),0.22-0.11(m,1H),0.10-0.03(m,1H)。

Isomer 2: the compound (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- [ 2-oxo-3- (2, 2-trifluoroethylamino) -1-pyridinyl]Propionamide (2.56 mg,12.3% yield, 96.3% purity). LCMS: rt=0.794 min; c (C) ₂₀ H ₂₄ F ₃ N ₅ O ₃ MS calculated: 439.18; MS experimental values: 440.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.04(dd,J＝1.5,7.0Hz,1H),6.61(d,J＝7.0Hz,1H),6.32(t,J＝7.2Hz,1H),5.52(t,J＝7.8Hz,1H),5.01(dd,J＝6.5,9.3Hz,1H),3.89(q,J＝9.3Hz,2H),3.37-3.32(m,2H),2.55-2.46(m,1H),2.36(m,1H),2.32-2.24(m,1H),1.99-1.93(m,2H),1.93-1.87(m,1H),1.87-1.78(m,1H),0.64-0.54(m,1H),0.46-0.34(m,2H),0.18-0.09(m,1H),0.07 -0.02(m,1H)。

Example 146 Synthesis of viral protease inhibitor Compound 468

Step 1:7- [ (1S) -1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] -6-oxo-1, 7-diazaspiro [4.4] nonane-1-carboxylic acid tert-butyl ester

At 0 ℃, 6-oxo-1, 7-diazaspiro [4.4]]To a solution of t-butyl nonane-1-carboxylate (0.5 g,2.08mmol,1 eq.) in toluene (7 mL) was added NaH (124.8 mg,3.12mmol,60% purity, 1.5 eq.). After stirring at 25℃for 1h, methyl (R) -2-bromo-3-cyclopropylpropionate (517.0 mg,2.50mmol,1.2 eq.) was added at 0℃and the mixture was stirred at 80℃for 8h. By adding H ₂ O (15 mL) to quench the reaction mixture and extract with EtOAc (15 mL x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 7- (3-cyclopropyl-1-methoxy-1-oxopropan-2-yl) -6-oxo-1, 7-diazaspiro [4.4] as a colorless oil]Nonane-1-carboxylic acid tert-butyl ester (600 mg, crude material).

Step 2:2- (1-Boc-6-oxo-1, 7-diazaspiro [4.4] non-7-yl) -3-cyclopropyl-propionic acid

To 2 (450.0 mg,1.23mmol,1 eq.) in H ₂ To a solution of O (1 mL) and MeOH (3 mL) was added NaOH (196.4 mg,4.91mmol,4 eq.). The mixture was stirred at 25℃for 1h. LC-MS showed complete exhaustion of 2 and 66% of the desired compound was detected. By adding H ₂ O (15 mL) to quench the reaction mixture. The pH of the mixture was adjusted to 5-6 with HCl (2M). And the mixture was then extracted with EtOAc (20 ml x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue, and 2- (1-t-butoxycarbonyl-6-oxo-1, 7-diazaspiro [4.4] as a colorless oil was obtained]Non-7-yl) -3-cyclopropyl-propionic acid (0.4 g, crude material).

468:7- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-oxo-1, 7-diazaspiro [4.4] nonane-1-carboxylic acid tert-butyl ester

To 2- (1-tert-butoxycarbonyl-6-oxo-1, 7-diazaspiro [4.4] at 0deg.C]To a solution of non-7-yl) -3-cyclopropyl-propionic acid (50.0 mg,0.14mmol,1 eq.) in THF (1 mL) was added Et ₃ N (14.3 mg,0.14mmol,19.7uL,1.0 eq.) and isobutyl chloroformate (21.3 mg,0.15mmol,20.4uL,1.1 eq.). The mixture was stirred at 25℃for 1h. Addition of (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (32.2 mg,0.17mmol,1.2 eq, HCl) and Et ₃ A solution of N (15.7 mg,0.15mmol,21.7uL,1.1 eq.) in DMF (1 mL) and the mixture stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini: 25 mm. Times.10 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH4HCO3)-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,9.5 min) to give 468 as a white solid (9.02 mg,13% yield).

LCMS：Rt＝0.821min；C ₂₅ H ₃₇ N ₅ O ₅ MS calculated: 487.28; MS experimental values: 388.1[ M-Boc+H ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.34-8.15(m,1H),5.72(d,J＝10.0Hz,1H),5.29-4.98(m,1H),4.95-4.81(m,1H),3.59-3.47(m,2H),3.46-3.19(m,4H),2.64-2.29(m,4H),2.28-2.16(m,1H),2.10-2.00(m,2H),1.97-1.84(m,4H),1.73-1.60(m,2H),1.53-1.38(m,9H),0.71-0.52(m,1H),0.51-0.38(m,2H),0.17-0.07(m,2H)。

EXAMPLE 147 Synthesis of viral protease inhibitor Compound 469

Isomer 1 and isomer 2:7- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-oxo-1, 7-diazaspiro [4.4] nonane-1-carboxylic acid tert-butyl ester; isomer 3: (5R) -7- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-oxo-1, 7-diazaspiro [4.4] nonane-1-carboxylic acid tert-butyl ester; isomer 3: (5S) -7- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-oxo-1, 7-diazaspiro [4.4] nonane-1-carboxylic acid tert-butyl ester

To 2- (1- (t-butoxycarbonyl) -6-oxo-1, 7-diazaspiro [4.4]]To a solution of non-7-yl) -3-cyclopropylpropionic acid (200 mg,0.56mmol,1 eq.) in DMF (2 mL) was added HATU (431.5 mg,1.13mmol,2.0 eq.) (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionitrile (129.1 mg,0.68mmol,1.2 eq., HCl) and DIPEA (146.6 mg,1.13mmol,197.7uL,2.0 eq.). The mixture was stirred at 25℃for 0.5h. TLC (dichloromethane: methanol=10/1, pma) indicated complete exhaustion of reactant 1 and formation of a new spot. The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with methylene chloride methanol=100/1 to 10/1 to give compound 469 (150 mg) as a white solid. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,9.5 min) to purify compound 469 (150 mg) to give a white solidCompound 469 (60 mg). By preparative SFC (column: DAICEL CHIRALPAK IC (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%) of compound 469 (60 mg) was purified to give three white solids, isomer 1 and isomer 2 (15 mg,30.7umol,5.42% yield), isomer 3 (8.46 mg,16.3umol,2% yield, 94% purity), isomer 4 (9.97 mg,18.2umol,3% yield, 89% purity).

Isomers 1 and 2: LCMS: rt=1.610, c ₂₅ H ₃₇ N ₅ O ₅ MS calculated: 487.28; MS experimental values: 488.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.20-4.98(m,1H),4.81-4.71(m,1H),3.61-3.41(m,3H),3.38-3.32(m,3H),2.61-2.41(m,2H),2.40-2.19(m,2H),2.18-1.66(m,9H),1.52-1.33(m,9H),0.78-0.57(m,1H),0.56-0.38(m,2H),0.25-0.04(m,2H)。

Isomer 3: LCMS: rt=1.631, c ₂₅ H ₃₇ N ₅ O ₅ MS calculated: 487.28; MS experimental values: 488.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.20-4.97(m,1H),4.53-4.32(m,1H),3.64-3.41(m,3H),3.31(s,3H),2.63-2.35(m,2H),2.35-2.12(m,2H),2.12-1.74(m,8H),1.73-1.52(m,1H),1.73-1.52(m,1H),1.50-1.35(m,9H),0.75(s,1H),0.62-0.36(m,2H),0.24-0.07(m,2H)。

Isomer 4: LCMS: rt=1.630, c ₂₅ H ₃₇ N ₅ O ₅ MS calculated: 487.28; MS experimental values: 488.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.23-5.01(m,1H),4.81-4.74(m,1H),3.64-3.37(m,3H),3.35(s,3H),2.67-2.42(m,2H),2.42-2.10(m,3H),2.10-1.68(m,8H),1.54-1.39(m,9H),0.68-0.57(m,1H),0.55-0.39(m,2H),0.24-0.05(m,2H)。

EXAMPLE 148 Synthesis of viral protease inhibitor Compound 471

7- (1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-e -2-yl) -6-oxo-1, 7-diazaspiro [4.4]A solution of tert-butyl nonane-1-carboxylate (90 mg,0.18mmol,1 eq.) in H2O (4 mL) was stirred at 100deg.C for 16hr. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:11% -41%,9.5 min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3-cyclopropyl-2- (6-oxo-1, 7-diazaspiro [4.4 ]]Non-7-yl) propanamide (2.41 mg,6.10umol,3% yield, 98% purity).

LCMS：Rt＝0.603min；C ₂₀ H ₂₉ N ₅ O ₃ MS calculated: 387.23; MS experimental values: 388.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ5.01(dd,J＝6.3,9.8Hz,1H),4.55(s,1H),3.57-3.47(m,2H),3.37-3.32(m,2H),3.18-3.08(m,1H),2.99-2.87(m,1H),2.61-2.48(m,1H),2.34-2.24(m,2H),2.13-2.00(m,2H),1.93-1.80(m,7H),1.65-1.56(m,1H),0.75-0.63(m,1H),0.56-0.45(m,2H),0.17(d,J＝3.5Hz,2H)。

Example 149 Synthesis of viral protease inhibitor Compounds 473

Step 1: (2R) -2-bromo-3-cyclopropyl-propionic acid

To (2R) -2-amino-3-cyclopropyl-propionic acid (3.5 g,27.10mmol,1 eq.) and NaBr (9.76 g,94.85mmol,3.05mL,3.5 eq.) at 0deg.C in 2.5M H ₂ SO ₄ NaNO-containing solution was added dropwise to the solution in (35 mL) ₂ (2.43 g,35.23mmol,1.3 eq.) H ₂ O (7 mL). The reaction mixture was stirred at 0℃for 1h and at 25℃for 6h. The mixture was diluted with water (60 mL) and the resulting mixture was extracted with DCM (80 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure to give (2R) -2-bromo-3-cyclopropyl-propionic acid as a colorless oil (7.4 g,crude material).

¹ H NMR(400MHz,CDCl ₃ )δ4.33(t,J＝7.4Hz,1H),1.99(dt,J＝2.1,7.1Hz,2H),0.91-0.79(m,1H),0.58-0.51(m,2H),0.22-0.15(m,2H)。

Step 2: (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester

To a solution of (2R) -2-bromo-3-cyclopropyl-propionic acid (7.4 g,38.33mmol,1 eq.) in MeOH (70 mL) was added HCl (12 m,7.40mL,2.32 eq.) and the reaction mixture was then stirred at 50 ℃ for 16h. TLC (petroleum ether: ethyl acetate=10:1, pma) showed consumption of starting material. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (10 mL) and the resulting mixture was extracted with ethyl acetate (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester (4.9 g,59.2% yield) as a colorless oil.

¹ H NMR(400MHz,CDCl ₃ )δ4.31(t,J＝7.4Hz,1H),3.80(s,3H),2.00-1.94(m,2H),0.86-0.75(m,1H),0.57-0.44(m,2H),0.22-0.09(m,2H)。

Step 3:2- (6-Boc-1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) -3-cyclopropyl-propionic acid

At 0 ℃, to 1-oxo-2, 6-diazaspiro [4.5]]To a solution of tert-butyl decane-6-carboxylate (500 mg,1.97mmol,1 eq.) in toluene (10 mL) was added NaH (94.37 mg,2.36mmol,60% purity, 1.2 eq.) and the mixture was then stirred at 25 ℃ for 0.5h. The reaction mixture was cooled to 0 ℃. Methyl (2R) -2-bromo-3-cyclopropyl-propionate (488.5 mg,2.36mmol,1.2 eq.) was added and the reaction mixture was warmed to 80 ℃ and stirred at 80 ℃ for 16h. LC-MS showed complete depletion of starting material and detected a major peak with the desired MS. TLC (petroleum ether: ethyl acetate=1:1, pma) showed consumption of starting material. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (10 mL) and the resulting mixture was extracted with ethyl acetate (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressureDrying to give 2- [1- (cyclopropylmethyl) -2-methoxy-2-oxo-ethyl ] as a pale yellow oil]-1-oxo-2, 6-diazaspiro [4.5]]Decane-6-carboxylic acid tert-butyl ester (480 mg, crude material). The aqueous material was acidified with HCl (0.5N) to ph=5 and the resulting mixture was extracted with ethyl acetate (20 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure to give 2- (6-t-butoxycarbonyl-1-oxo-2, 6-diazaspiro [4.5] as a pale yellow oil]Dec-2-yl) -3-cyclopropyl-propionic acid (120 mg, crude material).

Isomer 1:2- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1-oxo-2, 6-diazaspiro [4.5] decane-6-carboxylic acid tert-butyl ester; isomer 2:2- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1-oxo-2, 6-diazaspiro [4.5] decane-6-carboxylic acid tert-butyl ester

To 2- (6- (tert-Butoxycarbonyl) -1-oxo-2, 6-diazaspiro [4.5]]To a solution of dec-2-yl) -3-cyclopropylpropionic acid (0.1 g,0.27mmol,1 eq.) in DMF (1 mL) was added HATU (207.5 mg,0.54mmol,2.0 eq.) (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionitrile hydrochloride (62.1 mg,0.32mmol,1.2 eq., HCl) and DIPEA (52.9 mg,0.40mmol,71.3uL,1.5 eq.). The mixture was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:27% -57%,9.5 min) to give isomer 1 (18.9 mg,13% yield) and isomer 2 (2.54 mg,1.8% yield) as two white solids.

Isomer 1: LCMS: rt= 0.831min; c (C) ₂₆ H ₃₉ N ₅ O ₅ MS calculated: 501.30; MS experimental values: 502.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.20-5.01(m,1H),4.81-4.68(m,1H),3.90(td,J＝4.6,8.6Hz,1H),3.59-3.40(m,1H),3.34(d,J＝3.3Hz,3H),3.07-2.85(m,1H),2.64-2.45(m,1H),2.45-2.22(m,3H),2.21-2.07(m,1H),2.05-1.92(m,1H),1.90-1.62(m,7H),1.57(d,J＝10.5Hz,2H),1.51-1.39(m,9H),0.81-0.57(m,1H),0.55-0.33(m,2H),0.22-0.04(m,2H)。

Isomer 2: LCMS: rt=0.845 min; c (C) ₂₆ H ₃₉ N ₅ O ₅ MS calculated: 501.30; MS experimental values: 502.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.28-5.14(m,1H),5.28-5.14(m,1H),4.77(t,J＝7.7Hz,1H),3.93(br d,J＝13.1Hz,1H),3.51-3.38(m,1H),3.35-3.31(m,1H),3.35-3.31(m,2H),3.06-2.92(m,1H),2.61-2.48(m,1H),2.45-2.22(m,3H),2.19-2.08(m,1H),1.97(td,J＝8.2,13.7Hz,1H),1.89-1.65(m,7H),1.64-1.52(m,2H),1.49(s,9H),0.64-0.54(m,1H),0.53-0.30(m,2H),0.22-0.00(m,2H)。

Example 150 Synthesis of viral protease inhibitor Compound 475

Isomer 1 and isomer 2: (2R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (1-oxo-2, 6-diazaspiro [4.5] dec-2-yl) acrylamide; isomer 3: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5R) -1-oxo-2, 6-diazaspiro [4.5] dec-2-yl ] acrylamide; isomer 4: (2S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- [ (5S) -1-oxo-2, 6-diazaspiro [4.5] dec-2-yl ] propionamide

2- (1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1-oxo-2, 6-diazaspiro [4.5]]Decane-6-carboxylic acid tert-butyl ester (0.15 g,0.29mmol,1 eq.) in H ₂ The solution in O (5 mL) was stirred at 100deg.C for 16h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -31%,9.5 min) to give 475 isomer 1 and isomer as two white solids2 (6.00 mg,5% yield), and 475 isomer 3 and isomer 4 (24.65 mg). By preparative SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%, min) to purify 475 isomer 3 and isomer 4 to give 475 isomer 3 (5.53 mg,4% yield) and 475 isomer 4 (4.84 mg,3% yield) as two white solids.

475 isomer 1 and isomer 2: LCMS: rt=1.232 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ5.04-4.92(m,1H),4.67-4.60(m,1H),3.73-3.39(m,2H),3.37-3.32(m,2H),3.15-3.00(m,1H),2.88(d,J＝6.5Hz,1H),2.62-2.42(m,1H),2.40-2.15(m,6H),2.11-1.76(m,8H),1.68-1.51(m,1H),0.75-0.57(m,1H),0.57-0.39(m,2H),0.23-0.11(m,2H)。

Isomer 3: LCMS: rt=1.332 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ4.90-4.82(m,1H),4.47(dd,J＝6.5,9.0Hz,1H),3.46-3.34(m,1H),3.31-3.23(m,1H),3.20-3.15(m,3H),3.02-2.89(m,1H),2.64-2.48(m,1H),2.43-2.32(m,1H),2.27-2.01(m,3H),1.92-1.65(m,5H),1.55-1.39(m,5H),0.53-0.41(m,1H),0.39-0.23(m,2H),0.07--0.08(m,2H)。

Isomer 4: LCMS: rt=1.329 min; c (C) ₂₁ H ₃₁ N ₅ O ₃ MS calculated: 401.24; MS experimental values: 402.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ4.96(d,J＝3.0Hz,1H),4.66(dd,J＝6.0,9.5Hz,1H),3.65-3.56(m,1H),3.49-3.40(m,1H),3.37-3.32(m,3H),3.20-3.09(m,1H),2.82-2.69(m,1H),2.50(td,J＝8.1,16.3Hz,1H),2.41-2.31(m,2H),2.25(ddd,J＝6.3,9.5,13.9Hz,1H),2.10(td,J＝9.0,12.6Hz,1H),2.01-1.79(m,4H),1.71-1.57(m,5H),0.65-0.55(m,1H),0.54-0.37(m,2H),0.21-0.08(m,2H)。

EXAMPLE 151 Synthesis of viral protease inhibitor Compound 477

Step 1: (3S, 6S,8 aS) -3- (chlorocarbonyl) -5-oxohexahydro-2H-thiazolo [3,2-a ] pyridin-6-yl) carbamic acid (9H-fluoren-9-yl) methyl ester

At 0℃under N ₂ Downward (3R, 6S) -6- (9H-fluoren-9-ylmethoxycarbonylamino) -5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]To a solution of pyridine-3-carboxylic acid (200 mg,0.45mmol,1 eq.) in DCM (4 mL) was added (COCl) ₂ (86.8 mg,0.68mmol,59uL,1.5 eq.) and DMF (3.3 mg,45.6umol,3uL,0.1 eq.). The mixture was stirred at 0℃for 1h. The reaction mixture was concentrated under reduced pressure to give a residue. It was used in the next step without purification. Obtaining the compound N- [ (3S, 6S) -3-chlorocarbonyl-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ] as a yellow solid]Pyridin-6-yl]9H-fluoren-9-ylmethyl carbamate (200 mg, crude material).

(3R, 6S,8 aS) -3- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) carbamoyl) -5-oxohexahydro-2H-thiazolo [3,2-a ] pyridin-6-yl) carbamic acid (9H-fluoren-9-yl) methyl ester

To N- [ (3S, 6S) -3-chlorocarbonyl-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridin-6-yl]To a solution of 9H-fluoren-9-ylmethyl carbamate (200 mg,0.43mmol,1 eq.) in DMF (5 mL) was added TEA (132.8 mg,1.31mmol,0.18mL,3 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (99.6 mg,0.52mmol,1.2 eq, HCl). The mixture was stirred at 25℃for 1hr. TLC (petroleum ether/ethyl acetate=0:1, uv 254). Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography4g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient,at 30 mL/min) to give a yellow solid. By preparative HPLC (column: 3_Phenomenex Luna C18 75*30mm*3um; mobile phase: [ water (0.05% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8.5 min) of the purified residue. The compound N- [ (3R, 6S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid]Ethyl group]Carbamoyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridin-6-yl]9H-fluoren-9-ylmethyl carbamate (10.63 mg,18.4umol,4.2% yield, 99.7% purity). LCMS: rt=0.834 min; c (C) ₃₀ H ₃₁ N ₅ O ₅ S MS calculated: 573.66; MS experimental values: 574.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.82(d,J＝7.53Hz,2H),7.65-7.72(m,2H),7.37-7.45(m,2H),7.29-7.36(m,2H),5.02-5.11(m,3H),4.43(d,J＝6.78Hz,2H),4.23-4.30(m,1H),4.01(br dd,J＝11.29,6.78Hz,1H),3.42(dd,J＝11.54,7.78Hz,1H),3.19-3.30(m,3H),2.52-2.65(m,1H),2.17-2.43(m,4H),2.03-2.13(m,1H),1.84-1.97(m,2H),1.73-1.83(m,1H)。

Example 152 Synthesis of viral protease inhibitor Compound 479

To N- [ (3R, 6S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-5-oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridin-6-yl]To a solution of 9H-fluoren-9-ylmethyl carbamate (100 mg,0.17mmol,1 eq.) in MeOH (0.1 mL) was added NH ₃ (7M, 2.00mL,80.31 eq). The mixture was stirred at 25℃for 1.5h. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The aqueous phase was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:0% -23%,7.8 min) of the purified residue. The compound (3R, 6S) -6-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid]Ethyl group]-5-Oxo-2, 3,6,7,8 a-hexahydrothiazolo [3,2-a ]]Pyridine-3-carboxamide (16.59 mg,47.2umol,27.0% yield, 100% purity).

LCMS：Rt＝1.495min；C ₁₅ H ₂₁ N ₅ O ₃ S MS calculated: 351.42; MS experimental values: 352.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δppm 4.99(br dd,J＝10.63,5.63Hz,3H),3.33-3.45(m,4H),3.14-3.25(m,1H),2.58-2.71(m,1H),2.19-2.44(m,4H),1.75-2.00(m,4H)。

EXAMPLE 153 Synthesis of viral protease inhibitor Compounds 483

To (3S) -5-oxo-6- [ (2-oxo-2-phenoxy-ethyl) amino]To a solution of 2, 3-dihydro-1H-indolizine-3-carboxylic acid (100 mg,0.30mmol,1 eq.) in DCM (3 mL) was added HATU (138.9 mg,0.36mmol,1.2 eq.) and DIPEA (118.0 mg,0.91mmol,0.15mL,3 eq.) for 1H. Next, (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl is added to the mixture]Propionitrile (55.9 mg,0.29mmol,9.69e-1 eq, HCl) and the resulting mixture was stirred at 25℃for 15h. TLC (DCM/meoh=10:1). H for the reaction mixture ₂ Dilute with O (5 mL) and extract with ethyl acetate (5 mL x 3). The combined organic phases were washed with brine (5 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography4g/>Silica flash column, eluent: the residue was purified with a gradient of 0-10% DCM/MeOH ether at 20mL/min to give 2- [ [ (3S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-5-oxo-2, 3-dihydro-1H-indolizin-6-yl]Amino group]Phenyl acetate (35 mg,75.3umol,24.7% yield, 99.8% purity).

LCMS：Rt＝0.770min；C ₂₄ H ₂₅ N ₅ O ₅ MS calculated: 463.19; MS experimental values: 464.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.03(br s,1H),7.46-7.24(m,5H),6.32(br d,J＝7.6Hz,1H),5.19(s,2H),5.10-4.99(m,3H),3.34(br d,J＝3.3Hz,1H),3.24-3.06(m,2H),2.74-2.63(m,1H),2.62-2.45(m,2H),2.40-2.23(m,3H),1.97-1.80(m,2H)。

EXAMPLE 154 Synthesis of viral protease inhibitor Compound 489

Step 1: (1R, 2S,6R, 7R) -8-trimethylsilyloxy-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undec-8-ene-3, 5-dione

A solution of cyclohex-1, 5-dien-1-yloxy (trimethyl) silane (5.0 g,29.71mmol,5.50mL,1 eq.) and pyrrole-2, 5-dione (2.88 g,29.71mmol,1 eq.) in MTBE (50 mL) was stirred at 25℃for 16h. TLC (petroleum ether: ethyl acetate=2:1, i ₂ ). The reaction mixture was concentrated under reduced pressure. MTBA (15 mL) and PE (15 mL) were added and the suspension was then filtered to give the title compound as a white solid. Obtaining the compound (1R, 2S,6R, 7R) -8-trimethylsilyloxy-4-azatricyclo [5.2.2.0 ] as a white solid ^2,6 ]Undec-8-ene-3, 5-dione (5.2 g,65.9% yield)

Step 2: (1R, 2S,6R, 7R) -4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecane-3, 5, 8-triones

(1R, 2S,6R, 7R) -8-trimethylsilyloxy-4-azatricyclo [5.2.2.0 ] ^2,6 ]A solution of undec-8-ene-3, 5-dione (2.9 g,10.93mmol,1 eq.) in HCl/dioxane (25 mL) was stirred at 25℃for 16hr. TLC (petroleum ether: ethyl acetate=5:1). The reaction mixture was concentrated in vacuo. No purification was performed. The crude product was used in the next step without further purification. Obtaining the compound (1R, 2S,6R, 7R) -4-azatricyclo [5.2.2.0 ] as a white solid ^2,6 ]Undecane-3, 5,8Trione (2.16 g, crude material).

Step 3: (1R, 2S,6R, 7R) -4- [ (4-methoxyphenyl) methyl]-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecane-3, 5, 8-triones

To (1R, 2S,6R, 7R) -4-azatricyclo [5.2.2.0 ] ^2,6 ]To a solution of undecane-3, 5, 8-trione (2.1 g,11.18mmol,1 eq.) in DMF (20 mL) was added PMBCl (2.1 g,13.42mmol,1.83mL,1.2 eq.) and K ₂ CO ₃ (2.3 g,16.77mmol,1.5 eq.). The mixture was stirred at 25℃for 16h. LCMS showed detection of the desired compound. TLC (petroleum ether: ethyl acetate=1:1). H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography12g/>Silica flash column, eluent: gradient of 0-40% petroleum ether/ethyl acetate ether at 25 mL/min). Obtaining the compound (1R, 2S,6R, 7R) -4- [ (4-methoxyphenyl) methyl as a white solid]-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecane-3, 5, 8-trione (3.03 g,86.4% yield).

Step 4: (1R, 2S,6R, 7R) -8-amino-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecane-8-carbonitrile

To (1R, 2S,6R, 7R) -4- [ (4-methoxyphenyl) methyl]-4-azatricyclo [5.2.2.0 ] ^2,6 ]To a solution of undecane-3, 5, 8-trione (1.7 g,5.43mmol,1 eq.) in DCM (25 mL) was added NH ₃ (7M, 7.75mL,10 eq.) and Ti (i-PrO) ₄ (1.85 g,6.51mmol,1.92mL,1.2 eq.). The reaction mixture was stirred at 25℃for 2hr. TMSCN (807.3 mg,8.14mmol,1.02mL,1.5 eq.) was added and the solution stirred at 25℃for 16h. Ethyl acetate (100 mL) and H were added ₂ O (10 mL), the reaction mixture was filtered, and the filtrate was concentrated to reduced pressure. Obtaining the compound as white solid(1R, 2S,6R, 7R) -8-amino-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecane-8-carbonitrile (1.75 g, crude material).

Step 5: (2S) -2- (Benzylmethoxycarbonylamino) -3-cyclopropyl-propionic acid

To a solution of (2S) -2-amino-3-cyclopropyl-propionic acid (3.0 g,23.23mmol,1 eq.) in THF (45 mL) at 0deg.C was added Na ₂ CO ₃ (2M, 13.94mL,1.2 eq.). CbzCl (5.15 g,30.20mmol,4.29mL,1.3 eq.) was added and the reaction mixture was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and the resulting mixture was extracted with ethyl acetate (50 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionic acid as a colorless oil. The compound (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionic acid (3.2 g,10.21mmol,43.9% yield, 84% purity) was obtained as a colourless oil. ¹ H NMR(400MHz,CD ₃ OD)δ7.43-7.20(m,5H),5.09(s,2H),4.23(dd,J＝5.5,8.0Hz,1H),1.73-1.58(m,2H),0.86-0.72(m,1H),0.53-0.39(m,2H),0.20-0.02(m,2H)。

Step 6: n- [ (1S) -2- [ [ (1R, 2S,6R, 7R) -8-cyano-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzoic acid phenylmethyl ester

(1R, 2S,6R, 7R) -8-amino-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]A solution of undecane-8-carbonitrile (1.7 g,5.01mmol,1 eq.) and (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionic acid (1.45 g,5.51mmol,1.1 eq.) and pyridine (3.96 g,50.09mmol,4.04mL,10 eq.) in THF (35 mL) was stirred at 25℃for 15min. Drop wise addition of POCl at 0deg.C ₃ After (1.92 g,12.52mmol,1.16mL,2.5 eq.) the reaction mixture was stirred at 25℃for 2 hours. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and the resulting mixture was mixedThe material was extracted with ethyl acetate (80 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM: meoh=1:0 to 20:1) to give N- [ (1S) -2- [ [ (1 r,2S,6r,7 r) -8-cyano-4- [ (4-methoxyphenyl) methyl as a colorless oil]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Carbamate (2.4 g,72.9% yield, 89% purity).

Step 7: n- [ (1S) -2- [ [ (1R, 2S,6R, 7R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzoic acid phenylmethyl ester

To N- [ (1S) -2- [ [ (1R, 2S,6R, 7R) -8-cyano-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (500 mg,0.85mmol,1 eq.) in ACN (15 mL) and H ₂ CAN (1.41 g,2.57mmol,1.28mL,3 eq.) was added to a solution in O (5 mL), and the reaction mixture was then stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and the resulting mixture was extracted with ethyl acetate (50 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give N- [ (1S) -2- [ [ (1 r,2S,6r,7 r) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] as a white solid ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (260 mg,62.8% yield, 96% purity).

Step 8: (2S) -2-amino-N- [ (1R, 2S,6R, 7R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl ]-3-cyclopropyl-propionamide

At N ₂ Next, N- [ (1S) -2- [ [ (1R, 2S,6R, 7R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (200 mg, 0.4)To a solution of 3mmol,1 eq.) in THF (2 mL) was added Pd/C (100 mg,10% purity). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (15 psi) and stirred at 25℃for 16h. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. Obtaining (2S) -2-amino-N- [ (1R, 2S,6R, 7R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] as a colourless oil ^2,6 ]Undecan-8-yl]-3-cyclopropyl-propionamide (140 mg, crude material).

Isomer 1: n- [ (1S) -2- [ [ (1R, 2S,6R,7R, 8S) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.02,6] undec-8-yl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide; isomer 2: n- [ (1S) -2- [ [ (1S, 2R,6S,7S, 8S) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.02,6] undecan-8-yl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1R, 2S,6R, 7R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]To a solution of 3-cyclopropyl-propionamide (140 mg,0.42mmol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (81.01 mg,0.42mmol,1 eq) and DIPEA (109.5 mg,0.84mmol,147.62ul,2 eq) in DCM (4 mL) was added HATU (193.3 mg,0.50mmol,1.2 eq). The reaction mixture was stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (10 mL) and the resulting mixture was extracted with DCM (20 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness under reduced pressure. The residue was purified by preparative TLC (DCM: meoh=10:1) to give the crude product. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,9.5 min) to give isomer 1 (13.30 mg,6.0% yield, 97.4% purity) and isomer 2 (31.40 mg,14.6% yield, 99.5% purity) as two white solids.

Isomer 1: LCMS: rt=0.808 min; c (C) ₂₇ H ₂₉ N ₅ O ₅ MS meterCalculating: 503.22; MS experimental values: 504.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.28(s,1H),7.18-7.12(m,1H),7.03(d,J＝8.3Hz,1H),6.52(d,J＝7.5Hz,1H),4.56(dd,J＝4.4,9.9Hz,1H),3.93(s,3H),3.17(d,J＝2.5Hz,1H),3.02-2.97(m,1H),2.96-2.90(m,1H),2.41(d,J＝15.3Hz,1H),2.33(d,J＝2.3Hz,1H),2.22-2.10(m,1H),1.94(d,J＝15.3Hz,1H),1.88-1.63(m,5H),0.90-0.75(m,1H),0.56-0.40(m,2H),0.31-0.13(m,2H)。

Isomer 2: LCMS: rt= 0.806min; c (C) ₂₇ H ₂₉ N ₅ O ₅ MS calculated: 503.22; MS experimental values: 504.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.25(s,1H),7.18-7.11(m,1H),7.03(d,J＝8.3Hz,1H),6.51(d,J＝7.8Hz,1H),4.64-4.60(m,1H),3.93(s,3H),3.17(d,J＝2.0Hz,1H),3.00-2.93(m,1H),2.92-2.86(m,1H),2.43(d,J＝15.6Hz,1H),2.31(s,1H),2.23-2.11(m,1H),1.94(d,J＝15.6Hz,1H),1.84-1.61(m,5H),0.85-0.70(m,1H),0.55-0.40(m,2H),0.23-0.09(m,2H)。

Example 155 Synthesis of viral protease inhibitor Compound 491

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl propionate (240 mg,1.01mmol,1 eq., HCl), (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a mixture of propionic acid (412.2 mg,1.22mmol,1.2 eq, HCl) and TEA (410.4 mg,4.06mmol,0.56mL,4 eq) in DMF (3 mL) was added T ₃ P (1.2 g,2.03mmol,1.21mL,50% purity, 2 eq.). The mixture was stirred at 25℃for 16h. TLC (DCM: meOH=10:1/UV 254 nm). H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2)By anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography12g/>Silica flash column, eluent: 100-25% ethyl acetate/MeOH at 30 mL/min). Obtaining the compound (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (256 mg,0.48mmol,48.2% yield, 92.5% purity).

In a sealed tube, (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (246.3 mg,0.47mmol,92.5% purity, 1 eq.) in NH ₃ The mixture in (7M, 6.72mL,100 eq.) 7M in MeOH was stirred at 80deg.C for 36h. The reaction mixture was concentrated in vacuo. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (220 mg, crude material) which was used in the next step without further purification.

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]A mixture of 4-methoxy-1H-indole-2-carboxamide (250 mg,0.53mmol,1 eq.) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (444.0 mg,1.86mmol,3.5 eq.) in DCM (3 mL)Stirred at 25℃for 16h. LC-MS showed detection of the desired compound. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column Phenomenex Gemini-NX 80. Times.30 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,9.5 min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (83 mg,0.18mmol,34.2% yield, 99.0% purity).

Isomer 1: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide; isomer 2: n- [ (1S) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide; isomer 3: n- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide; isomer 3: n- [ (1R) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%, min) purification of N- [2- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ]]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (50 mg,0.11mmol,1 eq.) gives three fragments: isomer 1, a mixture of isomers 2 and 3, and isomer 4.

Isomer 1: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide (28.1 mg,62.2umol,56.2% yield, 100% purity) was obtained as a white solid.

LCMS：Rt＝0.755min；C ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.2[ M+H ] ⁺ ]。

Isomer 4: n- [ (1R) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide (6.1 mg,13.5umol,12.2% yield, 100% purity) was obtained as a white solid.

LCMS：Rt＝0.752min；C ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.2[ M+H ] ⁺ ]。

Isomer 3: n- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide (5.1 mg,11.3umol,25.6% yield, 100% purity) was obtained as a white solid.

LCMS：Rt＝0.754min；C ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, ms experiment: 452.1[ M+H ] ⁺ ]。

Isomer 2: n- [ (1S) -2- [ [ (1R) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide (6.3 mg,14.0umol,31.6% yield, 100% purity) was obtained as a white solid.

LCMS：Rt＝0.754min；C ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.22, MS practice

And (3) checking: 452.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.12(s,1H),7.01-6.96(m,1H),6.87(d,J＝8.3Hz,1H),6.35(d,J＝7.8Hz,1H),4.89(t,J＝7.2Hz,1H),4.43(dd,J＝6.3,8.3Hz,2H),3.77(s,3H),3.08-3.00(m,2H),2.32-2.22(m,1H),2.20-2.10(m,1H),2.27-2.07(m,1H),1.84-1.73(m,2H),1.72-1.62(m,2H),1.60-1.50(m,2H),1.43-1.34(m,1H),0.75-0.62(m,1H),0.40-0.27(m,2H),0.08--0.04(m,2H)。

EXAMPLE 156 Synthesis of viral protease inhibitor Compound 493

Step 1: (2R) -2- (Benzylmethoxycarbonylamino) -3-bromo-propionic acid methyl ester

To (2S) -2- (benzyloxycarbonylamino) -3-hydroxy-propionic acid methyl ester (10 g,39.49mmol,1 eq.) and CBr at 0deg.C ₄ (15.7 g,47.38mmol,1.2 eq.) in THF (120 mL)Adding PPh into the liquid ₃ (12.4 g,47.38mmol,1.2 eq.) in THF (20 mL). Next, the mixture was stirred at 25℃for 16hr. TLC (petroleum ether/ethyl acetate=5/1, i ₂ ). The reaction mixture was filtered and the filtrate concentrated in vacuo. Through flash silica gel chromatography 25gSilica flash column, eluent: 0-20% ethyl acetate/petroleum ether gradient at 30 mL/min) to give methyl (2R) -2- (benzyloxycarbonylamino) -3-bromo-propionate (8.2 g,65.6% yield) as a white solid.

Step 2: (2S) -3- (3-acetyl-2-oxo-imidazolidin-1-yl) -2- (benzyloxycarbonylamino) propionic acid methyl ester

To a solution of 1-acetylimidazolidin-2-one (1.3 g,10.31mmol,1 eq.) in DMA (10 mL) was added NaH (618.6 mg,15.47mmol,60% purity, 1.5 eq.) at 25deg.C and the mixture stirred at 45deg.C for 15min. Next, DMA (30 mL) containing methyl (2R) -2- (benzyloxycarbonylamino) -3-bromo-propionate (3.2 g,10.31mmol,1 eq.) was added to the mixture at 45℃and the resulting mixture was stirred at 45℃for 15min. LC-MS showed detection of the desired compound. TLC (petroleum ether: ethyl acetate=0:1) showed that a new spot was detected. H for the reaction mixture ₂ O (20 mL) was diluted and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography20g/>Silica flash column, eluent: gradient of 0-80% petroleum ether/ethyl acetate ether at 30 mL/min). Obtaining the compound (2S) -3- (3-acetyl-2-oxo-imidazolidin-1-yl) -2- (benzyloxycarbonylamino) as a yellow oil ) Methyl propionate (1.5 g,40.0% yield).

Step 3: benzyl N- [ 2-amino-2-oxo-1- [ (2-oxoimidazolidin-1-yl) methyl ] ethyl ] carbamate

A solution of methyl 3- (3-acetyl-2-oxo-imidazolidin-1-yl) -2- (benzyloxycarbonylamino) propanoate (2.0 g,5.50mmol,1 eq.) in ammonia (7M, 14.94mL,19 eq.) was stirred at 65℃for 16hr. TLC (DCM: meoh=10:1). The reaction mixture was filtered and concentrated in vacuo. Through flash silica gel chromatography25gSilica flash column, eluent: 0-30% DCM/MeOH ether gradient at 30 mL/min). Obtaining the compound N- [ 2-amino-2-oxo-1- [ (2-oxo-imidazolidin-1-yl) methyl as a white solid]Ethyl group]Benzyl carbamate (460 mg,27.4% yield).

Step 4: 2-amino-3- (2-oxoimidazolidin-1-yl) propanamides

At N ₂ Next, pd/C (0.2 g,10% purity) was added to a solution of 4 (450 mg,1.47mmol,1 eq.) in MeOH (3 mL). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (15 psi) and stirred at 25℃for 1h. TLC (dichloromethane: methanol=10/1, ninhydrin (ninhydro)). The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was used in the next step without further purification and 2-amino-3- (2-oxoimidazolidin-1-yl) propionamide (250 mg, crude material) was obtained as a white solid.

Step 5: (2S) -2-amino-3-cyclopropyl-propionic acid tert-butyl ester

To a solution of 2-amino-3- (2-oxoimidazolidin-1-yl) propionamide (0.3 g,2.3mmol,1 eq.) in tert-butyl acetate (4.33 g,37.2mmol,5mL,16.0 eq.) was slowly added HClO at 0deg.C ₄ (533.3 mg,3.7mmol,0.32mL,70% purity, 1.6 eq.). The mixture was stirred at 25℃for 15h. TLC (Petroleum)Ether ethyl acetate=2/1, ninhydrin). H for the reaction mixture ₂ O (10 mL) was diluted, followed by 1N HCl (8 mL). With 10% Na ₂ CO ₃ The pH of the mixture was adjusted to about 9 with aqueous solution and then extracted with DCM (3×15 ml). The combined organic layers were purified by Na ₂ SO ₄ Drying gave (2S) -2-amino-3-cyclopropyl-propionic acid tert-butyl ester (0.4 g, crude material) as a colorless oil.

Step 6: (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] propionic acid tert-butyl ester

To a solution of 4-methoxy-1H-indole-2-carboxylic acid (206.3 mg,1.08mmol,1 eq.) and HOBt (153.1 mg,1.1mmol,1.0 eq.) in DCM (6 mL) were added EDCI (223.5 mg,1.17mmol,1.0 eq.) and tert-butyl (2S) -2-amino-3-cyclopropyl-propanoate (200 mg,1.08mmol,1 eq.). The mixture was stirred at 25℃for 16h. TLC (petroleum ether: ethyl acetate=2/1, uv). The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=100/1 to 2/1) the residue was purified to give (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow solid]Tert-butyl propionate (150 mg,38% yield).

Step 7: (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] propionic acid

To (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a solution of tert-butyl propionate (100 mg,0.27mmol,1 eq.) in DCM (1 mL) was added TFA (7.7 g,67.5mmol,5.0mL,242.05 eq.) and the resulting mixture stirred at 25℃for 1h. TLC (petroleum ether: ethyl acetate=2/1, uv). The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=100/1 to 2/1) purification of the residue to give (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a white solid]Propionic acid (50 mg,59.2% yield).

Step 8: n- [ (1S) -2- [ [ 2-amino-2-oxo-1- [ (2-oxoimidazolidin-1-yl) methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -3-cyclopropyl-2- [ (4 ]methoxy-1H-indole-2-carbonyl) amino group]To a solution of propionic acid (50 mg,0.16mmol,1 eq.) in DMF (2 mL) was added HATU (94.3 mg,0.24mmol,1.5 eq.), 2-amino-3- (2-oxoimidazolidin-1-yl) propanamide (42.7 mg,0.24mmol,1.5 eq.) and DIPEA (53.4 mg,0.41mmol,72.0uL,2.5 eq.). The mixture was stirred at 25℃for 1h. TLC (dichloromethane: methanol=10/1, uv). The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=100/1 to 10/1) to give 10 (60 mg,79% yield) as a white solid.

Step 9: n- [ (1S) -2- [ [ 1-cyano-2- (2-oxoimidazolidin-1-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To a solution of 10 (60 mg,0.13mmol,1 eq.) in DCM (3.0 mL) was added the Buerger's reagent (93.9 mg,0.39mmol,3.0 eq.). The mixture was stirred at 25℃for 16hr. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:21% -51%,9.5 min) to give N- [ (1S) -2- [ [ 1-cyano-2- (2-oxoimidazolidin-1-yl) ethyl ] as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (9.72 mg,16% yield).

LCMS：Rt＝0.772min；C ₂₂ H ₂₆ N ₆ O ₄ MS calculated: 438.20; MS experimental values: 439.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.28(s,1H),7.19-7.12(m,1H),7.03(d,J＝8.3Hz,1H),6.52(d,J＝7.6Hz,1H),5.22-5.01(m,1H),4.59(s,1H),3.93(s,3H),3.62-3.52(m,4H),3.44-3.34(m,2H),1.92-1.78(m,1H),1.70(tt,J＝6.8,13.2Hz,1H),0.83(d,J＝6.0Hz,1H),0.61-0.40(m,2H),0.27-0.08(m,2H)。

EXAMPLE 157 Synthesis of viral protease inhibitor Compounds 495

Isomer 1: n- [ (1S) -2- [ [ (1R, 2S,6R,7R, 8S) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]Benzyl carbamate; isomer 2: n- [ (1S) -2- [ [ (1S, 2R,6S,7S, 8R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzoic acid phenylmethyl ester

N- [ (1S) -2- [ [ (1R, 2S,6R, 7R) -8-cyano-4- [ (4-methoxyphenyl) methyl]-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (200 mg,0.34mmol,1 eq.) ammonia; cerium (4+); nitric acid; tetranitrate (1.13 g,2.05mmol,1.02mL,6 eq.) in H ₂ A mixture of O (1 mL) and MeCN (3 mL) was degassed and replaced with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred under an atmosphere at 25℃for 16h. H for mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (40 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column: 3_Phenomenex Luna C18 75*30mm*3um; mobile phase: [ water (0.05% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8.5 min) to give N- [ (1S) -2- [ [ (1R, 2S,6R,7R, 8S) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] as a white solid ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (17.25 mg,35.6umol,10.4% yield, 96.1% purity) and N- [ (1S) -2- [ [ (1S, 2R,6S,7S, 8R) -8-cyano-3, 5-dioxo-4-azatricyclo [5.2.2.0 ] as a white solid ^2,6 ]Undecan-8-yl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (17.56 mg,36.37umol,10.63% yield, 96.2% purity).

Isomer 1: LCMS: rt=0.798 min; c (C) ₂₅ H ₂₈ N ₄ O ₅ MS calculated: 464.21; MS experimental values: 465.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.47-7.21(m,5H),5.17-5.08(m,2H),4.10(dd,J＝4.3,9.8Hz,1H),3.12(br d,J＝2.5Hz,1H),3.01-2.88(m,2H),2.42-2.28(m,2H),2.20-2.09(m,1H),1.89(br d,J＝15.3Hz,1H),1.80-1.73(m,2H),1.72-1.61(m,2H),1.56(br d,J＝7.5Hz,1H),0.82-0.67(m,1H),0.42-0.42(m,1H),0.48-0.38(m,1H),0.23-0.09(m,2H)。

Isomer 2: LCMS: rt=0.818 min; c (C) ₂₅ H ₂₈ N ₄ O ₅ MS calculated: 464.21; MS experimental values: 465.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.45-7.25(m,5H),5.18-5.09(m,2H),4.17(br dd,J＝6.0,7.6Hz,1H),3.35(s,1H),3.11-2.93(m,2H),2.42(br d,J＝15.6Hz,1H),2.31(br s,1H),2.23-2.12(m,1H),1.91(br d,J＝15.3Hz,1H),1.76(br d,J＝6.8Hz,2H),1.68(br d,J＝11.4Hz,1H),1.65-1.58(m,1H),1.56-1.45(m,1H),0.78-0.67(m,1H),0.44(d,J＝5.1Hz,2H),0.12(br s,2H)。

EXAMPLE 158 Synthesis of viral protease inhibitor Compounds 496

Step 1:4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] pentanoic acid methyl ester

To a solution of 4-methoxy-1H-indole-2-carboxylic acid (281.6 mg,1.47mmol,1 eq.) in DCM (1 mL) was added HATU (672.2 mg,1.77mmol,1.2 eq.), DIPEA (571.2 mg,4.42mmol,0.76mL,3 eq.) and methyl 2-amino-4, 4-difluoro-pentanoate (300 mg,1.47mmol,1 eq., HCl). The mixture was stirred at 25℃for 2h. TLC (petroleum ether: ethyl acetate=0:1). H for the reaction mixture ₂ O (10 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography4g/>Silica flash column, eluent: gradient of 0-30% petroleum ether/ethyl acetate, at 20 mL/min) to give 4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow solid]Methyl valerate (317 mg,1.04mmol,70.7% yield, 99.4% purity).

Step 2:4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] pentanoic acid

At 0℃to 4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a solution of methyl valerate (317 mg,1.05mmol,1 eq.) in THF (3 mL) and MeOH (1 mL) was added LiOH. H ₂ H of O (132.0 mg,3.15mmol,3 eq) ₂ O (2 mL). The mixture was stirred at 0deg.C for 20min. The pH of the reaction was adjusted to about 4 with 4M HCl. H for the reaction mixture ₂ Dilute with O (5 mL) and extract with ethyl acetate (5 mL x 3). The combined organic phases were washed with brine (5 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was used in the next step without further purification. Obtaining the compound 4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]Valeric acid (321 mg,93.7% yield).

Step 3: n- [1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3, 3-difluoro-butyl ] -4-methoxy-1H-indole-2-carboxamide

To 4, 4-difluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a solution of pentanoic acid (20 mg,61.2umol,1 eq.) in DCM (0.5 mL) was added (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (13.9 mg,73.5umol,1.2 eq, HCl), TEA (18.6 mg,0.18mmol,25.5uL,3 eq) and T ₃ P (50.7 mg,79.6umol,50% purity, 1.3 eq.) in DMF (0.2 mL). The mixture was stirred at 0℃for 2h. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:17% -47%,9.5 min) pureThe residue was taken up to give N- [1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Carbamoyl group]-3, 3-difluoro-butyl]-4-methoxy-1H-indole-2-carboxamide (2.54 mg,8.7% yield, 97.7% purity).

LCMS：Rt＝0.772min；C ₂₂ H ₂₅ F ₂ N ₅ O ₄ MS calculated: 461.19; MS experimental values: 462.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.24(d,J＝3.8Hz,1H),7.15(dt,J＝2.3,8.0Hz,1H),7.03(dd,J＝2.5,8.3Hz,1H),6.52(dd,J＝1.5,7.5Hz,1H),5.07-5.00(m,1H),4.84(br s,1H),3.93(d,J＝1.8Hz,3H),3.30-3.18(m,2H),2.67-2.57(m,1H),2.56-2.40(m,2H),2.37-2.25(m,2H),1.95-1.85(m,1H),1.85-1.76(m,1H),1.69(dt,J＝2.6,18.8Hz,3H)。

EXAMPLE 159 Synthesis of viral protease inhibitor Compound 501

To 2- [ [ (3S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl group]-5-oxo-2, 3-dihydro-1H-indolizin-6-yl]Amino group]To a solution of phenyl acetate (100 mg,0.21mmol,1 eq.) in THF (1 mL) and MeOH (0.3 mL) was added LiOH. H ₂ H of O (27.1 mg,0.64mmol,3 eq) ₂ O (0.5 mL). The mixture was stirred at 25℃for 4h. LC-MS and HPLC showed detection of the desired compound. The pH of the reaction was adjusted to about 1 with 4M HCl. H for the reaction mixture ₂ Dilute with O (5 mL) and extract with ethyl acetate (5 mL x 3). The combined organic phases were washed with brine (5 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: 3_Phenomenex Luna C18 75*30mm*3um; mobile phase: [ water (0.05% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:0% -30%,8.5 min) of the purified residue. The residue was checked by LCMS and HPLC. By preparative HPLC (column: 3_Phenomenex Luna C18 75*30mm*3um; mobile phase: [ water (0.05% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:0% -30%,8.5 min) of the purified residue. Obtaining the compound as white solid2- [ [ (3S) -3- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]Carbamoyl group]-5-oxo-2, 3-dihydro-1H-indolizin-6-yl]Amino group]Acetic acid (1.2 mg,1.27% yield, 98.9% purity, CHOOH).

LCMS：Rt＝0.643min；C ₁₈ H ₂₁ N ₅ O ₅ MS calculated: 387.15; MS experimental values: 388.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.49(br s,1H),8.03(s,1H),6.32(d,J＝7.5Hz,1H),5.09-5.03(m,2H),3.74(s,2H),3.34(br s,1H),3.26-3.18(m,2H),3.17-3.07(m,1H),2.74-2.64(m,1H),2.62-2.51(m,1H),2.40-2.26(m,3H),2.24-2.15(m,1H),1.97-1.88(m,1H),1.87-1.77(m,1H)。

EXAMPLE 160 Synthesis of viral protease inhibitor Compound 505

To 2- (3-amino-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a solution of 3-cyclopropyl-propionamide (100 mg,0.27mmol,1 eq.) in THF (1 mL) was added Boc2O (610.6 mg,2.80mmol,0.64mL,10 eq.). The mixture was stirred at 25℃for 16h. TLC (DCM: meoh=10:1). H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative TLC (SiO) ₂ DCM: meoh=20:1) the residue was purified to give N- [1- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-2-oxo-3-pyridinyl]Tert-butyl carbamate (12.62 mg,9.0% yield, 91.4% purity).

LCMS：Rt＝0.832min；C ₂₃ H ₃₁ N ₅ O ₅ MS calculated: 457.23; MS experimental values: 458.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.98(d,J＝6.3Hz,1H),7.39-7.31(m,1H),6.45-6.34(m,1H),5.56-5.39(m,1H),5.03(d,J＝6.8Hz,1H),3.34(s,1H),3.29-3.22(m,1H),2.57-2.43(m,1H),2.41-2.30(m,1H),2.29-2.20(m,1H),2.01-1.94(m,2H),1.92-1.72(m,2H),1.52(d,J＝2.5Hz,9H),0.62(dd,J＝7.4,12.3Hz,1H),0.50-0.36(m,2H),0.21-0.12(m,1H),0.09-0.02(m,1H)。

EXAMPLE 161 Synthesis of viral protease inhibitor Compound 504

The residue was further separated by SFC. By SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ OETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -30%, min) to further separate the residue.

Isomer 1: obtaining the compound N- [1- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-2-oxo-3-pyridinyl]Tert-butyl carbamate (2.47 mg,23.1% yield). LCMS: rt=0.837 min; c (C) ₂₃ H ₃₁ N ₅ O ₅ MS calculated: 457.23; MS experimental values: 458.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.98(d,J＝7.1Hz,1H),7.34(dd,J＝1.7,7.1Hz,1H),6.39(t,J＝7.2Hz,1H),5.56-5.31(m,1H),5.01(dd,J＝6.8,9.3Hz,1H),3.34(d,J＝2.8Hz,2H),2.56-2.44(m,1H),2.41-2.32(m,1H),2.32-2.24(m,1H),2.00-1.91(m,3H),1.89-1.82(m,1H),1.52(s,9H),0.59(s,1H),0.46-0.37(m,2H),0.15(d,J＝8.4Hz,1H),0.03(d,J＝11.3Hz,1H)。

Isomer 2: obtaining the compound N- [1- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-2-oxo-3-pyridinyl]Tert-butyl carbamate (2.71 mg,25.5% yield). LCMS: rt=0.837 min; c (C) ₂₃ H ₃₁ N ₅ O ₅ MS calculated: 457.23; MS experimental values: 458.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.97(d,J＝7.3Hz,1H),7.35(dd,J＝1.8,7.0Hz,1H),6.38(t,J＝7.3Hz,1H),5.42(dd,J＝7.0,8.5Hz,1H),5.44-5.40(m,1H),5.03-4.99(m,1H),3.30-3.25(m,2H),2.48(dq,J＝5.3,9.2Hz,1H),2.29-2.22(m,1H),2.32-2.22(m,1H),2.02-1.94(m,2H),1.91-1.85(m,1H),1.84-1.73(m,1H),1.51(s,9H),0.63(br d,J＝6.8Hz,1H),0.49-0.42(m,2H),0.18-0.13(m,1H),0.06(dd,J＝4.3,8.8Hz,1H)。

EXAMPLE 162 Synthesis of viral protease inhibitor Compounds 509

Step 1: (2S) -2- [ [ (2S) -2- [ [4- (difluoromethoxy) -1H-indole-2-carbonyl ] amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4-methyl-pentanoyl ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (160 mg,476.44umol,1 eq, HCl) and 4- (difluoromethoxy) -1H-indole-2-carboxylic acid (108.23 mg,476.44umol,1 eq) were added to a mixture of DMAP (174.62 mg,1.43mmol,3 eq) and EDCI (274.00 mg,1.43mmol,3 eq) in DCM (4 mL), DMF (1 mL) was added to the mixture and stirred at 25 ℃ for 14H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (neutral conditions, column Waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) of the purified residue. Obtaining the compound (2S) -2- [ [ (2S) -2- [ [4- (difluoromethoxy) -1H-indole-2-carbonyl ] as a white solid]Amino group]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg,294.98umol,61.91% yield). MS (ESI) M/z494.3[ M+H ]] ⁺

Step 2: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4- (difluoromethoxy) -1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ [4- (difluoromethoxy) -1H-indole-2-carbonyl]Amino group]-4-methyl-pentanoyl ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, 294.9)A mixture of 8umol,1 eq.) in ammonia (7.65 g,449.19mmol,7.50mL,1522.81 eq.) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give a residue. Obtaining the compound N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4- (difluoromethoxy) -1H-indole-2-carboxamide (100 mg,202.63umol,68.69% yield) and used in the next step. MS (ESI) m/z 494.3[ M+H ]] ⁺

Step 3: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3-methyl-butyl ] -4- (difluoromethoxy) -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of 4- (difluoromethoxy) -1H-indole-2-carboxamide (100 mg,202.63umol,1 eq.) in DCM (3 mL) was added at once the bergs reagent (193.16 mg,810.53umol,4 eq.). The mixture was stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (neutral conditions, column Waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) of the purified residue. Obtaining the compound N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Carbamoyl group]-3-methyl-butyl]-4- (difluoromethoxy) -1H-indole-2-carboxamide (30 mg,63.09umol,31.14% yield). MS (ESI) m/z 476.3[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.88(d,J＝1.8Hz,1H),8.93(d,J＝8.1Hz,1H),8.65(d,J＝7.7Hz,1H),7.78-7.67(m,1H),7.43(d,J＝1.5Hz,1H),7.35-7.27(m,1H),7.21-7.12(m,1H),6.82(d,J＝7.6Hz,1H),5.04-4.85(m,1H),4.56-4.40(m,1H),3.20-3.03(m,2H),2.42-2.04(m,3H),1.85-1.47(m,5H),1.00-0.84(m,6H)

EXAMPLE 163 Synthesis of viral protease inhibitor Compound 515

Step 1: 4-hydroxy-1H-indole-2-carboxylic acid

To a mixture of 4-methoxy-1H-indole-2-carboxylic acid (500 mg,2.62mmol,1 eq.) in DCM (10 mL) at 0deg.C was added BBr ₃ (1.31 g,5.23mmol,2 eq.). The mixture was stirred at 25℃for 16h. H for mixture ₂ O (30 mL) was diluted and extracted with DCM (60 mL, extracted as 30mL x 2). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 4-hydroxy-1H-indole-2-carboxylic acid (200 mg, crude material) as a red solid. MS (ESI) M/z176.1[ M-H]+

Step 2: 4-hydroxy-1H-indole-2-carboxylic acid methyl ester

4-hydroxy-1H-indole-2-carboxylic acid (200 mg,1.13mmol,1 eq.) was added to HCl/MeOH (4M, 10mL,35.43 eq.). The mixture was stirred at 70℃for 5h. The reaction mixture was concentrated under reduced pressure to give a crude product. Purification of crude material by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=9/1 to 8/1), 4-hydroxy-1H-indole-2-carboxylic acid methyl ester (170 mg,800.28umol,70.89% yield, 90% purity) was obtained as a yellow solid. MS (ESI) m/z 190.1[ M-H] ⁺

Step 3:4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid methyl ester

To a mixture of methyl 4-hydroxy-1H-indole-2-carboxylate (300 mg,1.57mmol,1 eq.) and 2-morpholinoethanol (205.83 mg,1.57mmol,192.37uL,1 eq.) in THF (4 mL) was added PPh ₃ (452.73 mg,1.73mmol,1.1 eq.) in N ₂ DIAD (317.30 mg,1.57mmol,305.10uL,1 eq.) was added at 0deg.C. The mixture was stirred at 25℃for 60min. H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 2). The combined organic layers were washed with 20mL brine, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (petroleum ether: ethyl acetate=0:1) to give methyl 4- (2-morpholinoethoxy) -1H-indole-2-carboxylate (200 mg,591.44umol,37.69% yield, 90% purity) as a yellow solid. MS (ESI)m/z304.9[M+H] ⁺

Step 4:4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid

To 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid methyl ester (200 mg,657.16umol,1 eq.) in THF (2 mL) and H at 25 ℃ ₂ LiOH.H was added to the mixture in O (1 mL) ₂ O (41.37 mg,985.74umol,1.5 eq.). The mixture was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to give a residue. The crude material was purified by HCl prep HPLC to give 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid (80 mg,261.79 mol,39.84% yield, 95% purity) as a white solid. MS (ESI) m/z 289.2[ M-H ]] ⁺

To 4- (2-morpholinoethoxy) -1H-indole-2-carboxylic acid (70 mg,241.12umol,1 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl at 0deg.C]Ethyl group]To a mixture of 3-cyclopropyl-propionamide (159.33 mg,241.12umol,40% purity, 1 eq.) in DCM (2 mL) was added DIEA (93.49 mg,723.36umol,125.99uL,3 eq.) and T in one portion ₃ P (230.16 mg,361.68umol,215.10uL,50% purity, 1.5 eq.). The mixture was stirred at 0℃for 2h. EDTA solution (2 mL) was added to the reaction mixture and stirred at 25℃for 10min, and then extracted with DCM (6 mL, extracted as 2 mL. Times.3). The combined organic layers were washed with brine (5 mL, washed with 5mL of 3) and concentrated under reduced pressure to give a residue. Purification of the residue by preparative HPLC (neutral conditions) gives N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4- (2-morpholinoethoxy) -1H-indole-2-carboxamide (13 mg,24.23 mol,10.05% yield). MS (ESI) m/z 537.3[ M+H ]] ⁺

Column:waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：20％-50％，8min

EXAMPLE 164 Synthesis of viral protease inhibitor Compound 519

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propanoate (230 mg,578.67umol,1 eq.) in HCl/MeOH (3 mL) was stirred at 25℃for 30min. The reaction mixture was concentrated under reduced pressure to give crude methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (170 mg,571.72umol,98.80% yield) as a white solid.

Step 2: (2S) -2- [ [ (2S) -2- [ (5-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (170 mg, 571.72. Mu. Mol,1 eq.) in DCM (2 mL) and DMF (0.5 mL) was added DMAP (209.54 mg,1.72mmol,3 eq.) in one portion. To the mixture were added 5-chloro-1H-indole-2-carboxylic acid (134.20 mg,686.06umol,1.2 eq.) and EDCI (328.80 mg,1.72mmol,3 eq.) and stirred at 25 ℃ for 2H. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product was purified by preparative TLC (SiO ₂ EA: meoh=10:1) to give (2S) -2- [ [ (2S) -2- [ (5-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (140 mg,294.78 mol,51.56% yield). MS (ESI) m/z 475.2[ M+H ]] ⁺

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (5-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (130 mg,273.72umol,1 eq.) in NH ₃ The mixture in MeOH (7M) (5 mL) was stirred at 80℃for 16h. The reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-chloro-1H-indole-2-carboxamide (100 mg,217.43umol,79.43% yield). MS (ESI) m/z 460.2[ M+H ]] ⁺

Step 4: 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-chloro-1H-indole-2-carboxamide (100 mg,217.43umol,1 eq.) in DCM (2 mL) in one portionGabor reagent (103.63 mg,434.85umol,2 eq.). The mixture was stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure to give a residue. Purification of the residue by preparative HPLC (neutral conditions) gave 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (33 mg,74.68umol,34.35% yield). MS (ESI) m/z 442.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.71(s,1H),8.85(d,J＝8.2Hz,1H),8.59(d,J＝7.5Hz,1H),7.71-7.56(m,2H),7.34(d,J＝8.6Hz,1H),7.19(s,1H),7.10(dd,J＝1.5,8.8Hz,1H),4.97-4.80(m,1H),4.48-4.30(m,1H),3.12-2.94(m,2H),2.36-2.21(m,1H),2.13-1.96(m,2H),1.83-1.54(m,3H),1.47-1.34(m,1H),0.82-0.65(m,1H),0.39-0.26(m,2H),0.19-0.04(m,2H)

EXAMPLE 165 Synthesis of viral protease inhibitor Compound 531

(2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.68mmol,1 eq.) in DCM (10 mL) and DMF (2.5 mL) was added DMAP (616.30 mg,5.04mmol,3 eq.) at 25℃in one portion. To the mixture were added 7-chloro-1H-indole-2-carboxylic acid (394.69 mg,2.02mmol,1.2 eq.) and EDCI (967.04 mg,5.04mmol,3 eq.) and the reaction was stirred at 25 ℃ for 2H. The reaction mixture was concentrated under reduced pressure to give a crude product. By column chromatography (SiO ₂ Purifying the crude material with petroleum ether/ethyl acetate=5/1 to 0/1 to give (2S) -2- [ [ (2S) in the form of a white solid2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (550 mg,1.16mmol,68.87% yield). MS (ESI) m/z 475.1[ M+H ]] ⁺

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (500 mg,1.05mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,66.49 eq.) was stirred at 60℃for 16h. The reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (440 mg,956.68umol,90.87% yield). MS (ESI) m/z 460.3[ M+H ]] ⁺

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 7-chloro-1H-indole-2-carboxamide (430 mg,934.94 mmol,1 eq.) in DCM (6 mL) was added the Bungeus reagent (445.61 mg,1.87mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 4h. The reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃₎ -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (180 mg,407.32umol,43.57% yield). MS (ESI) m/z 442.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.71(br s,1H),9.01(d,J＝7.9Hz,1H),8.72(d,J＝7.5Hz,1H),7.71(s,1H),7.63(dd,J＝0.7,7.9Hz,1H),7.34-7.25(m,2H),7.07(t,J＝7.8Hz,1H),5.00(q,J＝7.9Hz,1H),4.58-4.49(m,1H),3.13(quin,J＝9.2Hz,2H),2.42-2.31(m,1H),2.22-2.05(m,2H),1.89-1.64(m,3H),1.57-1.46(m,1H),0.89-0.75(m,1H),0.50-0.37(m,2H),0.25-0.07(m,2H)

EXAMPLE 166 Synthesis of viral protease inhibitor Compound 539

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (500 mg,1.75mmol,1 eq.) in HCl/MeOH (4M, 20mL,45.81 eq.) was stirred at 20deg.C for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (350 mg, crude, HCl) as a yellow solid.

To (2S, 4R) -1-tert-butoxycarbonyl-4-methyl-pyrrolidine-2-carboxylic acid (250 mg,1.09mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a solution of methyl propionate (304.45 mg,1.64mmol,1.5 eq.) in DCM (10 mL) was added dropwise T3P (1.04 g,1.64mmol,972.75uL,50% purity, 1.5 eq.) and Et ₃ N (662.02 mg,6.54mmol,910.62uL,6 eq.) and the reaction stirred at 20℃for 2h. By adding H at 0 ℃ ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with 40mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) purification residueThe remainder, the product (2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-pyrrolidin-3-yl) is obtained as a colourless oil]Methyl group]Ethyl group]Carbamoyl group]-4-methyl-pyrrolidine-1-carboxylic acid tert-butyl ester (320 mg,805.10umol,73.86% yield). MS (ESI) m/z 398.2[ M+H ]] ⁺ 。

(2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A solution of tert-butyl 4-methyl-pyrrolidine-1-carboxylate (260 mg,654.15umol,1 eq.) in HCl/MeOH (4M, 8mL,48.92 eq.) was stirred at 20℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product (2S) -2- [ [ (2S, 4R) -4-methylpyrrolidine-2-carbonyl ] as a colorless oil ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg, crude material, HCl). MS (ESI) m/z 298.2[ M+H ]] ⁺ 。

To (2S) -2- [ [ (2S, 4R) -4-methylpyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (200 mg,599.14umol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (229.09 mg,1.20mmol,2.0 eq) in DMF (2.0 mL) was added DMAP (219.59 mg,1.80mmol,3.0 eq) and EDCI (229.71 mg,1.20mmol,2 eq) and DCM (8.0 mL) and the mixture was stirred at 20 ℃ for 2H. By adding H at 0 ℃ ₂ O (50 mL) to quench the reaction mixture and then extract with DCM (40 mL x 3). The combined organic layers were washed with 60mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:1 to 0:1) the residue was purified to give the product (2S) -2- [ [ (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carbonyl as a yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]Methyl propionate (250 mg,494.14umol,82.47% yield, 93% purity). MS (ESI) m/z 471.3[ M+H ]] ⁺ 。

(2S) -2- [ [ (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (220 mg,434.84umol,93% purity, 1 eq.) in NH ₃ A solution in MeOH (7M, 20mL,321.96 eq.) was stirred at 60℃for 12h. The reaction mixture was concentrated under reduced pressure to give the product (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid]Methyl group]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (200 mg, crude). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。

To (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (100 mg,219.54umol,1 eq.) in DCM (5 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (313.90 mg,1.32mmol,6 eq.) and the mixture was stirred at 20 ℃ for 3H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -60%,8 min) to give the product (2S, 4 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methyl-pyrrolidine-2-carboxamide (33 mg,75.43umol,34.36% yield, 100% purity). MS (ESI) m/z 438.2[ M+H ]] ⁺ 。

EXAMPLE 167 Synthesis of viral protease inhibitor Compounds 547

Containing (1S, 2S, 5R) -3- (9H-fluoren-9-ylmethoxycarbonyl) -3-azabicyclo [3.2.0]Heptane-2-carboxylic acid (250 mg,687.94umol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionitrile (486.36 mg,825.52umol,26% purity, 1.2 eq.) DCM (3 mL) was added to T ₃ The solution was stirred in P (656.67 mg,1.03mmol,613.71uL,50% purity, 1.5 eq.) and DIEA (266.73 mg,2.06mmol,359.48uL,3 eq.) at 25℃for 2h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with ethyl acetate (30 mL. Times.3), and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. Obtaining (1S, 2S, 5R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-pyrrolidin-3-yl ] as a yellow solid]Ethyl group]Carbamoyl group]-3-azabicyclo [3.2.0]Heptane-3-carboxylic acid 9H-fluoren-9-ylmethyl ester (185 mg,371.06umol,53.94% yield, 100% purity). MS (ESI) m/z 499.2[ M+H ]] ⁺ 。

To (1S, 2S, 5R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Carbamoyl (C)Base group]-3-azabicyclo [3.2.0]To a solution of 9H-fluoren-9-ylmethyl heptane-3-carboxylate (440 mg,706.02umol,80% purity, 1 eq.) in DCM (4.5 mL) was added piperidine (60.11 mg,706.02umol,69.72ul,1 eq.) and the solution stirred at 25 ℃ for 1H. The solution was dried to remove DCM and a residue was obtained. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. (1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a yellow solid]Ethyl group]-3-azabicyclo [3.2.0]Heptane-2-carboxamide (165 mg,597.10umol,84.57% yield, 100% purity).

To N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-azabicyclo [3.2.0]To a solution of heptane-2-carboxamide (165.00 mg,597.10umol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (171.23 mg,895.66umol,1.5 eq.) in DCM (2 mL) was added T ₃ P (284.98 mg, 895.66. Mu.L, 266.34. Mu.L, 1.5 eq.) and DIEA (154.34 mg,1.19mmol, 208.01. Mu.L, 2 eq.) were stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with ethyl acetate (30 mL. Times.3), and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. The residue was purified by preparative HPLC (neutral conditions).

Column: waters Xbridge Prep OBD C18, 150×40mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：25％-45％，8min。

(1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.2.0]Heptane-2-carboxamide (98 mg,218.02umol,36.51% yield, 100% purity). ¹ H NMR(400MHz,DMSO-d6)δ＝11.57(br s,1H),8.79(br d,J＝7.4Hz,1H),7.69(br s,1H),7.17-6.95(m,3H),6.52(br d,J＝7.3Hz,1H),4.97(br d,J＝6.8Hz,1H),4.63(br d,J＝8.2Hz,1H),4.33-3.97(m,2H),3.89(br s,3H),3.28-2.79(m,4H),2.30-1.55(m,9H)。MS(ESI)m/z 450.3[M+H] ⁺ 。

Obtaining (1R, 2R, 5S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.2.0]Heptane-2-carboxamide (23 mg,51.17umol,8.57% yield, 100% purity). ¹ H NMR(400MHz,DMSO-d6)δ＝11.56(br s,1H),9.13-8.71(m,1H),7.83-7.44(m,1H),7.23-6.89(m,3H),6.77-6.36(m,1H),5.18-4.57(m,2H),4.32-3.94(m,2H),3.92-3.74(m,3H),3.71-3.40(m,1H),3.23-2.76(m,3H),2.32-1.47(m,9H)。MS(ESI)m/z 450.3[M+H] ⁺ 。

EXAMPLE 168 Synthesis of viral protease inhibitor Compounds 549

At 0℃under N ₂ Downward (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (283.01 mg,1.27mmol,1.2 eq, HCl) and (2S, 4R) -1-tert-butoxycarbonyl-4- (trifluoromethyl) pyrrolidine-2-carboxylic acid (300 mg,1.06mmol,1 eq), DIEA (684.44 mg,5.30mmol,922.43uL,5 eq.) in THF (3 mL) was added T ₃ P (1.01 g,1.59mmol,944.87uL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 1h. After completion, the residue was poured into saturated sodium bicarbonate solution (10 mL) and stirred for 1min. The aqueous phase was extracted with ethyl acetate (10 ml x 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give (2S, 4 r) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a pale yellow oil]Methyl group]Ethyl group]Carbamoyl group]-4- (trifluoromethyl) pyrrolidine-1-carboxylic acid tert-butyl ester (0.5 g, crude material) and used directly in the next step. MS (ESI) m/z 452.1[ M+H ] ] ⁺ 。

At 25℃under N ₂ Downward (2S, 4R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]To a mixture of tert-butyl 4- (trifluoromethyl) pyrrolidine-1-carboxylate (0.5 g,1.11mmol,1 eq.) was added HCl/MeOH (4M, 3mL,10.83 eq.). The mixture was stirred at 25℃for 15min. After completion, the reaction mixture was concentrated to give the crude product (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl as a pale yellow oil]-2- [ [ (2 s,4 r) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]Methyl propionate (450 mg, crude material, HCl). MS (ESI) m/z 352.1[ M+H ]] ⁺ 。

At 0℃under N ₂ Downward (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl]-2- [ [ (2 s,4 r) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]Methyl propionate (395.52 mg,1.02mmol,1.3 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (150 mg,784.59umol,1 eq) and DIPEA (507.01 mg,3.92mmol,683.31uL,5 eq) were added T in a mixture of THF (3 mL) and DCM (3 mL) ₃ P (748.92 mg,1.18mmol,699.93uL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was poured into saturated sodium bicarbonate solution (5 mL) and stirred for 2min. The aqueous phase was extracted with ethyl acetate (5 ml x 2). The combined organic phases were washed with brine (5 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The crude product was purified by preparative TLC (dichloromethane: methanol=10:1, r _f =0.43) to give (2S) -2- [ [ (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl as a pale yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (250 mg, crude material). MS (ESI) m/z 525.2[ M+H ]] ⁺ 。

At 25℃under N ₂ Downward (2S) -2- [ [ (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]NH was added at one time to a mixture of methyl propionate (250 mg,476.65umol,1 eq) ₃ MeOH (7M, 3mL,44.06 eq.). The mixture was stirred at 80℃for 12h. After completion, the reaction mixture was cooled to 25 ℃ and concentrated to give the crude product. The crude product was purified by preparative TLC (dichloromethane: methanol=10:1, r _f =0.3) to give (2S, 4 r) -1- (4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1- (nitroso-methyl) -2- [ (3S) -2-oxopyrrolidin-3-yl as a pale yellow solid]Ethyl group]-4- (trifluoromethyl) pyrrolidine-2-carboxamide (130 mg,247.51umol,51.93% yield, 97% purity). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

At 25℃under N ₂ Downward (2S, 4R) -1- (4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1- (nitroso-methyl) -2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]To a mixture of 4- (trifluoromethyl) pyrrolidine-2-carboxamide (120 mg,235.54umol,1 eq.) in DCM (6 mL) was added at once the Bungeus reagent (112.26 mg,471.07umol,2 eq.). The mixture was stirred at 25℃for 4.5h. After completion, the residue was poured into water (0.5 mL) and stirred for 10min. The reaction mixture was then concentrated to give the crude product. The crude product was purified by preparative HPLC (column Phenomenex Gemini-NX 80X 40mM X3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8 min) to give (2S, 4 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide (22.56 mg,45.90umol,19.49% yield, 100% purity). MS (ESI) m/z 492.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.12-7.21(m,1H),6.84-7.10(m,2H),6.50(br s,1H),4.94-5.26(m,1H),4.75(br s,1H),4.07-4.47(m,2H),3.79-4.01(m,3H),3.45(br s,1H),2.16-2.98(m,6H),1.62-2.02(m,2H),1.39(br s,1H)

EXAMPLE 169 Synthesis of viral protease inhibitor Compound 551

Step 1: (2S, 4R) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -4-methylsulfanyl-pyrrolidine-1-carboxylic acid 9H-fluoren-9-ylmethyl ester

At 25℃under N ₂ Downward (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]Propionamide (200.57 mg,782.35 mol,81% purity, 1 eq, HCl) and (2 s,4 r) -1- (9H-fluoren-9-ylmethoxycarbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxylic acid (300 mg,782.35 mol,1 eq) were added EDCI (299.96 mg,1.56mmol,2 eq) and DMAP (191.16 mg,1.56mmol,2 eq) in a mixture of DCM (4 mL) and DMF (2 mL) at one time. The mixture was stirred at 25 ℃ and for 1 hour. After completion. The aqueous phase was treated with ethyl acetate (30 ml x 3) and H ₂ O (40 mL) extraction. The combined organic phases were washed with brine (30 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give (2S, 4 r) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl group]Ethyl group]Carbamoyl group]-4-methylsulfanyl-pyrrolidine-1-carboxylic acid 9H-fluoren-9-ylmethyl ester (180 mg,322.00umol,41.16% yield, 96% purity). MS (ESI) m/z 537.3[ M+H ]] ⁺

Step 2: (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -4-methylsulfanyl-pyrrolidine-2-carboxamide

To (2S, 4R) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl at 20 ℃C]Methyl group]Ethyl group]Carbamoyl group]To a mixture of 9H-fluoren-9-ylmethyl-4-methylsulfanyl-pyrrolidine-1-carboxylate (180 mg,335.42umol,1 eq.) in DCM (2 mL) was added piperidine (344.8)8mg,4.05mmol,0.4mL,12.08 eq). The mixture was stirred at 20℃for 1h. After completion. By preparative TLC (SiO) ₂ The crude material was purified with DCM/meoh=5/1) to give (2S, 4 r) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]-4-methylsulfanyl-pyrrolidine-2-carboxamide (80 mg,127.23umol,37.93% yield, 50% purity).

¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.15(br d,J＝9.6Hz,1H),7.63(s,1H),7.53(s,1H),7.12(br s,1H),4.28(br s,1H),3.73(br t,J＝7.2Hz,1H),3.22-3.03(m,4H),2.99(br s,2H),2.78(br d,J＝7.2Hz,1H),2.28-1.86(m,8H),1.74-1.43(m,6H)。

Step 3: (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxamide

To (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl at 20 ℃C]Methyl group]Ethyl group]To a mixture of 4-methylsulfanyl-pyrrolidine-2-carboxamide (80 mg,254.45 mol,1 eq) and 4-methoxy-1H-indole-2-carboxylic acid (48.65 mg,254.45 mol,1 eq) in DCM (2 mL) and DMF (1 mL) was added EDCI (97.56 mg,508.90 mol,2 eq) and DMAP (62.17 mg,508.90 mol,2 eq) in one portion and stirred for 1H. After completion, the mixture was treated with N ₂ And (5) drying. The crude material was purified by preparative HPLC, column: phenomenex Gemini-NX C18 75 x 30mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,8min to obtain (2S, 4R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-pyrrolidin-3-yl) as white solid]Methyl group]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxamide (80 mg,164.08umol,64.48% yield, 100% purity). MS (ESI) m/z 488.3[ M+H ]] ⁺

Step 4: (2S, 4R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxamide

At 20 ℃, to (2S, 4R) -N- [ (1S) -2-amino-2-oxo group -1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a mixture of 1- (4-methoxy-1H-indole-2-carbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxamide (80 mg,164.08umol,1 eq.) in DCM (4 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (312.81 mg,1.31mmol,8 eq.) in one portion and stirred for 3H. After completion. N for crude material ₂ And (5) drying. The crude material was purified by preparative HPLC, column: waters Xbridge BEH C18, 100×30mm×10um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,10min, to give (2S, 4R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-methylsulfanyl-pyrrolidine-2-carboxamide (31.9 mg,67.94umol,41.40% yield, 100% purity). MS (ESI) m/z 470.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.38(br s,1H),8.85(br s,1H),7.46(br s,1H),7.17-7.09(m,1H),7.09-7.02(m,1H),6.91(br s,1H),6.52(d,J＝7.5Hz,1H),4.95(br d,J＝7.1Hz,1H),4.86-4.60(m,1H),4.27(br s,1H),3.90(s,4H),3.54(br s,1H),3.18-3.12(m,2H),2.50-2.39(br s,8H),1.89-1.61(m,2H)。

EXAMPLE 170 Synthesis of viral protease inhibitor Compounds 555

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (300 mg,1.05mmol,1 eq.) and HCl/EA (3 mL) was stirred at 25℃for 0.5h. After completion, the residue was concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Methyl propionate (200 mg, crude material, HCl). MS (ESI) m/z 187.1[ M+H ]] ⁺ 。

Step 2: (1S) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3, 4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (200 mg,898.19umol,1 eq, HCl), (1S) -2-tert-butoxycarbonyl-3, 4-dihydro-1H-isoquinoline-1-carboxylic acid (249.08 mg,898.19umol,1 eq) and TEA (454.44 mg,4.49mmol,625.09uL,5 eq) in DCM (2 mL) and DMF (1 mL) was cooled to 0deg.C. Adding T at 0deg.C ₃ After P (1.71 g,2.69mmol,1.60mL,50% purity, 3 eq.) the mixture was stirred for 1h and gradually warmed to 25 ℃. After completion, H is added to the mixture ₂ O (30 mL) and then extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product (1S) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-3, 4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (300 mg, crude material). MS (ESI) m/z 446.2[ M+H ] ] ⁺ 。

Step 3: (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl ] -2- [ [ (1S) -1,2,3, 4-tetrahydroisoquinoline-1-carbonyl ] amino ] propionic acid methyl ester

(1S) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A solution of tert-butyl 3, 4-dihydro-1H-isoquinoline-2-carboxylate (300 mg,673.39umol,1 eq.) in HCl/EA (4M) was stirred at 25℃for 1H. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -3- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]-2- [ [ (1S) -1,2,3, 4-tetrahydroisoquinoline-1-carbonyl]Amino group]Methyl propionate (210 mg, crude material). MS (ESI) m/z 346.2[ M+H ]] ⁺ 。

Step 4: (2S) -2- [ [ (1S) -2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -3- [ (3S) -2-oxopyrrolidin-3-yl]-2- [ [ (1S) -1,2,3, 4-tetrahydroisoquinoline-1-carbonyl]Amino group]Methyl propionate (190 mg,497.57 mol,1 eq, HCl), 4-methoxy-1H-indole-2-carboxylic acid (95.13 mg,497.57 mol,1 eq), EDCI (286.16 mg, 1.4)A solution of 9mmol,3 eq.) and DMAP (182.36 mg,1.49mmol,3 eq.) in DCM (4 mL) was stirred at 25℃for 1h. After completion, H is added to the mixture ₂ O (30 mL) and then extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product (2S) -2- [ [ (1S) -2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carbonyl ] as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (40.1 mg,73.46umol,14.76% yield, 95% purity). MS (ESI) m/z 519.2[ M+H ]] ⁺ 。

Step 5: (1S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carboxamide

(2S) -2- [ [ (1S) -2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (40 mg,77.14umol,1 eq.) and NH ₃ A solution of MeOH (7M, 10mL,907.48 eq.) was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (1S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid ]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carboxamide (35 mg, crude material). MS (ESI) m/z 504.2[ M+H ]] ⁺ 。

Step 6: (1S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carboxamide

(1S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carboxamide (35 mg,69.51umol,1 eq.) and methoxycarbonyl- (triethylammonium) sulfonyl-imide (82.82 mg,a solution of 347.53umol,5 eq.) in DCM (5 mL) was stirred for 5h at 25 ℃. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the product (1S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -3, 4-dihydro-1H-isoquinoline-1-carboxamide (6 mg,12.08umol,17.38% yield, 97.74% purity). MS (ESI) m/z 486.2[ M+H ] ] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.57-7.47(m,1H),7.40-7.25(m,1H),7.12-7.11(m,1H),7.10-6.99(m,2H),6.59-6.50(m,1H),6.82-6.61(m,1H),5.67(s,1H),5.03-4.96(m,1H),4.46(s,1H),4.05-3.95(m,1H),3.94-3.86(m,3H),3.37-3.32(m,1H),3.28-3.16(m,2H),3.05-2.90(m,2H),2.62(s,1H),2.44-2.20(m,2H),1.98-1.67(m,2H)

EXAMPLE 171 Synthesis of viral protease inhibitor Compound 557

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (500 mg,1.75mmol,1 eq.) in HCl/dioxane (4M, 8.73mL,20 eq.) was degassed and treated with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 20℃for 0.5h under an atmosphere. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl propionate (630 mg, crude material, HCl). MS (ESI) m/z 223.2[ M+H ]] ⁺ 。

To 2-tert-Butoxycarbonyl isoindoline-1-carboxylic acid (436.93 mg,1.66mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (630 mg,1.74mmol,61.58% purity, 1.05 eq., HCl) in DCM (5 mL) and DMF (5 mL) was added T ₃ P (1.58 g,2.49mmol,1.48mL,50% purity, 1.5 eq.) and TEA (1.01 g,9.96mmol,1.39mL,6 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (20 mL) to quench the reaction mixture and extract with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (15 mL), and dried over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives the product 1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Carbamoyl group]Isoindoline-2-carboxylic acid tert-butyl ester (720 mg, crude). MS (ESI) M/z432.2[ M+H] ⁺ 。

Step 3: (2S) -2- (isoindoline-1-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]A mixture of tert-butyl isoindoline-2-carboxylate (720 mg,1.67mmol,1 eq.) in HCl/dioxane (4M, 8.34mL,20 eq.) was degassed and purified with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred at 20℃for 0.5h under an atmosphere. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2- (isoindoline-1-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl as a brown oil]Methyl propionate (770 mg, crude material, HCl). MS (ESI) m/z 332.3[ M+H ]] ⁺ 。

Step 4: (2S) -2- (2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

4-methoxy-1H-indole-2-carboxylic acid (287.43 mg,1.50mmol,1 eq.) 2S-2- (isoindoline-1-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (770 mg,1.65mmol,79% purity, 1.1 eq., HCl), DMAP (367.34 mg,3.01mmol,2 eq.), EDCI (576.42 mg,3.01mmol,2 eq.) in DCM (8 mL) and DMF2.7 mL) and deaerated with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred under an atmosphere at 20℃for 1h. After completion, by adding H ₂ O (25 mL) to quench the reaction mixture and then extract with DCM (10 mL x 3). The combined organic layers were washed with brine (15 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By neutral preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,10 min) to give the product (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl) as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (isomer 1:150mg,297.30umol,19.78% yield). MS (ESI) m/z 505.3[ M+H ]] ⁺ 。

To give (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carbonyl ] as a white solid ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (isomer 2:140mg,277.48umol,18.46% yield). MS (ESI) m/z 505.3[ M+H ]] ⁺ 。

Step 5.1: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

Step 5.2: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl ] carbonyl group) Isoindoline-1-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (140 mg,277.48umol,1 eq.) in MeOH/NH ₃ The solution in (7M, 792.81uL,20 eq.) was stirred at 45℃for 24h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (110 mg, crude). MS (ESI) m/z 490.3[ M+H ]] ⁺ 。

Step 6.1: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (125 mg,255.35 mol,1 eq.) in DCM (8 mL) was added the Bolus reagent (273.84 mg,1.15mmol,4.5 eq.). The mixture was stirred at 30℃for 20h. After completion, by adding H ₂ O (0.5 mL) to quench the reaction mixture, and then concentrated under reduced pressure to give a residue. By neutral prep HPLC (column: waters Xbridge BEH C18 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (31.50 mg,66.81umol,26.16% yield, 100% purity). MS (ESI) m/z 472.3[ M+H ]] ⁺ 。

Step 6.2: n- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]To a solution of 2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (105 mg,214.49umol,1 eq.) in DCM (6 mL) was added the berges reagent (204.47 mg,857.98umol,4 eq.). The mixture was stirred at 30℃for 7h. After completion, by adding H ₂ O (0.5 mL) to quench the reaction mixture, and then concentrated under reduced pressure to give a residue. By neutral preparative HPLC (column: waters Xbridge Prep OBD C18 150X 40mm X10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) isoindoline-1-carboxamide (34.83 mg,73.72umol,34.37% yield, 99.791% purity). MS (ESI) m/z 472.3[ M+H ] ] ⁺ 。

EXAMPLE 172 Synthesis of viral protease inhibitor Compounds 577

Step 1: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (350 mg,1.22mmol,1 eq.) in MeOH (1 mL) was added dropwise HCl/MeOH (4M, 10mL,32.72 eq.) and the resulting mixture was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a colorless oil]Methyl propionate (240 mg, crude, HCl). MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclobutylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (200 mg,898.19umol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclobutyl-propionic acid (218.53 mg,898.19umol,1 eq.) in DCM (5 mL) was added Et ₃ N (545.33 mg,5.39mmol,750.11uL,6.0 eq.) and T ₃ P (1.71 g,2.69mmol,1.60mL,50% purity, 3.0 eq.). The mixture was stirred at 25℃for 2h. By adding H at 0 ℃ ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (20 mL x 3). The combined organic layers were washed with 30mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclobutyl-propionyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (200 mg,486.04umol,54.11% yield). MS (ESI) m/z 412.1[ M+H ]] ⁺

Step 3: (S) -2- ((S) -2-amino-3-cyclobutylpropionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclobutyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (180 mg,437.43umol,1 eq.) in MeOH (1 mL) was added dropwise HCl/MeOH (4M, 12.00mL,109.73 eq.) and the resulting mixture was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give (2S) -2- [ [ (2S) -2-amino-3-cyclobutyl-propionyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, HCl). MS (ESI) m/z 312.2[ M+H ] ] ⁺

Step 4: (S) -2- ((S) -3-cyclobutyl-2- (4-methoxy-1H-indole-2-carboxamide) propionylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclobutyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg, 431.24. Mu. Mol,1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (82.45 mg, 431.24. Mu. Mol,1 eq.) inTo a solution of DMAP (105.37 mg,862.47 mol,2.0 eq.) in DMF (1.5 mL), EDCI (165.34 mg,862.47 mol,2.0 eq.) and DCM (6 mL) were added. The mixture was stirred at 25℃for 2h. By adding H at 0 ℃ ₂ O (40 mL) was used to quench the reaction mixture and extracted with DCM (20 mL x 3). The combined organic layers were washed with 30mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=0/1) to give (2S) -2- [ [ (2S) -3-cyclobutyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow oil]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (120 mg,247.66umol,57.43% yield). MS (ESI) m/z 485.2[ M+H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclobutyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclobutyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (100 mg,206.38umol,1 eq.) in NH ₃ A solution in MeOH (7M, 10mL,339.18 eq.) was stirred at 80℃for 6h. The reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclobutylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (90 mg, crude). MS (ESI) m/z 470.1[ M+H ]] ⁺

Step 7: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclobutyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclobutylmethyl) -2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (90 mg,191.68umol,1 eq.) in DCM (2 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (228.40 mg,958.40umol,5.0 eq.) and the mixture was stirred at 25 ℃2h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclobutylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (18.04 mg,39.95umol,20.84% yield, 100% purity). MS (ESI) m/z 452.3[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δppm 7.26(d,J＝0.7Hz,1H),7.11-7.18(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.6Hz,1H),5.05(dd,J＝10.1,5.9Hz,1H),4.41(dd,J＝8.6,6.2Hz,1H),3.93(s,3H),3.25-3.30(m,2H),2.61(dd,J＝8.7,5.3Hz,1H),2.42-2.53(m,1H),2.25-2.39(m,2H),2.06-2.18(m,2H),1.73-2.01(m,8H)。

EXAMPLE 173 Synthesis of viral protease inhibitor Compound 589

Step 1: (1- (bicyclo [3.1.0] hex-3-yl) -2- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -2-oxoethyl) carbamic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of propionitrile (89.1 mg,0.47mmol,1.2 eq, HCl), HATU (223.3 mg,0.58mmol,1.5 eq) and DIEA (151.8 mg,1.18mmol,0.20mL,3 eq) in DCM (2 mL) was stirred at 25℃for 0.5h and then 2- (3-bicyclo [ 3.1.0) was added to the reaction mass]Hexyl) -2- (t-butoxycarbonylamino) acetic acid (100 mg,0.39mmol,1 eq). The mixture was stirred at 25℃for 2h. LCMS detected the desired compound. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography4g />Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 20 mL/min). Obtaining the compound N- [1- (3-bicyclo [3.1.0] as a colorless oil]Hexyl) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Amino group]-2-oxo-ethyl]Tert-butyl carbamate (150 mg,0.23mmol,58.8% yield, 60% purity).

Step 2: 2-amino-2- (bicyclo [3.1.0] hex-3-yl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) acetamide

To a solution of tert-butyl N- [1- (3-bicyclo [3.1.0] hexyl) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -2-oxo-ethyl ] carbamate (140 mg,0.21mmol,60% purity, 1 eq.) in EtOAc (0.1 mL) was added HCl/EtOAc (4M, 0.84mL,15.62 eq.). The mixture was stirred at 25℃for 2h. The reaction mixture was concentrated under reduced pressure to give a residue. It was used in the next step without purification. The compound 2-amino-2- (3-bicyclo [3.1.0] hexyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] acetamide (110 mg, crude material, HCl) was obtained as a white solid.

Step 3: n- (1- (bicyclo [3.1.0] hex-3-yl) -2- (((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) amino) -2-oxoethyl) -4-methoxy-1H-indole-2-carboxamide

A mixture of 4-methoxy-1H-indole-2-carboxylic acid (52.1 mg,0.27mmol,1.2 eq.), HATU (129.6 mg,0.34mmol,1.5 eq.) and DIEA (88.1 mg,0.68mol,0.11mL,3 eq.) in DCM (2 mL) was stirred at 25℃for 0.5H and then 2-amino-2- (3-bicyclo [ 3.1.0) was added to the reaction mixture]Hexyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Acetamide (110 mg,0.22mmol,60% purity, 1 eq.). The reaction mixture was stirred at 25℃for 2h. TLC (petroleum ether/ethyl acetate=0:1, uv 254) indicated complete exhaustion of starting material and formation of new spots. LCMS detected the desired compound. Reaction toH is added to the mixture ₂ O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography4g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 30 mL/min) to give 50mg of 46% of the desired compound. Next, the mixture was purified by preparative HPLC (column Phenomenex Gemini-NX 80X 40mm X3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,9.5 min). Obtaining the compound N- [1- (3-bicyclo [ 3.1.0) as a white solid]Hexyl) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]Amino group]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (7 mg,15.1umol,6.6% yield, 100% purity).

LCMS：Rt＝0.813min；C ₂₅ H ₂₉ N ₅ O ₄ MS calculated: 463.53; MS experimental values: 464.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.22-7.31(m,1H),7.11-7.18(m,1H),7.00-7.06(m,1H),6.51(d,J＝7.63Hz,1H),4.93-5.01(m,2H),4.16-4.34(m,1H),3.93(s,3H),3.24-3.29(m,1H),2.45-2.67(m,1H),2.25-2.38(m,2H),2.00-2.17(m,2H),1.76-1.95(m,3H),1.60-1.71(m,1H),1.27-1.45(m,3H),0.74(br d,J＝5.00Hz,1H),0.13-0.38(m,2H)。

EXAMPLE 174 Synthesis of viral protease inhibitor Compound 590

Step 2: (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid methyl ester

At 0 ℃, 4-methyl-3-nitroTo a solution of base-1H-pyridin-2-one (1 g,6.49mmol,1 eq.) in DMF (15 mL) was added NaH (363.3 mg,9.08mmol,60% purity, 1.4 eq.) and the reaction mixture stirred at 25℃for 0.5H. Next, (2R) -2-bromo-3-cyclopropyl-propionic acid methyl ester (1.34 g,6.49mmol,1 eq.) was added to the reaction at 0deg.C. At N ₂ The mixture was stirred at 25℃for 16h. H for mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography24g/>Silica flash column, eluent: 0-50% ethyl acetate/petroleum ether gradient, at 35 mL/min) to give methyl (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanoate (867 mg,47.4% yield) as a yellow solid.

Step 3: (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid

Methyl (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanoate (867 mg,3.09mmol,1 eq.) LiOH. H ₂ O (519.2 mg,12.37mmol,4 eq.) in THF (6 mL), meOH (2 mL), H ₂ The mixture in O (2 mL) was degassed and replaced with N ₂ Purge 3 times, and then at N ₂ The mixture was stirred under an atmosphere at 25℃for 1h. LCMS showed detection of one peak with the expected MS. Adding H to the mixture ₂ O (5 mL), and then 2M HCl (4 mL) was added to the mixture to adjust the pH of the mixture to about 6-7. The mixture was extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to give (2S) as a yellow solid-3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid (791 mg,94.8% yield).

Step 4: n- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide

To a solution of (2S) -3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propionic acid (791 mg,2.97mmol,1 eq.) in DCM (10 mL) was added HATU (1.36 g,3.57mmol,1.2 eq.), DIPEA (1.15 g,8.91mmol,1.55mL,3 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]Propionitrile (676.0 mg,3.57mmol,1.2 eq. HCl). The mixture was stirred at 25℃for 2h. H for mixture ₂ O (20 mL) was quenched and extracted with DCM (40 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography24g/>Silica flash column, eluent: gradient of 0-10% DCM/MeOH ether at 35 mL/min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Ethyl group]-3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanamide (838 mg,64.5% yield).

LCMS：Rt＝0.741min；C ₁₉ H ₂₃ N ₅ O ₅ MS calculated: 401.17; MS experimental values: 402.1[ M+H ] ⁺ ]。

Step 5:2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -3-cyclopropyl-propionamide

To N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-2- (4-methyl-3-nitro-2-oxo-1-pyridinyl) propanTo a solution of the amide (838 mg,2.09mmol,1 eq.) in THF (10 mL) was added Pd/C (566.5 mg,0.53mmol,10% purity). At H ₂ The mixture was stirred at 25℃for 1h. The mixture was filtered and concentrated under reduced pressure to give 2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-3-cyclopropyl-propionamide (616 mg,1.43mmol,68.7% yield, 86.5% purity).

Step 6: n- [1- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methyl-2-oxo-3-pyridinyl ] carbamic acid tert-butyl ester

2- (3-amino-4-methyl-2-oxo-1-pyridinyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl]Ethyl group]-3-cyclopropyl-propionamide (100 mg,0.26mmol,1 eq.) in Boc ₂ Mixtures in O (1 mL) and THF (1 mL), and then in N ₂ The mixture was stirred under an atmosphere at 66 ℃ for 16h. The mixture was concentrated under reduced pressure. H for mixture ₂ O (20 mL) was quenched and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:26% -56%,7.8 min) to give N- [1- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-4-methyl-2-oxo-3-pyridinyl]Tert-butyl carbamate (44.33 mg,33.5% yield).

LCMS：Rt＝0.798min；C ₃₄ H ₅₁ N ₅ O ₁₀ MS calculated: 471.55; MS experimental values: 472.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.53(dd,J＝1.5,7.3Hz,1H),6.36-6.27(m,1H),5.56-5.35(m,1H),5.18-4.97(m,1H),3.35-3.32(m,1H),3.29-3.25(m,1H),2.52(tq,J＝4.8,9.3Hz,1H),2.45-2.22(m,2H),2.18(d,J＝5.0Hz,3H),2.06-1.92(m,2H),1.91-1.71(m,2H),1.48(d,J＝2.5Hz,9H),0.69-0.56(m,1H),0.50-0.37(m,2H),0.19-0.01(m,2H)。

EXAMPLE 175 Synthesis of viral protease inhibitor Compounds 591

By SFC (conditions: column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%, min) further isolation of N- [1- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ]]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methyl-2-oxo-3-pyridinyl]Tert-butyl carbamate (42 mg,89.0umol,1 eq.) gives N- [1- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methyl-2-oxo-3-pyridinyl]Tert-butyl carbamate (8.32 mg,19.8% yield).

Isomer 1: LCMS: rt=0.803 min; c (C) ₃₄ H ₅₁ N ₅ O ₁₀ MS calculated: 471.55; MS experimental values: 472.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.53(d,J＝7.3Hz,1H),6.32(d,J＝7.3Hz,1H),5.50(t,J＝7.8Hz,1H),5.01(dd,J＝7.0,9.0Hz,1H),3.35-3.32(m,1H),2.56-2.45(m,1H),2.42-2.23(m,2H),2.19(s,3H),2.00-1.92(m,2H),1.92-1.78(m,2H),1.49(s,9H),0.65-0.55(m,1H),0.46-0.36(m,2H),0.20-0.01(m,2H)。

Isomer 2: LCMS: rt=0.794 min; c (C) ₃₄ H ₅₁ N ₅ O ₁₀ MS calculated: 471.55; MS experimental values: 472.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,CD ₃ OD)δ7.53(d,J＝7.0Hz,1H),6.31(d,J＝7.3Hz,1H),5.41(t,J＝7.8Hz,1H),5.10-4.97(m,1H),3.30-3.25(m,2H),2.52(dq,J＝5.5,9.2Hz,1H),2.33-2.19(m,2H),2.18(s,3H),2.05-1.90(m,2H),1.89-1.71(m,2H),1.48(s,9H),0.70-0.58(m,1H),0.52-0.35(m,2H),0.20-0.04(m,2H)。

Example 176 Synthesis of viral protease inhibitor Compound 611

Step 1: (2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -4-phenyl-pyrrolidine-1-carboxylic acid tert-butyl ester

To a solution of (2S, 4S) -1-tert-butoxycarbonyl-4-phenyl-pyrrolidine-2-carboxylic acid (100 mg,0.34mmol,1 eq.) and DMAP (125.8 mg,1.03mmol,3 eq.) in DCM (0.7 mL) was added EDCI (78.9 mg,0.41mmol,1.2 eq.) and then (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (81.2 mg,0.34mmol,1 eq. HCl) in DMF (0.7 mL). The reaction mixture was stirred at 25℃for 2h. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (10 mL) was quenched and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give (2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Carbamoyl group]-4-phenyl-pyrrolidine-1-carboxylic acid tert-butyl ester (100 mg,60.4% yield).

LCMS：Rt＝0.826min；C ₂₅ H ₃₅ N ₃ O ₆ MS calculated: 473.25; MS experimental values: 474.1[ M+H ] ⁺ ]。

Step 2: (2S) -3- [ (3S) -2-oxo-3-piperidinyl ] -2- [ [ (2S, 4S) -4-phenylpyrrolidine-2-carbonyl ] amino ] propionic acid methyl ester

(2S, 4S) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]A mixture of tert-butyl-4-phenyl-pyrrolidine-1-carboxylate (90 mg,0.17mmol,1 eq. HCl) in 2M HCl/EtOAc (6 mL) and then N ₂ The mixture was stirred for 3h at 25℃under an atmosphere. LCMS showed detection of one peak with the expected MS. The mixture was concentrated under reduced pressure to give (2S) -3- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]Methyl propionate (70 mg,83.3% yield, HCl).

Step 3: (2S) -2- [ [ (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To a solution of 4-methoxy-1H-indole-2-carboxylic acid (40.5 mg,0.21mmol,1.5 eq.) and DMAP (51.8 mg,0.42mmol,3 eq.) in DCM (0.5 mL) was added EDCI (32.5 mg,0.16mmol,1.2 eq.) and then (2S) -3- [ (3S) -2-oxo-3-piperidinyl was added]-2- [ [ (2 s,4 s) -4-phenylpyrrolidine-2-carbonyl group]Amino group]A solution of methyl propionate (58 mg,0.14mmol,1 eq. HCl) in DMF (0.5 mL). The reaction mixture was stirred at 0℃for 1h. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (20 mL) was quenched and then extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give (2S) -2- [ [ (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carbonyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (30 mg,36.6% yield).

LCMS：Rt＝1.730min；C ₃₀ H ₃₄ N ₄ O ₆ MS calculated: 546.25; MS experimental value：547.1[M+H ⁺ ]。

Step 4: (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide

In a sealed tube, a solution of methyl (2S) -2- [ [ (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (30 mg,54.8umol,1 eq) and NH3 (7 m,6mL,765.2 eq) and MeOH (6 mL). The mixture was stirred at 60℃for 16h. LCMS showed detection of one peak with the expected MS. The mixture was concentrated under reduced pressure to give the compound (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide (29 mg,99.40% yield) as a yellow solid.

Step 5: (2S, 4S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide

To (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 0deg.C]Methyl group]Ethyl group]To a solution of 1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide (29 mg,54.5umol,1 eq.) in DCM (1 mL) was added the berges reagent (39.0 mg,0.16mmol,3 eq.). The mixture was stirred at 25℃for 16hr. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (5 mL) was quenched and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column: welch Xtime C18X 25mm X5 um; mobile phase: [ water (0.225% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:37% -67%,9.5 min) to give the compound (2S, 4S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide (1.9 mg,6.6% yield).

LCMS：Rt＝1.730min；C ₃₀ H ₃₄ N ₄ O ₆ MS calculated: 546.25; MS experimentValue: 547.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.38-7.31(m,1H),7.30-7.23(m,4H),7.19-7.13(m,1H),7.10-7.04(m,1H),6.95(d,J＝8.5Hz,1H),6.41(br d,J＝7.5Hz,1H),5.17-5.02(m,1H),4.43-4.20(m,1H),3.96-3.76(m,4H),3.74-3.41(m,1H),3.18-3.11(m,1H),3.01-2.55(m,2H),2.51-2.20(m,3H),2.15-1.62(m,4H),1.55-1.27(m,2H)。

EXAMPLE 177 Synthesis of viral protease inhibitor Compounds 619

Step 1: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-methoxy-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (120.0 mg,0.40mmol,1 eq.) and 5-methoxy-1H-indole-2-carboxylic acid (77.4 mg,0.40mmol,1 eq.) in DMF (2 mL) was added HATU (184.7 mg,0.48mmol,1.2 eq.) and DIEA (104.6 mg,0.8mmol,0.14mL,2 eq.). The mixture was stirred at 25 ℃ for 0.5h, and then the reaction mixture was concentrated under reduced pressure to remove DMF. Residue with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (25 mL x 3). The combined organic layers were washed with brine (10 ml x 3), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography4g/>Silica flash column, eluent: 0-10% methanol/dichloromethane at 20 mL/min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl group]-5-methoxy-1H-indole-2-carboxamide (180.0 mg,94.6% yield).

Step 2: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-methoxy-1H-indole-2-carboxamide

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 5-methoxy-1H-indole-2-carboxamide (180.0 mg,0.38mmol,1 eq.) in DCM (0.5 mL) was added the Buerger's reagent (274.0 mg,1.15mmol,3 eq.). After stirring the mixture at 25 ℃ for 16h, the reaction mixture was concentrated under reduced pressure to remove DCM. Residue with H ₂ O (5 mL) was diluted and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (5 ml x 3), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:24% -54%,7.8 min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-methoxy-1H-indole-2-carboxamide (37.3 mg,82.6umol,5.4 yield).

LCMS：Rt＝0.785min；C ₂₄ H ₂₉ N ₅ O ₄ MS calculated: 451.52; MS experimental values: 452.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.32(d,J＝8.8Hz,1H),7.13-7.06(m,2H),6.89(dd,J＝2.4,8.9Hz,1H),5.16-5.10(m,1H),4.55(t,J＝7.4Hz,1H),3.81(s,3H),3.26-3.20(m,2H),2.54-2.41(m,2H),2.04-1.85(m,3H),1.84-1.77(m,1H),1.74-1.62(m,2H),1.56-1.47(m,1H),0.95-0.79(m,1H),0.60-0.47(m,2H),0.19(br dd,J＝4.8,10.0Hz,2H)。

EXAMPLE 178 Synthesis of viral protease inhibitor Compound 621

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of benzyl carbamate (400 mg,0.92mmol,1 eq.) in MeOH (5 mL) was added Pd (200 mg,10% purity) and H ₂ (0.92 mmol). The mixture was stirred at 25℃for 1h at 15 psi. LCMS showed detection of one peak with the expected MS. The mixture was filtered to obtain a filtrate. The mixture was concentrated under reduced pressure to give (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (274 mg,99.5% yield).

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-methoxy-1H-pyrrolo [3,2-b ] pyridine-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (137 mg,0.46mmol,1 eq.) and 5-methoxy-1H-pyrrolo [3, 2-b)]To a solution of pyridine-2-carboxylic acid (88.8 mg,0.46mmol,1 eq.) in DMF (2 mL) was added DIPEA (119.4 mg,0.92mmol,0.16mL,2 eq.) and HATU (210.9 mg,0.55mmol,1.2 eq.). The mixture was stirred at 25℃for 1h. LCMS showed detection of one peak with the expected MS. The mixture was concentrated under reduced pressure. Through flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropane)Methyl) -2-oxo-ethyl]-5-methoxy-1H-pyrrolo [3,2-b]Pyridine-2-carboxamide (144 mg,63.1% yield).

LCMS：Rt＝0.675min；C ₂₃ H ₃₀ N ₆ O ₅ MS calculated: 470.23; MS practice

And (3) checking: 471.1[ M+H ] ⁺ ]。

Step 3: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-methoxy-1H-pyrrolo [3,2-b ] pyridine-2-carboxamide

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-methoxy-1H-pyrrolo [3,2-b]To a solution of pyridine-2-carboxamide (44 mg,93.5umol,1 eq.) in DCM (1 mL) was added the Buerger's reagent (66.86 mg,0.28mmol,3 eq.). The mixture was stirred at 25℃for 12hr. LCMS showed detection of one peak with the expected MS. H for mixture ₂ O (10 mL) was quenched and extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,7.8 min) to give the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-methoxy-1H-pyrrolo [3,2-b]Pyridine-2-carboxamide (12.08 mg,9.3% yield).

LCMS：Rt＝0.727min；C ₂₃ H ₂₈ N ₆ O ₄ MS calculated: 452.22; MS experimental values: 453.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.79-7.74(m,1H),7.17(s,1H),6.72(d,J＝9.0Hz,1H),5.17-5.04(m,1H),4.56(t,J＝7.4Hz,1H),3.97-3.96(m,1H),3.95(s,2H),3.26-3.19(m,2H),2.56-2.40(m,2H),2.02-1.87(m,3H),1.85-1.78(m,1H),1.76-1.63(m,2H),1.59-1.46(m,1H),0.90-0.77(m,1H),0.59-0.46(m,2H),0.27-0.07(m,2H)。

Example 179 Synthesis of viral protease inhibitor Compound 623

Step 1: methoxy-1H-pyrrolo [3,2-c ] pyridine-2-carboxylic acid

To 4-methoxy-1H-pyrrolo [3,2-c]To a solution of methyl pyridine-2-carboxylate (150 mg,0.72mmol,1 eq.) in THF (1 mL) was added LiOH. H ₂ O (30.5 mg,0.72mmol,1 eq.) and MeOH (0.5 mL). The mixture was stirred at 25℃for 16h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The aqueous layer was acidified with concentrated HCl and extracted with DCM. The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The crude product was wet-milled with petroleum ether at 25℃for 60min. Obtaining the compound 4-methoxy-1H-pyrrolo [3,2-c ] as a white solid]Pyridine-2-carboxylic acid (120 mg,84.9% yield, 99% purity).

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (150 mg,0.50mmol,1 eq.) in DMF (1 mL) was added HATU (192.4 mg,0.50mmol,1 eq.), 4-methoxy-1H-pyrrolo [3, 2-c) ]Pyridine-2-carboxylic acid (106.9 mg,0.55mmol,1.1 eq.) and DIEA (130.8 mg,1.01mmol,0.17mL,2 eq.) were stirred at 25℃for 16hr. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1-(Cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (110 mg, crude).

Step 3: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-pyrrolo [3,2-c]To a solution of pyridine-2-carboxamide (110 mg,0.23mmol,1 eq.) in DCM (1 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (167.1 mg,0.70mmol,3 eq.). The mixture was stirred at 25℃for 24h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,9.5 min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (40.69 mg,38.4% yield, 100% purity).

LCMS：Rt＝1.387min；C ₂₃ H ₂₈ N ₆ O ₄ MS calculated: 452.51; MS experimental values: 453.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.77(d,J＝6.0Hz,1H),7.34(s,1H),7.05(d,J＝6.3Hz,1H),4.47(dd,J＝4.0,11.8Hz,1H),4.57(dd,J＝6.0,8.3Hz,1H),4.05(s,3H),3.28-3.17(m,2H),2.47-2.35(m,1H),2.28(ddd,J＝4.4,12.0,14.0Hz,1H),2.08-1.95(m,1H),1.90-1.77(m,3H),1.77-1.63(m,2H),1.62-1.48(m,1H),0.96-0.78(m,1H),0.59-0.42(m,2H),0.26-0.11(m,2H)。

EXAMPLE 180 Synthesis of viral protease inhibitor Compound 625

Step 1: (2S) -2- (Benzylmethoxycarbonylamino) -3-cyclopropyl-propionic acid

To a solution of (2S) -2-amino-3-cyclopropyl-propionic acid (5 g,38.71mmol,1 eq.) was added NaOH (1M, 135.4mL,3.5 eq.) and benzyl chloroformate (8.5 g,50.33mmol,7.1mL,1.3 eq.) and stirred at 25℃for 2hr. TLC (petroleum ether/ethyl acetate=1:1, pma). H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The aqueous layer was acidified with concentrated HCl and extracted with DCM. The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The compound (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionic acid (7.7 g,68.1% yield, 90% purity) was obtained as a white solid.

Step 2: (S) -2- ((S) -2- (((benzyloxy) carbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionic acid (3.5 g,13.29mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (3.15 g,13.29mmol,1 eq., HCl) in DMF (60 mL) was added TEA (4.04 g,39.88mmol,5.55mL,3 eq.) and T ₃ P (8.46 g,13.29mmol,7.91mL,50% purity, 1 eq.). The mixture was stirred at 25℃for 2hr. TLC (petroleum ether/ethyl acetate=0:1, i ₂ ). LCMS detected the desired compound. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica rapidQuick column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 40 mL/min). The compound (2S) -2- [ [ (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionyl was obtained as a colorless oil ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (4.5 g,9.49mmol,71.4% yield, 94% purity).

Step 3: benzyl (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamate

To stirred (2S) -2- [ [ (2S) -2- (benzyloxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (4.5 g,10.10mmol,1 eq.) in MeOH (10 mL) was added NH ₃ (7M, 50mL,34.65 eq.) in solution. The mixture was stirred in a sealed tube at 80℃for 24h. TLC (DCM/MeOH=10:1, I) ₂ ). LCMS detected the desired compound. The reaction mixture was concentrated under reduced pressure to give a residue. Through flash silica gel chromatography24g/>Silica flash column, eluent: 0-5% DCM/MeOH at 40 mL/min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Benzyl carbamate (3.6 g,7.69mmol,76.17% yield, 92% purity).

Step 4: (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropylpropionamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of benzyl carbamate (800 mg,1.86mmol,1 eq.) in MeOH (3 mL) was added Pd/C (100 mg,1.86mmol,10% purity, 1 eq.). At H ₂ The mixture was stirred at 25℃for 2h (15 psi). LCMS refers toShowing complete depletion of starting material and detection of the desired compound. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The compound (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl was obtained as a colorless oil]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (550 mg,1.86mmol,99.87% yield).

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

1H-pyrrolo [3,2-c]A mixture of pyridine-2-carboxylic acid (90.2 mg,0.55mmol,1.1 eq), HATU (288.6 mg,0.75mmol,1.5 eq.) and DIPEA (196.2 mg,1.52mmol,0.26mL,3 eq.) in DMF (2 mL) was stirred at 25℃for 0.5h and then (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ (3S) -2-oxo-3-piperidinyl was added to the reaction ]Methyl group]Ethyl group]3-cyclopropyl-propionamide (150 mg,0.50mmol,1 eq.). The resulting mixture was stirred at 25℃for 2hr. TLC (DCM/meoh=5:1, uv 254) indicated complete exhaustion of starting material and formation of new spots. LCMS detected the desired compound. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-20% MeOH/DCM at 30 mL/min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (200 mg,0.43mmol,86.1% yield, 96% purity).

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-1H-pyrrolo [3,2-c]To a solution of pyridine-2-carboxamide (120 mg,0.27mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (194.7 mg,0.81mmol,3 eq.). At N ₂ The mixture was stirred at 25℃for 16h. LCMS detected the desired compound. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:7% -37%,9.5 min) to give-20 mg of crude product. By preparative HPLC (column: welch Xtimate C18X 25mm X5 um; mobile phase: [ water (0.05% ammonium hydroxide, v/v) -MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:0% -60%,7.8 min) of the purified residue. Obtaining the compound N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (2.37 mg,5.4umol,1.9% yield, 96.8% purity).

LCMS：Rt＝1.321min；C ₂₂ H ₂₆ N ₆ O ₃ MS calculated: 422.48; MS experimental values: 423.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ9.03(s,1H),8.43(br d,J＝6.27Hz,1H),8.25(d,J＝6.27Hz,1H),7.14-7.26(m,1H),5.16(t,J＝8.16Hz,1H),4.64(br d,J＝2.01Hz,1H),3.25-3.29(m,2H),2.41-2.60(m,2H),1.93-2.09(m,2H),1.71-1.91(m,4H),1.49-1.63(m,1H),0.88(br s,1H),0.46-0.53(m,2H),0.12-0.26(m,2H)。

Example 181 Synthesis of viral protease inhibitor Compounds 669

Step 1: 4-chloro-1H-pyrrolo [2,3-c ] pyridine-2-carboxylic acid

To 4-chloro-1H-pyrrolo [2,3-c]To a solution of ethyl pyridine-2-carboxylate (300 mg,1.34mmol,1 eq.) in THF (5 mL) and MeOH (2 mL) was added LiOH. H ₂ O (280.2 mg,6.68mmol,5 eq.) and H ₂ O (2 mL). The mixture was stirred at 25℃for 16h. The reaction mixture was concentrated under reduced pressure to remove MeOH and THF. Next, the pH of the residue was adjusted (neutralized) to about 6-7 with 2m hcl, filtered and then the filter cake was concentrated under reduced pressure to give a residue. Obtaining 4-chloro-1H-pyrrolo [2,3-c ] as a white solid]Pyridine-2-carboxylic acid (190 mg,0.96mmol,72.3% yield).

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-chloro-1H-pyrrolo [2,3-c ] pyridine-2-carboxamide

To 4-chloro-1H-pyrrolo [2,3-c]Pyridine-2-carboxylic acid (70 mg,0.35mmol,1 eq.) and (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (105.5 mg,0.35mmol,1 eq.) in DMF (2 mL) was added T ₃ P (226.5 mg,0.35mmol,0.21mL,50% purity, 1 eq.) and TEA (108.0 mg,1.07mmol,0.14mL,3 eq.). The mixture was stirred at 25℃for 2h. TLC (DCM/meoh=5:1, uv 254). Addition of H to the reaction mixture ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-15% MeOH/DCM at 30 mL/min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropyl)Methyl) -2-oxo-ethyl]-4-chloro-1H-pyrrolo [2,3-c]Pyridine-2-carboxamide (80 mg,0.16mmol,46.8% yield, 99% purity).

Step 3: 4-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrolo [2,3-c ] pyridine-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-1H-pyrrolo [2,3-c]To a solution of pyridine-2-carboxamide (60 mg,0.12mmol,1 eq.) in DCM (2 mL) was added Bugess (60.2 mg,0.25mmol,2 eq.). At N ₂ The mixture was stirred at 25℃for 16h. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: welch Xtimate C18X 25mm X5 um; mobile phase: [ water (0.05% ammonium hydroxide, v/v) -MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:53% -83%,7.8 min). The compound 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrolo [2,3-c]Pyridine-2-carboxamide (9.41 mg,19.8umol,15.7% yield, 96.6% purity).

LCMS：Rt＝1.895min；C ₂₂ H ₂₅ ClN ₆ O ₃ MS calculated: 456.93; MS experimental values: 457.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.72(s,1H),8.13(s,1H),7.35-7.39(m,1H),5.14(dd,J＝10.04,6.02Hz,1H),4.56(t,J＝7.53Hz,1H),3.22-3.28(m,2H),2.40-2.57(m,2H),1.88-2.05(m,3H),1.82(td,J＝9.16,4.27Hz,1H),1.68(dd,J＝14.43,7.15Hz,1H),1.47-1.58(m,1H),1.31(t,J＝7.28Hz,1H),0.80-0.91(m,1H),0.48-0.57(m,2H),0.15-0.26(m,2H)。

EXAMPLE 182 Synthesis of viral protease inhibitor Compounds 633

Step 1: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-chloro-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (100.0 mg,0.33mmol,1 eq.) and 5-chloro-1H-indole-2-carboxylic acid (66.0 mg,0.33mmol,1 eq.) in DMF (2 mL) was added HATU (153.9 mg,0.40mmol,1.2 eq.) and DIEA (87.2 mg,0.67mmol,0.11mL,2 eq.). The mixture was stirred at 25℃for 0.5h. The reaction mixture was concentrated under reduced pressure to remove DMF. Residue with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (25 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography4g/>Silica flash column, eluent: 0-10% methanol/dichloromethane at 20 mL/min). Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-chloro-1H-indole-2-carboxamide (150.0 mg,90.9% yield).

Step 2: 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 5-chloro-1H-indole-2-carboxamide (129.0 mg,0.27mmol,1 eq.) in DCM (2.5 mL) was added the Buerger's reagent (259.4 mg,1.09mmol,4 eq.). At 25 DEG CThe mixture was stirred for 3h. The reaction mixture was concentrated under reduced pressure to remove DCM. Residue with H ₂ O (15 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:31% -61%,7.8 min). The compound 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (40.2 mg,30.2% yield).

LCMS：Rt＝0.832min；C ₂₃ H ₂₆ ClN ₅ O ₃ MS calculated: 455.94; MS (MS)

Experimental values: 456.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.61(d,J＝1.8Hz,1H),7.41(d,J＝8.8Hz,1H),7.19(dd,J＝2.0,8.8Hz,1H),7.14(s,1H),5.13(br dd,J＝6.1,10.2Hz,1H),4.57-4.52(m,1H),3.24-3.20(m,1H),2.56-2.40(m,2H),2.05-1.84(m,4H),1.80-1.59(m,3H),1.57-1.42(m,1H),0.85(br s,1H),0.54(br d,J＝8.3Hz,2H),0.19(br dd,J＝5.1,9.9Hz,2H)。

EXAMPLE 183 Synthesis of viral protease inhibitor Compound 635

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of benzyl carbamate (400 mg,0.92mmol,1 eq.) in MeOH (5 mL) was added Pd (200 mg,10% purity) and H ₂ (0.92 mmol). The mixture was stirred at 25℃for 1h at 15 psi. Filtration The mixture gave a filtrate and the reaction was concentrated under reduced pressure to give (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (274 mg,0.92mmol,99.5% yield).

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (137 mg,0.46mmol,1 eq.) and 6-chloro-1H-indole-2-carboxylic acid (90.4 mg,0.46mmol,1 eq.) in DMF (2 mL) was added DIPEA (119.4 mg,0.92mmol,0.16mL,2 eq.) and HATU (210.9 mg,0.55mmol,1.2 eq.). The mixture was stirred at 25℃for 1h. Concentrating the mixture under reduced pressure and purifying by flash silica gel chromatography12g/>Silica flash column, eluent: the residue was purified with 0-10% DCM/MeOH at 30mL/min to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group ]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-1H-indole-2-carboxamide (200 mg,89.0% yield).

LCMS：Rt＝0.780min；C ₂₃ H ₂₈ ClN ₅ O ₄ MS calculated: 473.18; MS experimental values: 474.1[ M+H ] ⁺ ]。

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-1H-indole-2-carboxamide (47.5 mg,0.1mmol,1 eq.) in DCM (1 mL)) To the solution in (b) was added a primary Gibbs reagent (71.6 mg,0.3mmol,3 eq.). The mixture was stirred at 25℃for 12h. The mixture was concentrated under reduced pressure and purified by preparative HPLC (column Phenomenex Gemini-NX 80 x 40mm x 3um; mobile phase: [ water (0.05% nh ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:31% -61%,7.8 min) to give 6-chloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (64.33 mg,34.7% yield).

LCMS：Rt＝0.832min；C ₂₃ H ₂₆ ClN ₅ O ₃ The method comprises the steps of carrying out a first treatment on the surface of the MS calculated: 455.17; MS experimental values: 456.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,DMSO-d ₆ )δ11.73(br s,1H),8.95(br d,J＝8.0Hz,1H),8.66(br d,J＝7.5Hz,1H),7.66(d,J＝8.5Hz,1H),7.53(br s,1H),7.44(s,1H),7.31(s,1H),7.05(dd,J＝1.8,8.5Hz,1H),5.11-4.96(m,1H),4.52-4.42(m,1H),3.09(br s,2H),2.34-2.21(m,2H),1.89-1.75(m,3H),1.74-1.65(m,1H),1.56(br s,1H),1.51-1.29(m,2H),0.79(br s,1H),0.42(br d,J＝7.0Hz,2H),0.23-0.01(m,2H)。

EXAMPLE 184 Synthesis of viral protease inhibitor Compound 637

Step 1:4, 7-dichloro-2- (trichloromethyl) -1H-benzimidazole

To a solution of 3, 6-dichlorobenzene-1, 2-diamine (0.3 g,1.69mmol,1 eq.) in AcOH (12.57 g,209.2mmol,11.97mL,123.8 eq.) was added methyl 2, 2-trichloroethyliminoate (313.0 mg,1.77mmol,0.21mL,1.05 eq.) at 0deg.C. The mixture was stirred at 25℃for 16h. LC-MS showed 48% 1 residual and 43% of the desired compound was detected. H for the reaction mixture ₂ O (40 mL) was diluted and filtered to give 2 (300 mg, crude) as a white solid.

Step 2:4, 7-dichloro-1H-benzimidazole-2-carboxylic acid

At 0deg.C, naOH (0.8 g,20.0mmol,20.2 eq.) was added to H ₂ To a solution of O (10 mL) was added 4, 7-dichloro-2- (trichloromethyl) -1H-benzo [ d ]]Imidazole (0.3 g,985.58umol,1 eq). The mixture was stirred at 25℃for 1h. The pH of the mixture was adjusted to 2-3 with HCl (2M) and the mixture was then filtered to give 4, 7-dichloro-1H-benzo [ d ] as a white solid]Imidazole-2-carboxylic acid (0.2 g, crude material).

To (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropylpropionamide (130 mg,0.43mmol,1 eq.) and 4, 7-dichloro-1H-benzo [ d ]]To a solution of imidazole-2-carboxylic acid (101.3 mg,0.43mmol,1.0 eq.) in DMF (3 mL) was added HATU (250.1 mg,0.65mmol,1.5 eq.) and DIPEA (113.3 mg,0.87mmol,0.15mL,2.0 eq.). The mixture was stirred at 25℃for 1h. TLC (dichloromethane: methanol=10/1, uv). The reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=100/1 to 10/1) to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 7-dichloro-1H-benzo [ d ] as a white solid]Imidazole-2-carboxamide (0.2 g,0.39mmol,89% yield).

To N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 7-dichloro-1H-benzo [ d ] ]To a solution of imidazole-2-carboxamide (100.00 mg,0.19mmol,1 eq.) in DCM (3.0 mL) was added the Buerger's reagent (140.3 mg,0.58mmol,3.0 eq.). The mixture was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80X 40mm 3um; flow)The phases are as follows: [ Water (0.05% NH) ₃ H ₂ O+10mM NH4HCO3)-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified for 7.8min, 20% -50% to give compound 637 (22.11 mg,22% yield) as a white solid.

LCMS：Rt＝0.824min；C ₂₂ H ₂₄ Cl ₂ N ₆ O ₃ MS calculated: 490.13; MS experimental values: 491.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.30(s,2H),5.22-5.09(m,1H),4.60(t,J＝7.1Hz,1H),3.27-3.19(m,2H),2.56-2.37(m,2H),2.06-1.88(m,3H),1.87-1.79(m,1H),1.73(td,J＝7.2,14.0Hz,2H),1.60-1.44(m,1H),0.96-0.75(m,1H),0.54(d,J＝6.9Hz,2H),0.21(dd,J＝4.8,10.4Hz,2H)。

EXAMPLE 185 Synthesis of viral protease inhibitor Compound 639

Step 1: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-1H-indole-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (70 mg,0.23mmol,1 eq.) in DMF (1 mL) was added HATU (89.8 mg,0.23mmol,1 eq.), 7-chloro-1H-indole-2-carboxylic acid (50.8 mg,0.25mmol,1.1 eq.) and DIEA (61.0 mg,0.47mmol,82.2uL,2 eq.). The mixture was stirred at 25℃for 16h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:16% -46%,9.5 min) of the purified residue. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2- [ ] as a white solidOxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (67 mg,0.12mmol,53.8% yield, 90% purity).

Step 2: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (60 mg,0.12mmol,1 eq.) in DCM (1 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (90.5 mg,0.37mmol,3 eq.). The mixture was stirred at 25℃for 6h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:26% -56%,7.8 min) of the purified residue. Obtaining 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (12.34 mg,21.3% yield, 100% purity).

LCMS：Rt＝2.130min；C ₂₃ H ₂₆ ClN ₅ O ₃ MS calculated: 455.94; MS experimental values: 456.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.58(d,J＝8.0Hz,1H),7.32-7.21(m,2H),7.07(t,J＝7.8Hz,1H),5.14(dd,J＝6.0,10.0Hz,1H),4.57(t,J＝7.4Hz,1H),3.28-3.16(m,2H),2.56-2.28(m,2H),2.05-1.88(m,3H),1.87-1.78(m,1H),1.77-1.61(m,2H),1.59-1.44(m,1H),0.92-0.80(m,1H),0.60-0.49(m,2H),0.26-0.14(m,2H)。

Example 185a Synthesis of viral protease inhibitor Compounds 639 and 639A

To (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (1.07 g,4.65mmol,1.1 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1 g,4.22mmol,1 eq., HCl) in DCM (10 mL) was added DMAP (1.55 g,12.67mmol,3 eq.), EDCI (1.62 g,8.45mmol,2 eq.) and the resulting solution was stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (30 mL) was diluted, extracted with ethyl acetate (30 mL. Times.3), and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By column chromatography (SiO ₂ The residue was purified with DCM/meoh=30/1 to 10/1 to give (2S) -2- [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.92mmol,68.97% yield, 100% purity). MS (ESI) m/z 412.3[ M+H ]] ⁺ 。

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A solution of tert-butyl carbamate (580 mg,1.46mmol,1 eq.) in HCl/MeOH (4M, 10.00mL,7.93 eq.) was stirred at 25℃for 1h. After completion, the solution was concentrated to give (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (380 mg, crude material). MS (ESI) m/z 297.2[ M+H ]] ⁺ 。

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (380 mg,1.28mmol,1 eq.) in DCM (3 mL) was added 7-chloro-1H-indole-2-carboxylic acid (275.88 mg,1.41mmol,1.1 eq.), T3P (1.22 g,1.93mmol,1.14mL,50% purity, 1.5 eq.) and DIEA (331.44 mg,2.56mmol,446.68uL,2 eq.) and the mixture stirred at 25℃for 2H. After completion, the solution is treated with H ₂ O (20 mL) was diluted, extracted with DCM (30 mL. Times.3) and the combined organic phases were taken up in Na ₂ SO ₄ Drying, filtration and concentration gave the crude material. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (350 mg,738.47 mol,57.59% yield, 100% purity). MS (ESI) M/z474.3[ M+H ]] ⁺ 。

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (350 mg,738.47 mmole, 1 eq.) in DCM (4 mL) was added the Buerger's reagent (527.94 mg,2.22mmol,3 eq.) and the solution was stirred at 25℃for 6H. After completion, drying using a blow dryerThe DCM was removed to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mm X10 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH4HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give the desired compound as a white solid, which was purified by SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:33% -33%,8 min) to give 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (250 mg,530.89umol,74.25% yield, 96.82% purity). MS (ESI) m/z456.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.58(d,J＝7.9Hz,1H),7.35-7.20(m,2H),7.06(t,J＝7.8Hz,1H),5.22-5.05(m,1H),4.57(t,J＝7.5Hz,1H),3.27-3.14(m,2H),2.61-2.34(m,2H),2.09-1.61(m,6H),1.59-1.43(m,1H),0.98-0.76(m,1H),0.55(dd,J＝1.3,8.2Hz,2H),0.31-0.09(m,2H)。

Obtaining 7-chloro-N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (45 mg,98.70umol,13.37% yield, 100% purity). MS (ESI) M/z456.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.59(dd,J＝0.9,7.9Hz,1H),7.32-7.21(m,2H),7.07(t,J＝7.8Hz,1H),5.12-5.02(m,1H),4.59(dd,J＝6.4,7.9Hz,1H),3.21(dd,J＝4.6,7.7Hz,2H),2.44-2.23(m,2H),2.09-1.62(m,6H),1.60-1.47(m,1H),0.94-0.78(m,1H),0.62-0.43(m,2H),0.27-0.11(m,2H)。

EXAMPLE 186 Synthesis of viral protease inhibitor Compounds 643

Will T ₃ P (2.69 g,4.22mmol,2.51mL,50% purity, 2 eq.) was added to (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,2.11mmol,1 eq., HCl), (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (570.0 mg,2.32mmol,1.1 eq.) and TEA (855.0 mg,8.45mmol,1.18mL,4 eq.) in DMF (5 mL). The mixture was then stirred at 70℃for 16h. TLC (petroleum ether: ethyl acetate=0:1/PMA). H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography20g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (436 mg,0.99mmol,47.2% yield, 97.9% purity) and confirmed by LC-MS, SFC and HNMR.

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (300 mg,0.70mmol,1 eq.) in HCl/dioxane (4M, 175.42uL,1 eq.) was stirred at 25℃for 2h. The reaction mixture was filtered to give ethyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (250 mg, crude material, HCl) as a white solid.

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (310 mg,0.85mmol,1 eq. HCl), 4-methoxy-1H-indole-2-carboxylic acid (179.1 mg,0.93mmol,1.1 eq.), HATU (647.8 mg,1.70mmol,2 eq.) and DIPEA (440.4 mg,3.41mmol,0.60mL,4 eq.) in DCM (4 mL) was stirred at 25℃for 2H. TLC (PE: EA=0:1/UV 254 nm). H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow oil]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (4571 mg,0.68mmol,80.1% yield, 75.8% purity).

To (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH was added to a mixture of methyl propionate (400 mg,0.79mmol,1 eq.) in the form of a mixture of NH and NH ₃ (7M, 11.42mL,100 eq.) and then stirring the resulting mixture at 80℃for 16h. TLC (DCM: meOH=10:1/UV 254 nm). The reaction mixture was concentrated in vacuo and purified by flash silica gel chromatography @12g/>Silica flash column, eluent: 0-50% ethyl acetate/MeOH at 30 mL/min). Obtaining N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (295 mg,0.60mmol,75.1% yield, 98.9% purity).

Methoxycarbonyl- (triethylammonium) sulfonyl-imide (284.6 mg,1.19mmol,2 eq.) was added to N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) at 25 ℃ ]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]A mixture of 4-methoxy-1H-indole-2-carboxamide (290 mg,0.59mmol,1 eq.) in DCM (3 mL). Subsequently, the mixture was stirred at 25℃for 16h. Next, methoxycarbonyl- (triethylammonium) sulfonyl-imide (142.3 mg,0.59mmol,1 eq.) was added to the mixture and the mixture was stirred at 25℃for an additional 16h. H for the reaction mixture ₂ O (10 mL) was diluted and the mixture was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column: welch Xtimate C18X 25mm X5 um; mobile phase: [ water (0.05% ammonium hydroxide, v/v) -MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -85%,9.5 min) of the purified residue. Obtaining N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (28.1 mg,59.3umol,9.9% yield, 98.7% purity).

LCMS：Rt＝0.832min；C ₂₅ H ₃₃ N ₅ O ₄ MS calculated: 467.25, ms experimental value: 468.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.26-7.22(m,1H),7.18-7.12(m,1H),7.05-7.00(m,1H),6.51(d,J＝7.5Hz,1H),5.08(dd,J＝6.3,9.8Hz,1H),4.67-4.63(m,1H),3.93(s,3H),3.21-3.15(m,2H),2.47-2.38(m,2H),1.98-1.72(m,6H),1.70-1.58(m,1H),1.54-1.43(m,1H),1.02(s,8H),1.04-1.01(m,2H)。

EXAMPLE 187 Synthesis of viral protease inhibitor Compounds 653

/>

To N- [ (1S) -1- [ [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]Carbamoyl group]-3-methyl-butyl]To a mixture of (4-methoxy-1H-indole-2-carboxamide (1 g,1.81mmol,80% purity, 1 eq.) in EtOH (20 mL) was added 2-glycine (271.74 mg,3.62mmol,20.52uL,2 eq.), znCl ₂ (1M, 18.10uL,0.01 eq.). The mixture was stirred at 25 ℃ for 30min, and then TMSCN (359.14 mg,3.62mmol,452.89ul,2 eq.) was added. The mixture was stirred at 40℃for 6h. After the reaction was completed, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. Preparative HPLC by HCl (column Phenomenex luna C, 18, 80, 40mm, 3um; mobile phase: [ water (0.04% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -45%,7 min) to obtain a mixture. By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,10 min) to give the compound 2- [ [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Amino group]Acetic acid (125 mg,235.87umol,13.03% yield, 99.363% purity) and 2- [ [ (2S) -1-cyano-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group ]-4-methyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Amino group]Acetic acid (205 mg,373.82umol,20.65% yield, 96.023% purity). MS (ESI) m/z 527.3[ M+H ]] ⁺ 。

isomer 2: ¹ H NMR(400MHz,DMSO-d6)δ＝11.57(d,J＝2.0Hz,1H),8.39(d,J＝7.8Hz,1H),8.20(d,J＝9.5Hz,1H),7.54(s,1H),7.37(d,J＝1.6Hz,1H),7.16-6.94(m,2H),6.50(d,J＝7.6Hz,1H),4.53-4.36(m,1H),4.18-4.01(m,1H),3.88(s,3H),3.77(d,J＝8.8Hz,1H),3.43-3.33(m,2H),3.15-2.96(m,2H),2.38-2.25(m,1H),2.08-2.01(m,1H),1.91-1.47(m,6H),0.91(dd,J＝6.4,14.8Hz,6H)。

example 188 Synthesis of viral protease inhibitor Compounds 655

/>

To a solution of (2 s,4 r) -1-tert-butoxycarbonyl-4-hydroxy-pyrrolidine-2-carboxylic acid (5 g,21.62mmol,1 eq.) in THF (75 mL) was added 2-tert-butyl-1, 3-diisopropyl-isourea (6.50 g,32.43mmol,1.5 eq.) at 25 ℃ and the solution was then stirred at 60 ℃ for 2.5h. To the mixture was added 2-tert-butyl-1, 3-diisopropyl-isourea (6.50 g,32.43mmol,1.5 eq.) and then stirred at 60 ℃ for 14h. After completion, the reaction mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give di-tert-butyl (2 s,4 r) -4-hydroxypyrrolidine-1, 2-dicarboxylate (4.3 g,14.22mmol,65.75% yield, 95% purity) as a colorless oil. MS (ESI) m/z 288.2[ M+H ]] ⁺

Step 2: (2S, 4S) -4-bromopyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

CBr is added to a solution of di-tert-butyl (2S, 4R) -4-hydroxypyrrolidine-1, 2-dicarboxylate (4 g,13.92mmol,1 eq.) in DCM (40 mL) at 25 ℃ ₄ (14.08 g,42.46mmol,3.05 eq.). The mixture was cooled to 0 ℃ and PPh was carefully added ₃ (11.32 g,43.15mmol,3.1 eq.). The reaction was stirred at 25℃for 15h. After completion, ethanol (4 mL) was added and the solution was stirred for 2h. MTBE (40 mL) was added dropwise to precipitate the phosphine oxide, which was filtered off and taken up inThe filter cake was washed with DCM (30 ml x 2). The filtrate was concentrated under reduced pressure to give a brown oil. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=100:0 to 10:1) to give (2 s,4 s) -4-bromopyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester (1.5 g,4.07mmol,29.23% yield, 95% purity) as a pale yellow oil.

A mixture of phenylsulfanyl copper (1.58 g,9.14mmol,6.4 eq.) in anhydrous THF (30 mL) was cooled to-70℃and treated by careful addition of t-BuLi (1.3M, 7.03mL,6.4 eq.). The yellow mixture was stirred for 30min, and a solution of pre-cooled (-20 ℃) di-tert-butyl (2S, 4S) -4-bromopyrrolidine-1, 2-carboxylate (500 mg,1.43mmol,1 eq.) in anhydrous THF (5 mL) was added. The reaction was stirred at-70 ℃ for 5h and then at N ₂ The temperature is raised to 25 ℃ and maintained for 15 hours. After completion, by pouring saturated NH ₄ Aqueous Cl (30 mL) was used to quench the reaction. The aqueous mixture was vigorously stirred for 30min. The solid was filtered off and the phases were separated. The aqueous phase was extracted with MTBE (10 ml x 3) and the combined organic phases were extracted with saturated NaHCO ₃ Aqueous (10 mL) and brine (10 mL) washed with Na ₂ SO ₄ Drying and concentrating under reduced pressure to obtain crude material. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=100:0 to 10:1) to give di-tert-butyl (2 s,4 s) -4- (tert-butyl) pyrrolidine-1, 2-dicarboxylate (290 mg,797.05umol,55.83% yield, 90% purity) as an off-white solid.

Step 4: (2S, 4S) -4- (tert-butyl) pyrrolidine-2-carboxylic acid

To (2S, 4S) -4-tert-butylpyrroleAlkane-2-carboxylic acid (158 mg, 760.72. Mu. Mol,1 eq. HCl) in THF (1 mL) and H ₂ K was added to the mixture in O (1 mL) ₂ CO ₃ (315.41 mg,2.28mmol,3 eq.) and Boc ₂ O (199.23 mg, 912.87. Mu.L, 209.72. Mu.L, 1.2 eq.). At N ₂ The reaction was stirred at 25℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give (2 s,4 s) -1- (tert-butoxycarbonyl) -4- (tert-butyl) pyrrolidine-2-carboxylic acid (650 mg, crude substance) as a yellow solid.

To a solution of (2S, 4S) -1- (tert-butoxycarbonyl) -4- (tert-butyl) pyrrolidine-2-carboxylic acid (630 mg,696.51umol,30% purity, 1 eq) in DCM (6 mL) and DMF (3 mL) was added TEA (422.88 mg,4.18mmol,581.68uL,6 eq), (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl)]Methyl propionate (186.11 mg,835.82umol,1.2 eq., HCl). After adding T3P (1.33 g,2.09mmol,1.24mL,50% purity, 3 eq.) at 0deg.C, the mixture was stirred at 25deg.C for 1h. After completion, the mixture was quenched with water (10.0 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10.0 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether Ethyl acetate=10:1 to 0:1) the residue was purified to give (2S, 4S) -4- (tert-butyl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester (240 mg,546.02umol,78.39% yield) as a yellow solid. MS (ESI) m/z 440.3[ M+H ]] ⁺ 。

A solution of (2S, 4S) -4- (tert-butyl) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester (230 mg,523.27umol,1 eq) in HCl/MeOH (4M, 2.3mL,17.58 eq) was stirred at 25℃for 1h. After completion ofThe reaction mixture was concentrated under reduced pressure to give methyl (S) -2- ((2S, 4S) -4- (tert-butyl) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (196 mg, crude material, HCl) as a pale yellow solid. MS (ESI) M/z340.2[ M+H] ⁺ 。

To a solution of methyl (S) -2- ((2S, 4S) -4- (tert-butyl) pyrrolidin-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (196 mg,521.43umol,1 eq, HCl) in DCM (2 mL) and DMF (1 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (99.69 mg,521.43umol,1 eq), DMAP (127.41 mg,1.04mmol,2 eq) and then EDCI (199.92 mg,1.04mmol,2 eq) at 0 ℃. The mixture was then stirred at 25℃for 1h. After completion, the mixture was quenched with water (10.0 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10.0 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Dichloromethane: methanol = 10:1 to 4:1) the residue was purified to give methyl (S) -2- ((2S, 4S) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (250 mg,414.56umol,79.50% yield, 85% purity) as a yellow solid. MS (ESI) m/z 513.3[ M+H ]] ⁺ 。

Methyl (S) -2- ((2S, 4S) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (235 mg,389.68umol,85% purity, 1 eq.) in NH ₃ A solution in MeOH (7M, 5 mL) was stirred at 40℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butyl) -1- (4-methoxy) o-f-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (193 mg, crude). MS (ESI) m/z 498.3[ M+H ] ] ⁺ 。

To a solution of (2S, 4S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4- (tert-butyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (193 mg,329.69umol,85% purity, 1 eq) in DCM (3 mL) was added the bergs reagent (235.71 mg,989.08umol,3 eq) and then the reaction stirred at 25 ℃ for 4H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified 30% -55%,10 min) to give (2S, 4S) -4- (tert-butyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (59.58 mg,124.24umol,37.68% yield, 100% purity) as a white solid. MS (ESI) m/z 480.2[ M+H ]] ⁺ 。

EXAMPLE 189 Synthesis of viral protease inhibitor Compounds 659

To a solution of (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (5 g,20.38mmol,1 eq.) in THF (100 mL) was slowly added drop wise BH at 0deg.C ₃ -Me ₂ S (10M, 4.08mL,2.0 eq.) and then stirring the mixture at 20deg.C for 15h. The reaction mixture was added to MeOH (40 mL) and stirred for 20min. After concentrating the mixture, the residue was taken up in NaHCO ₃ Aqueous solution (150 mL) was diluted and extracted with DCM (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 1:1) the residue was purified to give N- [ (1S) -1- (hydroxymethyl) -3, 3-dimethyl-butyl as a colorless oil]Tert-butyl carbamate (2.5 g,10.81mmol,53.02% yield).

Step 2: (S) - (4, 4-dimethyl-1-oxopent-2-yl) carbamic acid tert-butyl ester

To N- [ (1S) -1- (hydroxymethyl) -3, 3-dimethyl-butyl]To a solution of tert-butyl carbamate (2.4 g,10.37mmol,1 eq.) in DCM (40 mL) was added Dess-Martin periodate (Dess-Martin periodinane) (5.72 g,13.49mmol,4.18mL,1.3 eq.) and stirred at 0deg.C for 1h, and the mixture was then warmed to 20deg.C and stirred for 1h. By adding 60mL of H at 0deg.C ₂ O quench the reaction mixture and then drop wise addition of NaHCO at 0deg.C ₃ The pH of the mixture was adjusted to about 8 and extracted with EtOAc (40 ml x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1 to 1:1) the residue was purified to give N- [ (1S) -1-formyl-3, 3-dimethyl-butyl as a colorless oil]Tert-butyl carbamate (1.6 g,6.98mmol,67.25% yield).

To N- [ (1S) -1-formyl-3, 3-dimethyl-butyl]Carbamic acid tert-butyl ester (0.8 g,3.49mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (1.17 g,5.23mmol,1.5 eq. HCl) in DCE (20 mL) was added Et ₃ N (529.52 mg,5.23mmol,728.36uL,1.5 eq.) and NaBH (OAc) ₃ (2.22 g,10.47mmol,3 eq.). The reaction was stirred at 20℃for 2h. By addition of NaHCO at 0deg.C ₃ The reaction mixture was quenched with aqueous solution (100 mL) and stirred for 0.5h, and then extracted with DCM (60 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=0:1 to 1:3) the residue was purified to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (450 mg,1.13mmol,32.29% yield). MS (ESI) m/z 400.3[ M+H ]] ⁺

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentyl ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (168 mg, 50)To a solution of 0.20 mol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (95.63 mg,500.20 mol,1 eq) in DMF (1 mL) was added DMAP (183.32 mg,1.50mmol,3.0 eq), EDCI (191.78 mg,1.00mmol,2 eq) and DCM (3 mL). The mixture was stirred at 20℃for 2h. By adding 40mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (20 ml x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) to give (2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow oil]-4, 4-dimethyl-pentyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (150 mg,301.54 mol,60.28% yield, 95% purity). MS (ESI) m/z 473.2[ M+H ]] ⁺

(2S) -2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group ]-4, 4-dimethyl-pentyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (130 mg,275.09umol,1 eq.) in NH ₃ A solution in MeOH (7M, 15mL,381.70 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with ethyl acetate meoh=50:3 to give the product N- [ (1S) -1- [ [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl) as a yellow solid]Methyl group]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (60 mg,131.13umol,47.67% yield). MS (ESI) M/z458.3[ M+H] ⁺

To N- [ (1S) -1- [ [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl [ (2-amino-3-carbonyl)]Methyl group]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxyTo a solution of the base-1H-indole-2-carboxamide (50 mg,109.27umol,1 eq.) in EtOAc (2 mL) was added dropwise T ₃ P (2.14 g,3.36mmol,2mL,50% purity, 30.77 eq.) and then stirring the resulting mixture at 65℃for 12h. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,8 min) and purified the residue by SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -25%,20 min) to give N- [ (1S) -1- [ [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]Methyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (4.4 mg,9.92umol,29.07% yield, 99.1% purity). MS (ESI) m/z 440.2[ M+H ]] ⁺ 。

Example 190 Synthesis of viral protease inhibitor Compounds 667

Step 1: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4-fluoro-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

To a solution of compound (S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanoic acid (300.0 mg,1.20mmol,1.0 eq.) and compound (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (313.3 mg,1.32mmol,1.1 eq., HCl) in DMF (3 mL) was added T ₃ P (1.53 g,2.41mmol,1.43mL,50% purity, 2.0 eq.) and TEA (487.1 mg,4.81mmol,0.67mL,4.0 eq.). The mixture was stirred at 80℃for 16h. TLC (petroleum ether/ethyl acetate=0/1, pma). At the position of The mixture was concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12gSilica flash column, eluent: the residue was purified with 0-1% MeOH in DCM at 25mL/min to give methyl (S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (620 mg,1.15mmol,95.5% yield, 80% purity) as a yellow solid.

LCMS：Rt＝0.773min；C ₂₀ H ₃₄ FN ₃ O ₆ MS calculated: 431.24; MS experimental values: 432.2[ M+H ] ⁺ ]。

Step 2: (2S) -2- [ [ (2S) -2-amino-4-fluoro-4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

A mixture of the compound methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (520 mg,1.21mmol,1 eq.) in HCl/EtOAc (4 mL) was stirred at 25deg.C for 0.5h. The mixture was concentrated under reduced pressure to give the compound methyl (S) -2- ((S) -2-amino-4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (550 mg, crude material, HCl, yellow oil) and used in the next step.

Step 3: (2S) -2- [ [ (2S) -4-fluoro-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -4-methyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To a solution of compound 4-methoxy-1H-indole-2-carboxylic acid (200 mg,1.05mmol,1 eq.) in DCM (5 mL) was added HATU (477.3 mg,1.26mmol,1.2 eq.) and DIEA (540.8 mg,4.18mmol,0.73mL,4 eq.). The mixture was stirred at 25℃for 0.5h. To the mixture was added the compound methyl (S) -2- ((S) -2-amino-4-fluoro-4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (461.8 mg,1.26mmol,1.2 eq, HCl). The mixture was stirred at 25℃for 16h. TLC (DCM/meoh=10/1, uv). H for the reaction mixture ₂ O(15mL) diluted and extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-100% ethyl acetate/petroleum ether gradient, at 30 mL/min) to give methyl (S) -2- ((S) -4-fluoro-2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (480 mg,83.9% yield) as a yellow solid.

LCMS：Rt＝0.794min；C ₂₅ H ₃₃ FN ₄ O ₆ MS calculated: 504.24; MS experimental values: 505.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CDCl ₃ )δ9.56-9.81(m,1H),8.24(br s,1H),7.23-7.06(m,3H),7.01(d,J＝8.28Hz,1H),6.49(d,J＝7.78Hz,1H),6.17(br s,1H),4.95-4.82(m,1H),4.60-4.51(m,1H),3.94(s,3H),3.80-3.60(m,5H),3.16(br d,J＝7.28Hz,2H),3.00-2.77(m,1H),1.98(br d,J＝6.02Hz,2H),1.92-1.83(m,2H),1.77(br s,2H),1.51-1.44(m,6H)。

Step 4: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3-fluoro-3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

In a 30mL sealed tube, the compound methyl (S) -2- ((S) -4-fluoro-2- (4-methoxy-1H-indole-2-carboxamido) -4-methylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propionate (1 g,1.98mmol,1 eq.) was reacted in NH ₃ The solution in (7M in MeOH, 14.16mL,50 eq.) was stirred at 80℃for 16h. TLC (DCM/meoh=10/1, uv). The reaction mixture was concentrated under reduced pressure to give a residue. Through flash silica gel chromatography20g/>Silica flash column, eluent: the residue was purified with 0-15% MeOH in ethyl acetate at 30mL/min to give the compound N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (370 mg,0.74mmol,37.2% yield, 97.6% purity) as a yellow solid.

LCMS：Rt＝0.743min；C ₂₄ H ₃₂ FN ₅ O ₅ MS calculated: 489.24; MS experimental values: 490.2[ M+H ] ⁺ ]。

Step 5: n- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3-fluoro-3-methyl-butyl ] -4-methoxy-1H-indole-2-carboxamide

To a solution of compound N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4-fluoro-4-methyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide (350 mg,0.71mmol,1 eq) in DCM (6 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (170.4 mg,0.71mmol,1 eq) at 0 ℃. The mixture was stirred at 25℃for 0.5h. Methoxycarbonyl- (triethylammonium) sulfonyl-imide (170.4 mg,0.71mmol,1 eq.) was added to the mixture at 0deg.C. The mixture was stirred at 25℃for 0.5h. Methoxycarbonyl- (triethylammonium) sulfonyl-imide (170.4 mg,0.71mmol,1 eq.) was added to the mixture at 0deg.C. The mixture was stirred at 25℃for 16h. H for mixture ₂ O (20 mL) was diluted and extracted with ethyl acetate (40 mL x 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was checked by LCMS. By preparative HPLC (column: phenomenex Gemini-NX 80. Times.40 mm. Times.3 um; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:28% -58%,7.8 min) to give N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4-fluoro-4-methyl-1-oxo) amino) as a white solidPentan-2-yl) -4-methoxy-1H-indole-2-carboxamide (95 mg,28.1% yield).

LCMS：Rt＝0.780min；C ₂₄ H ₃₀ FN ₅ O ₄ MS calculated: 471.23; MS experimental values: 472.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ7.24-7.20(m,1H),7.18-7.11(m,1H),7.07-7.01(m,1H),6.51(d,J＝7.78Hz,1H),5.13-5.01(m,1H),4.81-4.71(m,1H),3.93(s,3H),3.18(dd,J＝7.40,5.14Hz,2H),2.49-2.34(m,2H),2.32-2.11(m,2H),2.00-1.87(m,2H),1.83-1.73(m,1H),1.72-1.60(m,1H),1.54-1.37(m,7H)。

EXAMPLE 191 Synthesis of viral protease inhibitor Compound 681

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,2.11mmol,1.1 eq, HCl), 2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid (684.45 mg,1.92mmol,1 eq.) in DMF (15 mL) was added DIPEA (744.57 mg,5.76mmol,1.00mL,3 eq.) and HATU (730.19 mg,1.92mmol,1 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (80 mL) to quench the two batches and extract with ethyl acetate (40 mL x 3). The combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.35 g, crude material). MS (ESI) m/z 539.3[ M+H ]] ⁺ 。

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (650 mg,1.21mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 3.45mL,20 eq.) was stirred at 65℃for 17h. After completion, the two batches of the reaction mixture were concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a colorless oil]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (1.22 g, crude). MS (ESI) m/z 524.3[ M+H ] ] ⁺ 。

Step 3:

n- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a solution of decane-3-carboxamide (1.22 g,2.33mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (1.39 g,5.82mmol,2.5 eq.). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (3 mL) to quench the reaction mixture and then concentrate under reduced pressure to give a residue. By preparative HPLC (column: agela DuraShell C, 250X 70mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:43% -63%,20 min) to give the desired compound (490 mg) as a white solid, which was purified by SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:58% -58%,10 min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide isomer 1 (201.77 mg,394.36umol,16.93% yield, 98.820% purity). MS (ESI) M/z506.3[ M+H ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.26(br s,1H)8.50-8.85(m,1H)7.23(br s,1H)7.00-7.16(m,2H)6.89(br s,1H)6.52(br d,J＝7.46Hz,1H)4.86-5.06(m,1H)4.48-4.79(m,1H)3.80-3.98(m,4H)3.59(br d,J＝4.65Hz,1H)3.09(br s,2H)2.15-2.31(m,3H)1.73-2.01(m,2H)1.67(br dd,J＝12.17,8.62Hz,2H)1.33-1.61(m,12H)。

Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5 ]]Decane-3-carboxamide isomer 2 (200.95 mg,394.35umol,16.93% yield, 99.222% purity). MS (ESI) m/z 506.3[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.27(br s,1H)8.61(br d,J＝1.22Hz,1H)7.02-7.26(m,3H)6.91(br s,1H)6.53(d,J＝7.46Hz,1H)4.91-5.06(m,1H)4.62(br s,1H)3.82-3.98(m,4H)3.52-3.75(m,1H)3.09(br s,2H)2.09-2.28(m,3H)1.63-1.92(m,4H)1.33-1.62(m,12H)。

EXAMPLE 192 Synthesis of viral protease inhibitor Compounds 711

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (900 mg,1.81mmol,80% purity, 1 eq.) in HCl/MeOH (4M, 12.00mL,26.50 eq.) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a white oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (600 mg, crude material, HCl). MS (ESI) m/z 298.1[ M+H ]] ⁺ 。

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (600 mg,1.80mmol,1 eq. HCl) in DCM (7 mL) and DMF (0.5 mL)To the mixture of (1) was added 4,5,6, 7-tetrahydro-1H-indole-2-carboxylic acid (415.68 mg,2.52mmol,1.4 eq.), TEA (1.09 g,10.78mmol,1.50mL,6 eq.) and T3P (1.72 g,2.70mmol,1.60mL,50% purity, 1.5 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) and TLC (SiO ₂ The residue was purified with DCM: meoh=10:1) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- (4, 5,6, 7-tetrahydro-1H-indole-2-carbonylamino) propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (350 mg,787.36umol,43.80% yield). MS (ESI) m/z 445.3[ M+H ]] ⁺ 。

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4,5,6, 7-tetrahydro-1H-indole-2-carboxamide (290 mg,675.19umol,1 eq.) in T3P (3 mL,50% purity) and acetic acidThe mixture in ethyl ester (3 mL) was stirred at 40℃for 16h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters X bridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4,5,6, 7-tetrahydro-1H-indole-2-carboxamide (61.92 mg,150.48umol,22.29% yield, 100% purity). MS (ESI) m/z 412.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝10.96(br s,1H),9.00-8.77(m,1H),7.89-7.66(m,2H),6.60(br s,1H),5.04-4.81(m,1H),4.48-4.28(m,1H),3.24-3.04(m,2H),2.47-1.96(m,7H),1.81-1.61(m,7H),1.40(br dd,J＝6.6,13.1Hz,1H),0.74(br s,1H),0.38(br s,2H),0.22--0.03(m,2H)。

EXAMPLE 193 Synthesis of viral protease inhibitor Compounds 715

Step 1: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

To 4-chloro-1H-pyrrolo [3,2-c]To a solution of pyridine-2-carboxylic acid (110.5 mg,0.56mmol,1 eq.) in DMF (2 mL) was added HATU (256.6 mg, 0).67mmol,1.2 eq), DIEA (218.0 mg,1.69mmol,0.29ml,3 eq.) and (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]3-cyclopropyl-propionamide (200 mg,0.67mmol,1.2 eq). The mixture was stirred at 25℃for 16hr. LC-MS showed detection of the desired compound. TLC (DCM/meoh=10:1). H for the reaction mixture ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 ml x 3), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Through flash silica gel chromatography4g/>Silica flash column, eluent: the residue was purified with a gradient of 0-20% DCM/MeOH ether at 20mL/min to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (214 mg,76.2% yield).

Step 2: 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

At 0℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-1H-pyrrolo [3,2-c]To a solution of pyridine-2-carboxamide (214 mg,0.45mmol,1 eq.) in DCM (3 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (322.1 mg,1.35mmol,3 eq.). The mixture was stirred at 25℃for 16h. TLC (DCM: meoh=10:1). The reaction mixture was filtered and concentrated in vacuo. Through flash silica gel chromatography 12g/>Silica flash column, eluent: gradient of 0-10% petroleum ether/ethyl acetate at 25 mL/min) to give 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrolo [3,2-c]Pyridine-2-carboxamide (80 mg,36.5% yield).

LCMS：Rt＝1.356min；C ₂₂ H ₂₅ ClN ₆ O ₃ MS calculated: 456.17; MS experimental values: 457.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.75-8.65(m,1H),7.53-7.43(m,1H),7.41-7.31(m,1H),5.14(br d,J＝9.5Hz,1H),4.54(br t,J＝7.2Hz,1H),3.24(br s,2H),2.56-2.41(m,2H),2.02-1.87(m,2H),1.86-1.61(m,4H),1.59-1.45(m,1H),0.85(br s,1H),0.54(br d,J＝8.0Hz,2H),0.23-0.15(m,2H)。

EXAMPLE 194 Synthesis of viral protease inhibitor Compound 639

7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-indole-2-carboxamide

Step 1:

(S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (1.45 g,6.34mmol,1.5 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1 g,4.22mmol,1 eq., HCl), DMAP (1.55 g,12.67mmol,3 eq.) in DCM (10 mL) was added EDCI (2.43 g,12.67mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 50mL of H at 20 ℃ ₂ O, and then extracted with DCM (50 ml×3). The combined organic layers were washed with brine (50 ml x 2), dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 1/2 to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.5 g,3.10mmol,73.34% yield, 85% purity). MS (ESI) m/z 412.2[ M+H ]] ⁺ 。

(2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (29.06 g,126.75mmol,1.5 eq.) 2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (20 g,84.50mmol,1 eq., HCl) and DMAP (25.81 g,211.24mmol,2.5 eq.) in DCM (200 mL) was added followed by EDCI (32.40 g,168.99mmol,2.0 eq.). The mixture was stirred at 20deg.C for 2hr. The reaction mixture was quenched by addition of 300mL h2o at 0 ℃ and extracted with 300mL DCM (100 mL x 3). The combined organic layers were washed with 100ml 0.5n HCl and brine (100 ml x 2), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 1/2) the residue was purified. Obtaining (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl group as a white solid ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (23 g,54.83mmol,64.89% yield, 98.1% purity).

Step 2:

(S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (1.5 g,3.65mmol,1 eq.) in HCl/MeOH (4M, 15.00mL,16.46 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.27 g, crude material, HCl). MS (ESI) m/z 312.2[ M+H ]] ⁺ 。

Step 3:

(S) -2- ((S) -2- (7-chloro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

At 0 ℃, to (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.27 g,3.65mmol,1 eq., HCl), 7-chloro-1H-indole-2-carboxylic acid (714.17 mg,3.65mmol,1 eq.) and DMAP (1.34 g,10.95mmol,3 eq.) in DCM (13 mL) was added EDCI (1.75 g,9.13mmol,2.5 eq.) and the mixture stirred at 20deg.C for 2H. After completion, the reaction mixture was poured into H at 20 ℃ ₂ O (20 mL) and then extracted with DCM (25 mL x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 1/0 to give (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.3 g,2.53mmol,69.18% yield, 95% purity). MS (ESI) m/z 489.2[ M+H ]] ⁺ 。

Step 4:

n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.3 g,2.66mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 26mL,68.45 eq.) was stirred at 65℃for 16h. The reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-7-chloro-1H-indole-2-carboxamide (1.26 g, crude). MS (ESI) m/z 474.2[ M+H ]] ⁺ 。

Step 5:

To N- [ (1)S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (1.26 g,2.66mmol,1 eq.) in DCM (13 mL) was added the Buerger's reagent (1.58 g,6.65mmol,2.5 eq.). The mixture was stirred at 25℃for 7h. After completion, the reaction mixture was concentrated with N2 to remove the solvent. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 0/1) to give 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (950 mg, crude). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。

Example 195 Synthesis of viral protease inhibitor Compounds 717

At N ₂ In the atmosphere, N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of benzyl carbamate (600 mg,1.39mmol,1 eq.) in THF (1 mL) was added Pd/C. Degassing the suspension and using H ₂ Purging 3 times. The mixture is put in H ₂ (15 Psi or atmospheric pressure) was stirred at 25℃for 2h. Pd/C was filtered and the reaction concentrated under reduced pressure to give a residue. The compound (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl was obtained as a colorless oil]Methyl group]Ethyl group]-3-cyclopropyl-propionamide (400 mg, crude material).

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-chloro-1H-pyrrolo [2,3-c ] pyridine-2-carboxamide

To (2S) -2-amino-N- [ (1S) -2-amino-2-oxo-o-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 3-cyclopropyl-propionamide (250 mg,0.84mmol,1 eq.) in DMF (1 mL) was added HATU (320.7 mg,0.84mmol,1 eq.) 5-chloro-1H-pyrrolo [2, 3-c)]Pyridine-2-carboxylic acid (182.4 mg,0.92mmol,1.1 eq.) and DIEA (218.0 mg,1.69mmol,0.29mL,2 eq.) were stirred at 25℃for 16hr. TLC (DCM/MeOH=10:1, I) ₂ ). H for the reaction mixture ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-7% DCM/MeOH at 35 mL/min). Obtaining N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-chloro-1H-pyrrolo [2,3-c]Pyridine-2-carboxamide (260 mg,64.3% yield, 99.2% purity).

Step 3: 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-pyrrolo [2,3-c ] pyridine-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-chloro-1H-pyrrolo [2,3-c]To a solution of pyridine-2-carboxamide (100 mg,0.21mmol,1 eq.) in DCM (1 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (150.5 mg,0.63mmol,3 eq.). The mixture was stirred at 25℃for 24h. H for the reaction mixture ₂ O (30 mL) was diluted and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Gemini-NX 80X 40mm 3um; flow)The phases are as follows: [ Water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,9.5 min). Obtaining 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrolo [2,3-c]Pyridine-2-carboxamide (94 mg,96.7% yield, 99% purity).

LCMS：Rt＝0.754min；C ₂₂ H ₂₅ ClN ₆ O ₃ MS calculated: 456.93; MS experimental values: 457.1[ M+H ] ⁺ ]。

¹ H NMR(400MHz,CD ₃ OD)δ8.58(s,1H),7.72-7.65(m,1H),7.20(s,1H),5.13(dd,J＝6.1,10.2Hz,1H),4.58-4.52(m,1H),3.28-3.16(m,2H),2.59-2.39(m,2H),2.07-1.87(m,3H),1.87-1.79(m,1H),1.78-1.62(m,2H),1.60-1.44(m,1H),0.91-0.78(m,1H),0.58-0.47(m,2H),0.26-0.13(m,2H)。

EXAMPLE 196 Synthesis of the viral protease inhibitor Compound (2S) -1-hydroxy-2- ((2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propane-1-sulfonic acid potassium salt

Step 1: [ (2S) -1-hydroxy-2- [ [ (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propyl ] sulfonyloxy potassium

At 45℃under N ₂ Downward (2S, 4S) -N- [ (1S) -1-formyl-2- [ (3S) -2-oxopyrrolidin-3-yl ]Ethyl group]To a mixture of 1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carboxamide (40 mg,79.59umol,1 eq.) in THF (0.5 mL) was added K-containing ₂ S ₂ O ₅ (8.67 mg,39.00umol,0.49 eq.) H ₂ O (0.1 mL). The mixture was stirred at 45 ℃ for 3h, and then at N ₂ The mixture was stirred at 25℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure and then wet-milled with THF (1 mL) at 25 ℃ for 1h to give an off-white colorSolid [ (2S) -1-hydroxy-2- [ [ (2S, 4S) -1- (4-methoxy-1H-indole-2-carbonyl) -4-phenyl-pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Propyl group]Potassium sulfonyloxy (14.09 mg,20.36umol,25.58% yield, 90% purity). MS (ESI) m/z 585.3[ M-36.8] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.68-11.40(m,1H),8.08-7.64(m,0.5H),7.52-7.43(m,0.5H),7.41-7.29(m,4H),7.28-7.19(m,1H),7.15-6.97(m,2H),6.96-6.76(m,1H),6.55-6.37(m,1H),5.48-5.27(m,1H),5.25-4.92(m,0.5H),4.80-4.60(m,0.5H),4.46-4.07(m,2H),4.02-3.93(m,0.5H),3.93-3.63(m,5H),3.62-3.36(m,1H),3.18-3.02(m,1H),2.94-2.69(m,0.5H),2.35-2.17(m,3H),2.10-1.87(m,1H),1.82-1.27(m,3H)。

EXAMPLE 197 Synthesis of (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionitrile hydrochloride

Step 1: (2S, 4R) -2- ((tert-Butoxycarbonyl) amino) -4- (cyanomethyl) glutarate dimethyl ester

at-78deg.C, at N ₂ To a solution of dimethyl 2- (t-butoxycarbonylamino) glutarate (10 g,36.32mmol,1 eq.) in THF (150 mL) was added LiHMDS (1M, 83.55mL,2.3 eq.). The mixture was stirred at-78℃for 1.5h. Next, 2-bromoacetonitrile (6.54 g,54.49mmol,3.63mL,1.5 eq.) was added dropwise to the reaction at-78deg.C. The mixture was stirred at-78℃for 2.5h. TLC (PE: EA=3:1, I) ₂ ). The reaction was completed, and pre-cooled methanol (15 mL) and glacial acetic acid (12 mL) were added in order to quench the reaction. The reaction was warmed to 25 ℃ and the solvent distilled off under reduced pressure. Through flash silica gel chromatography120g/>Silica flash column, eluent: gradient of 0-20% ethyl acetate/petroleum ether at 80 mL/min) purification residueAnd the remainder. Dimethyl (4R) -2- (t-butoxycarbonylamino) -4- (cyanomethyl) glutarate (15 g,47.72mmol,43.79% yield) was obtained as a yellow oil.

Step 2: (S) -2- ((tert-Butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoic acid methyl ester

(4R) -2- (tert-Butoxycarbonylamino) -4- (cyanomethyl) glutaric acid dimethyl ester (15 g,47.72mmol,1 eq.) was dissolved in MeOH (250 mL) and CoCl was added at 0deg.C ₂ .6H ₂ O (6.81 g,28.63mmol,0.6 eq.). After slow addition of NaBH4 (10.83 g,286.32mmol,6 eq.) in portions, the reaction was carried out at 25℃for 12h. TLC (petroleum ether/ethyl acetate=1:2, i ₂ ). After the reaction was completed, 100mL of saturated ammonium chloride solution was added to quench the reaction. The organic phase was collected by filtration, the solvent was distilled off under reduced pressure and extracted with EtOAc (150 ml x 3), and the organic phase was collected. The organic phase was washed with saturated brine (100 mL). The organic phase was treated with anhydrous Na ₂ SO ₄ Dried, the filtrate was collected by filtration and the solvent was evaporated under reduced pressure. Through flash silica gel chromatography40g/>Silica flash column, eluent: 0-80% ethyl acetate/petroleum ether gradient at 40 mL/min). Methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (7 g,24.45mmol,51.2% yield, 100% purity) was obtained as a white solid.

Step 3: (S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamic acid tert-butyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (2 g,6.99mmol,1 eq.) in MeOH (10 mL) was added NH ₃ (7M, 24.00mL,24.05 eq.). The mixture was stirred in a sealed tube at 60℃for 16h. The reaction mixture was concentrated under reduced pressure to give a residue. Obtaining the compound ((S) -1-amino-1) as a yellow solid-tert-butyl oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamate (1.8 g,6.63mmol,94.9% yield).

Step 4: (S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) carbamic acid tert-butyl ester

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ]Methyl group]Ethyl group]To a solution of tert-butyl carbamate (1 g,3.69mmol,1 eq.) in DCM (10 mL) was added the Buerger reagent (3.51 g,14.74mmol,4 eq.). At N ₂ The mixture was stirred at 25℃for 1h. Addition of H to the reaction mixture ₂ O (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through flash silica gel chromatography12g/>Silica flash column, eluent: 0-70% ethyl acetate/petroleum ether gradient at 30 mL/min). Obtaining the compound N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]Tert-butyl carbamate (900 mg,3.23mmol,87.7% yield, 91% purity).

Step 5: (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionitrile hydrochloride

To a solution of tert-butyl N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamate (600 mg,2.37mmol,1 eq.) in EtOAc (20 mL) was added HCl/EtOAc (4M, 4.00mL,6.75 eq.). The mixture was stirred at 25℃for 2h. LCMS showed starting material was consumed and the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The compound (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ] propionitrile (440 mg,2.32mmol,97.9% yield, HCl) was obtained as a white solid.

EXAMPLE 198 Synthesis of viral protease inhibitor Compounds 842

Step 1: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (5, 7-dichloro-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 5, 7-dichloro-1H-indole-2-carboxylic acid (1 g,4.35mmol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl [ (2S) -2 at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (1.35 g,3.89mmol,8.95e-1 eq., HCl) in DCM (24 mL) was added DMAP (1.59 g,13.04mmol,3 eq.) and EDCI (1.67 g,8.69mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and extracted with ethyl acetate (20 mL x 4). The combined organic layers were washed with brine (40 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (5, 7-dichloro-1H-indole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.29mmol,52.74% yield). MS (ESI) m/z 521.0[ M-H ] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5, 7-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (5, 7-dichloro-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.29mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 30 mL) was stirred at 55deg.C for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5, 7-dichloro-1H-indole-2-carboxamide (1 g,1.97mmol,85.79% yield). MS (ESI) m/z 508.2[ M+H ]] ⁺

Step 3:5, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 5, 7-dichloro-1H-indole-2-carboxamide (260 mg,475.61umol,93% purity, 1 eq.) in DCM (5 mL) was added the bergs reagent (226.68 mg,951.23umol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: phenomenex Gemini-NX C18 75X 30mM 3um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,8 min) to give 5, 7-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (100 mg,203.92umol,42.88% yield). MS (ESI) m/z 490.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.98(br s,1H),9.00(d,J＝7.9Hz,1H),8.77(d,J＝7.7Hz,1H),7.75(d,J＝1.8Hz,1H),7.54(br s,1H),7.41(d,J＝1.8Hz,1H),7.26(s,1H),5.07(q,J＝8.0Hz,1H),4.55-4.47(m,1H),3.16-3.02(m,2H),2.30-2.20(m,2H),1.90-1.65(m,4H),1.63-1.33(m,3H),0.87-0.75(m,1H),0.50-0.36(m,2H),0.24-0.07(m,2H)。

Example 199 Synthesis of viral protease inhibitor Compounds 852

Step 1: 7-bromo-5-fluoro-1H-indole-2-carboxylic acid

To 7-bromo-5-fluoro-1H-indole-2-carboxylic acid ethyl ester (800 mg,2.80mmol,1 eq.) in THF (8 mL) and H at 40 ℃ ₂ LiOH H was added to the solution in O (4 mL) ₂ O (117.34 mg,2.80mmol,1 eq.). The mixture was stirred at 40℃for 16h.After the reaction was complete, the mixture was concentrated in vacuo and then pH was adjusted to about 1 with 1M HCl (10 mL) and extracted with ethyl acetate (10 mL x 3) to give 7-bromo-5-fluoro-1H-indole-2-carboxylic acid (700 mg, crude material) as a yellow solid. MS (ESI) m/z 256.0[ M-H] ⁺

Step 2: (S) -2- ((S) -2- (7-bromo-5-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]To a solution of methyl propionate (800 mg,2.30mmol,1 eq, HCl) and 7-bromo-5-fluoro-1H-indole-2-carboxylic acid (700 mg,2.76mmol,1.2 eq) in DCM (10 mL) was added DMAP (561.96 mg,4.60mmol,2 eq) and EDCI (881.79 mg,4.60mmol,2 eq) was then added to the mixture. After stirring at 20 ℃ for 2h, the mixture was poured into water (30 mL) and extracted with DCM (10 mL x 3) and dried over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and passed over a column (SiO ₂ PE/ea=1:0 to 0:1) to yield methyl (S) -2- ((S) -2- (7-bromo-5-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1 g,1.75mmol,76.09% yield, 96.5% purity) as a pale yellow solid. MS (ESI) m/z 551.1[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-5-fluoro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-bromo-5-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1 g,1.81mmol,1 eq.) was reacted in NH ₃ A solution in MeOH (30 mL, 7M) was stirred at 30℃for 16h. The mixture was concentrated in vacuo. After the reaction was completed, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-5-fluoro-1H-indole-2-carboxamide (800 mg, crude material) as a pale yellow solid. MS (ESI) m/z 536.2[ M+H ] ] ⁺

Step 4: 7-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-fluoro-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-5-fluoro-1H-indole-2-carboxamide (800 mg,1.81mmol,1 eq.) in DCM (10 mL) was added the bergs reagent (1.30 g,5.44mmol,3 eq.) and the mixture stirred at 30 ℃ for 4H. After the reaction was completed, the reaction mixture was quenched with water (1 mL) and N was used ₂ Dried and purified by preparative HPLC (column: welch Xtime C18X 70mM #10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,20 min) to give 7-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-fluoro-1H-indole-2-carboxamide (740 mg,1.33mmol,53.51% yield, 98% purity) as a white solid. MS (ESI) m/z 518.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 9.01(d,J＝7.7Hz,1H),8.81(d,J＝7.5Hz,1H),7.59-7.48(m,2H),7.45(dd,J＝2.4,9.0Hz,1H),7.26(s,1H),5.07(q,J＝7.8Hz,1H),4.57-4.46(m,1H),3.14-3.01(m,2H),2.31-2.19(m,2H),1.90-1.64(m,4H),1.63-1.34(m,3H),0.85-0.75(m,1H),0.49-0.37(m,2H),0.24-0.06(m,2H)。

Example 200 Synthesis of viral protease inhibitor Compound 876

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (1.3 g,5.49mmol,1 eq, HCl) in DCM (20 mL) was added (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (1.62 g,6.59mmol,1.2 eq), DMAP (1.68 g,13.73mmol,2.5 eq) and EDCI (2.11 g,10.98mmol,2 eq.). After stirring for 1h at 20 ℃, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE: ea=6/1-4/1 to give the product (2S) -2- [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.9 g,4.00mmol,72.82% yield, 90% purity). MS (ESI) m/z 428.3[ M+H ]] ⁺ 。

Step 2: (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1.71 g,5.69mmol,1 eq.) in HCl/MeOH (4M, 20.00mL,14.05 eq.) was stirred at 20deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl as a white oil ]Methyl propionate (1.35 g, crude material, HCl). MS (ESI) m/z 328.3[ M+H ]] ⁺ 。

Step 3: (2S) -2- [ [ (2S) -2- [ [4- [2- (2-methoxyethoxy) ethoxy ] -1H-indole-2-carbonyl ] amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1.35 g,3.71mmol,1 eq. HCl) in DCM (20 mL) was followed by the addition of 4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxylic acid (1.24 g,4.45mmol,1.2 eq.), DMAP (1.13 g,9.28mmol,2.5 eq.) and EDCI (1.42 g,7.42mmol,2 eq.) were stirred at 20℃for 1.5H. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ PE: EA=8/1-4/1) the residue was purified to give (2S) -2- [ [ (2S) -2- [ [4- [2- (2-methoxyethoxy) ethoxy ] as a yellow solidBase group]-1H-indole-2-carbonyl]Amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (2.1 g,2.85mmol,76.92% yield, 80% purity). MS (ESI) m/z 589.4[ M+H ] ] ⁺ 。

Step 4:2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [ (2S) -2- [ [4- [2- (2-methoxyethoxy) ethoxy ] ethoxy]-1H-indole-2-carbonyl]Amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.52 g, parallel 4 batches, 706.65umol,80% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 8mL,79.25 eq.) was stirred at 80℃for 16h. After completion, the mixture was concentrated under reduced pressure to give a residue, and then dissolved with DCM (30 ml x 3). The reaction was concentrated under reduced pressure to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide (1.3 g, crude). MS (ESI) m/z 574.4[ M+H ]] ⁺ 。

Step 5: n- [1- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -4- [2- (2-methoxyethoxy) ethoxy ] -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4- [2- (2-methoxyethoxy) ethoxy ]]To a mixture of 1H-indole-2-carboxamide (1.1 g,1.92mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (1.37 g,5.76mmol,3 eq.). The resulting mixture was stirred at 30℃for 3h. After completion, the mixture was quenched with water (1 mL) and N was used ₂ And (5) drying. By preparative HPLC (column: waters X bridge C, 150X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) was purified and passed through SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:53% -53%,10 min) for further separationObtaining N- [1- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide isomer 1 (250.32 mg,450.49umol,23.46% yield). MS (ESI) m/z 556.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.29(s,1H),7.17-7.10(m,1H),7.07-7.01(m,1H),6.52(d,J＝7.5Hz,1H),5.08(dd,J＝6.2,9.9Hz,1H),4.64(dd,J＝4.2,8.6Hz,1H),4.29-4.23(m,2H),3.93(dd,J＝4.0,5.3Hz,2H),3.79-3.74(m,2H),3.62-3.54(m,2H),3.37(s,3H),3.23-3.14(m,2H),2.49-2.37(m,2H),2.00-1.41(m,7H),1.03(s,9H)。

Obtaining N- [1- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide isomer 2 (27.92 mg,50.25umol,2.62% yield). MS (ESI) m/z 556.3[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.29(d,J＝0.9Hz,1H),7.17-7.11(m,1H),7.04(d,J＝8.4Hz,1H),6.52(d,J＝7.5Hz,1H),5.08(dd,J＝5.8,8.0Hz,1H),4.68(dd,J＝4.0,8.8Hz,1H),4.30-4.23(m,2H),3.93(dd,J＝3.9,5.2Hz,2H),3.80-3.73(m,2H),3.62-3.56(m,2H),3.37(s,3H),3.22-3.13(m,2H),2.45-2.28(m,2H),2.01-1.76(m,5H),1.71-1.49(m,2H),1.02(s,9H)。

Obtaining N- [1- [ [ 1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide isomer 3 (31.42 mg,56.54umol,2.95% yield). MS (ESI) m/z 556.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.30(d,J＝0.9Hz,1H),7.13(d,J＝7.7Hz,1H),7.08-7.01(m,1H),6.53(d,J＝7.3Hz,1H),5.01(s,1H),4.65(s,1H),4.30-4.23(m,2H),3.93(dd,J＝4.0,5.3Hz,2H),3.81-3.73(m,2H),3.63-3.55(m,2H),3.37(s,3H),3.21(br d,J＝4.6Hz,2H),2.49-2.37(m,1H),2.34-2.23(m,1H),1.97-1.88(m,2H),1.87-1.63(m,4H),1.58-1.45(m,1H),1.02(s,9H)。

EXAMPLE 201 Synthesis of viral protease inhibitor Compounds 880

A mixture of 2-chloro-3-methoxy-benzaldehyde (4 g,23.45mmol,1 eq.) and NaOMe (2.53 g,46.90mmol,2 eq.) with MeOH (20 mL) was cooled to-10deg.C and then a mixture of methyl azide acetate (5.49 g,46.90mmol,2 eq.) in MeOH (50 mL) was added dropwise. The mixture was stirred at 25 ℃ for 16h and a white solid was observed. After completion, the reaction mixture was filtered to give compound (Z) -2-azido-3- (2-chloro-3-methoxy-phenyl) prop-2-enoic acid methyl ester (3 g,10.09mmol,43.02% yield, 90% purity) as a white solid. MS (ESI) m/z 267.0[ M+H ]] ⁺

Step 2: 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester

To a solution of methyl (Z) -2-azido-3- (2-chloro-3-methoxy-phenyl) prop-2-enoate (1 g,3.74mmol,1 eq.) in THF (30 mL) was added bis (trifluoromethylsulfonyloxy) iron (2.64 g,7.47mmol,2 eq.) and the mixture stirred at 80 ℃ for 48h. After completion, the reaction was concentrated in vacuo and purified by addition of H ₂ O (100 mL) and then extracted with DCM (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 5/1) to give 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester (140 mg,584.17umol,15.64% yield) as a brown solid. MS (ESI) m/z 240.0[ M+H ]] ⁺

Step 3: 4-chloro-5-methoxy-1H-indole-2-carboxylic acid

To 4-chloro-5-methoxy-1H-indole-2-carboxylic acid methyl ester (0.55 g,2.29mmol,1 eq.) in THF (5 mL), H ₂ LiOH H was added to the solution in O (2.5 mL) ₂ O (96.31 mg,2.29mmol,1 eq.) and the mixture was stirred at 60℃for 2h. At the endAfter completion, the pH of the reaction mixture was adjusted to-3 with HCl. The mixture was extracted with ethyl acetate (100 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtration, and concentration gave 4-chloro-5-methoxy-1H-indole-2-carboxylic acid (340 mg, crude material) as a brown solid. MS (ESI) M/z226.0[ M+H ]] ⁺

Step 4: (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5 ]A solution of tert-butyl decane-2-carboxylate (1.3 g,2.88mmol,1 eq.) in HCl/MeOH (15 mL) was stirred at 25℃for 1h. After completion, by adding NaHCO ₃ (200 mL) the mixture was quenched and then extracted with DCM (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (2-azaspiro [ 4.5) as a yellow solid]Decane-3-carboxamido) propionic acid methyl ester (1.1 g, crude material). MS (ESI) m/z 352.2[ M+H ]] ⁺

Step 5: (2S) -2- (2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (2-azaspiro [ 4.5)]To a solution of methyl decane-3-carboxamido) propionate (934.56 mg,2.66mmol,1 eq.) in DCM (20 mL) was added 4-chloro-5-methoxy-1H-indole-2-carboxylic acid (600 mg,2.66mmol,1 eq.), EDCI (1.02 g,5.32mmol,2 eq.) and DMAP (974.62 mg,7.98mmol,3 eq.). After stirring the mixture at 25℃for 1H, the mixture was prepared by adding H ₂ O (200 mL) quenched the reaction and then extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (2S) -2- (2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamido) -3- ((S) -2-oxo-pyrroleMethyl alkan-3-yl) propionate (1.2 g,1.93mmol,72.57% yield, 89.9% purity). MS (ESI) m/z 559.2[ M+H ]] ⁺

Step 6: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- (2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [ 4.5)]Decane-3-formylamino) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester (1.2 g,2.15mmol,1 eq.) in NH ₃ The solution in (7M, 30mL,97.83 eq.) was stirred at 40℃for 8h. After completion, the reaction was concentrated in vacuo to give the crude product N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (1.15 g, crude). MS (ESI) m/z 544.2[ M+H ]] ⁺

Step 7: (2S) -2- (2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (1.15 g,2.11mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (1.51 g,6.34mmol,3 eq.) and the mixture stirred at 25℃for 2h. After completion, the sample was purified by preparative HPLC (column: waters Xbridge C18 150X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give 2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (400 mg,760.42umol,35.97% yield). MS (ESI) m/z 526.2[ M+H ]] ⁺

Step 8: (2S) -2- (2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

By SFC (column: DAICEL CHI)Ralpaas (250 mm x 30mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O MEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:43% -43%,8 min) isolation of 2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5 ]Decane-3-carboxamide gives 2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (170 mg,323.18umol,42.50% yield, 100% purity). MS (ESI) m/z 526.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.80-11.70(m,1H),9.07-8.78(m,1H),7.72-7.50(m,1H),7.41-7.33(m,1H),7.21-7.12(m,1H),6.92-6.57(m,1H),5.00-4.89(m,1H),4.82-4.46(m,1H),3.88-3.81(m,4H),3.73-3.38(m,1H),3.17-2.90(m,2H),2.40-2.20(m,2H)2.17-2.05(m,2H),1.82-1.64(m,2H),1.61-1.51(m,2H),1.49-1.27(m,9H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.52(br s,1H),8.65(br s,1H),7.40-7.39(m,2H),7.16-7.13(m,1H),6.86(br s,1H),4.94(br s,1H),4.59(br s,1H),3.90-3.68(m,5H),3.15-3.06(m,2H),2.26-2.05(m,4H),1.80(br s,1H),1.68(br s,1H),1.56-1.52(m,3H),1.45-1.40(m,8H)

Obtaining 2- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (170 mg,323.18umol,42.50% yield, 100% purity). MS (ESI) m/z 526.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.79-11.65(m,1H),9.10-8.87(m,1H),7.75-7.55(m,1H),7.43-7.27(m,1H),7.21-7.08(m,1H),6.93-6.58(m,1H),4.99-4.94(m,1H),4.69-4.44(m,1H),3.92-3.79(m,4H),3.77-3.67(m,1H),3.31-3.06(m,2H),2.48-2.34(m,1H),2.46-2.34(m,1H),2.20-2.05(m,2H),1.97-1.64(m,2H),1.63-1.52(m,2H),1.50-1.29(m,9H)

1H NMR(400MHz,DMSO-d ₆ )δ＝11.52(br s,1H),8.75(br s,1H),7.57-7.34(m,2H),7.15-7.13(m,1H),6.84(br s,1H),4.91(br s,1H),4.61(br s,1H),3.86-3.68(m,5H),3.17-3.09(m,2H),2.43-2.02(m,4H),1.81(br s,1H),1.67(br s,1H),1.53(br s,3H),1.45-1.41(m,8H)

Example 202 Synthesis of viral protease inhibitor Compounds 882

Step 1: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]A mixture of tert-butyl decane-2-carboxylate (1.5 g,3.32mmol,1 eq.) in HCl/MeOH (4M, 20mL,24.08 eq.) was stirred at 20deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product (2S) -2- (2-azaspiro [ 4.5) as a white oil ]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.29 g, crude material, HCl). MS (ESI) m/z 352.2[ M+H ]] ⁺ 。

Step 2: (2S) -2- [ [2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (1.24 g,3.20mmol,1 eq, HCl) in DCM (15 mL) was added 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (865.52 mg,3.84mmol,1.2 eq), DMAP (976.35 mg,7.99mmol,2.5 eq) and EDCI (1.23 g,6.39mmol,2 eq). The resulting mixture was stirred at 20 ℃ for 1h, and then the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ PE: EA=8/1-5/1) purification of the residue to give (2S) -2-azaspiro [2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow oil]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.6 g,2.46mmol,77.00% yield, 86%) Purity). MS (ESI) m/z 559.3[ M+H ]] ⁺ 。

Step 3: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (400 mg, parallel 4 batches, 615.33umol,86% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,227.52 eq.) was stirred at 50deg.C for 16h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid]Methyl group]Ethyl group]-2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.3 g, crude). MS (ESI) M/z544.3[ M+H ]] ⁺ 。

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ]Methyl group]Ethyl group]-2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.2 g,2.21mmol,1 eq.) in DCM (15 mL) was added the Buerger reagent (1.58 g,6.62mmol,3 eq.). After stirring for 1h at 30 ℃, the mixture was quenched with water (1 mL) and N was used ₂ And (5) drying. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) was purified and passed through SFC (column: DAICEL CHIRALPAK IC (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,9 min) to give 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide isomer 1 (378.42 mg,719.39umol,32.62% yield, 100% purity). MS (ESI) m/z 526.2[ [M+H] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.20-7.13(m,1H),7.11(s,1H),6.59-6.42(m,1H),5.11-5.02(m,1H),4.80-4.58(m,1H),3.99-3.89(m,3H),3.89-3.82(m,1H),3.77-3.38(m,1H),3.28(br s,1H),2.99-2.66(m,1H),2.52-2.25(m,3H),2.17-1.69(m,3H),1.65-1.26(m,11H)。

Obtaining 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide isomer 2 (367.22 mg,698.10umol,31.65% yield, 100% purity). MS (ESI) m/z 526.2[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.18(d,J＝8.2Hz,1H),7.14(s,1H),6.54(d,J＝8.3Hz,1H),5.03(dd,J＝6.0,10.1Hz,1H),4.63(dd,J＝7.8,9.7Hz,1H),3.99-3.88(m,4H),3.76(d,J＝10.3Hz,1H),3.30-3.23(m,1H),2.53-2.40(m,1H),2.39-1.96(m,3H),1.95-1.70(m,3H),1.68-1.38(m,11H)。

EXAMPLE 203 Synthesis of viral protease inhibitor Compound 886

Step 1: (2S) -3- ((S) -2-oxopyrrolidin-3-yl) -2- (6-azaspiro [3.4] octane-7-carboxamido) propionic acid methyl ester hydrochloride

A solution of 7- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (1.5 g,3.54mmol,1 eq.) in HCl/MeOH (4M, 37.50mL,42.35 eq.) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- (6-azaspiro [3.4] octane-7-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionate (1.2 g, crude material, HCl) as a white solid.

Step 2: (2S) -2- (6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

At 0 ℃, to (2S) -2- (6-azaspiro [3.4]]Octane-7-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.1 g,3.06mmol,1 eq, HCl) and 4-chloro-5-methoxy-1H-indole-2-carboxylic acid (700 mg,3.10mmol,1.01 eq) in DMF (7 mL) and DCM (30 mL) were added DMAP (1.12 g,9.17mmol,3 eq) and EDCI (1.17 g,6.11mmol,2 eq) and the mixture was then stirred at 20 ℃ for 2H. After completion, by adding 100mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (50 ml x 3). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) to give (2S) -2- [ [6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] as a yellow solid]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.2 g,2.19mmol,71.71% yield, 97% purity). MS (ESI) m/z 531.2[ M+H ]] ⁺ 。

Step 3: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -2- [ [6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.2 g,2.26mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 50mL,154.87 eq.) was stirred at 25℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow solid ]Methyl group]Ethyl group]-6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]Octane-7-carboxamide (1.1 g, crude). MS (ESI) m/z 516.2[ M+H ]] ⁺ 。

Step 4:6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-6-(4-chloro-5-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]To a solution of octane-7-carboxamide (1.1 g,2.13mmol,1 eq.) in DCM (40 mL) was added the Buerger's reagent (1.27 g,5.33mmol,2.5 eq.) and the mixture was then stirred at 40℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Titank C Bulk 250. Times.70 mM 10u; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified from 6% -36%,20 min) to give the desired compound as a white solid (500 mg,47% yield, 99% purity) which was purified by SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-MeOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,7 min) to give 6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid ]Ethyl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 1 (232.45 mg,466.79umol,21.90% yield, 100% purity). MS (ESI) m/z 498.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.85-11.68(m,1H),9.09-8.67(m,1H),7.74-7.42(m,1H),7.42-7.32(m,1H),7.21-7.10(m,1H),7.01-6.46(m,1H),5.02-4.40(m,2H),4.11-3.65(m,5H),3.20-2.90(m,2H),2.36-1.63(m,13H)

Obtaining 6- (4-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 2 (232.89 mg,467.68umol,21.94% yield, 100% purity). MS (ESI) m/z 498.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.79-11.70(m,1H),9.19-8.76(m,1H),7.76-7.60(m,1H),7.42-7.29(m,1H),7.20-7.08(m,1H),6.96-6.48(m,1H),5.04-4.37(m,2H),4.05-3.78(m,5H),3.18-2.92(m,2H),2.43-1.79(m,13H)

Example 204 Synthesis of viral protease inhibitor Compound 888

/>

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A solution of methyl propionate (20 g,69.85mmol,1 eq.) in HCl/MeOH (200 mL) was stirred at 20deg.C for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl propionate (13 g, crude material, HCl). MS (ESI) m/z 187.1[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl ]To a solution of methyl propionate (13 g,58.38mmol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (14.32 g,58.38mmol,1 eq.) in DCM (200 mL) was added DMAP (21.40 g,175.15mmol,3 eq.) and then EDCI (33.58 g,175.15mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding 100mL H ₂ The reaction mixture was quenched with O and then extracted with 100mL DCM (50 mL x 2). The combined organic layers were washed with 100mL HCl (1M) (50 mL x 2), then with 100mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1 to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (23 g,50.62mmol,86.70% yield, 91% purity). MS (ESI) m/z 414.3[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl ]A solution of methyl propionate (23 g,55.62mmol,1 eq.) in HCl/MeOH (200 mL) was stirred at 20deg.C for 2h. After completion, at the time of subtractionThe reaction mixture was concentrated under reduced pressure to remove the solvent to give (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl as a yellow solid]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (19 g, crude material, HCl). MS (ESI) m/z 314.2[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]To a solution of methyl propionate (1 g,2.86mmol,1 eq, HCl) and 7-chloro-1H-indole-2-carboxylic acid (559.10 mg,2.86mmol,1 eq) in DCM (40 mL) was added DMAP (1.05 g,8.58mmol,3 eq). After EDCI (1.64 g,8.58mmol,3 eq.) was added, the resulting mixture was stirred at 20℃for 2h. After completion, by adding H ₂ The reaction mixture was quenched with O (30 mL) and then extracted with 40mL DCM (20 mL x 2). The combined organic layers were washed with 30mL HCl (1M) (15 mL x 2), the combined organic layers were washed with brine (40 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (930 mg,1.84mmol,64.28% yield, 97% purity). MS (ESI) M/z491.2[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (900 mg,1.83mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 30mL,114.56 eq.) was stirred at 20℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo as a white solidPyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-7-chloro-1H-indole-2-carboxamide (770 mg, crude). MS (ESI) m/z 476.2[ M+H ] ] ⁺ 。

Step 6: 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] carbamoyl ] -3, 3-dimethyl-butyl ] -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 7-chloro-1H-indole-2-carboxamide (760 mg,1.60mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (761.03 mg,3.19mmol,2 eq.). The mixture was stirred at 25 ℃ for 2h, and then the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -55%,10 min) to give 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl) as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (426 mg,919.31umol,57.57% yield, 100% purity). MS (ESI) m/z 458.2[ M+H ]] ⁺ 。

1H NMR(400MHz,DMSO-d ₆ )δ＝11.70(s,1H),9.01(d,J＝7.8Hz,1H),8.72(d,J＝8.1Hz,1H),7.74-7.58(m,2H),7.37-7.22(m,2H),7.07(t,J＝7.8Hz,1H),4.98(q,J＝7.8Hz,1H),4.65-4.52(m,1H),3.19-3.03(m,2H),2.42-2.27(m,1H),2.20-2.06(m,2H),1.82(d,J＝7.4Hz,1H),1.75-1.64(m,3H),0.95(s,9H)。

EXAMPLE 205 Synthesis of viral protease inhibitor Compound 898

Step 1: (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester hydrochloride

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (23 g,55.89mmol,1 eq.)) A solution in HCl/MeOH (4M, 230mL,16.46 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (20 g, crude material, HCl). MS (ESI) m/z 312.1[ M+H ]] ⁺ 。

Step 2: (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g,2.30mmol,1 eq, HCl) and 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (622.71 mg,2.76mmol,1.2 eq) in DMF (5 mL) and DCM (20 mL) were added DMAP (842.95 mg,6.90mmol,3 eq) and EDCI (881.79 mg,4.60mmol,2 eq) and the mixture was then stirred at 20 ℃ for 2H. After completion, by adding H at 0deg.C ₂ O (100 mL) to quench the reaction mixture and then extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,1.83mmol,79.59% yield, 95% purity). MS (ESI) m/z 519.2[ M+H ]] ⁺ 。

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.9 g,1.73mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 36.00mL,145.32 eq.) was stirred at 25℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give a product in the form ofN- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-4-methoxy-1H-indole-2-carboxamide (0.8 g, crude material). MS (ESI) m/z 504.2[ M+H ] ] ⁺ 。

Step 4: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-4-methoxy-1H-indole-2-carboxamide (0.8 g,1.59mmol,1 eq.) in DCM (30 mL) was added the berg reagent (945.70 mg,3.97mmol,2.5 eq.) and the mixture was then stirred at 40 ℃ for 2H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: phenomenex Gemini C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:28% -48%,20 min) to give the product 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (0.21 g,432.13umol,27.22% yield, 100% purity). MS (ESI) m/z 486.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.70(br d,J＝1.6Hz,1H),8.97(d,J＝7.9Hz,1H),8.65(d,J＝7.5Hz,1H),7.53(br s,1H),7.28(s,1H),7.21(d,J＝8.3Hz,1H),6.56(d,J＝8.3Hz,1H),5.07(q,J＝7.8Hz,1H),4.56-4.43(m,1H),3.89(s,3H),3.15-3.02(m,2H),2.30-2.22(m,2H),1.87-1.68(m,4H),1.59-1.39(br s,3H),0.86-0.77(m,1H),0.48-0.38(m,2H),0.23-0.08(m,2H)

Example 206 Synthesis of viral protease inhibitor Compounds 902

Step 1: (Z) -3-bromo-2- (hydroxyimino) propionic acid ethyl ester

At N ₂ Next, 3-bromo-2-oxo-propionic acid ethyl ester (60 g,307.67mmol,38.46mL,1 eq.) was added to CHCl ₃ NH-containing was added to the solution in (250 mL) ₂ H of OH HCl (23.52 g,338.44mmol,1.1 eq) ₂ O (250 mL). The mixture was stirred at 25℃for 16h. Reactant H ₂ O (500 mL) was quenched and then extracted with DCM (300 mL x 4). The combined organic phases were washed with brine (400 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give ethyl (Z) -3-bromo-2- (hydroxyimino) propionate (51 g, crude material) as a yellow solid. MS (ESI) m/z 210.3[ M+H ]] ⁺ 。

Step 2:1- (cyclohexylidene methyl) pyrrolidine

A mixture of cyclohexane formaldehyde (15 g,133.73mmol,16.09mL,1 eq), pyridine (11.41 g,160.47mmol,13.40mL,1.2 eq) in toluene (300 mL) was heated at 130℃for 14h and water was removed by a Dean-Stark trap. The reaction mixture was concentrated under reduced pressure at 50 ℃ to give a residue, which gave 1- (cyclohexylenemethyl) pyrrolidine (20 g, crude material) as a yellow oil. MS (ESI) m/z 166.2[ M+H ]] ⁺ 。

Step 3: 1-hydroxy-2-oxa-3-azaspiro [5.5] undec-3-ene-4-carboxylic acid ethyl ester

at-10deg.C, at N ₂ To a solution of 1- (cyclohexylmethyl) pyrrolidine (20 g,121.01mmol,1 eq.) in THF (200 mL) was added dropwise a solution of (2Z) -3-bromo-2-hydroxyimino-propionic acid ethyl ester (25.42 g,121.01mmol,1 eq.) in THF (200 mL). After 1h, at-10℃under N ₂ TEA (12.24 g,121.01mmol,16.84mL,1 eq.) was added dropwise. At N ₂ The reaction mixture was stirred at 25℃for 12h. HCl (36%, 2.2 eq, 26mL of H at 3.5 volumes) was added drop wise to the reaction at 25 ℃ ₂ O) and stirred at 25 ℃ for 1h. By adding H at 25 ℃ ₂ O(350mL)The reaction mixture was quenched and extracted with ethyl acetate (200 ml x 3). The combined organic layers were washed with brine (300 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 1/1) the residue was purified to give 1-hydroxy-2-oxa-3-azaspiro [5.5 ] as a yellow oil]Undec-3-ene-4-carboxylic acid ethyl ester (15 g,58.44mmol,48.29% yield, 94% purity). MS (ESI) m/z 242.2[ M+H ]] ⁺ 。

Step 4: 2-azaspiro [4.5] decane-3-carboxylic acid ethyl ester

In Ar ₂ Next, 1-hydroxy-2-oxa-3-azaspiro [5.5 ]]To a solution of ethyl undec-3-ene-4-carboxylate (15 g,62.17mmol,1 eq.) in EtOH (150 mL) was added Raney Nickel (10.65 g,124.34mmol,2 eq.). The suspension was degassed in vacuo and purified by H ₂ (125.58 mg,62.17mmol,1 eq.) several times. The mixture is put in H ₂ (125.58 mg,62.17mmol,1 eq.) at 50psi, for 18h at 50 ℃. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=4/1 to ethyl acetate/methanol=10/1) to give 2-azaspiro [4.5] as a yellow substance]Decane-3-carboxylic acid ethyl ester (6 g,22.72mmol,36.54% yield, 80% purity) and 2-azaspiro [4.5] as yellow oil]Decane-3-carboxylic acid ethyl ester (3 g,5.11mmol,8.22% yield, 36% purity). MS (ESI) m/z 212.2[ M+H ]] ⁺ 。

Step 5: 2-azaspiro [4.5] decane-2, 3-dicarboxylic acid 2-tert-butyl 3-ethyl ester

At 0℃to 2-azaspiro [4.5]]To a solution of ethyl decane-3-carboxylate (6 g,28.40mmol,1 eq.) in DCM (60 mL) was added TEA (5.75 g,56.79mmol,7.90mL,2 eq.) and Boc ₂ O (7.44 g,34.07mmol,7.83mL,1.2 eq.). The mixture was stirred at 20℃for 12h. By adding H ₂ O (300 mL) quenched the reaction mixture and extracted with ethyl acetate (150 mL x 3). The combined organic layers were washed with brine (200 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (S)iO ₂ Petroleum ether/ethyl acetate=1/0 to 9/1) to give 2-azaspiro [4.5] as a yellow oil]Decane-2, 3-dicarboxylic acid 2-tert-butyl 3-ethyl ester (6 g,19.27mmol,67.85% yield, N/A purity). MS (ESI) m/z 312.2[ M+H ] ] ⁺ 。

Step 6:2- (tert-Butoxycarbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid

To 2-azaspiro [4.5]]2-tert-butyl 3-ethyl decane-2, 3-dicarboxylic acid (7 g,22.48mmol,1 eq.) in H ₂ To a solution of O (14 mL) and MeOH (56 mL) was added LiOH.H ₂ O (1.89 g,44.96mmol,2 eq.). The mixture was stirred at 40℃for 12h. The reaction mixture was concentrated under reduced pressure to remove MeOH. Residue with H ₂ O (80 mL) was diluted and extracted with ethyl acetate.

Step 7:3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (6.25 g,26.40mmol,1.1 eq. HCl) and 2-tert-butoxycarbonyl-2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid (6.8 g,24.00mmol,1 eq.) in DCM (90 mL) was added DMAP (5.86 g,48.00mmol,2 eq.) and EDCI (6.90 g,36.00mmol,1.5 eq.). The mixture was stirred at 25℃for 2h. The reaction was quenched with 0.5M HCl (200 mL) and then extracted with DCM (100 mL x 3). The combined organic phases were washed with brine (150 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid ]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (9 g,18.36mmol,76.53% yield, 95% purity). MS (ESI) m/z 466.2[ M+H ]] ⁺

Step 8: (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5]Decane-2-carboxylic acid tert-butyl esterA mixture of butyl ester (1.5 g,2.90mmol,90% purity, 1 eq.) in HCl/MeOH (4M, 20mL,27.59 eq.) was cooled to 0deg.C and then stirred at 25deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [ 4.5) as a white solid]Decane-3-carboxamido) propionic acid methyl ester (1.5 g, crude material, HCl). MS (ESI) M/z366.1[ M+H] ⁺ 。

Step 9: (2S) -2- (2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [ 4.5)]To a mixture of methyl decane-3-carboxamido) propionate (1.5 g,3.55mmol,1 eq, HCl) in DCM (30 mL) and DMF (10 mL) was added 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (959.94 mg,4.25mmol,1.2 eq) followed by DMAP (1.30 g,10.64mmol,3 eq) and EDCI (1.36 g,7.09mmol,2 eq) at 0deg.C. The resulting mixture was stirred at 25 ℃ for 2h, and then the reaction mixture was quenched with water (10 mL) at 0 ℃. After extraction with DCM (10 mL x 3), the combined organic layers were washed with brine (10 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=100:1 to 10:1 to give (2S) -2- [ [2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow oil]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.91 g,3.00mmol,84.60% yield, 90% purity). MS (ESI) m/z 573.3[ M+H ]] ⁺ 。

Step 10: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.91 g,3.00mmol,90% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 17.79mL,41.52 eq.) was stirred at 80℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give a product in the form ofN- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (1.3 g, crude). MS (ESI) m/z 558.3[ M+H ]] ⁺ 。

Step 11:2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.3 g,1.98mmol,1 eq.) in DCM (25 mL) was added the Bungeus reagent (1.42 g,5.94mmol,3 eq.). After stirring at 25℃for 3h, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ (]<30 ℃ C.). By preparative HPLC (column: phenomenex Titank C mass (250X 100mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -75%,20 min) to give 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamide (350 mg,648.09umol,32.73% yield). MS (ESI) m/z 540.1[ M+H ]] ⁺ 。

Step 12:2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: waters Xbridge BEH C (100. Times.30 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -60%,8 min) purification of 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ]Decane-3-carboxamide (350.00 mg,550.87umol,95% purity, 1 eq) gave 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 1 (62.40 mg,112.77umol,20.47% yield, 97.6% purity). MS (ESI) m/z540.2[ M+H ]] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.23-6.79(m,2H),6.58-6.39(m,1H),5.11(dd,J＝5.7,10.6Hz,1H),4.77-4.52(m,1H),4.03-3.76(m,4H),3.74-3.37(m,1H),3.47-2.89(m,2H),2.65-2.10(m,3H),2.09-1.27(m,16H)。

Obtaining 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 2 (131.81 mg,244.07umol,44.31% yield, 100% purity). MS (ESI) M/z540.2[ M+H] ⁺ 。

¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.22-6.84(m,2H),6.59-6.44(m,1H),5.07-4.95(m,1H),4.69-4.50(m,1H),4.02-3.81(m,4H),3.80-3.43(m,1H),3.23-3.02(m,2H),2.54-2.13(m,3H),2.11-1.36(m,16H)。

Obtaining 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 3 (34.64 mg,64.14umol,11.64% yield, 100% purity). MS (ESI) M/z540.2[ M+H] ⁺ 。

1H NMR (400 MHz, methanol-d) ₄ )δ＝7.31-6.74(m,2H),6.63-6.43(m,1H),5.29-4.96(m,1H),4.87-4.58(m,1H),3.91(br d,J＝9.0Hz,4H),3.80-3.38(m,1H),3.29-3.02(m,2H),2.64-2.13(m,3H),2.10-1.35(m,16H)。

Obtaining 2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- (1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide isomer 4 (5.66 mg,10.45umol,1.90% yield, 99.7% purity). MS (ESI) m/z540.2[ M+H ] ] ⁺ 。

1H NMR (400 MHz, methanol-d) ₄ )δ＝7.31-6.77(m,2H),6.62-6.46(m,1H),5.17-4.91(m,1H),4.75-4.56(m,1H),4.04-3.80(m,4H),3.73(d,J＝10.4Hz,1H),3.28-3.01(m,2H),2.55-2.44(m,1H),2.44-2.25(m,2H),2.08-1.40(m,16H)。

(50 ml x 2). Aqueous HCl was added to the aqueous phase to adjust to ph=2 and extracted with EA (90 mL x 3), and the combined organic layers were washed with brine (90 mL), over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to give a white solid2-Boc-2-azaspiro [4.5 ]]Decane-3-carboxylic acid (6.1 g, crude material). MS (ESI) m/z 284.2[ M+H ]] ⁺

EXAMPLE 207 Synthesis of viral protease inhibitor Compound 906

Step 1: (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester

at-10deg.C, to CH ₃ To a solution of ONa (2.53 g,46.90mmol,2 eq.) in MeOH (40 mL) was added a mixture of 4-chloro-2-methoxy-benzaldehyde (4 g,23.45mmol,1 eq.) and methyl 2-azidoacetate (5.40 g,46.90mmol,2 eq.) in MeOH (15 mL). After stirring for 16H at 20℃the solution was treated with H ₂ O (60 mL) was diluted and concentrated and extracted with ethyl acetate (50 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ Petroleum ether ethyl acetate=20:1 to 3:1) to give the product (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (3.3 g,12.33mmol,52.58% yield) as a white solid. MS (ESI) m/z 268.1[ M+H ] ] ⁺

Step 2: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (3000 mg,11.21mmol,1 eq.) in xylene (30 mL) was stirred at 170℃for 4h. After completion, the solution was concentrated to give a crude material. Through the column (SiO) ₂ Petroleum ether ethyl acetate=10:1 to 1:10) to give the product 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester as a white solid (1500 mg,6.26mmol,55.84% yield). MS (ESI) m/z 240.1[ M+H ]] ⁺

Step 3: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid

To 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (1500 mg,6.26mmol,1 eq.) in THF (15 mL) and H ₂ LiOH.H was added to the solution in O (15 mL) ₂ O (787.95 mg,18.78mmol,3 eq.). After stirring for 2h at 65 ℃, the solution was concentrated and usedEthyl acetate (50 ml x 2) and the aqueous layer was adjusted to ph=4-5 with HCl (concentrate) and extracted with ethyl acetate (80 ml x 3) and purified over Na ₂ SO ₄ Drying and concentrating to obtain crude material. The crude material was used directly in the next step. 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (1070 mg,4.74mmol,75.77% yield) was as a brown solid. MS (ESI) m/z 226.2[ M+H ] ] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (500 mg,1.44mmol,1 eq, HCl) in DCM (10 mL) and DMF (10 mL) was added DMAP (351.22 mg,2.87mmol,2 eq), 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (372.98 mg,1.65mmol,1.15 eq) and EDCI (551.13 mg,2.87mmol,2 eq). After stirring for 2H at 20℃the mixture was taken up in H ₂ O (30 mL) was diluted and extracted with ethyl acetate (50 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ Petroleum ether ethyl acetate=10:1 to EA: meOH=10:1) the crude material was purified to give (2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,963.41umol,67.02% yield). MS (ESI) m/z 519.3[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,963.41umol,1 eq.) in NH ₃ A solution in MeOH (7M, 20mL,145.32 eq.) was stirred at 60℃for 17h. The solution was concentrated to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-4-methoxy1H-indole-2-carboxamide (485 mg, crude). The crude material was used directly in the next step. MS (ESI) m/z 504.3[ M+H ]] ⁺

Step 6: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 6-chloro-4-methoxy-1H-indole-2-carboxamide (470 mg,932.58 mol,1 eq.) in DCM (25 mL) was added a Bungeus reagent (666.72 mg,2.80mmol,3 eq.). After stirring at 20 ℃ for 3h, the solution was washed with brine (50 mL) and N was used ₂ Drying to obtain crude material. Purification of the crude material by preparative HPLC (neutral) gave 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (210 mg,432.13 mol,46.34% yield). MS (ESI) m/z 486.3[ M+H ]] ⁺

Preparative HPLC conditions: column: waters Xbridge C18 is 150 x 50mm x 10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; b%:30% -60%,10min

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.72(s,1H),8.91(br d,J＝8.2Hz,1H),8.58(br d,J＝7.3Hz,1H),7.53(br s,1H),7.38(s,1H),7.03(s,1H),6.56(s,1H),5.17-4.93(m,1H),4.53-4.31(m,1H),3.91(s,3H),3.09(br s,2H),2.37-2.15(m,2H),1.89-1.27(m,7H),0.80(br s,1H),0.40(br s,1H),0.23-0.10(m,2H)。

Example 208 Synthesis of viral protease inhibitor Compound 1511

Step 1: (S) -2-amino-2-methylpentanedioic acid dimethyl ester

A mixture of (2S) -2-amino-2-methyl-glutaric acid (1 g,6.21mmol,1 eq.) in HCl/MeOH (4M, 10mL,6.45 eq.) was stirred inStirring for 2h at 80 ℃. After completion, the reaction mixture was concentrated under reduced pressure to give dimethyl (S) -2-amino-2-methylpentanoate (1.4 g, crude material) as a yellow oil. MS (ESI) m/z 190.2[ M+H ]] ⁺ 。

Step 2: (S) -2- (((Benzylmethoxy) carbonyl) amino) -2-methylpentanedioic acid dimethyl ester

To a mixture of dimethyl (S) -2-amino-2-methylpentanoate (1.1 g,4.87mmol,1 eq. HCl) in DCM (11 mL) at 0deg.C was added K ₂ CO ₃ (2.02 g,14.62mmol,3 eq.) and CbzCl (914.69 mg,5.36mmol,762.24uL,1.1 eq.). After stirring for 14h at 20 ℃, the reaction mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 2:1) the residue was purified to give dimethyl (S) -2- (((benzyloxy) carbonyl) amino) -2-methylglutarate (920 mg,2.85mmol,58.37% yield) as a yellow oil. MS (ESI) m/z 324.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- (((Benzylmethoxy) carbonyl) amino) -4- (cyanomethyl) -2-methylpentanedioic acid dimethyl ester

At N ₂ To a mixture of dimethyl (S) -2- (((benzyloxy) carbonyl) amino) -2-methylglutarate (920 mg,2.42mmol,85% purity, 1 eq.) in anhydrous THF (18.4 mL) was added dropwise LiHMDS (1M, 5.32mL,2.2 eq.) at-65 to-55 ℃ in atmosphere for 0.5h. After stirring for another 1h at-65 to-55 ℃, 2-bromoacetonitrile (435.14 mg,3.63mmol,241.75ul,1.5 eq.) was added dropwise to the mixture solution for 0.5h while keeping the temperature below-65 to-55 ℃. At N ₂ The reaction mixture was stirred at-65 to-55℃for 1h. After completion, the reaction mixture was quenched with MeOH (2.8 mL) containing pre-cooled (EtOH with dry ice) and a THF solution of acetic acid (0.46 mL HOAc/3.7mL THF) pre-cooled (EtOH with dry ice) to at-60 ℃. After stirring for a further 30min at-60 ℃, the cooling bath was removed and replaced with an aqueous bath. The reaction mixture was warmed to 0.+ -. 5 ℃ and then concentrated under reduced pressure at 30 ℃ to give Dark brown solid. The residue obtained was dissolved in ethyl acetate (37 mL) and washed with brine (18 mL x 2). The organic phase was purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 2:1) the residue was purified to give dimethyl (2S) -2- (((benzyloxy) carbonyl) amino) -4- (cyanomethyl) -2-methylglutarate (740 mg,1.84mmol,75.99% yield, 90% purity) as a yellow oil. MS (ESI) m/z 363.1[ M+H ]] ⁺ 。

Step 4: (2S) -2- (((benzyloxy) carbonyl) amino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a stirred solution of dimethyl (2S) -2- (((benzyloxy) carbonyl) amino) -4- (cyanomethyl) -2-methylglutarate (740 mg,1.84mmol,90% purity, 1 eq.) in MeOH (34 mL) at 0deg.C was added CoCl ₂ .6H ₂ O (262.37 mg,1.10mmol,0.6 eq.) and then NaBH was added to the mixture in 4 portions at 0deg.C ₄ (319 mg,11.08mmol,6.03 eq.) for 1h and then stirring the black mixture at 25℃for 2h. After completion, at 0deg.C with NH ₄ The mixture was quenched with aqueous Cl (41 mL), filtered through celite, then extracted with DCM (41 mL x 3) and the organic layer was dried over Na ₂ SO ₄ Dried, filtered, and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 1:1) to give methyl (2S) -2- (((benzyloxy) carbonyl) amino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionate (320 mg,957.03umol,52.07% yield) as a white solid. MS (ESI) m/z 335.2[ M+H ]] ⁺ 。

Step 5: (2S) -2-amino-2-methyl-3- (2-oxopyrrolidin-3-yl) propionic acid methyl ester

At N ₂ Next, methyl (2S) -2- (((benzyloxy) carbonyl) amino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propanoate (320 mg,957.03umol,1 eq.) was added to H ₂ Pd/C (160 mg,10% purity) was added to a mixture of O (1.5 mL) and T-BuOH (6 mL). The resulting mixture was degassed and purified with H ₂ Purge 3 times, and then mixCompounds of formula H ₂ (15 Psi) at 25℃for 2h. After completion, the mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give methyl (2S) -2-amino-2-methyl-3- (2-oxopyrrolidin-3-yl) propionate (140 mg, crude) as a white solid. MS (ESI) m/z 201.1[ M+H ]] ⁺ 。

Step 6: (2S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (2S) -2-amino-2-methyl-3- (2-oxopyrrolidin-3-yl) propionate (140 mg,699.18 mol,1 eq.) in DCM (2 mL) and DMF (1 mL) was added (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (192.36 mg,839.02 mol,1.2 eq.), TEA (212.25 mg,2.10mmol,291.95ul,3 eq.). Adding T at 0deg.C ₃ After P (667.40 mg,1.05mmol,623.74uL,50% purity, 1.5 eq.) the mixture was stirred at 25℃for 1h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give methyl (2S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionate (280 mg,612.41umol,87.59% yield, 90% purity) as a yellow oil. MS (ESI) m/z 412.3[ M+H ]] ⁺ 。

Step 7: (2S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propanoate (260 mg, 568.66. Mu. Mol,90% purity, 1 eq.) in HCl/MeOH (4M, 2.6mL,18.29 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propanoate (200 mg, crude material, HCl) as a yellow solid. MS (ESI) m/z 312.2[ M+H ] ] ⁺ 。

Step 8: (2S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propanamido) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (2S) -2- ((S) -2-amino-3-cyclopropylpyrrolidin-3-yl) propionate (200 mg,546.23 mol,95% purity, 1 eq, HCl) in DCM (4 mL) and DMF (2 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (125.32 mg,655.48 mol,1.2 eq), DMAP (200.20 mg,1.64mmol,3 eq) and EDCI (209.43 mg,1.09mmol,2 eq) at 0 ℃. The mixture was stirred at 25 ℃ for 1h, and then the reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give methyl (2S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propionylamino) -2-methyl-3- (2-oxopyrrolidin-3-yl) propionate (300 mg,451.97umol,82.74% yield, 73% purity) as a yellow oil. MS (ESI) m/z 485.3[ M+H ]] ⁺ 。

Step 9: n- ((2S) -1- (((2S) -1-amino-2-methyl-1-oxo-3- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

Methyl (2S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propanamido) -2-methyl-3- (2-oxopyrrolidin-3-yl) propanoate (280.00 mg,421.84umol,73% purity, 1 eq.) was taken in NH ₃ A solution in MeOH (7M, 6mL,99.56 eq.) was stirred at 80℃for 86h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give N- ((2S) -1- (((2S) -1-amino-2-methyl-1-oxo-3- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide as a yellow solid (35 mg,70.82umol,16.79% yield, 95% purity). MS (ESI) m/z 470.3[ M+H ]] ⁺ 。

Step 10: n- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((2S) -1- (((2S) -1-amino-2-methyl-1-oxo-3- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (30 mg,60.70umol,95% purity, 1 eq) in DCM (1 mL) was added the bergs reagent (43.40 mg,182.10umol,3 eq) and then stirred for 6H at 25 ℃. After completion, by adding H at 20℃ ₂ O (0.1 mL) to quench the reaction mixture and then under reduced pressure<Concentrated at 20 ℃ to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified for 20% -50%,10 min) to give N- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (25 mg,52.60umol,86.66% yield, 95% purity) as a white solid. MS (ESI) M/z452.2[ M+H] ⁺ 。

Step 11: n- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,7 min) purification of N- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (25 mg,52.60umol,95% purity, 1 eq) gave N- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 1 (2.35 mg,5.10umol,9.69% yield, 97.9% purity) as a white solid. MS (ESI) m/z 452.1[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.26(d,J＝0.7Hz,1H),7.19-7.11(m,1H),7.03(d,J＝8.2Hz,1H),6.51(d,J＝7.7Hz,1H),4.59(t,J＝7.3Hz,1H),3.93(s,3H),3.38-3.32(m,2H),2.77-2.66(m,1H),2.54-2.45(m,1H),2.40(dd,J＝5.1,14.3Hz,1H),2.07(dd,J＝7.3,14.3Hz,1H),2.02-1.91(m,1H),1.86(td,J＝7.1,14.0Hz,1H),1.74(s,3H),1.68(td,J＝7.1,14.1Hz,1H),0.93-0.79(m,1H),0.59-0.44(m,2H),0.26-0.14(m,2H)。

N- ((2S) -1- (((2S) -2-cyano-1- (2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 2 (2.08 mg,4.53umol,8.62% yield, 98.4% purity) was obtained as a white solid. MS (ESI) m/z 452.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.26(s,1H),7.20-7.13(m,1H),7.03(d,J＝8.2Hz,1H),6.52(d,J＝7.7Hz,1H),4.62(dd,J＝6.4,7.9Hz,1H),3.93(s,3H),3.28-3.16(m,2H),2.73-2.61(m,1H),2.39(td,J＝6.6,12.8Hz,1H),2.27(dd,J＝7.4,14.9Hz,1H),2.01-1.92(m,1H),1.92-1.78(m,2H),1.76-1.66(m,4H),0.89-0.78(m,1H),0.55-0.44(m,2H),0.24-0.14(m,2H)

EXAMPLE 209 Synthesis of viral protease inhibitor Compound 749

Step 1: n- (3-hydroxy-1, 1-dimethyl-propyl) carbamic acid benzyl ester

To a solution of 3-amino-3-methyl-butan-1-ol (3.5 g,33.93mmol,1 eq.) in IPA (60 mL) was added 60mL saturated NaHCO ₃ (64.80 g,771.37mmol,30mL,22.74 eq.) in pH=11 buffer, adjusted with 4M NaOH (4M, 30mL,3.54 eq.). The reaction mixture was cooled to 0 ℃ and then benzyl 2, 5-dioxopyrrolidine-1-carboxylate (7.91 g,33.93mmol,1 eq.) was added. The reaction mixture was stirred at 20℃for 16h. After completion, the reaction mixture was filtered and then concentrated under reduced pressure to remove IPA. Residue with H ₂ O (50 mL) was diluted and extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=10/1 to 5/1) to give benzyl N- (3-hydroxy-1, 1-dimethyl-propyl) carbamate (5 g,20.02mmol,59.00% yield, 95% purity) as a colorless oil. MS (ESI) m/z 238.1[ M+H ]] ⁺

Step 2: n- (1, 1-dimethyl-3-oxo-propyl) carbamic acid benzyl ester

To a solution of benzyl N- (3-hydroxy-1, 1-dimethyl-propyl) carbamate (2.2 g,9.27mmol,1 eq.) in DCM (1 mL) was added DMP (4.72 g,11.13mmol,3.44mL,1.2 eq.). The reaction mixture was stirred at 25℃for 2h. After completion, the reaction mixture was taken up with H ₂ O (100 mL) was diluted and extracted with ethyl acetate (100 mL x 2). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=20/1 to 10/1) to give N- (1, 1-dimethyl-3-oxo-propyl) carbamate (1.2 g,4.59mmol,49.51% yield, 90% purity) as a colorless oil. MS (ESI) m/z 236.1[ M+H ]] ⁺

Step 3: (Z) -5- (Benzylmethoxycarbonylamino) -2- [ (2S) -2- (tert-butoxycarbonylamino) -3-methoxy-3-oxo-propyl ] -5-methyl-hex-2-enoic acid

At-60 ℃ under N ₂ To a solution of dimethyl (2S) -2- (t-butoxycarbonylamino) glutarate (1.4 g,5.09mmol,1 eq.) in THF (15 mL) was added dropwise a solution of LiHMDS (1M, 10.68mL,2.1 eq.). After stirring at-60℃for 0.5h, benzyl N- (1, 1-dimethyl-3-oxo-propyl) carbamate (1.20 g,5.09mmol,1 eq.) in THF (10 mL) was added below-60℃and the reaction mixture stirred at-60℃for 3h. After completion, the reaction mixture was quenched by addition of THF (20 mL) containing 5mL AcOH at 0 ℃ and concentrated under reduced pressure to give a residue. Neutral prep HPLC (column: welch Xtime C18X 70mM #10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,20 min) to obtain white residue(Z) -5- (Benzyloxycarbonylamino) -2- [ (2S) -2- (t-butoxycarbonylamino) -3-methoxy-3-oxo-propyl ] as a solid]-5-methyl-hex-2-enoic acid (230 mg,456.60umol,8.98% yield, 95% purity). MS (ESI) m/z 379.1[ M+H-100 ]] ⁺

Step 4: (2Z, 4S) -2- [3- (Benzylmethoxycarbonylamino) -3-methyl-butylene ] -4- (t-butoxycarbonylamino) glutaric acid dimethyl ester

To (Z) -5- (benzyloxycarbonylamino) -2- [ (2S) -2- (tert-butoxycarbonylamino) -3-methoxy-3-oxo-propyl ]K is added to a mixture of 5-methyl-hex-2-enoic acid (250 mg,522.43umol,1 eq.) in DMF (2.5 mL) ₂ CO ₃ (144.41 mg,1.04mmol,2 eq.) and CH ₃ I (222.46 mg,1.57mmol,97.57uL,3 eq.). The mixture was stirred at 25℃for 1h. After completion, by adding H at 0deg.C ₂ O (10 mL) to quench the reaction mixture, and then H was used ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 2). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the residual compound (2Z, 4S) -2- [3- (benzyloxycarbonylamino) -3-methyl-butylene) as a colorless oil]Dimethyl 4- (t-butoxycarbonylamino) glutarate (230 mg,420.25umol,80.44% yield, 90% purity). The residue was used directly in the next step. MS (ESI) m/z 393.2[ M+H-100 ]] ⁺

Step 5: (4S) -2- (3-amino-3-methyl-butyl) -4- (tert-Butoxycarbonylamino) glutaric acid dimethyl ester

To (2Z, 4S) -2- [3- (benzyloxycarbonylamino) -3-methyl-butylene]To a mixture of dimethyl 4- (t-butoxycarbonylamino) glutarate (230 mg,466.95umol,1 eq.) in i-PrOH (10 mL) was added Pd/C (300 mg,466.95umol,10% purity, 1 eq.). At H ₂ (50 Psi) the mixture was stirred at 50℃for 5h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give the residual compound dimethyl (4S) -2- (3-amino-3-methyl-butyl) -4- (tert-butoxycarbonylamino) glutarate (120 mg,299.63umol,64.17% yield, 90% purity) as a colorless oil and used directly in the next step. MS (ESI) M/z361.2[ M+H ] ⁺

Step 6: (2S) -2- (tert-Butoxycarbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To (4S) -2- (3-amino-3-methyl-butyl) -4- (tert-butoxycarbonylamino) glutaric acid dimethyl ester (120 mg,332.92umol,1 eq.) in MeOH (0.5 mL) and CHCl ₃ KOAc (65.35 mg,665.84umol,2 eq.) was added to the mixture in (0.05 mL). The mixture was stirred at 80℃for 16h. After completion, the residue was taken up with 5mL H ₂ Dilute with O and extract with ethyl acetate (5 ml x 2). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the residual compound methyl (2S) -2- (tert-butoxycarbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (100 mg,274.05umol,82.32% yield, 90% purity) as a colorless oil and used directly. MS (ESI) m/z 329.2[ M+H ]] ⁺

Step 7: (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (100 mg,304.50umol,1 eq.) was added HCl/MeOH (4M, 76.13uL,1 eq.). The resulting mixture was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the residual compound methyl (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (80 mg,287.07umol,94.27% yield, 95% purity, HCl) as a colorless oil.

Step 8: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To a mixture of methyl (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (80 mg,302.17umol,1 eq., HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (69.28 mg,302.17umol,1 eq.) in DCM (2 mL) and DMF (1 mL) was added DMAP (73.83 mg,604.35umol,2 eq.) and EDCI (115.85 mg,604.35umol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with H ₂ Dilute with O (20 mL) and use ethyl acetate (20 mL x 2) And (5) extracting. The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=10/1 to 3/1 to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a colorless oil]Amino group]-methyl 3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propanoate (110 mg,225.23 mol,74.54% yield, 90% purity). MS (ESI) m/z 440.3[ M+H ]] ⁺

Step 9: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propanoate (110 mg,250.26umol,1 eq.) was added HCl/MeOH (4M, 7.33mL,117.21 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the residual compound (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester (90 mg,239.43umol,95.67% yield, HCl) as a colorless oil.

Step 10: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To 7-chloro-1H-indole-2-carboxylic acid (46.83 mg,239.43umol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]To a mixture of methyl 3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (90 mg,239.43umol,1 eq., HCl) in DCM (4 mL) and DMF (2 mL) was added EDCI (91.80 mg,478.86umol,2 eq.) and DMAP (58.50 mg,478.86umol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 40mL EA (20 mL x 2). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ Petroleum ether: ethyl acetate=0:1) to give the residue as a white solidCompound (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-methyl 3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (100 mg,183.75umol,76.74% yield, 95% purity). MS (ESI) m/z 517.3[ M+H ]] ⁺

Step 11: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-methyl 3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propanoate (100 mg,193.42umol,1 eq.) in NH ₃ A solution in MeOH (7M, 10.00mL,361.91 eq.) was stirred at 55deg.C for an additional 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl as a white solid ]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (100 mg,185.26umol,95.78% yield, 93% purity). MS (ESI) M/z502.2[ M+H] ⁺

Step 12: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 7-chloro-1H-indole-2-carboxamide (80 mg,159.36umol,1 eq.) in DCM (5 mL) was added the Bungeus reagent (75.95 mg,318.72umol,2 eq.). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was taken up with H ₂ O (5 mL) was diluted and extracted with DCM (5 mL x 2). The combined organic layers were concentrated using a blow dryer to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to obtain a white solid mixed product(65 mg). By SFC (column: REGIS (s, s) WHELK-O1 (250 mm. Times.30 mm,5 um); mobile phase: [0.1% NH) ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,8 min) to give the compound 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (35 mg,72.32umol,45.38% yield, 100% purity) and 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (25 mg,51.65umol,32.41% yield, 100% purity). MS (ESI) m/z 484.2[ M+H ]] ⁺

Isomer 1:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.86-11.59(m,1H),9.00(d,J＝8.0Hz,1H),8.72(d,J＝7.6Hz,1H),7.63(d,J＝8.0Hz,1H),7.49(s,1H),7.31(d,J＝7.5Hz,1H),7.28-7.23(m,1H),7.07(t,J＝7.8Hz,1H),5.09(q,J＝8.0Hz,1H),4.61-4.46(m,1H),2.30-2.08(m,2H),1.88-1.67(m,3H),1.64-1.38(m,4H),1.17-1.03(m,6H),0.89-0.70(m,1H),0.51-0.36(m,2H),0.28-0.01(m,2H)。

isomer 2:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.73(s,1H),9.04(d,J＝7.4Hz,1H),8.74(d,J＝7.7Hz,1H),7.63(d,J＝7.9Hz,1H),7.51(s,1H),7.31(d,J＝7.5Hz,1H),7.26(s,1H),7.07(t,J＝7.7Hz,1H),5.02(q,J＝7.4Hz,1H),4.61-4.52(m,1H),2.32(td,J＝6.8,13.7Hz,1H),2.20-2.06(m,1H),1.88-1.49(m,7H),1.12(d,J＝8.0Hz,6H),0.88-0.70(m,1H),0.52-0.34(m,2H),0.26-0.05(m,2H)。

EXAMPLE 210 Synthesis of viral protease inhibitor Compounds 928

Step 1: (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To 2- (7-chloro-1H-indole-2-carbonyl)) -2-azaspiro [4.5]]Decane-3-carboxylic acid (354.36 mg,982.06umol,1 eq.) and methyl (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (260 mg,982.06umol,1 eq., HCl) were added to a mixture of HATU (448.09 mg,1.18mmol,1.2 eq.) in DMF (10 mL) and DMF (5 mL) containing DIEA (380.78 mg,2.95mmol,513.17uL,3 eq.) at 0deg.C. The mixture was stirred at 0deg.C for 30min. After completion, the reaction mixture was quenched with 50mL H ₂ O was diluted and extracted with 100mL EA (50 mL x 2). The combined organic layers were washed with 50mL brine (50 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=10/1 to 0/1) to give the compound (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-methyl 3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (550 mg,866.74umol,88.26% yield, 90% purity). MS (ESI) M/z571.3[ M+H] ⁺

Step 2: n- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl ] -2-oxo-ethyl ] -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 25 ℃]Decane-3-carbonyl]Amino group]NH was added to a mixture of methyl-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (550 mg,963.04umol,1 eq) ₃ MeOH (7M, 137.58uL,1 eq.). The mixture was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the residual compound N- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl as a white solid ]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (520 mg,841.58umol,87.39% yield, 90% purity) and the residue was used directly in the next step. MS (ESI) m/z 556.3[ M+H ]] ⁺

Step 3:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

At the temperature of 25 deg.c,to N- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (500 mg,899.13umol,1 eq.) in DCM (10 mL) was added the Bungeus reagent (428.53 mg,1.80mmol,2 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was quenched with 10mL H ₂ O was diluted and extracted with 20mL DCM (10 mL x 2). The combined organic layers were washed with 10mL brine (10 mL x 1) and dried over N2 to give a residue. By neutral prep HPLC (column: waters Xbridge C18. Times.50 mm. Times.10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,10 min) of the purified residue. MS (ESI) M/z538.2[ M+H ]] ⁺

Isomers 1 and 2:

2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide (100 mg,185.10umol,20.59% yield, 99.6% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.10(br s,1H),8.70(br d,J＝16.5Hz,1H),7.62(br s,1H),7.38-6.82(m,4H),4.98(br s,1H),4.60(br s,1H),3.83(br d,J＝10.1Hz,1H),3.62(br s,1H),2.31-1.96(m,3H),1.94-1.26(m,16H),1.22-1.01(m,6H)

Isomer 3:

2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide (50 mg,92.92umol,10.33% yield, 100% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.12(br s,1H),9.01-8.62(m,1H),7.83-7.52(m,1H),7.49-6.65(m,4H),4.94(br d,J＝5.7Hz,1H),4.61(br s,1H),4.00-3.33(m,2H),2.35-1.99(m,3H),1.91-1.28(m,16H),1.20-1.07(m,6H)

Isomer 4:

2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide (50 mg,90.69umol,10.09% yield, 97.6% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.54-10.62(m,1H),8.96-8.58(m,1H),7.63(br d,J＝7.3Hz,1H),7.39-6.91(m,4H),4.94(q,J＝6.8Hz,1H),4.60(br s,1H),3.92-3.46(m,2H),2.31-2.01(m,3H),1.76-1.29(m,16H),1.14(d,J＝18.3Hz,6H)

EXAMPLE 211 Synthesis of viral protease inhibitor Compounds 930

Step 1: 2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

2-azaspiro [4.5]]A mixture of decane-3-carboxylic acid (400 mg,1.82mmol,1 eq., HCl) in HCl/MeOH (4M, 6mL,13.18 eq.) was stirred at 70℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give 2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid methyl ester (400 mg,1.71mmol,94.00% yield, HCl). MS (ESI) m/z 198.2[ M+H ]] ⁺ 。

Step 2:2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid methyl ester

At 0℃to 2-azaspiro [4.5]]To a mixture of decane-3-carboxylic acid methyl ester (400 mg,1.71mmol,1 eq. HCl) and 7-chloro-1H-indole-2-carboxylic acid (334.74 mg,1.71mmol,1 eq.) in DCM (6 mL) was added DIEA (663.54 mg,5.13mmol,894.26uL,3 eq.) and T in one portion ₃ P (816.78 mg,2.57mmol,763.34uL,1.5 eq.). The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative TLC (SiO) ₂ Petroleum ether ethyl acetate=0:1) to give 2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid methyl ester (350 mg,933.68umol,54.56% yield). MS (ESI) m/z 375.1[ M+H ]] ⁺

Step 3:2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid

To 2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 25 ]]Decane-3-carboxylic acid methyl ester (350 mg,933.68umol,1 eq.) in THF (2 mL) and H ₂ LiOH.H was added at once to the mixture in O (2 mL) ₂ O (78.36 mg,1.87mmol,2 eq.). The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was made acidic with 1M HCl and extracted with ethyl acetate (10 ml x 3). The combined organic layers were washed with brine (15 ml x 1), dried over Na ₂ SO ₄ Dried and filtered and concentrated under reduced pressure to give 2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carboxylic acid (280 mg,775.98umol,83.11% yield). MS (ESI) m/z 361.0[ M+H ] ] ⁺

Step 4: (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To 2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] at 0deg.C]Decane-3-carboxylic acid (250 mg,692.84umol,1 eq.) and (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (225.82 mg,900.69umol,1.3 eq, HCl) in DCM (4 mL) was added T in one portion ₃ P (661.35 mg,1.04mmol,618.08uL,50% purity, 1.5 eq.) and TEA (210.32 mg,2.08mmol,289.30uL,3 eq.). The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative TLC (SiO) ₂ The crude material was purified with PE ethyl acetate=0:1) to give (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (270 mg,484.67umol,69.95% yield). MS (ESI) M/z557.1[ M+H] ⁺

Step 5: n- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl ] -2-oxo-ethyl ] -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (250 mg,448.77umol,1 eq.) in NH ₃ Mixing in MeOH (7M, 5mL,77.99 eq)The mixture was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl as a white solid]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (240 mg,442.75umol,98.66% yield). MS (ESI) m/z 542.2[ M+H ]] ⁺

Step 6:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl at 25 ℃]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (250 mg,392.02umol,85% purity, 1 eq.) in DCM (5 mL) was added the primary buchner reagent (186.84 mg,784.03umol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: waters Xbridge Prep OBD C: 150 x 40mM x 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl as a white solid]-2-azaspiro [4.5]]Decane-3-carboxamide (100 mg,190.82umol,48.68% yield). MS (ESI) m/z 524.2[ M+H ]] ⁺

Step 7:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,10 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl as a white solid]-2-azaspiro [4.5]]Decane-3-carboxamide (2 mg,3.82umol,2.00% yield), 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl)) Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (2 mg,3.82umol,2.00% yield), 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl]-2-azaspiro [4.5]]Decane-3-carboxamide (30 mg,57.25umol,30.00% yield), 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ]-2-azaspiro [4.5]]Decane-3-carboxamide (5 mg,9.54umol,5.00% yield), 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl]-2-azaspiro [4.5]]Decane-3-carboxamide (20 mg,38.16umol,20.00% yield) and 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl]-2-azaspiro [4.5]]Decane-3-carboxamide (15 mg). MS (ESI) m/z 524.2[ M+H ]] ⁺

Isomer 1:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69-11.44(m,1H),8.95(br d,J＝7.9Hz,1H),7.87-7.75(m,1H),7.68-7.43(m,1H),7.33-7.20(m,1H),7.14(s,1H),7.11-6.97(m,1H),4.99-4.75(m,1H),4.50(t,J＝8.6Hz,1H),3.83(br d,J＝10.4Hz,1H),3.66(d,J＝10.6Hz,1H),2.75-2.63(m,1H),2.36-2.12(m,2H),1.99(dd,J＝8.5,12.2Hz,1H),1.83-1.69(m,1H),1.60(br dd,J＝9.9,11.9Hz,1H),1.55-1.28(m,11H),1.17-1.06(m,3H),1.05-0.91(m,3H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.07(br s,1H),8.73(br d,J＝7.5Hz,1H),7.75-7.47(m,2H),7.28(d,J＝7.5Hz,1H),7.07(br t,J＝7.7Hz,2H),4.91(br d,J＝7.3Hz,1H),4.59(br s,1H),3.84(d,J＝10.1Hz,1H),3.63(br s,1H),2.30-1.92(m,3H),1.78(br s,1H),1.72-1.63(m,1H),1.60-1.33(m,12H),1.18(s,3H),1.09(s,3H)

isomer 2:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.56(br s,1H),9.00-8.79(m,1H),7.82(s,1H),7.68-7.48(m,1H),7.32-7.22(m,1H),7.15(s,1H),7.12-6.99(m,1H),4.98-4.71(m,1H),4.50(t,J＝8.7Hz,1H),3.90-3.77(m,1H),3.73-3.60(m,1H),2.47-2.39(m,1H),2.24(br dd,J＝7.9,12.3Hz,1H),2.17-2.08(m,1H),1.98(br dd,J＝8.4,11.9Hz,1H),1.83-1.68(m,1H),1.61-1.51(m,2H),1.50-1.36(m,7H),1.35-1.21(m,3H),1.15(s,3H),1.08-0.97(m,3H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.09(br s,1H),8.66(br s,1H),7.65-7.53(m,2H),7.26(d,J＝7.7Hz,1H),7.12-6.97(m,2H),4.89(br d,J＝5.7Hz,1H),4.57(br s,1H),3.88-3.56(m,2H),2.28-1.96(m,3H),1.85-1.60(m,2H),1.58-1.22(m,12H),1.16(s,3H),1.07(s,2H),1.09-0.99(m,1H)

isomer 3:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.66-11.45(m,1H),8.94(d,J＝8.2Hz,1H),7.92-7.75(m,1H),7.64(d,J＝7.9Hz,1H),7.29(d,J＝7.3Hz,1H),7.17-7.12(m,1H),7.11-6.98(m,1H),5.00-4.73(m,1H),4.50(br t,J＝8.6Hz,1H),3.83(br d,J＝10.4Hz,1H),3.72-3.62(m,1H),2.75-2.63(m,1H),2.31-2.12(m,2H),2.08-1.94(m,1H),1.80-1.57(m,2H),1.54-1.36(m,8H),1.35-1.18(m,3H),1.17-1.07(m,3H),1.07-0.92(m,3H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.07(br s,1H),8.86-8.69(m,1H),7.70-7.55(m,2H),7.28(d,J＝7.5Hz,1H),7.07(br t,J＝7.6Hz,2H),4.98-4.85(m,1H),4.60(br s,1H),3.84(d,J＝10.6Hz,1H),3.64(s,1H),2.29-1.96(m,3H),1.77(br s,1H),1.73-1.63(m,1H),1.61-1.32(m,12H),1.20-1.14(m,3H),1.13-1.06(m,3H)

isomer 4:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69-11.53(m,1H),9.11-8.97(m,1H),7.98-7.85(m,1H),7.68-7.45(m,1H),7.33-7.20(m,1H),7.15(s,1H),7.12-6.96(m,1H),4.97-4.72(m,1H),4.70-4.48(m,1H),3.83(br d,J＝10.4Hz,1H),3.72-3.59(m,1H),2.70-2.54(m,1H),2.35-2.12(m,3H),2.01-1.53(m,3H),1.53-1.39(m,6H),1.39-1.27(m,4H),1.20-1.01(m,6H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.13(br s,1H),8.85(br s,1H),7.74-7.57(m,2H),7.28(br d,J＝7.7Hz,1H),7.18-6.96(m,2H),4.90(br s,1H),4.62(br s,1H),3.85(br d,J＝10.4Hz,1H),3.64(s,1H),2.31-2.22(m,1H),2.14(br s,2H),1.89-1.75(m,1H),1.73-1.64(m,1H),1.60-1.28(m,12H),1.20(s,3H),1.14(s,3H)

isomer 5:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(br s,1H),9.02-8.77(m,1H),7.82(s,1H),7.69-7.47(m,1H),7.32-7.22(m,1H),7.15(s,1H),7.11-6.98(m,1H),4.98-4.71(m,1H),4.50(t,J＝8.5Hz,1H),3.87-3.77(m,1H),3.74-3.59(m,1H),2.47-2.40(m,1H),2.35-2.20(m,1H),2.19-2.08(m,1H),1.98(dd,J＝8.5,12.5Hz,1H),1.89-1.70(m,1H),1.69-1.52(m,2H),1.51-1.39(m,6H),1.38-1.28(m,4H),1.15(s,3H),1.07-0.97(m,3H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.12(br s,1H),8.65(br s,1H),7.67-7.52(m,2H),7.28(d,J＝7.7Hz,1H),7.14-6.92(m,2H),4.91(br d,J＝7.1Hz,1H),4.59(br s,1H),3.83(br d,J＝11.0Hz,1H),3.63(s,1H),2.31-2.20(m,1H),2.19-1.96(m,2H),1.81(br s,1H),1.68(br d,J＝10.6Hz,1H),1.61-1.34(m,12H),1.18(s,3H),1.09(s,3H)

isomer 6:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69-11.50(m,1H),9.10-8.98(m,1H),7.97-7.88(m,1H),7.68-7.45(m,1H),7.33-7.19(m,1H),7.15(s,1H),7.11-6.97(m,1H),4.96-4.71(m,1H),4.69-4.47(m,1H),3.83(br d,J＝10.1Hz,1H),3.66(d,J＝10.4Hz,1H),2.69-2.54(m,1H),2.39-2.12(m,3H),1.97-1.56(m,2H),1.55-1.47(m,3H),1.42(br s,4H),1.38-1.28(m,4H),1.21-1.01(m,6H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.12(br s,1H),8.84(br d,J＝7.3Hz,1H),7.74-7.56(m,2H),7.28(br d,J＝7.5Hz,1H),7.07(br t,J＝7.6Hz,2H),4.89(br s,1H),4.61(br s,1H),3.84(br d,J＝10.4Hz,1H),3.62(br s,1H),2.29-2.06(m,3H),1.85-1.61(m,2H),1.59-1.33(m,12H),1.20(s,3H),1.14(s,3H)

step 8:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,10 min) to obtain white solid

2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl as a white solid]-2-azaspiro [4.5]]Decane-3-carboxamide (18 mg,34.35umol,60.00% yield) and 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ]-2-azaspiro [4.5 ]]Decane-3-carboxamide (4 mg,7.63umol,13.33% yield). MS (ESI) m/z 524.2[ M+H ]] ⁺

Isomer 1:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.68-11.45(m,1H),8.95(br d,J＝7.9Hz,1H),7.86-7.76(m,1H),7.68-7.44(m,1H),7.33-7.20(m,1H),7.14(s,1H),7.11-6.97(m,1H),4.98-4.77(m,1H),4.50(br t,J＝8.5Hz,1H),3.83(br d,J＝10.1Hz,1H),3.66(br d,J＝10.1Hz,1H),2.76-2.63(m,1H),2.36-2.10(m,2H),2.05-1.94(m,1H),1.82-1.56(m,2H),1.54-1.18(m,11H),1.17-1.06(m,3H),1.05-0.92(m,3H)。

isomer 2:

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.58(br s,1H),9.10-8.90(m,1H),7.89(s,1H),7.70-7.44(m,1H),7.30(d,J＝7.5Hz,1H),7.16(s,1H),7.12-6.99(m,1H),4.94-4.82(m,1H),4.51(t,J＝8.6Hz,1H),3.81(br d,J＝10.4Hz,1H),3.70(br d,J＝10.4Hz,1H),2.30-2.10(m,2H),2.03(dd,J＝8.5,12.0Hz,1H),1.81-1.65(m,1H),1.62-1.18(m,13H),1.16-1.01(m,6H)。

example 212 Synthesis of viral protease inhibitor Compounds 820

Step 1: 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid ethyl ester (800 mg,3.37mmol,1 eq.) in THF (10 mL) and H ₂ LiOH H was added to the solution in O (5 mL) ₂ O (283.03 mg,6.74mmol,2 eq.) and then the mixture was stirred at 30℃for 10h. After completion, the pH of the reaction mixture was adjusted to about 3 with aqueous HCl (1M). The mixture was extracted with EtOAc (100 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtering, concentrating to give the product 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid as a white solid (680 mg,crude material). MS (ESI) m/z 210.0[ M+H ]] ⁺

Step 2: (S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamide) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To a solution of 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (0.68 g,3.25mmol,1 eq.) in DCM (20 mL) was added methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.24 g,3.58mmol,1.1 eq., HCl), EDCI (1.25 g,6.50mmol,2 eq.), DMAP (1.19 g,9.75mmol,3 eq.) and the mixture was stirred at 25 ℃ for 1H. After completion, by adding H ₂ O (200 mL) to quench the reaction and then extract with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ EtOAc: meoh=10:1) to give the product methyl (S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamido) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.15 g,2.11mmol,65.01% yield, 92.35% purity) as a white solid. MS (ESI) m/z 503.2[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamide) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.08 g,2.15mmol,1 eq.) was reacted in NH ₃ The solution in (7M in MeOH, 60mL,195.43 eq.) was stirred at 50deg.C for 48h. After completion, the reaction was concentrated in vacuo to give the crude product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (1.06 g, crude material) as a white solid. MS (ESI) m/z 488.2[ M+H ] ] ⁺

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (1.03 g,2.11mmol,1 eq.) in DCM (60 mL) was added the bergs reagent (1.51 g,6.34mmol,3 eq.) and the mixture stirred at 25 ℃ for 2H. After completion, the reaction was concentrated in vacuo and purified by preparative HPLC (column Waters Xbridge Prep OBD C18 150 x 40mm x 10um; mobile phase: [ water (0.05% nh ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (400 mg,845.40umol,40.01% yield, 99.23% purity) as a white solid. MS (ESI) M/z470.1[ M+H] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.95-8.94(m,1H),8.57-8.55(m,1H),7.54(br s,1H),7.36-7.33(m,1H),6.95-6.90(m,1H),6.43-6.40(m,1H),5.09-5.04(m,1H),4.52-4.41(m,1H),3.87(s,3H),3.15-3.03(m,2H),2.33-2.19(m,2H),1.89-1.75(m,3H),1.72-1.69(m,1H),1.64-1.52(m,1H),1.51-1.34(m,2H),0.86-0.76(m,1H),0.47-0.37(m,2H),0.24-0.15(m,1H),0.14-0.06(m,1H)。

EXAMPLE 213 Synthesis of viral protease inhibitor Compounds 838

Step 1: (Z) -2-azido-3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid methyl ester

A mixture of NaOMe (3.41 g,63.07mmol,2 eq.) in MeOH (40 mL) was cooled to-10deg.C, and then a mixture of 4-chloro-2-fluoro-benzaldehyde (5 g,31.53mmol,1 eq.) and methyl 2-azidoacetate (7.26 g,63.07mmol,2 eq.) with MeOH (10 mL) was added dropwise. The mixture was stirred at 20℃for 18h. After completion, by adding H at 25 DEG C ₂ O (20 mL) to quench the reaction mixture,with 100mL H ₂ O was diluted and extracted with ethyl acetate (100 ml x 2). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0) to give methyl (Z) -2-azido-3- (4-chloro-2-fluoro-phenyl) prop-2-enoate (4 g,14.87mmol,47.14% yield, 95% purity) as a white solid.

Step 2: 6-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester

A mixture of (Z) -2-azido-3- (4-chloro-2-fluoro-phenyl) prop-2-enoic acid methyl ester (4 g,15.65mmol,1 eq.) in xylene (20 mL) was stirred at 170℃for 5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The crude product was wet triturated with petroleum ether ethyl acetate=10:1 to give 6-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester as a white solid (2 g,8.35mmol,53.35% yield, 95% purity).

Step 3: 6-chloro-4-fluoro-1H-indole-2-carboxylic acid

To 6-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester (1.4 g,6.15mmol,1 eq.) in THF (10 mL) and H ₂ LiOH H was added to the mixture in O (5 mL) ₂ O (516.20 mg,12.30mmol,2 eq.). After stirring at 60℃for 2H, the pH of the reaction mixture was adjusted to 3 with HCl (1M) and then with H ₂ O (30 mL) was diluted and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound 6-chloro-4-fluoro-1H-indole-2-carboxylic acid (1.3 g,5.78mmol,94.01% yield, 95% purity) was obtained as a white solid.

Step 4: (2S) -2- [ [ (2S) -2- [ (6-chloro-4-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 6-chloro-4-fluoro-1H-indole-2-carboxylic acid (600 mg,2.81mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (977.10 mg,2.81mmol,1 eq.)EDCI (1.08 g,5.62mmol,2 eq.) and DMAP (686.36 mg,5.62mmol,2 eq.) are added to a mixture of DCM (2 mL) and DMF (1 mL). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was taken up with H ₂ O (100 mL) was diluted and extracted with ethyl acetate (100 mL x 2). The combined organic layers were washed with brine (100 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=8/1 to 0/1) to give the compound (2S) -2- [ [ (2S) -2- [ (6-chloro-4-fluoro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.1 g,1.95mmol,69.52% yield, 90% purity). MS (ESI) m/z 507.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-4-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-4-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,1.97mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 20mL,70.97 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-4-fluoro-1H-indole-2-carboxamide (950 mg,1.74mmol,88.11% yield, 90% purity). MS (ESI) m/z 492.2[ M+H ] ] ⁺

Step 6: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-4-fluoro-1H-indole-2-carboxamide (500 mg,1.02mmol,1 eq.) in DCM (20)mL) was added to the mixture. The mixture was stirred at 25℃for 4h. After completion, the reaction mixture was taken up with H ₂ O (5 mL) was diluted and extracted with DCM (20 mL x 2). The combined organic layers were concentrated using a blow dryer to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,10 min) to give 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-fluoro-1H-indole-2-carboxamide (120 mg,253.20umol,24.91% yield, 100% purity). MS (ESI) m/z 474.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.08(br s,1H),8.94(d,J＝8.0Hz,1H),8.73(d,J＝7.5Hz,1H),7.61-7.23(m,3H),6.99(d,J＝10.1Hz,1H),5.06(q,J＝8.1Hz,1H),4.57-4.37(m,1H),3.18-2.98(m,2H),2.37-2.17(m,2H),1.89-1.26(m,7H),0.89-0.65(m,1H),0.51-0.32(m,2H),0.27-0.01(m,2H)。

Example 214 Synthesis of viral protease inhibitor Compound 848

Step 1: 6-bromo-3-fluoro-1H-indole-2-carboxylic acid methyl ester

To a mixture of methyl 6-bromo-1H-indole-2-carboxylate (2 g,7.87mmol,1 eq.) in ACN (84 mL) at 25℃was added NaHCO in one portion ₃ (36.42 g,433.52mmol,16.86mL,55.07 eq.). Select F (3.07 g,8.66mmol,1 eq.) was added and stirred at 80℃for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions, column Agela DuraShell C, 250, 50mm, 10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -55%,20 min) to give methyl 6-bromo-3-fluoro-1H-indole-2-carboxylate (500 mg,1.84mmol,23.35% yield) as a yellow solid.

Step 2: 6-bromo-3-fluoro-1H-indole-2-carboxylic acid

To 6-bromo-3-fluoro-1H-indole-2-carboxylic acid methyl ester (500 mg,1.84mmol,1 eq.) in THF (5 mL) and H at 25 ℃ ₂ LiOH.H was added at once to the mixture in O (5 mL) ₂ O (154.22 mg,3.68mmol,2 eq.). The mixture was stirred at 60℃for 2h. After completion, the reaction mixture was made acidic with 1M HCl and extracted with ethyl acetate (6 ml x 3). The combined organic layers were washed with brine (9 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 6-bromo-3-fluoro-1H-indole-2-carboxylic acid (440 mg,1.71mmol,92.78% yield) as a yellow solid. (ESI) m/z 256.0[ M-H ] ⁺

Step 3: (2S) -2- [ [ (2S) -2- [ (6-bromo-3-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 6-bromo-3-fluoro-1H-indole-2-carboxylic acid (440 mg,1.71mmol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (654.28 mg,1.88mmol,1.1 eq., HCl) in DCM (8 mL) and DMF (2 mL) was added DMAP (626.73 mg,5.13mmol,3 eq.) and EDCI (655.61 mg,3.42mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and extracted with ethyl acetate (15 mL x 4). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (6-bromo-3-fluoro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (510 mg,924.91umol,54.09% yield). (ESI) m/z 551.1[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-bromo-3-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-bromo)-3-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (510 mg,924.91umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,75.68 eq.) was stirred at 55deg.C for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-bromo-3-fluoro-1H-indole-2-carboxamide (500 mg, crude). MS (ESI) m/z 536.2[ M+H ]] ⁺

Step 5: 6-bromo-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -3-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 6-bromo-3-fluoro-1H-indole-2-carboxamide (500 mg,745.72umol,80% purity, 1 eq.) in DCM (8 mL) was added the berges reagent (533.14 mg,2.24mmol,3 eq.) in one portion. The mixture was stirred at 25℃for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: phenomenex Gemini-NX C18 75X 30mm 3um; mobile phase: [ water (10 mM NH4HCO 3) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -65%,8 min) to give 6-bromo-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-3-fluoro-1H-indole-2-carboxamide (170 mg,327.95umol,43.98% yield). MS (ESI) m/z 518.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.70(s,1H),8.98(d,J＝7.9Hz,1H),7.84(dd,J＝3.1,7.3Hz,1H),7.65-7.58(m,2H),7.55(br s,1H),7.26(dd,J＝1.5,8.6Hz,1H),5.09(q,J＝8.1Hz,1H),4.57-4.49(m,1H),3.13-3.05(m,2H),2.30-2.20(m,2H),1.82(dt,J＝6.6,14.0Hz,3H),1.77-1.67(m,1H),1.64-1.51(m,2H),1.47-1.35(m,1H),0.81-0.70(m,1H),0.48-0.37(m,2H),0.21-0.07(m,2H)

EXAMPLE 215 Synthesis of viral protease inhibitor Compounds 862

Step 1: 6-cyano-1H-indole-2-carboxylic acid

To 6-cyano-1H-indole-2-carboxylic acid methyl ester (1 g,5.00mmol,1 eq.) in H ₂ LiOH.H was added to a solution of O (4 mL) and THF (8 mL) ₂ O (358.88 mg,14.99mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give 6-cyano-1H-indole-2-carboxylic acid (805 mg, crude material) as a white solid. MS (ESI) m/z 187.0[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S) -2- [ (6-cyano-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 6-cyano-1H-indole-2-carboxylic acid (776.06 mg,4.17mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.45 g,4.17mmol,1 eq., HCl) in DCM (50 mL) was added DMAP (1.53 g,12.51mmol,3 eq.) and EDCI (2.40 g,12.51mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (10 mL x 2). The combined organic layers were washed with 20mL HCl (1M) (10 mL x 2), then brine (20 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1, dichloromethane: methanol=10:1, (UV 254 nm)) to give (2S) -2- [ [ (2S) -2- [ (6-cyano-1H-indole-2-carbonyl) amino as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.3 g,2.56mmol,61.46% yield, 94.5% purity). MS (ESI) m/z 480.2[ M+H ]] ⁺

Step 3: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-cyano-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-cyano-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.50mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 20mL,55.94 eq.) was stirred at 50deg.C for 16h. The reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-cyano-1H-indole-2-carboxamide (1.1 g, crude). MS (ESI) m/z 465.2[ M+H ]] ⁺

Step 4: 6-cyano-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 6-cyano-1H-indole-2-carboxamide (1 g,2.15mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (1.03 g,4.31mmol,2 eq.). The mixture was stirred at 20℃for 6h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give 6-cyano-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (414.8 mg,929.00umol,43.15% yield, 100% purity). MS (ESI) m/z 447.2[ M+H ] ] ⁺ 。

1H NMR(400MHz,DMSO-d ₆ )δ＝12.08(s,1H),8.96(d,J＝8.4Hz,1H),8.82(d,J＝7.8Hz,1H),7.89-7.81(m,2H),7.53(s,1H),7.45-7.33(m,2H),5.07(q,J＝8.2Hz,1H),4.54-4.46(m,1H),3.17-3.01(m,2H),2.35-2.20(m,2H),1.91-1.65(m,4H),1.63-1.32(m,3H),0.88-0.73(m,1H),0.50-0.35(m,2H),0.25-0.07(m,2H)

EXAMPLE 216 Synthesis of viral protease inhibitor Compounds 866

Step 1: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 5-dichloro-1H-pyrrole-2-carbonyl) amino ] propanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

To 4, 5-dichloro-1H-pyrrole-2-carboxylic acid (300 mg,1.67mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (637.74 mg,1.83mmol,1.1 eq., HCl) in DCM (8 mL) and DMF (2 mL) was added DIEA (430.84 mg,3.33mmol,580.64uL,2 eq.), HOBt (450.44 mg,3.33mmol,2 eq.) and EDCI (639.05 mg,3.33mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and extracted with ethyl acetate (15 mL x 4). The combined organic layers were washed with brine (30 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 5-dichloro-1H-pyrrole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]Methyl propionate (550 mg,1.16mmol,69.71% yield). MS (ESI) m/z 473.1[ M+H ]] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 5-dichloro-1H-pyrrole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 5-dichloro-1H-pyrrole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg,1.06mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,132.54 eq.) was stirred at 40℃for 16 h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4, 5-dichloro-1H-pyrrole-2-carboxamide (480 mg,1.05mmol,99.14% yield). MS (ESI) m/z 458.1[ M+H ]] ⁺

Step 3:4, 5-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-pyrrole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4, 5-dichloro-1H-pyrrole-2-carboxamide (480 mg,555.05umol,53% purity, 1 eq.) in DCM (8 mL) was added at once the primary (396.82 mg,1.67mmol,3 eq.). The mixture was stirred at 25℃for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: phenomenex Gemini-NX C18 75X 30mm 3um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -50%,8 min) to give 4, 5-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-pyrrole-2-carboxamide (108 mg,245.27umol,44.19% yield). MS (ESI) m/z 440.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.74(br s,1H),8.88(d,J＝8.2Hz,1H),8.22(br d,J＝7.1Hz,1H),7.53(br s,1H),7.05(s,1H),5.10-5.00(m,1H),4.44-4.33(m,1H),3.15-3.02(m,2H),2.30-2.17(m,2H),1.91-1.65(m,4H),1.55(br dd,J＝3.5,9.9Hz,1H),1.47-1.32(m,2H),0.82-0.70(m,1H),0.45-0.34(m,2H),0.21-0.02(m,2H)

EXAMPLE 217 Synthesis of viral protease inhibitor Compounds 872

Step 1: (S) -2- ((S) -3-cyclopropyl-2- (1H-pyrazole-5-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To 1H-pyrazole-5-carboxylic acid (500 m)g,4.46mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]To a solution of methyl propionate (1.24 g,3.57mmol,0.8 eq, HCl) in DCM (40 mL) was added DMAP (1.09 g,8.92mmol,2 eq) and EDCI (1.71 g,8.92mmol,2 eq) and the mixture was then stirred at 20deg.C for 2h. After completion of the reaction, the mixture was poured into water (40 mL) and extracted with DCM (15 ml×3) and dried over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -40%,10 min) to give methyl (S) -2- ((S) -3-cyclopropyl-2- (1H-pyrazole-5-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (550 mg,1.26mmol,28.25% yield, 92.9% purity) as a white solid. MS (ESI) m/z 406.2[ M+H ]] ⁺

Step 2: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrazole-5-carboxamide

Methyl (S) -2- ((S) -3-cyclopropyl-2- (1H-pyrazole-5-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (500 mg,1.23mmol,1 eq.) was reacted in NH ₃ A solution in MeOH (20 mL, 7M) was stirred at 50deg.C for 24h. After the reaction was completed, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrazole-5-carboxamide (500 mg, crude material) as a pale yellow solid. MS (ESI) m/z 391.2[ M+H ] ] ⁺

Step 3: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrazole-5-carboxamide and methyl (5- (((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamoyl) -1H-pyrazol-1-yl) sulfonyl carbamate

To N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1To a solution of H-pyrazole-5-carboxamide (500 mg,1.28mmol,1 eq.) in DCM (8 mL) was added the Buerger's reagent (915.53 mg,3.84mmol,3 eq.) and the mixture was then stirred at 30℃for 4H. After the reaction was complete, the reaction mixture was quenched with water (1 mL) and N was used ₂ Dried and then purified by preparative HPLC (column Waters Xbridge Prep OBD C, 150 x 40mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,8 min) to give N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrazole-5-carboxamide (compound 872, 160mg,425.76umol,33.25% yield, 99.1% purity) as a white solid. MS (ESI) m/z 373.1[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 13.57-13.18(m,1H),8.91(d,J＝8.0Hz,1H),7.92(d,J＝8.0Hz,1H),7.83(s,1H),7.52(s,1H),7.02-6.59(m,1H),5.05(q,J＝7.9Hz,1H),4.48(q,J＝7.4Hz,1H),3.16-3.02(m,2H),2.31-2.17(m,2H),1.89-1.65(m,4H),1.63-1.32(m,3H),0.71(d,J＝6.4Hz,1H),0.40(d,J＝8.0Hz,2H),0.09(dd,J＝4.6,14.9Hz,2H)。

Methyl (5- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamoyl) -1H-pyrazol-1-yl) sulfonyl carbamate (20 mg,425.76umol,33.25% yield, 99.1% purity) was obtained as a white solid. MS (ESI) m/z 510.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 9.06(s,1H),8.91(d,J＝8.0Hz,1H),8.03(s,1H),7.77(s,1H),7.52(s,1H),7.21(s,1H),7.08(s,1H),7.01-6.92(m,1H),6.77-6.32(m,2H),5.05(q,J＝7.9Hz,1H),4.56-4.40(m,1H),3.47(s,3H),3.16-3.01(m,2H),2.30-2.15(m,2H),1.89-1.65(m,4H),1.63-1.31(m,3H),0.77-0.65(m,1H),0.45-0.32(m,2H),0.21-0.00(m,2H)。

EXAMPLE 218 Synthesis of viral protease inhibitor Compound 731

Step 1: (Z) -3-bromo-2- (hydroxyimino) propionic acid ethyl ester

At N ₂ Next, 3-bromo-2-oxo-propionic acid ethyl ester (167 g,428.18mmol,107.05mL,50% purity, 1 eq.) was added to CHCl ₃ NH-containing was added to the solution in (800 mL) ₂ H of OH HCl (32.73 g,471.00mmol,1.1 eq) ₂ O (800 mL). The mixture was stirred at 25℃for 16h. After completion, the reaction was extracted with DCM (1000 ml x 4). The combined organic phases were washed with brine (2000 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give ethyl (Z) -3-bromo-2- (hydroxyimino) propionate (440 g, crude material) as a white solid. MS (ESI) m/z 210.3[ M+H ]] ⁺ 。

Step 2:1- (cyclohexylidene methyl) pyrrolidine

A mixture of cyclohexane formaldehyde (100 g,891.51mmol,107.30mL,1 eq.) and pyrrolidine (82.43 g,1.16mol,96.74mL,1.3 eq.) in toluene (1.6L) was heated to 130℃for 14h and then water was removed by a dean-Stark trap. After completion, the reaction mixture was concentrated under reduced pressure at 55 ℃ to give 1- (cyclohexylenemethyl) pyrrolidine (420 g, crude material) as a yellow oil. MS (ESI) m/z 166.2[ M+H ] ] ⁺ 。

at-20deg.C, at N ₂ To a solution of 1- (cyclohexylmethyl) pyrrolidine (140 g,847.08mmol,1 eq.) in THF (1000 mL) was added dropwise a solution of (2Z) -3-bromo-2-hydroxyimino-propionic acid ethyl ester (177.91 g,847.08mmol,1 eq.) in THF (1000 mL). After 1h, at-20℃under N ₂ TEA (85.72 g,847.08mmol,117.90mL,1 eq.) was added dropwise. At N ₂ The reaction mixture was stirred at 25℃for 12h. After completion, the residue was poured into HCl (2 m,2500 ml) and stirred for 30min and extracted with ethyl acetate (1500 ml x 4). The combined organic layers were washed with brine (2000 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 1/1)The residue was purified to give 1-hydroxy-2-oxa-3-azaspiro [5.5] as a yellow oil]Undec-3-ene-4-carboxylic acid ethyl ester (200 g,497.34mmol,19.57% yield, 60% purity). MS (ESI) m/z 242.2[ M+H ]] ⁺ 。

Step 4: 2-azaspiro [4.5] decane-3-carboxylic acid ethyl ester

In Ar ₂ Next, 1-hydroxy-2-oxa-3-azaspiro [5.5]]To a solution of ethyl undec-3-ene-4-carboxylate (20 g,49.73mmol,60% purity, 1 eq.) in EtOH (150 mL) was added Raney nickel (12.00 g,140.07mmol,2.82 eq.). The suspension was degassed in vacuo and purified by H ₂ (100.46 mg,49.73mmol,1 eq.) several times. The mixture is put in H ₂ (100.46 mg,49.73mmol,1 eq.) at 50psi, for 18h at 50 ℃. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=20/1 to ethyl acetate: meOH=10/1) purification of the residue gives 2-azaspiro [4.5] as a yellow oil]Decane-3-carboxylic acid ethyl ester (35 g,165.64mmol,33.31% yield). MS (ESI) m/z 212.2[ M+H ]] ⁺ 。

At 0℃to 2-azaspiro [4.5]]To a solution of decane-3-carboxylic acid ethyl ester (35 g,132.51mmol,80% purity, 1 eq.) in DCM (350 mL) was added Boc ₂ O (34.70 g,159.02mmol,36.53mL,1.2 eq.) and TEA (26.82 g,265.03mmol,36.89mL,2 eq.). The mixture was stirred at 25℃for 14h. After completion, by adding H ₂ O (400 mL) to quench the reaction mixture and extract with ethyl acetate (200 mL x 3). The combined organic layers were washed with brine (300 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 10/1) purification of the residue to give 2-azaspiro [4.5] as a yellow oil ]Decane-2, 3-dicarboxylic acid 2-tert-butyl 3-ethyl ester (40 g,95.05mmol,71.73% yield, 74% purity). MS (ESI) m/z 312.2[ M+H ]] ⁺ 。

Step 6:2- (tert-Butoxycarbonyl) -2-azaspiro [4.5] decane-3-carboxylic acid

To 2-azaspiro [4.5]]2-tert-butyl 3-ethyl decane-2, 3-dicarboxylic acid (40 g,128.45mmol,1 eq.) in H ₂ To a solution of O (120 mL) and MeOH (480 mL) was added LiOH.H ₂ O (16.17 g,385.34mmol,3 eq.). The mixture was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH. Residue with H ₂ O (800 mL) was diluted and extracted with ethyl acetate (500 mL x 2). HCl (aqueous solution) was added to the aqueous phase to adjust the pH to 2 and extracted with ethyl acetate (900 ml x 3). The combined organic layers were washed with brine (900 mL), and dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to give 2-tert-butoxycarbonyl-2-azaspiro [4.5] as a yellow oil]Decane-3-carboxylic acid (35 g, crude material). MS (ESI) m/z 284.2[ M+H ]] ⁺

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (22.97 g,97.05mmol,1.1 eq. HCl) and 2-tert-butoxycarbonyl-2-azaspiro [4.5] ]To a solution of decane-3-carboxylic acid (25 g,88.23mmol,1 eq.) in DCM (400 mL) was added DMAP (21.56 g,176.45mmol,2 eq.) and EDCI (25.37 g,132.34mmol,1.5 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction was quenched with 0.5M HCl (400 mL) and then extracted with DCM (150 mL x 3). The combined organic phases were washed with brine (150 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/1 to 0/1) to give 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (27 g,47.84mmol,54.23% yield, 82.5% purity). MS (ESI) m/z 466.2[ M+H ]] ⁺

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperaquineBoydo group]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]A solution of tert-butyl decane-2-carboxylate (27 g,47.84mmol,82.5% purity, 1 eq.) in HCl/MeOH (300 mL). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove HCl/MeOH, and DCM (150 mL) was added (three times), concentrated under reduced pressure to give (2S) -2- (2-azaspiro [ 4.5) as a yellow solid ]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (25 g, crude material, HCl). MS (ESI) m/z 366.3[ M+H ]] ⁺

Step 9: (2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (25 g,62.20mmol,1 eq, HCl) and 7-chloro-1H-indole-2-carboxylic acid (13.38 g,68.42mmol,1.1 eq) in DCM (400 mL) was added EDCI (17.89 g,93.30mmol,1.5 eq) and DMAP (15.20 g,124.40mmol,2 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction was quenched with 0.5M HCl (400 mL) and then extracted with DCM (300 mL x 2). The combined organic phases were washed with brine (400 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/1 to 0/1) to give (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (25 g,44.19mmol,71.05% yield, 96% purity). MS (ESI) m/z 543.3[ M+H ] ] ⁺

Step 10: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] in a 100mL autoclave]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (5 g,8.84mmol,96% purity, 1 eq.) in NH ₃ The solution in (7M in MeOH, 57.60mL,45.62 equivalents) (15 Psi) was stirred at 65℃for 16h. At the endAfter completion, the reaction mixture was concentrated under reduced pressure to remove NH ₃ MeOH, and DCM (300 mL) was added (three times), concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (24 g, crude). MS (ESI) m/z 528.3[ M+H ]] ⁺

Step 11:2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (12 g,22.73mmol,1 eq.) in DCM (200 mL) was added the Buerger reagent (11.91 g,50.00mmol,2.2 eq.). The mixture was stirred at 25℃for 2h. By adding H at 20 ℃ ₂ O (10 mL) to quench the reaction mixture, and then N ₂ The solvent was removed. By preparative HPLC (column: phenomenex Titank C Bulk 250. Times.100 mM 10u; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,20 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (17 g,99.29% purity). MS (ESI) m/z 510.3[ M+H ]] ⁺

Step 12:2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

Isomer 1:

by SFC (conditions: column: REGIS (s, s) WHELK-O1 (250 mm. Times.50 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,9.5 min) to further isolate the desired compound to give 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (6.1 g,11.96mmol,26.31% yield). MS (ESI) m/z 510.3[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.64-11.51(m,1H),8.98-8.86(m,1H),7.70-7.38(m,2H),7.32-7.21(m,1H),7.16-6.69(m,2H),5.08-4.47(m,2H),3.88-3.76(m,1H),3.70-3.60(m,1H),3.27-2.93(m,2H),2.35(br s,3H),1.88-1.31(m,16H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.28-11.09(m,1H),8.82-8.62(m,1H),7.73-7.52(m,1H),7.37-7.22(m,2H),7.20-6.96(m,2H),5.08-4.86(m,1H),4.70-4.46(m,1H),3.88-3.78(m,1H),3.70-3.51(m,1H),3.14-3.09(m,2H),2.40-2.13(m,3H),1.87-1.37(m,16H)。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.67-7.46(m,1H),7.32-7.22(m,1H),7.14-6.81(m,2H),5.16-4.97(m,1H),4.83-4.58(m,1H),3.98-3.81(m,1H),3.76-3.38(m,1H),3.27-2.98(m,2H),2.67-2.20(m,3H),2.05-1.43(m,16H)。

Isomer 2:

obtaining 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Ethyl group ]-2-azaspiro [4.5 ]]Decane-3-carboxamide (7 g,13.72mmol,30.20% yield). MS (ESI) m/z 510.3[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.59-11.47(m,1H),8.98-8.77(m,1H),7.69-7.63(m,1H),7.54-7.46(m,1H),7.32-7.23(m,1H),7.18-6.68(m,2H),5.06-4.84(m,1H),4.80-4.47(m,1H),3.90-3.78(m,1H),3.74-3.61(m,1H),3.28-3.00(m,2H),2.33-2.09(m,1H),2.08-2.06(m,1H),1.88-1.32(m,16H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.26-11.02(m,1H),8.74-8.57(m,1H),7.76-7.51(m,1H),7.32-7.21(m,2H),7.17-6.93(m,2H),5.07-4.87(m,1H),4.73-4.51(m,1H),3.87-3.79(m,1H),3.73-3.52(m,1H),3.08(s,2H),2.29-2.12(m,3H),1.86-1.38(m,16H)。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.72-7.52(m,1H),7.33-7.19(m,1H),7.14-6.79(m,2H),5.09-4.92(m,1H),4.70-4.54(m,1H),3.99-3.89(m,1H),3.83-3.40(m,1H),3.22-3.00(m,2H),2.57-2.12(m,3H),2.01-1.40(m,16H)。

EXAMPLE 219 Synthesis of viral protease inhibitor Compound 900

Step 1: (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (1 g,3.24mmol,85% purity, 1.1 eq., HCl) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (918.04 mg,2.95mmol,1 eq.) in DMF (15 mL) was added PyBOP (1.53 g,2.95mmol,1 eq.). TEA-containing DMF (5 mL) was added (895.02 mg,8.85mmol,1.23mL,3 eq.) and the mixture was then stirred at-40℃for 2h. After completion, the mixture was quenched with water (60 mL) and extracted with DCM (20 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) to give methyl (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.5 g,2.02mmol,68.62% yield, 70% purity) as a brown gum. MS (ESI) m/z 519.2[ M+H ] ] ⁺

Step 2: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (350 mg,674.39umol,1 eq.) was reacted with NH ₃ A solution in MeOH (7M, 4 mL) was stirred at 50℃for 20h. After completion, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (1.3 g, crude material) as a brown gum. MS (ESI) m/z 502.1[ M-H] ⁺

Step 3: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (1.3 g,2.58mmol,1 eq.) in DCM (20 mL) was added the bergs reagent (1.84 g,7.74mmol,3 eq.) at 30 ℃. The resulting mixture was stirred at 30℃for 2h. After completion, the mixture was quenched with water (2 mL) and purified by blowing N ₂ To be dried. The mixture was purified by prep HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-methoxy-1H-indole-2-carboxamide (260 mg,535.02umol,20.74% yield, 100% purity) as a white solid. MS (ESI) m/z 486.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.57(s,1H),8.99(d,J＝8.0Hz,1H),8.65(d,J＝7.6Hz,1H),7.53(s,1H),7.15(dd,J＝2.2,11.3Hz,2H),7.00(d,J＝2.2Hz,1H),5.07(q,J＝8.0Hz,1H),4.58-4.44(m,1H),3.84-3.72(m,3H),3.17-3.00(m,2H),2.30-2.20(m,2H),1.91-1.65(m,4H),1.64-1.33(m,3H),0.87-0.73(m,1H),0.50-0.35(m,2H),0.26-0.05(m,2H)。

Example 220 Synthesis of viral protease inhibitor Compound 908

Step 1: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]A solution of tert-butyl decane-2-carboxylate (1.5 g,3.22mmol,1 eq.) in HCl/MeOH (15 mL) was stirred at 20deg.C for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give a white solid(2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.25 g, crude material, HCl). MS (ESI) m/z 366.2[ M+H ]] ⁺

Step 2: (2S) -2- [ [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.25 g,3.10mmol,1 eq, HCl) and 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (700 mg,3.10mmol,1 eq) in DCM (40 mL) was added DMAP (1.14 g,9.31mmol,3 eq). After EDCI (1.78 g,9.31mmol,3 eq.) was added, the mixture was stirred for 1h at 20 ℃. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (15 mL x 2). The combined organic layers were washed with HCl (1M) (10 mL x 2), and then the combined organic layers were washed with brine (20 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=3/1 to 0/1, dichloromethane: methanol=10:1, (UV 254 nm)) the residue was purified to give (2S) -2-azaspiro [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,1.96mmol,63.24% yield, 93.7% purity). MS (ESI) m/z 573.2[ M+H ]] ⁺

Step 3: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.1 g,1.92mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 60mL,218.81 eq.) was stirred at 20deg.C for 16h. The reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-2- (6-chloro-4-methoxy-1)H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.05 g, crude). MS (ESI) m/z 558.2[ M+H ]] ⁺

Step 4:2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (1.04 g,1.86mmol,1 eq.) in DCM (20 mL) was added the Buerger reagent (888.20 mg,3.73mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -70%,10 min) to give 2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (500 mg,886.03umol,47.54% yield, 95.7% purity). MS (ESI) m/z 540.2[ M+H ]] ⁺ 。

Step 5:2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

Isomer 1:

by SFC (column: REGIS (s, s) WHELK-O1 (250 mm. Times.30 mm,5 um); mobile phase: [0.1% NH) ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,6 min) isolation of 2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (500 mg) gave 2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (66.7 mg,123.01umol,13.29% yield, 99.6% purity). MS (ESI) m/z 540.2[ M+H ]] ⁺ 。

1H NMR(400MHz,DMSO-d ₆ )δ＝11.76-11.61(m,1H),8.88(d,J＝8.4Hz,1H),7.67-7.33(m,1H),7.14-6.86(m,2H),6.67-6.48(m,1H),5.06-4.87(m,1H),4.49(t,J＝8.8Hz,1H),3.92(s,2H),3.88-3.80(m,1H),3.66(d,J＝10.3Hz,1H),3.33(s,6H),2.38-2.17(m,2H),2.03-0.83(m,14H)

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.46(s,1H),8.71(s,1H),7.27(s,1H),7.09(s,2H),6.55(s,1H),4.97(s,1H),4.61(s,1H),3.92(s,2H),3.85(d,J＝10.4Hz,1H),3.61(s,1H),3.08(s,6H),2.38-2.12(m,2H),2.01-1.02(m,14H)

Isomer 2:

obtaining 2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (111.6 mg,206.65umol,22.32% yield, 100% purity). MS (ESI) m/z 540.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.76-11.64(m,1H),8.81(d,J＝8.4Hz,1H),7.53-7.41(m,1H),7.06(s,1H),6.97(s,1H),6.65-6.51(m,1H),5.04-4.86(m,1H),4.58-4.38(m,1H),3.92(s,2H),3.84(d,J＝9.8Hz,1H),3.76-3.57(m,1H),3.33(s,6H),2.24-2.11(m,2H),1.88-1.10(m,14H)

1H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.48(s,1H),8.64(s,1H),7.23(s,1H),7.09(s,1H),6.93(s,1H),6.56(s,1H),4.97(s,1H),4.59(s,1H),3.93(s,2H),3.85(d,J＝10.8Hz,1H),3.65(s,1H),3.08(s,6H),2.20(s,2H),2.01-1.23(m,14H)

EXAMPLE 221 Synthesis of viral protease inhibitor Compound 1057

Step 1: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ [4- [2- (2-methoxyethoxy) ethoxy ] -1H-indole-2-carbonyl ] amino ] propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

At 25℃to 4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxylic acid (500 mg,1.79mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (684.99 mg,1.97mmol, 1).1 eq, HCl) to a mixture of DCM (9 mL) and DMF (3 mL) was added DMAP (656.15 mg,5.37mmol,3 eq.) and EDCI (686.39 mg,3.58mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was taken up with H ₂ O (10 mL) was diluted and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (15 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=3/1 to 0/1 to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ [4- [2- (2-methoxyethoxy) ethoxy ] ethoxy as a yellow solid ]-1H-indole-2-carbonyl]Amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,1.48mmol,82.91% yield). MS (ESI) m/z 573.3[ M+H ]] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4- [2- (2-methoxyethoxy) ethoxy ] -1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ [4- [2- (2-methoxyethoxy) ethoxy ] ethoxy]-1H-indole-2-carbonyl]Amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,1.41mmol,95% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 25mL,124.10 eq.) was stirred at 55deg.C for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide (781 mg,1.22mmol,86.41% yield, 87% purity). MS (ESI) m/z 558.3[ M+H ]] ⁺

Step 3: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4- [2- (2-methoxyethoxy) ethoxy ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃C]Methyl group]Ethyl group]Amino group]-1-(Cyclopropylmethyl) -2-oxo-ethyl]-4- [2- (2-methoxyethoxy) ethoxy ]]To a mixture of 1H-indole-2-carboxamide (730 mg,1.14mmol,87% purity, 1 eq.) in DCM (10 mL) was added the Bungeus reagent (542.82 mg,2.28mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: waters Xbridge Prep OBD C: 150 x 40mm x 10um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4- [2- (2-methoxyethoxy) ethoxy ]]-1H-indole-2-carboxamide (230 mg,426.22umol,37.42% yield). MS (ESI) m/z 540.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.56(s,1H),8.89(d,J＝8.2Hz,1H),8.58(d,J＝7.5Hz,1H),7.52(br s,1H),7.39(d,J＝1.5Hz,1H),7.11-6.99(m,2H),6.50(d,J＝7.5Hz,1H),5.10-5.01(m,1H),4.49-4.41(m,1H),4.21(t,J＝4.4Hz,2H),3.86-3.79(m,2H),3.63(dd,J＝3.7,5.7Hz,2H),3.49(dd,J＝3.7,5.5Hz,2H),3.26(s,3H),3.13-3.03(m,2H),2.36-2.20(m,2H),1.90-1.76(m,3H),1.75-1.65(m,1H),1.62-1.50(m,1H),1.49-1.34(m,2H),0.88-0.75(m,1H),0.48-0.33(m,2H),0.24-0.07(m,2H)

Example 221a Synthesis of viral protease inhibitor Compounds 822

Step 1: (Z) -2-azido-3- (2-chloro-3-fluoro-phenyl) prop-2-enoic acid methyl ester

A mixture of NaOMe (3.41 g,63.07mmol,2 eq.) in MeOH (30 mL) was cooled to-10deg.C, and a mixture of 2-chloro-3-fluoro-benzaldehyde (5 g,31.53mmol,1 eq.) and ethyl 2-azidoacetate (8.14 g,63.07mmol,7.21mL,2 eq.) in MeOH (100 mL) was added dropwise and the mixture stirred at 25deg.C for 18h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue, which was used 60mL H2O was diluted and extracted with 90mL EA (30 mL. Times.3). The combined organic layers were washed with 45mL brine (45 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=1/0) to give methyl (Z) -2-azido-3- (2-chloro-3-fluoro-phenyl) prop-2-enoate (2.5 g,9.78mmol,31.01% yield) as a yellow solid.

Step 2: 4-chloro-5-fluoro-1H-indole-2-carboxylic acid methyl ester

A mixture of (Z) -2-azido-3- (2-chloro-3-fluoro-phenyl) prop-2-enoic acid methyl ester (2.3 g,9.00mmol,1 eq.) in xylene (25 mL) was stirred at 170℃for 1h. After completion, the reaction mixture was filtered to give methyl 4-chloro-5-fluoro-1H-indole-2-carboxylate (1.4 g,6.15mmol,68.36% yield) as a white solid.

Step 3: 4-chloro-5-fluoro-1H-indole-2-carboxylic acid

To 4-chloro-5-fluoro-1H-indole-2-carboxylic acid methyl ester (1.4 g,6.15mmol,1 eq.) in THF (7 mL) and H at 25 ℃ ₂ LiOH.H was added at once to the mixture in O (7 mL) ₂ O (516.20 mg,12.30mmol,2 eq.). The mixture was stirred at 60℃for 1 hour. After completion, the reaction mixture was made acidic with 1M HCl solution and extracted with 45mL EA (15 mL x 3). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 4-chloro-5-fluoro-1H-indole-2-carboxylic acid (1 g,4.68mmol,76.12% yield) as a white solid. (ESI) M/z211.9[ M-H] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (4-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 4-chloro-5-fluoro-1H-indole-2-carboxylic acid (500 mg,2.34mmol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl [ (2S) -2 at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (895.68 mg,2.57mmol,1.1 eq., HCl) in DCM (10 mL) and DMF (3 mL) was added DMAP (857.96 mg,7.02mmol,3 eq.) and EDCI (897.50 mg,4.68mmol,2 eq.) in one portion. Stirring at 25deg.CThe mixture was stirred for 2 hours. After completion, the reaction mixture was diluted with 30mL h2o and extracted with 60mL EA (20 mL x 3). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] as a white solid ]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (800 mg,1.58mmol,67.41% yield). MS (ESI) m/z 505.0[ M-H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-5-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-5-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (800 mg,1.58mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,88.72 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-5-fluoro-1H-indole-2-carboxamide (730 mg,1.35mmol,85.57% yield, 91% purity). MS (ESI) M/z492.2[ M+H ]] ⁺

Step 6: 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-1H-indole-2-carboxamide

To a mixture of N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-5-fluoro-1H-indole-2-carboxamide (730 mg,1.26mmol,85% purity, 1 eq.) in DCM (20 mL) was added at once the bergs reagent (1.05 g,4.41mmol,3.5 eq.) at 25 ℃. The mixture was stirred at 25℃for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude material. The crude material was purified by preparative HPLC (neutral conditions; column: phenomenex Gemini-NX C18X 30mm X3 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN ]; B%:25% -55%,8 min) to give 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-1H-indole-2-carboxamide (300 mg,633.01umol,50.19% yield) as a white solid. MS (ESI) m/z 474.1[ M+H ]

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.06(br s,1H),8.94(d,J＝8.2Hz,1H),8.81(d,J＝7.5Hz,1H),7.54(br s,1H),7.47(s,1H),7.40(dd,J＝4.0,9.0Hz,1H),7.23(t,J＝9.4Hz,1H),5.11-5.03(m,1H),4.51-4.42(m,1H),3.09(br s,2H),2.31-2.20(m,2H),1.92-1.76(m,3H),1.76-1.64(m,1H),1.56(br d,J＝3.3Hz,1H),1.51-1.33(m,2H),0.88-0.76(m,1H),0.49-0.35(m,2H),0.26-0.05(m,2H)

EXAMPLE 222 Synthesis of viral protease inhibitor Compounds 824

Step 1: (Z) -2-azido-3- (2-chloro-4-fluorophenyl) acrylic acid methyl ester

A solution of NaOMe (13.63 g,252.27mmol,4 eq.) in MeOH (50 mL) was then added at-10deg.C with MeOH (50 mL) containing 2-chloro-4-fluoro-benzaldehyde (10 g,63.07mmol,1 eq.) and methyl 2-azidoacetate (30.49 g,264.89mmol,4.2 eq.). The mixture was stirred at 20℃for 18h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give methyl (Z) -2-azido-3- (2-chloro-4-fluoro-phenyl) prop-2-enoate (7 g, crude material) as a yellow solid.

Step 2: 4-chloro-6-fluoro-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (2-chloro-4-fluoro-phenyl) prop-2-enoic acid methyl ester (6 g,23.47mmol,1 eq.) in xylene (70 mL). The mixture was stirred at 170℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 8/1) to give a yellow solid4-chloro-6-fluoro-1H-indole-2-carboxylic acid methyl ester (2 g,8.79mmol,37.44% yield) in the form of a solid. MS (ESI) M/z228.1[ M+H ]] ⁺ 。

Step 3: 4-chloro-6-fluoro-1H-indole-2-carboxylic acid

To 4-chloro-6-fluoro-1H-indole-2-carboxylic acid methyl ester (2 g,8.79mmol,1 eq.) in THF (20 mL) and H ₂ LiOH.H was added to the solution in O (10 mL) ₂ O (1.11 g,26.36mmol,3 eq.). The mixture was stirred at 50℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. Next, 1M HCl was added, adjusted to ph=3, followed by filtration and concentration under reduced pressure to give 4-chloro-6-fluoro-1H-indole-2-carboxylic acid (1.6 g, crude material) as a yellow solid. MS (ESI) m/z 214.0[ M+H ]] ⁺ 。

Step 4: (S) -2- ((S) -2- (4-chloro-6-fluoro-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

4-chloro-6-fluoro-1H-indole-2-carboxylic acid (1 g,4.68mmol,1 eq), (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (1.63 g,4.68mmol,1 eq.) and DMAP (1.72 g,14.05mmol,3 eq.) in DCM (10 mL) was added followed by EDCI (1.80 g,9.36mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 35mL of H at 20 ℃ ₂ O, and then extracted with DCM (35 ml x 3). The combined organic layers were washed with 1M HCl (40 ml x 2), then brine (40 ml x 2), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-6-fluoro-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.1 g,2.17mmol,46.35% yield, 100% purity). MS (ESI) m/z 507.2[ M+H ]] ⁺ 。

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-chloro-6-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-6-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.06 g,2.09mmol,1 eq.) in NH ₃ Solution in MeOH (7M, 20mL,66.96 eq.). The mixture was stirred at 20℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-6-fluoro-1H-indole-2-carboxamide (1 g, crude). MS (ESI) m/z 492.2[ M+H ] ] ⁺ 。

Step 6: 4-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 4-chloro-6-fluoro-1H-indole-2-carboxamide (480 mg,1.99mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (949.46 mg,3.98mmol,2 eq.). The mixture was stirred at 30℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mm. Times.10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -60%,10 min) to give 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-fluoro-1H-indole-2-carboxamide (375 mg,791.26umol,39.72% yield, 100% purity). MS (ESI) m/z 474.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.04(s,1H),8.94(d,J＝8.1Hz,1H),8.77(d,J＝7.5Hz,1H),7.53(s,1H),7.45(s,1H),7.15(d,J＝9.4Hz,2H),5.07(d,J＝7.9Hz,1H),4.46(d,J＝5.7Hz,1H),3.16-2.98(m,2H),2.26(d,J＝9.0Hz,2H),1.97-1.63(m,4H),1.46(s,3H),0.81(dd,J＝5.7,7.7Hz,1H),0.41(dd,J＝3.5,7.5Hz,2H),0.26-0.03(m,2H)。

EXAMPLE 223 Synthesis of viral protease inhibitor Compound 828

Step 1: 7-chloro-6-fluoro-1H-indole

At-40 ℃ under N ₂ To a mixture of 2-chloro-1-fluoro-3-nitro-benzene (10 g,56.97mmol,1 eq.) in THF (100 mL) was added dropwise magnesium bromide (1 m,199.38mL,3.5 eq.). At N ₂ The mixture was stirred at-40℃for 2h. After completion, by adding NH ₄ The reaction was quenched with Cl (500 mL) and then extracted with EtOAc (300 mL x 2). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ EtOAc: meoh=10:1) to give the product 7-chloro-6-fluoro-1H-indole (4.8 g,25.47mmol,44.72% yield, 90% purity) as a yellow oil. MS (ESI) m/z 170.0[ M+H ]] ⁺

Step 2: 7-chloro-6-fluoro-indole-1-carboxylic acid tert-butyl ester

At 20℃under N ₂ Boc was added to a mixture of 7-chloro-6-fluoro-1H-indole (4.8 g,28.30mmol,1 eq.) in DCM (50 mL) ₂ O (6.80 g,31.14mmol,7.15mL,1.1 eq.), TEA (3.44 g,33.97mmol,4.73mL,1.2 eq.) and DMAP (691.60 mg,5.66mmol,0.2 eq.). The mixture was stirred at 20℃for 1.5h. After completion, the reaction mixture was poured into water (50 mL) and extracted with DCM (40 mL x 2). The combined organic layers were concentrated under reduced pressure and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=50/1 to 20/1) to give the product 7-chloro-6-fluoro-indole-1-carboxylic acid tert-butyl ester (6 g,22.25mmol,78.60% yield) as a white solid. MS (ESI) M/z270.0[ M+H ] ⁺

Step 3: 1-Boc-7-chloro-6-fluoro-indole-2-carboxylic acid

At-60 ℃ under N ₂ To a mixture of 7-chloro-6-fluoro-indole-1-carboxylic acid tert-butyl ester (2.3 g,8.53mmol,1 eq.) in THF (25 mL) was added LDA (2 m,7.25mL,1.7 eq.). The mixture was stirred at-60℃for 2h, then the solution was addedWas added to solid carbon dioxide (18.77 g,426.50mmol,50 eq.) and left to stand at 20℃for 1h. After completion, at N ₂ The reaction mixture was poured into water (100 mL) and stirred for 10min. 1M HCl was added to the aqueous phase at 0deg.C to pH 3-4 and extracted with ethyl acetate (50 mL. Times.3). The combined organic phases were washed with brine (80 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. Wet milling with petroleum ether ethyl acetate=50:1 (100 mL) afforded the product 1-tert-butoxycarbonyl-7-chloro-6-fluoro-indole-2-carboxylic acid (1.5 g,4.78mmol,56.06% yield) as a white powder. MS (ESI) m/z 314.0[ M+H ]] ⁺

Step 4: 7-chloro-6-fluoro-1H-indole-2-carboxylic acid

A solution of 1-tert-butoxycarbonyl-7-chloro-6-fluoro-indole-2-carboxylic acid (4.3 g,13.71mmol,1 eq.) in HCl/EtOAc (4M, 50mL,14.59 eq.) was stirred at 30℃for 40h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product 7-chloro-6-fluoro-1H-indole-2-carboxylic acid (2.9 g, crude material) as a white solid. MS (ESI) m/z 212.0[ M+H ] ] ⁺

Step 5: (2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 7-chloro-6-fluoro-1H-indole-2-carboxylic acid (0.7 g,3.28mmol,1.5 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl [ (2S) -2]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (759.97 mg,2.18mmol,1 eq., HCl) in DCM (7 mL) were added EDCI (837.67 mg,4.37mmol,2 eq.) and DMAP (800.77 mg,6.55mmol,3 eq.). The solution was stirred at 20℃for 1h. After completion, by adding H ₂ O (40 mL) to quench the mixture and extract with DCM (10 mL x 4). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure, and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/1 to 0/1 and then DCM: meOH=5:1) to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole-2-carbonyl) amino as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g,1.50mmol, 68.62%)Yield, 95% purity). MS (ESI) m/z 505.1[ M+H ]] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-6-fluoro-1H-indole-2-carboxamide

Containing (2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH of methyl propionate (0.78 g,1.54mmol,1 eq.) ₃ MEOH (15 mL) was stirred at 50deg.C for 24h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-6-fluoro-1H-indole-2-carboxamide (0.75 g, crude). MS (ESI) m/z 492.2[ M+H ]] ⁺

Step 7: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To DCM (10 mL) of 7-chloro-6-fluoro-1H-indole-2-carboxamide (0.7 g,1.31mmol,92% purity, 1 eq.) was added a Bolus reagent (935.91 mg,3.93mmol,3 eq.). The mixture was stirred at 30℃for 12h. After completion, the reaction was quenched with water (2 mL) and quenched with N ₂ Blow dried and purified by preparative HPLC (column: waters Xbridge C18 150X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give the product 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-fluoro-1H-indole-2-carboxamide (0.23 g,480.45umol,36.70% yield, 99% purity). MS (ESI) m/z 474.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.99(d,J＝7.9Hz,1H),8.68(d,J＝7.5Hz,1H),7.66(dd,J＝4.9,8.7Hz,1H),7.53(br s,1H),7.29(s,1H),7.18-7.07(m,1H),5.07(q,J＝7.9Hz,1H),4.59-4.42(m,1H),3.18-3.04(m,2H),2.32-2.18(m,2H),2.07(s,1H),1.93-1.76(m,3H),1.71(dt,J＝4.0,8.9Hz,1H),1.63-1.34(m,3H),0.89-0.74(m,1H),0.53-0.36(m,2H),0.24-0.16(m,1H),0.15-0.06(m,1H)。

EXAMPLE 224 Synthesis of viral protease inhibitor Compounds 830

Step 1: 7-chloro-5-fluoro-1H-indole-1-carboxylic acid tert-butyl ester

At N ₂ To a solution of 7-chloro-5-fluoro-1H-indole (4.5 g,26.54mmol,1 eq.) and TEA (3.22 g,31.84mmol,4.43mL,1.2 eq.) in DCM (20 mL) was added DMAP (648.36 mg,5.31mmol,0.2 eq.) and Boc ₂ O (6.37 g,29.19mmol,6.71mL,1.1 eq.) followed by stirring the mixture at 20deg.C for 2h. After completion, by adding 100mL H ₂ O quench the reaction mixture and then extract with 150ml DCM (50 ml x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 15:1) to give the product 7-chloro-5-fluoro-indole-1-carboxylic acid tert-butyl ester (6 g,21.13mmol,79.65% yield, 95% purity) as a yellow oil.

Step 2:1- (tert-Butoxycarbonyl) -7-chloro-5-fluoro-1H-indole-2-carboxylic acid

At-60 ℃ under N ₂ To a mixture of 7-chloro-5-fluoro-indole-1-carboxylic acid tert-butyl ester (3 g,11.12mmol,1 eq.) in THF (40 mL) was added LDA (2 m,7.23mL,1.3 eq.). The mixture was stirred at-60 ℃ for 1.5h, then the above solution was added to solid carbon dioxide (24.48 g,556.18mmol,50 eq.) and allowed to stand at 20 ℃ for 0.5h. After completion, at N ₂ The reaction mixture was poured into ice water (100 mL) and stirred for 10min. To the aqueous phase was added 1M HCl to pH 3-4 at 0 ℃ and extracted with ethyl acetate (60 ml x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give the product 1-tert-butoxycarbonyl-7-chloro-5-fluoro-indole-2-carboxylic acid (1.8 g,5.74mmol,51.58% yield, N/a purity) as a white solid.

Step 3: 7-chloro-5-fluoro-1H-indole-2-carboxylic acid

A solution of 1-tert-butoxycarbonyl-7-chloro-5-fluoro-indole-2-carboxylic acid (1 g,3.19mmol,1 eq.) in HCl/EtOAc (4M, 40.00mL,50.19 eq.) and the mixture was then stirred at 20deg.C for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the product 7-chloro-5-fluoro-1H-indole-2-carboxylic acid (660 mg, crude material, HCl) as a yellow solid.

Step 4: (S) -2- ((S) -2- (7-chloro-5-fluoro-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

7-chloro-5-fluoro-1H-indole-2-carboxylic acid (660 mg,2.64mmol,1 eq. HCl) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (1.07 g,3.43mmol,1.3 eq.) in DMF (5 mL) and DCM (20 mL) and then DMAP (967.38 mg,7.92mmol,3 eq.) and EDCI (1.01 g,5.28mmol,2 eq.) were added and the mixture was stirred at 20deg.C for 2h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (770 mg,1.41mmol,53.52% yield, 93% purity). MS (ESI) m/z 507.2[ M+H ] ] ⁺ 。

Step 5: (S) -2- ((S) -2- (7-chloro-5-fluoro-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

(2S) -2- [ [ (2S) -2- [ (7-chloro-5- ]fluoro-1H-indole-2-carbonyl) amino]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (770 mg,1.52mmol,1 eq.) in NH3/MeOH (7M, 40.00mL,184.35 eq.) and then the mixture was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-5-fluoro-1H-indole-2-carboxamide (720 mg, crude). MS (ESI) M/z492.2[ M+H ]] ⁺ 。

Step 6: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-5-fluoro-1H-indole-2-carboxamide (660 mg,1.34mmol,1 eq.) in DCM (15 mL) was added the bergs reagent (639.44 mg,2.68mmol,2 eq.) and the mixture was then stirred at 30 ℃ for 4.5H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product 7-chloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-fluoro-1H-indole-2-carboxamide (232.57 mg,490.73umol,36.58% yield, 100% purity). MS (ESI) m/z 474.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.87(s,1H),9.00(d,J＝7.9Hz,1H),8.74(br d,J＝7.6Hz,1H),7.53(br s,1H),7.47(dd,J＝2.2,9.3Hz,1H),7.33(dd,J＝2.2,9.3Hz,1H),7.26(s,1H),5.07(br d,J＝7.8Hz,1H),4.51(s,1H),3.15-3.04(m,2H),2.25(br t,J＝8.7Hz,2H),1.88-1.75(m,3H),1.74-1.67(m,1H),1.39-1.57(s,3H),0.86-0.76(m,1H),0.48-0.37(m,2H),0.23-0.07(m,2H)

EXAMPLE 225 Synthesis of viral protease inhibitor Compound 832

Step 1: (Z) -2-azido-3- (5-chloro-2-fluorophenyl) acrylic acid methyl ester

A solution of NaOMe (3.41 g,63.06mmol,2 eq.) in MeOH (50 mL) was then added at-10deg.C with 5-chloro-2-fluoro-benzaldehyde (5 g,31.53mmol,1 eq.) and ethyl 2-azidoacetate (8.14 g,63.06mmol,7.21mL,2 eq.) in MeOH (50 mL). The mixture was stirred at 20℃for 18h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 5:1) to give the product (Z) -2-azido-3- (5-chloro-2-fluoro-phenyl) prop-2-enoic acid methyl ester (3.7 g,13.75mmol,43.61% yield, 95% purity) as a white solid.

Step 2: 7-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (5-chloro-2-fluoro-phenyl) prop-2-enoic acid methyl ester (3.7 g,14.47mmol,1 eq.) in xylene (40 mL) and stirring the mixture at 170℃for 1.5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was wet-milled with PE: ea=20:1 (100 mL) for 10min at 20 ℃ to give the product 7-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester (1.6 g,6.68mmol,46.14% yield, 95% purity) as a white solid. MS (ESI) m/z 228.1[ M+H ]] ⁺ 。

Step 3: 7-chloro-4-fluoro-1H-indole-2-carboxylic acid

7-chloro-4-fluoro-1H-indole-2-carboxylic acid methyl ester (1.5 g,6.59mmol,1 eq.) in THF (10 mL) and H ₂ A solution in O (5 mL) was then added LiOH (315.64 mg,13.18mmol,2 eq.) and the mixture stirred at 60℃for 2h. After completion, by adding 100mL of H at 0deg.C ₂ O quenches the reaction mixture, and thenHCl (1M) was added dropwise to a pH of 3-4 and extracted with EA (50 ml x 3). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 7-chloro-4-fluoro-1H-indole-2-carboxylic acid (1.4 g, crude material) as a white solid.

Step 4: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-fluoro-1H-indole-2-carboxamide

7-chloro-4-fluoro-1H-indole-2-carboxylic acid (100 mg,468.18 mol,1.30 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]A solution of 3-cyclopropyl-propionamide (200 mg,359.26umol,50% purity, 1 eq.) in DMF (2 mL) and DCM (5 mL) and then DMAP (131.67 mg,1.08mmol,3 eq.) and EDCI (137.74 mg,718.52umol,2 eq.) were added, followed by stirring the mixture at 20℃for 2h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Waters Xbridge BEH C18100. 30 mM. 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,8 min) to give the product 7-chloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-fluoro-1H-indole-2-carboxamide (112.98 mg,238.39umol,66.36% yield, 100% purity). MS (ESI) m/z 474.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.12(br s,1H),9.10-8.97(m,1H),8.79(d,J＝7.2Hz,1H),7.55(br s,1H),7.36-7.33(m,1H),7.33-7.26(m,1H),6.90(dd,J＝8.6,9.6Hz,1H),5.13-4.98(m,1H),4.58-4.47(m,1H),3.14-3.03(m,2H),2.30-2.17(m,2H),1.88-1.67(m,4H),1.61-1.38(m,3H),0.86-0.77(m,1H),0.48-0.38(m,2H),0.24-0.18(m,1H),0.14-0.08(m,1H)

EXAMPLE 226 Synthesis of viral protease inhibitor Compounds 840

Step 1: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (1 g,2.87mmol,1 eq. HCl) and 4, 6-dichloro-1H-indole-2-carboxylic acid (661.37 mg,2.87mmol,1 eq.) in DCM (40 mL) was added followed by DMAP (1.05 g,8.62mmol,3 eq.) and then EDCI (1.65 g,8.62mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding 30mL H ₂ O quench the reaction mixture and then extract with 40mL DCM (20 mL x 2). The combined organic layers were washed with 30mL HCl (1M) (15 mL x 2), the combined organic layers were washed with 30mL brine, and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1 to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-indole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,1.85mmol,64.46% yield, 97% purity). MS (ESI) m/z 523.1[ M+H ] ] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 6-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 6-dichloro-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (960 mg,1.83mmol,1 eq.) in NH ₃ Solution in MeOH (7M, 20mL,76.33 eq.). The mixture was stirred at 50℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxoSubstituted-ethyl]-4, 6-dichloro-1H-indole-2-carboxamide (820 mg, crude). MS (ESI) m/z 508.1[ M+H ]] ⁺

Step 3:4, 6-dichloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A solution of 4, 6-dichloro-1H-indole-2-carboxamide (800 mg,1.57mmol,1 eq.) in DCM (15 mL) was added followed by the addition of the Buerger's reagent (749.98 mg,3.15mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,10 min) to give 4, 6-dichloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (213.1 mg,434.56umol,27.62% yield, 100% purity). MS (ESI) m/z 490.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.11(s,1H),8.95(d,J＝8.4Hz,1H),8.84(d,J＝7.4Hz,1H),7.53(s,1H),7.44(d,J＝17.4Hz,2H),7.24(d,J＝1.8Hz,1H),5.13-5.01(m,1H),4.52-4.41(m,1H),3.19-3.00(m,2H),2.35-2.18(m,2H),1.97-1.63(m,4H),1.61-1.33(m,3H),0.88-0.75(m,1H),0.51-0.32(m,2H),0.25-0.05(m,2H)

EXAMPLE 227 synthetic viral protease inhibitor Compound 856

Step 1:4- (trifluoromethyl) -1H-indole-2-carboxylic acid

To 4- (trifluoromethyl) -1H-indole-2-carboxylic acid ethyl ester (800 mg,3.11mmol,1 eq.) in THF (10 mL), H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (261.02 mg,6.22mmol,2 eq.) andand the mixture was stirred at 25℃for 8h. The reaction mixture was adjusted to pH 3 with HCl (1M in water). The mixture was extracted with EtOAc (100 x 3 ml). The combined organic layers were purified by Na ₂ SO ₄ Drying, filtration, and concentration gave the product 4- (trifluoromethyl) -1H-indole-2-carboxylic acid (700 mg, crude material) as a white solid. MS (ESI) M/z230.0[ M+H] ⁺

Step 2: (S) -2- ((S) -3-cyclopropyl-2- (4- (trifluoromethyl) -1H-indole-2-carboxamide) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To a solution of 4- (trifluoromethyl) -1H-indole-2-carboxylic acid (650 mg,2.84mmol,1 eq.) in DCM (20 mL) was added methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (986.64 mg,2.84mmol,1 eq., HCl), DMAP (1.04 g,8.51mmol,3 eq.) EDCI (1.09 g,5.67mmol,2 eq.) and the mixture stirred at 25 ℃ for 1H. After completion, by adding H ₂ O (200 mL) to quench the reaction and then extract with EtOAc (100 mL x 3). The combined organic layers were washed with (100 mL brine), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ EtOAc/meoh=1/0 to 10/1) to give the product methyl (S) -2- ((S) -3-cyclopropyl-2- (4- (trifluoromethyl) -1H-indole-2-carboxamido) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (800 mg,1.50mmol,52.83% yield, 97.87% purity) as a yellow solid. MS (ESI) m/z 523.2[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4- (trifluoromethyl) -1H-indole-2-carboxamide

A solution of methyl (S) -2- ((S) -3-cyclopropyl-2- (4- (trifluoromethyl) -1H-indole-2-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (700 mg,1.34mmol,1 eq.) in ammonia (7M, 40mL,209.01 eq.) was stirred at 50deg.C for 10H. After completion, the reaction was concentrated in vacuo to give the crude product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxo) as a white solid Propyl-2-yl) -4- (trifluoromethyl) -1H-indole-2-carboxamide (690 mg, crude). MS (ESI) m/z 508.2[ M+H ]] ⁺

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4- (trifluoromethyl) -1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4- (trifluoromethyl) -1H-indole-2-carboxamide (640 mg,1.32mmol,1 eq.) in DCM (30 mL) was added the bergs reagent (943.82 mg,3.96mmol,3 eq.) and the mixture stirred at 25 ℃ for 4H. After completion, the reaction was concentrated in vacuo and purified by preparative HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give the product N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4- (trifluoromethyl) -1H-indole-2-carboxamide (200 mg,408.59umol,30.95% yield, 100% purity) as a white solid. MS (ESI) m/z 490.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.14(br s,1H),8.97-8.95(m,1H),8.88-8.86(m,1H),7.75 7.71(m,1H),7.54(s,2H),7.45-7.43(m,1H),7.37-7.31(m,1H),5.11-5.03(m,1H),4.52-4.45(m,1H),3.15-3.04(m,2H),2.35-2.21(m,2H),1.93-1.76(m,3H),1.76-1.64(m,1H),1.62-1.51(m,1H),1.49-1.34(m,2H),0.84-0.81(m,1H),0.48-0.36(m,2H),0.26-0.07(m,2H)。

EXAMPLE 228 Synthesis of viral protease inhibitor Compound 896

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (2.6 g,9.08mmol,1 eq.) in HCl/MeOH (4M, 30mL,13.21 eq.) in a mixture of two different solventsThe mixture was stirred at 20℃for 1.5h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl propionate (2 g, crude material, HCl). MS (ESI) m/z 187.1[ M+H ]] ⁺ 。

Step 2:3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

(2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (2 g,8.98mmol,1 eq, HCl) in DCM (20 mL) and DMF (2 mL) was followed by the addition of 2-tert-butoxycarbonyl-2-azaspiro [4.5]]Decane-3-carboxylic acid (2.80 g,9.88mmol,1.1 eq.), T3P (11.43 g,17.96mmol,10.68mL,50% purity, 2 eq.) and TEA (5.45 g,53.89mmol,7.50mL,6 eq.) were stirred at 20℃for 3h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE: ea=4/1 to 0/1 to give the product 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (2.5 g,4.43mmol,49.31% yield, 80% purity). MS (ESI) m/z 452.3[ M+H ]] ⁺ 。

Step 3: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]A mixture of tert-butyl decane-2-carboxylate (2.1 g,3.72mmol,80% purity, 1 eq.) in HCl/MeOH (4M, 25mL,26.88 eq.) was stirred at 20℃for 3h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product (2S) -2- (2-azaspiro [ 4.5) as a white oil]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Propionic acidMethyl ester (1.4 g, crude material, HCl). MS (ESI) m/z 352.2[ M+H ]] ⁺ 。

Step 4: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl ] propionic acid methyl ester

(2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]A mixture of methyl propionate (1.4 g,3.61mmol,1 eq, HCl) in DCM (20 mL) was added followed by 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (1.06 g,4.69mmol,1.3 eq), DMAP (1.10 g,9.02mmol,2.5 eq) and EDCI (1.38 g,7.22mmol,2 eq) and stirred at 20deg.C for 1H. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ PE: EA=2/1-0/1) purification of the residue, giving the product (2S) -2- [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.5 g,2.68mmol,74.34% yield). MS (ESI) m/z 559.2[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl ] methyl ] ethyl ] -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl ]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.46 g,2.61mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,53.61 eq.) was stirred at 30℃for 20h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.35 g, crude). MS (ESI) m/z 544.2[ M+H ]] ⁺ 。

Step 6:2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.35 g,2.11mmol,85% purity, 1 eq.) in DCM (15 mL) was added a Bungeus reagent (1.51 g,6.33mmol,3 eq.) which was stirred for 1h at 30 ℃. After completion, the mixture was quenched with water (1 mL) and with N ₂ And (5) blow-drying. By preparative HPLC (column: waters X bridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) was purified and passed through SFC (column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,12 min) to give the product 2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (322.82 mg,613.70umol,29.10% yield). MS (ESI) M/z526.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.12(d,J＝1.7Hz,1H),7.02(s,1H),6.97(br d,J＝1.8Hz,1H),5.12-5.00(m,1H),4.62(dd,J＝7.9,9.7Hz,1H),3.92(br d,J＝10.3Hz,1H),3.86-3.33(m,5H),3.30-3.26(m,1H),2.77-2.55(m,1H),2.52-2.23(m,3H),1.98-1.67(m,3H),1.62-1.41(m,10H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.07(br d,J＝1.1Hz,1H),8.72(br d,J＝7.5Hz,1H),7.44(br d,J＝0.7Hz,1H),7.12(br s,1H),6.97(s,2H),4.92(br s,1H),4.60(br s,1H),3.85-3.77(m,4H),3.61(br s,1H),3.14(br s,2H),2.43-2.21(m,2H),2.20-1.89(m,2H),1.80(br s,1H),1.72-1.58(m,2H),1.57-1.35(m,10H)。

The product 2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl was obtained as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (289.32 mg,550.01umol,26.08% yield). MS (ESI) M/z526.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.12(d,J＝2.0Hz,1H),7.04(s,1H),6.99-6.93(m,1H),5.06-4.97(m,1H),4.63(dd,J＝7.9,9.5Hz,1H),3.94(br d,J＝10.4Hz,1H),3.88-3.68(m,4H),3.30-2.73(m,2H),2.68-2.10(m,4H),1.94-1.69(m,3H),1.62-1.40(m,10H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.46-10.49(m,1H),8.67(br d,J＝6.6Hz,1H),7.44(br s,1H),7.21-7.07(m,1H),6.98(s,2H),5.06-4.83(m,1H),4.59(br dd,J＝2.1,4.1Hz,1H),3.80(s,4H),3.70-3.44(m,1H),3.22-3.10(m,2H),2.25(s,4H),1.82(br s,1H),1.68(br d,J＝10.4Hz,2H),1.59-1.33(m,10H)。

EXAMPLE 229 Synthesis of viral protease inhibitor Compound 1059

Step 1: (2S) -2-amino-3- (2, 2-difluorocyclopropyl) propanoic acid

A mixture of (2S) -2-amino-3- (2, 2-difluorocyclopropyl) propanoic acid (630 mg,3.13mmol,1 eq. HCl) in HCl/MeOH (4M, 6mL,7.68 eq.) was stirred at 80℃for 2h. After completion, the mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- (2, 2-difluorocyclopropyl) propanoate (700 mg, crude material, HCl) as a yellow oil.

Step 2: (2S) -3- (2, 2-Difluorocyclopropyl) -2- (4-methoxy-1H-indole-2-carboxamido) propionic acid methyl ester

To a solution of methyl (2S) -2-amino-3- (2, 2-difluorocyclopropyl) propanoate (700 mg,3.25mmol,1 eq., HCl) and 4-methoxy-1H-indole-2-carboxylic acid (930.98 mg,4.87mmol,1.5 eq.) in DCM (15 mL) and DMF (3 mL) were added DMAP (793.21 mg,6.49mmol,2 eq.) and EDCI (1.24 g,6.49mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) to give the residue as yellow(2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as an oily substance]Methyl propionate (1 g,2.84mmol,87.43% yield). MS (ESI) m/z 353.1[ M+H ]] ⁺

Step 3: (2S) -3- (2, 2-Difluorocyclopropyl) -2- (4-methoxy-1H-indole-2-carboxamide) propionic acid

To (2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Methyl propionate (1 g,2.84mmol,1 eq.) in THF (10 mL) and H ₂ LiOH.H was added to the solution in O (3 mL) ₂ O (357.31 mg,8.51mmol,3 eq.). The mixture was stirred at 20℃for 16h. After completion, by adding H ₂ O (30 mL) to quench the mixture, and then aqueous HCl (1M) was added to adjust the pH to 3-4 and extracted with EA (20 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow solid]Propionic acid (1 g, crude material). MS (ESI) m/z 339.1[ M+H ]] ⁺

Step 4: (2S) -2- ((2S) -3- (2, 2-Difluorocyclopropyl) -2- (4-methoxy-1H-indole-2-carboxamide) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionic acid (1 g,2.96mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (887.81 mg,3.75mmol,1.27 eq., HCl) in DCM (15 mL) and DMF (3 mL) was added DMAP (722.23 mg,5.91mmol,2 eq.) and EDCI (1.13 g,5.91mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) to give (2S) -2- [ [ (2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperaquineBoydo group]Methyl propionate (1 g,1.92mmol,64.99% yield). MS (ESI) m/z 521.2[ M+H ]] ⁺

Step 5: n- ((2S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

Containing (2S) -2- [ [ (2S) -3- (2, 2-difluorocyclopropyl) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH of methyl propionate (1 g,1.92mmol,1 eq) ₃ MeOH (7M, 15mL,54.66 eq.) was stirred at 80℃for 16h. After completion, the mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a brown solid]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (1 g, crude). MS (ESI) m/z 506.2[ M+H ] ] ⁺

Step 6: n- ((2S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (1 g,1.98mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (1.41 g,5.93mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -50%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (0.6 g,1.23mmol,62.22% yield). MS (ESI) m/z 488.2[ M+H ]] ⁺

Step 7: n- ((2S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:46% -46%,7 min) isolation of N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (0.6 g,1.23 mmol) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide isomer 1 (210 mg,429.91umol,34.93% yield, 99.8% purity). MS (ESI) m/z 488.2[ M+H ]] ⁺

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27-7.26(m,1H),7.17-7.15(m,1H),7.13-7.04(m,1H),6.52-6.50(m,1H),5.14-5.09(m,1H),4.61-4.56(m,1H),3.93(s,3H),3.23-3.21(m,2H),2.46-2.42(m,2H),1.96-1.95(m,1H),1.93-1.92(m,3H),1.85-1.70(m,3H),1.56-1.44(m,2H),1.22-1.12(m,1H)

Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide isomer 2 (210 mg,429.05umol,34.86% yield, 99.6% purity). MS (ESI) m/z 488.2[ M+H ]] ⁺

1H NMR(400MHz,MeOD-d ₄ )δ＝7.32-7.24(m,1H),7.20-7.11(m,1H),7.13-7.05(m,1H),6.53-6.51(m,1H),5.14-5.00(m,1H),4.66-4.61(m,1H),3.94(s,3H),3.20-3.19(m,2H),2.43-2.25(m,2H),1.95-1.90(m,4H),1.85-1.63(m,3H),1.56-1.44(m,2H),1.22-1.03(m,1H)

Example 230 Synthesis of viral protease inhibitor Compound 1155

Step 1: (2S, 4S) -4-methoxy-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -4- (trifluoromethyl) pyrrolidine-1-carboxylic acid tert-butyl ester

To (2S, 4S) -1-tert-butoxycarbonyl-4-methoxy-4- (trifluoromethyl) pyrrolidine-2-carboxylic acid (0.5 g,1.60mmol,1.2 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (314.82 mg,1.33mmol,1 eq., HCl), EDCI (509.94 mg,2.66mmol,2 eq.) in DCM (5 mL) was added DMAP (487.48 mg,3.99mmol,3 eq.) and the mixture was stirred at 20deg.C for 2h. After completion, by adding H ₂ O (100 mL) to quench the reaction mixture and extract with DCM (15 mL x 4). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/1 to 0/1 and then DCM: meOH=5:1) to give the product (2S, 4S) -4-methoxy-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-tert-butyl 4- (trifluoromethyl) pyrrolidine-1-carboxylate (0.9 g,1.27mmol,95.60% yield, 70% purity). MS (ESI) m/z 496.2[ M+H ]] ⁺

Step 2: (2S) -2- [ [ (2S, 4S) -4-methoxy-4- (trifluoromethyl) pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoic acid methyl ester

(2S, 4S) -4-methoxy-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]Carbamoyl group]A solution of tert-butyl 4- (trifluoromethyl) pyrrolidine-1-carboxylate (0.8 g,1.61mmol,1 eq.) in HCl/MeOH (4M, 9mL,22.30 eq.) was stirred at 20deg.C for 1h. After completion, the reaction was concentrated under reduced pressure to give the crude (2S) -2- [ [ (2S, 4S) -4-methoxy-4- (trifluoromethyl) pyrrolidine-2-carbonyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.65 g, crude material, HCl). MS (ESI) m/z 396.1[ M+H ]] ⁺

Step 3: (2S) -2- [ [ (2S, 4S) -4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- [ [ (2S, 4S) -4-methoxy-4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Propionic acid methyl esterTo a solution of ester (0.65 g,1.51mmol,1 eq., HCl), 4-methoxy-1H-indole-2-carboxylic acid (345.32 mg,1.81mmol,1.2 eq.) in DCM (5 mL) was added DMAP (551.67 mg,4.52mmol,3 eq.) and EDCI (577.10 mg,3.01mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (80 mL) to quench the reaction and extract with DCM (10 mL x 4). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/1 to 0/1) to give the product (2S) -2- [ [ (2S, 4S) -4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl as a yellow oil]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.65 g, crude material). MS (ESI) m/z 569.2[ M+H ]] ⁺

Step 4: (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide

(2S) -2- [ [ (2S, 4S) -4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.53 g,932.21umol,1 eq.) in NH ₃ A solution in MeOH (3 mL) was stirred at 50deg.C for 28h. After completion, the reaction was concentrated under reduced pressure to give the crude product (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow solid]Methyl group]Ethyl group]-4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide (0.5 g, crude material). MS (ESI) m/z 554.2[ M+H ] ] ⁺

Step 5: (2S, 4S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide

To (2S, 4S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of-4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide (0.5 g,812.96umol,90% purity, 1 eq.) in DCM (8 mL) was added a berger reagent (581.21)mg,2.44mmol,3 eq) and the mixture was stirred at 30 ℃ for 4h. After completion, the mixture was quenched with water (1.5 mL) and quenched with N ₂ Blow dried and purified by preparative HPLC (column Waters Xbridge Prep OBD C18.40.5.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -50%,8 min) to give the product (2S, 4S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-4-methoxy-1- (4-methoxy-1H-indole-2-carbonyl) -4- (trifluoromethyl) pyrrolidine-2-carboxamide (0.21 g,392.15umol,48.24% yield, 100% purity). MS (ESI) m/z 536.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.25-7.13(m,1H),7.05(br d,J＝8.2Hz,2H),6.63-6.40(m,1H),5.36-4.89(m,2H),4.47-4.04(m,2H),4.02-3.79(m,3H),3.45(br s,3H),3.26-2.90(m,2H),2.86-2.50(m,2H),2.49-2.14(m,2H),2.04-1.04(m,5H)。

EXAMPLE 231 Synthesis of viral protease inhibitor Compounds 1053

Step 1: (2S, 4E) -4- (dimethylaminomethylene) -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester

A mixture of (2S) -5-oxopyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (282 g,1.16mol,1 eq.) 1-tert-butoxy-N, N, N ', N' -tetramethyl-methyldiamine (303.06 g,1.74mol,359.08mL,1.5 eq.) in DME (282 mL) was stirred at 75deg.C for 3h. After completion, the mixture was cooled to 0 ℃ and then filtered, and the filter cake was concentrated under reduced pressure to give the product (2 s,4 e) -4- (dimethylaminomethylene) -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (272 g, crude material) as a white solid.

Step 2: (2S) -4-methylene-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester

To (2S, 4E) -4- (dimethylaminomethylene) -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (70 g,234.64mmol, 1.)Amount) DIBAL-H (1M, 703.91mL,3 eq.) was added to a solution in THF (700 mL). The mixture was stirred at-78℃for 2h. After completion, by addition to saturated NH ₄ The reaction mixture was quenched in Cl (2500 mL) and then extracted with EA (1000 mL x 3). The combined organic layers were washed with brine (2000 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 1/1) to obtain the product. The product (2S) -4-methylene-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (35 g,137.11mmol,58.44% yield) was obtained as a white solid.

Step 3: (2S) -2- (tert-Butoxycarbonylamino) -4-methylene-glutarate dimethyl ester

To a solution of (2S) -4-methylene-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (25 g,97.94mmol,1 eq.) in THF (250 mL) was added lithium methoxide (1M, 117.52mL,1.2 eq.) at-40 ℃. The solution was stirred at-40℃for 2h. After completion, the solution is treated with NH ₄ Cl (70 mL) was quenched and concentrated, and extracted with EA (80 mL x 2) and concentrated to give crude (2S) -2- (tert-butoxycarbonylamino) -4-methylene-glutarate dimethyl ester (24 g, crude material) as a yellow oil and used directly in the next step. MS (ESI) M/z188.1[ M+H-100 ]] ⁺

Step 4: (2S) -2- (tert-Butoxycarbonylamino) -4- (2-methyl-2-nitro-propyl) glutaric acid dimethyl ester

To a solution of (2S) -dimethyl 2- (tert-butoxycarbonylamino) -4-methylene-glutarate (34 g,118.34mmol,1 eq.) and 2-nitropropane (11.60 g,130.17mmol,11.69mL,1.1 eq.) in ACN (350 mL) was added DBU (21.62 g,142.01mmol,21.40mL,1.2 eq.). The solution was stirred at 20℃for 2h. After completion, the solution was concentrated to give a crude material. Through the column (SiO) ₂ The crude material was purified with PE: ea=20:1 to 1:1 to give the product compound (2S) -2- (tert-butoxycarbonylamino) -4- (2-methyl-2-nitro-propyl) glutarate dimethyl ester (30 g,79.70mmol,67.35% yield) as a white solid. MS (ESI) m/z 277.1[ M+H-100 ]] ⁺

Step 5: (2S) -2- (tert-Butoxycarbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

At N ₂ Next, pd/C (24.54 g,20.72mmol,10% purity, 0.3 eq) was added to a solution of dimethyl (2S) -2- (tert-butoxycarbonylamino) -4- (2-methyl-2-nitro-propyl) glutarate (26 g,69.08mmol,1 eq.) in IPA (250 mL). The suspension was degassed in vacuo and purified by H ₂ Purging several times. The mixture is put in H ₂ (15 psi) and stirred at 40℃for 15h. After completion, the mixture was filtered and concentrated to give the crude compound. Through the column (SiO) ₂ PE: ea=20:1 to 0:1) to give the product methyl (2S) -2- (tert-butoxycarbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (14.6 g,46.44mmol, 67.23%) as a white solid and was purified by SFC (column: DAICEL CHIRALPAK IC (250 mm. Times.50 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ OIPA]The method comprises the steps of carrying out a first treatment on the surface of the B%: purification was continued for 11.5min at 30% -30% to give methyl BB7 (2S) -2- (tert-butoxycarbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (4.8 g,15.27mmol,32.65% yield) as a white solid. MS (ESI) m/z 315.2[ M+H ] ] ⁺

Step 6: (2S) -2-amino-3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (500 mg,1.59mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 20℃for 2h. After completion, the solution was concentrated to dryness to give methyl (2S) -2-amino-3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (398 mg, crude material, HCl) as a white solid and was used directly in the next step.

Step 7: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (2S) -2-amino-3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (370 mg,1.48mmol,1 eq. HCl) in DCM (10 mL) and DMF (5 mL) was added DMAP (360.58 mg,2.95mmol,2 eq.) and (2S) -2- (tert-butoxy)Carbonylamino) -3-cyclopropyl-propionic acid (372.18 mg,1.62mmol,1.1 eq.) and EDCI (565.80 mg,2.95mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the mixture was treated with H ₂ O (60 mL) was diluted and extracted with EA (60 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ The crude material was purified with pe:ea=10:1 to 0:1 to give the compound (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a yellow oil]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (550 mg,1.29mmol,87.59% yield). MS (ESI) m/z 426.2[ M+H ]] ⁺

Step 8: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propanoate (540 mg,1.27mmol,1 eq.) in HCl/MeOH (15 mL) was stirred at 20℃for 1h. After completion, the solution was concentrated to dryness to give methyl (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (456 mg, crude material, HCl) as a white solid.

Step 9: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]To a solution of methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (450 mg,1.24mmol,1 eq., HCl) in DCM (10 mL) and DMF (5 mL) was added DMAP (303.85 mg,2.49mmol,2 eq.) and 7-chloro-1H-indole-2-carboxylic acid (243.24 mg,1.24mmol,1 eq.) and EDCI (476.79 mg,2.49mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (60 mL) was diluted and extracted with EA (70 mL x 3) and washed with brine (100 mL x 2) and concentrated to give the crude material. Through the column (SiO) ₂ The crude material was purified with PE: ea=3:1 to 0:1 to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (550 mg,1.09mmol,87.93% yield). MS (ESI) m/z 503.2[ M+H ]] ⁺

Step 10: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (550 mg,1.09mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 7.81mL,50 eq.) was stirred at 60℃for 17h. After completion, the solution was concentrated to dryness to give crude material. The crude material was used directly in the next step. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl as a white solid]-2-oxo-ethyl ]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (530 mg, crude). MS (ESI) m/z 488.2[ M+H ]] ⁺

Step 11: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (530 mg,1.09mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (517.65 mg,2.17mmol,2 eq.). The mixture was stirred at 20℃for 2.5h. After the reaction was completed, the solution was treated with H ₂ O (30 mL) and washed with N ₂ The organic phase was dried to give a crude material. By neutral prep HPLC (Waters Xbridge C18 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -70%,10 min) to purify the crude material to obtain the product. MS (ESI) m/z 470.2[ M+H ]] ⁺

7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide (330 mg,702.18umol,64.65% yield) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d ₆ ) Offset = 11.56 (br s, 1H), 8.89 (d, J = 7.9hz, 1H), 8.60 (d, J = 7.5hz, 1H), 7.70 (s, 1H), 7.50 (d, J = 7.9hz, 1H), 7.19 (d, J = 7.6hz, 1H), 7.12 (s, 1H), 6.95 (t, J = 7.8hz, 1H), 4.95-4.76 (m, 1H), 4.46-4.20 (m, 1H), 2.52-2.44 (m, 1H), 2.12-1.99 (m, 1H), 1.88 (dd, J = 8.6,12.2hz, 1H), 1.76-1.60 (m, 2H), 1.45-1.33 (m, 2H), 1.03 (s, 3H), 0.95 (s, 3H), 0.46-4.20 (m, 1H), 2.52-2.44 (m, 1H), 2.12-1.76 (m, 1H), 0.38 (m, 0.25-0H).

EXAMPLE 232 Synthesis of viral protease inhibitor Compounds 1111

Step 1: (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (20 mg,60.90umol,1 eq.) was added HCl/MeOH (4M, 5mL,328.40 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester (16 mg,57.41umol,94.27% yield, 95% purity, HCl) was obtained as a colorless oil.

Step 2:3- [ [ (1S) -1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

To 2-Boc-2-azaspiro [4.5]]To a mixture of decane-3-carboxylic acid (171.25 mg,604.35umol,1 eq.) and methyl (2S) -2-amino-3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (160 mg,604.35umol,1 eq., HCl) in DMF (3 mL) and DCM (6 mL) was added EDCI (231.71 mg,1.21mmol,2 eq.) and DMAP (147.67 mg,1.21mmol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 50mL DCM (25 mL x 2). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=7/1 to 1/1) the residue was purified to give the compound 3- [ [ (1S) -1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl as a colorless oil]-2-methoxy-2-oxo-ethyl]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (195 mg,395.03umol,65.36% yield, N/A purity).

Step 3: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester

To 3- [ [ (1S) -1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester (170 mg,344.38umol,1 eq.) was added HCl/MeOH (4 m,17.00ml,197.45 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound methyl (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionate (145 mg,320.36umol,93.03% yield, 95% purity, HCl) was obtained as a colorless oil.

Step 4: (2S) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (64.47 mg,337.23umol,1 eq.) and (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) propionic acid methyl ester (145 mg,337.23umol,1 eq., HCl) to a mixture of DCM (6 mL) and DMF (3 mL) was added DMAP (82.40 mg,674.45umol,2 eq.) and EDCI (129.29 mg,674.45umol,2 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was taken up with 30mL H ₂ O was diluted and extracted with 100mL EA (50 mL x 2). The combined organic layers were washed with 50mL brine (50 mL x 1), dried over Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure to obtain residueAnd the remainder. The residue was purified by preparative TLC (DCM: meoh=10:1) to give compound (2S) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow oil]Decane-3-carbonyl]Amino group]Methyl propionate (200 mg,335.28umol,99.42% yield, 95% purity). MS (ESI) m/z 567.3[ M+H ]] ⁺

Step 5: n- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl ] -2-oxo-ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To (2S) -3- (6, 6-dimethyl-2-oxo-3-piperidinyl) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carbonyl]Amino group]NH was added to a mixture of methyl propionate (200 mg,352.93umol,1 eq) ₃ MeOH (7M, 50.42uL,1 eq.). The mixture was stirred at 30℃for 48h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and the residue was used directly in the next step. Obtaining the compound N- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl as a white solid]-2-oxo-ethyl]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (190 mg,309.96umol,87.83% yield, 90% purity). MS (ESI) m/z 552.3[ M+H ]] ⁺

Step 6: n- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-1- [ (6, 6-dimethyl-2-oxo-3-piperidinyl) methyl]-2-oxo-ethyl]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]To a mixture of decane-3-carboxamide (190 mg,344.41umol,1 eq.) in DCM (2 mL) was added the Bungeus reagent (164.15 mg,688.81umol,2 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was quenched with 5mL H ₂ O was diluted and extracted with 10mL DCM (5 mL x 2). The combined organic layers were concentrated by blow-drying to give a residue. By preparative HPLC (neutral conditions) (column: waters Xbridge BEH C, 100X 30mm X10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) purification residueThe residue gave the desired compound (80 mg) as a white solid. By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [ Neu-ETOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,7 min) to give isomers 1, 2, 3 and 4. By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [ Neu-MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,15 min) and separating the mixture (isomers 2 and 3). MS (ESI) m/z 534.2[ M+H ]] ⁺

Isomer 1: the compound N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide (13 mg,24.36umol,7.07% yield, 100% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.29(br s,1H),8.69(br s,1H),7.28-6.72(m,4H),6.52(d,J＝7.7Hz,1H),4.99(br s,1H),4.78-4.46(m,1H),3.95-3.82(m,4H),3.73-3.40(m,1H),2.34-2.04(m,3H),1.89-1.31(m,16H),1.13(br d,J＝10.6Hz,6H)

Isomer 2: the compound N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide (17 mg,31.86umol,9.25% yield, 100% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.31(br s,1H),8.64(br s,1H),7.25-6.75(m,4H),6.52(d,J＝7.7Hz,1H),4.99(q,J＝8.0Hz,1H),4.61(br s,1H),4.01-3.80(m,4H),3.66(br s,1H),2.30-1.94(m,3H),1.92-1.31(m,16H),1.12(d,J＝6.4Hz,6H)

Isomer 3: the compound N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide (14 mg,26.23umol,7.62% yield, 100% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.30(br s,1H),8.92-8.52(m,1H),7.41-6.74(m,4H),6.52(br d,J＝7.3Hz,1H),4.94(br s,1H),4.63(br s,1H),4.03-3.78(m,4H),3.73-3.44(m,1H),2.35-2.04(m,3H),1.93-1.32(m,16H),1.14(s,6H)

Isomer 4: the compound N- [ (1S) -1-cyano-2- (6, 6-dimethyl-2-oxo-3-piperidinyl) ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide (28 mg,52.23umol,15.16% yield, 99.538% purity) was obtained as a white solid.

¹ H NMR(400MHz,DMSO-d6)δ＝11.31(br s,1H),8.69(br s,1H),7.27-6.77(m,4H),6.53(d,J＝7.5Hz,1H),4.94(br s,1H),4.61(br s,1H),4.06-3.83(m,4H),3.66(br s,1H),2.32-1.98(m,3H),1.83-1.32(m,16H),1.13(d,J＝18.1Hz,6H)

EXAMPLE 233 Synthesis of viral protease inhibitor Compound 3069

Step 1: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (400 mg,905.88umol,1 eq.) in HCl/MeOH (5 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (330 mg, crude material, HCl) as a white solid. MS (ESI) m/z 342.2[ M+H ] ] ⁺ 。

Step 2: (6S, 9S, 12S) -6- (tert-butyl) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-9-neopentyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triazatride-13-oic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (330 mg,873.23umol,1 eq, HCl) and (S) -2- ((t-butoxycarbonyl) amino) -3, 3-dimethylbutyric acid (201.97 mg,873.23umol,1 eq) in DCM (10 mL) was added DMAP (320.05 mg,2.62mmol,3 eq) and EDCI (502.20 mg,2.62mmol,3 eq). Stirring and mixing at 30deg.CAnd (3) carrying out object 2h. After completion, by adding H ₂ O (10 mL) was used to quench the reaction mixture and was then extracted with DCM (5 mL x 2). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (6 s,9s,12 s) -6- (tert-butyl) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-9-neopentyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triaza-tridec-13-oic acid methyl ester (450 mg, crude material) as a white solid. MS (ESI) m/z 555.4[ M+H ]] ⁺ 。

Step 3: ((S) -1- (((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester

(6S, 9S, 12S) -6- (tert-butyl) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-9-neopentyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triazatride-13-oic acid methyl ester (400 mg,721.09umol,1 eq.) in NH ₃ A solution in MeOH (7M, 8.00mL,77.66 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give tert-butyl ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamate (430 mg, crude material) as a white solid. MS (ESI) M/z540.4[ M+H ]] ⁺ 。

Step 4: (S) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) -4, 4-dimethylpentanamide

A solution of tert-butyl ((S) -1- (((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamate (410 mg,759.67umol,1 eq) in HCl/EtOAc (4M, 189.92uL,1 eq) was stirred at 25℃for 1h. After completion, the resulting solution was concentrated in vacuo (40 ℃ C.) to give To (S) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -4, 4-dimethylpentanamide (340 mg, crude material) as a white solid. MS (ESI) m/z 440.3[ M+H ]] ⁺

Step 5: (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide

To a solution of (S) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -4, 4-dimethylpentanamide (300 mg,682.45umol,1 eq) in DCM (3 mL) were added TFAA (86.00 mg,409.47umol,56.95ul,0.6 eq) and DIPEA (264.61 mg,2.05mmol,356.61ul,3 eq) and the mixture was stirred at 0 ℃ for 1h. LCMS showed most of the starting material remained and TFAA (28.67 mg,136.49umol,18.98ul,0.2 eq.) was then added and the mixture was stirred for an additional 1h. LCMS showed little starting material remaining and TFAA (43.00 mg,204.74umol,28.48ul,0.3 eq.) was then added and stirred for an additional 30min. After completion, the resulting solution was poured into H ₂ O (10 mL) in NaHCO ₃ Adjust to pH-8 and then extract with EtOAc (10 ml x 2). The combined organic phases were taken up in Na ₂ SO ₄ Dried, filtered and concentrated to give the crude product (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide (290 mg, crude) as a white solid. MS (ESI) m/z 536.3[ M+H ]] ⁺

Step 6: (S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide

To (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylvaleramide (290 mg,541.45umol,1 eq.) in DCM aid3 mL) was added with the Buerger reagent (258.06 mg,1.08mmol,2 eq.) followed by stirring the mixture at 25℃for 3h. After completion, the resulting solution was treated with H ₂ O (0.3 mL) was quenched and then concentrated in vacuo (25 ℃ C.). By preparative HPLC (column Phenomenex Gemini-NX C18 75X 30mM X3 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give (S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butyrylamino) -4, 4-dimethylvaleramide (75.94 mg,146.72umol,27.10% yield, 100% purity) as a white solid. MS (ESI) m/z 518.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝4.96-4.91(m,1H),4.46-4.37(m,2H),2.78-2.64(m,1H),2.37-2.27(m,1H),2.22-2.14(m,1H),1.95-1.84(m,1H),1.74-1.59(m,3H),1.29(s,3H),1.23(s,3H),1.00-0.96(m,18H)

EXAMPLE 234 Synthesis of viral protease inhibitor Compound 3129

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 0deg.C]To a mixture of methyl propionate (1 g,3.80mmol,90% purity, 1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (1.03 g,4.18mmol,1.1 eq) in DCM (10 mL) was added DMAP (1.16 g,9.51mmol,2.5 eq), EDCI (1.46 g,7.60mmol,2 eq) followed by stirring the mixture at 20 ℃ for 2h. After completion, by adding H ₂ O (5 mL) to quench the reaction mixture, and then H was used ₂ O (10 mL) was diluted and extracted with 60mL DCM (30 mL. Times.2). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ DCM: meoh=0:1 to 10:1) The residue was purified to give methyl (S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.14 g,2.32mmol,61.01% yield, 87% purity) as a yellow solid. MS (ESI) m/z 428.2[ M+H ]] ⁺ 。

Methyl (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1 g,2.03mmol,87% purity, 1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 15mL,51.60 eq.) was stirred at 60℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate (1 g, crude material) as a yellow solid. MS (ESI) m/z 413.2[ M+H ]] ⁺ 。

A solution of tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate (1 g,2.01mmol,83% purity, 1 eq.) in HCl/EtOAc (4M, 8.30mL,16.50 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylvaleramide (800 mg, crude material) as a yellow solid. MS (ESI) M/z313.2[ M+H ] ⁺ 。

Step 4: (S) -1- (((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester

To (S) -2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide at 0deg.CTo a mixture of (700 mg,1.57mmol,78% purity, 1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butyric acid (433.33 mg,1.87mmol,1.2 eq) in DCM (10 mL) was added DMAP (478.01 mg,3.91mmol,2.5 eq), EDCI (600.05 mg,3.13mmol,2 eq) followed by stirring the mixture at 20 ℃ for 2h. After completion, the reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with 1N HCl (10 mL), then brine (20 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=0:1 to 10:1) to give tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamate (400 mg,684.83umol,43.76% yield, 90% purity) as a yellow solid. MS (ESI) m/z 526.4[ M+H ] ] ⁺ 。

Step 5: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -4, 4-dimethylpentanamide

A mixture of tert-butyl ((S) -1- (((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamate (300 mg,513.62umol,90% purity, 1 eq) in HCl/EtOAc (4M, 2.70mL,21.03 eq) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -4, 4-dimethylpentanamide (240 mg, crude material, HCl) as a yellow solid. MS (ESI) m/z 426.4[ M+H ]] ⁺ 。

Step 6: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide

To (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) at 0 ℃CPropan-2-yl) -2- ((S) -2-amino-3, 3-dimethylbutylamino) -4, 4-dimethylvaleramide (280 mg,484.82umol,80% purity, 1 eq, HCl) in DCM (3 mL) was added DIEA (187.98 mg,1.45mmol,253.34uL,3 eq) and TFAA (152.74 mg,727.23umol,101.15uL,1.5 eq) followed by stirring the mixture at 20℃for 1h. After completion, the reaction mixture was diluted with water (3 mL) and extracted with DCM (3 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=100:1 to 10:1 to give (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide (280 mg, crude material) as a yellow solid. MS (ESI) M/z522.3[ M+H] ⁺ 。

Step 7: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoylamino) -4, 4-dimethylpentanamide

To a mixture of (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butyrylamino) -4, 4-dimethylvaleramide (270 mg,414.13umol,80% purity, 1 eq.) in DCM (3 mL) was added the bergs reagent (197.38 mg,828.27umol,2 eq.) and stirred for 2h at 25 ℃. After completion, by adding H at 20℃ ₂ O (0.4 mL) to quench the reaction mixture, and then under reduced pressure<Concentrated at 30 ℃ to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give (S) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butyrylamino) -4, 4-dimethylpentanamide (104.51 mg,207.54umol,50.12% yield, 100% purity) as a white solid. MS (ESI) M/z504.2[ M+H] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.09(br s,1H),8.95(d,J＝8.1Hz,1H),8.37(d,J＝7.8Hz,1H),7.52(br s,1H),5.02-4.87(m,1H),4.39(br s,1H),4.33(br d,J＝6.5Hz,1H),3.15-3.00(m,2H),2.37-2.29(m,1H),2.22-2.12(m,1H),1.85-1.66(m,3H),1.64-1.44(m,3H),1.42-1.30(m,1H),0.96-0.78(m,18H)

EXAMPLE 235 Synthesis of viral protease inhibitor Compound 3065

Step 1: (6S, 9S, 12S) -6- (tert-butyl) -9- (cyclopropylmethyl) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triazatride-13-oic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (404.97 mg,1.75mmol,1.2 eq.) in DCM (3 mL) was added (600 mg,1.46mmol,88% purity, 1 eq., HCl), DMAP (534.77 mg,4.38mmol,3 eq.) and EDCI (559.43 mg,2.92mmol,2 eq.) followed by stirring the mixture at 20 ℃ for 2h. After completion, the reaction mixture was quenched by addition to water (1 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. Through the column (SiO) ₂ DCM: meoh=100:1 to 10:1) the residue was purified to give (6 s,9s,12 s) -6- (tert-butyl) -9- (cyclopropylmethyl) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triaza-tridec-13-oic acid methyl ester (550 mg,969.97umol,66.48% yield, 95% purity) as a white solid. MS (ESI) m/z 539.3[ M+H ]] ⁺ 。

Step 2: (S) -1- (((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester

(6S, 9S, 12S) -6- (tert-butyl) -9- (cyclopropylmethyl)) -12- (((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) methyl) -2, 2-dimethyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triazatridec-13-oic acid methyl ester (550 mg,969.97umol,95% purity, 1 eq) in NH ₃ The mixture in MeOH (7M, 5mL,39.70 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester (520 mg, crude material) as a white solid. MS (ESI) m/z 524.3[ M+H ] ] ⁺ 。

Step 3: (S) -2-amino-N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -3, 3-dimethylbutyramide

A mixture of tert-butyl ((S) -1- (((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) amino) -3, 3-dimethyl-1-oxobutan-2-yl) carbamate (500 mg,954.81umol,1 eq.) in HCl/EA (4M, 10 mL) was stirred at 25℃for 1h. After completion, the mixture was concentrated under reduced pressure to give (S) -2-amino-N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -3, 3-dimethylbutyramide (350 mg, crude material, HCl) as a white solid. MS (ESI) m/z 424.2[ M+H ]] ⁺ 。

Step 4: (S) -N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanamide

To a mixture of (S) -2-amino-N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -3, 3-dimethylbutyramide (300 mg,586.94umol,90% purity, 1 eq, HCl) in DCM (1 mL) was added TFAA (160.26 mg,763.03umol,106.13ul,1.3 eq) at 0 ℃ ) And DIEA (227.57 mg,1.76mmol,306.70ul,3 eq.) followed by stirring at 20 ℃ for 1h. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ]<30 ℃ C.). By preparative HPLC (column: phenomnex C18.times.30 mm.times.3 um; mobile phase: [ water (NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,12 min) to give (S) -N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanamide (40.23 mg,77.08umol,13.13% yield, 96.1% purity) as a white solid. MS (ESI) m/z 502.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.16-9.03(m,1H),8.98-8.85(m,1H),8.41-8.30(m,1H),7.94-7.80(m,1H),5.00-4.82(m,1H),4.47-4.37(m,1H),4.28-4.20(m,1H),2.22-2.11(m,1H),2.02-1.93(m,1H),1.80-1.62(m,2H),1.54(dd,J＝10.1,12.3Hz,1H),1.39-1.28(m,1H),1.25-1.20(m,1H),1.20-1.16(m,3H),1.10(s,3H),0.93(s,9H),0.76-0.62(m,1H),0.46-0.29(m,2H),0.17-0.01(m,2H)

EXAMPLE 236 Synthesis of viral protease inhibitor Compound 3071a

Step 1: (2S) -2- [ [6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butyryl ] -6-azaspiro [3.4] octane-7-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butanoic acid (206.60 mg,893.27umol,1.1 eq.) and (2S) -2- (6-azaspiro [3.4]]Octane-7-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (350 mg,812.06umol,90% purity, 1 eq, HCl) in DCM (10 mL) was added DMAP (297.63 mg,2.44mmol,3 eq) and EDCI (311.35 mg,1.62mmol,2 eq). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 45mL DCM (15 mL x 3). MergingThe organic layer was washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [6- [ (2S) -2- (t-butoxycarbonylamino) -3, 3-dimethyl-butyryl ] as a white solid]-6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (300 mg,494.06umol,60.84% yield, 93% purity). MS (ESI) m/z 565.3[ M+H ]] ⁺

Step 2: n- [ (1S) -1- [7- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carbonyl ] -2, 2-dimethyl-propyl ] carbamic acid tert-butyl ester

(2S) -2- [ [6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butanoyl]-6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (300 mg,531.24umol,1 eq.) in NH ₃ A solution in MeOH (7M, 15.00mL,197.65 eq.) was stirred at 50℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -1- [7- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] as a white solid ]Methyl group]-2-oxo-ethyl]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carbonyl group]-2, 2-dimethyl-propyl]Tert-butyl carbamate (290 mg, crude material). MS (ESI) m/z 550.3[ M+H ]] ⁺

Step 3:6- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -6-azaspiro [3.4] octane-7-carboxamide

A mixture of tert-butyl N- [ (1S) -1- [7- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carbonyl ] -2, 2-dimethyl-propyl ] carbamate (200 mg,338.37umol,93% purity, 1 eq) in HCl/EtOAc (4M, 4.65mL,54.97 eq) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give 6- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -6-azaspiro [3.4] octane-7-carboxamide (176 mg, crude material, HCl) as a yellow solid.

Step 4: n- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -6-azaspiro [3.4] octane-7-carboxamide

To 6- [ (2S) -2-amino-3, 3-dimethyl-butyryl at 0deg.C]-N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-6-azaspiro [3.4]]To a mixture of octane-7-carboxamide (120 mg,246.89 mol,1 eq, HCl) in DCM (2 mL) was added DIPEA (95.73 mg,740.67 mol,129.01uL,3 eq) and TFAA (103.71 mg,493.78 mol,68.68uL,2 eq) and the mixture was stirred at 0deg.C for 1h. After completion, by adding 4mL NaHCO at 25 DEG C ₃ The reaction mixture was quenched and extracted with 6mL DCM (2 mL x 3). The combined organic layers were washed with 6mL brine (3 mL x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-oxo-ethyl]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4]]Octane-7-carboxamide (130 mg, crude). MS (ESI) m/z 546.3[ M+H ]] ⁺

Step 5: n- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4 ]]To a mixture of octane-7-carboxamide (180 mg,170.59umol,50% purity, 1 eq.) in DCM (5 mL) was added the Bunges reagent (121.96 mg,511.77umol,3 eq.) and the mixture was stirred at 25℃for 4h. After completion, the reaction mixture was concentrated under reduced pressure to giveCrude product. By preparative HPLC (neutral conditions; column Waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give the desired product (50 mg) as a white solid, which was purified by SFC (condition: column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -35%,15 min) to give N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide (isomer 1, 20mg,37.53umol,22.00% yield, 99% purity). MS (ESI) m/z 528.2[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.36-9.14(m,1H),8.83(d,J＝8.2Hz,1H),7.82(s,1H),4.98-4.84(m,1H),4.51(s,1H),4.18(t,J＝8.0Hz,1H),3.87(d,J＝9.9Hz,1H),3.49(d,J＝10.1Hz,1H),2.62-2.56(m,1H),2.29-2.09(m,2H),2.06-1.80(m,7H),1.77-1.67(m,2H),1.58-1.50(m,1H),1.20-1.06(m,6H),1.05-0.87(m,9H)

Obtaining N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4]]Octane-7-carboxamide (isomer 2, 20mg,37.30umol,21.87% yield, 98.4% purity). MS (ESI) m/z 528.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.54-9.35(m,1H),8.56(d,J＝8.6Hz,1H),7.88(s,1H),5.04-4.87(m,1H),4.67(br d,J＝8.2Hz,1H),4.19(t,J＝7.4Hz,1H),3.83(d,J＝10.4Hz,1H),3.60(d,J＝10.1Hz,1H),2.52(br s,1H),2.25(dd,J＝8.3,12.5Hz,1H),2.10-1.68(m,10H),1.58-1.44(m,1H),1.23-1.08(m,6H),0.97(s,9H)。

EXAMPLE 237 Synthesis of viral protease inhibitor Compound 3039a

Step 1: 6-azaspiro [3.4] octane-7-carboxylic acid methyl ester

A solution of 6-tert-butoxycarbonyl-6-azaspiro [3.4] octane-7-carboxylic acid (0.3 g,1.18mmol,1 eq.) in 4M HCl/MeOH (5 mL). Subsequently, the reaction was stirred at 80℃for 4h. After completion, the reaction was concentrated to dryness in vacuo to give methyl 6-azaspiro [3.4] octane-7-carboxylate (240 mg, crude material, HCl) as a brown solid. The crude product was used directly in the next step.

Step 2:6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butyryl ] -6-azaspiro [3.4] octane-7-carboxylic acid methyl ester

To 6-azaspiro [3.4]]To a solution of methyl octane-7-carboxylate (240 mg,1.17mmol,1 eq, HCl) in DCM (5 mL) was added (2S) -2- (t-butoxycarbonylamino) -3, 3-dimethyl-butanoic acid (323.85 mg,1.40mmol,1.2 eq), DMAP (285.11 mg,2.33mmol,2 eq). Next, EDCI (447.37 mg,2.33mmol,2 eq.) was added to the reaction at 20deg.C. Subsequently, the reaction was stirred at 20℃for 3h. After completion, the reaction mixture was quenched by addition of 30mL of 1n HCl at 20 ℃ and then diluted with 20mL of EtOAc and extracted with 60mL of EtOAc (20 mL x 3). The combined organic layers were washed with 40mL saturated NaHCO ₃ (20 mL x 2) washing with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=20/1 to 15/1) to give 6- [ (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butyryl as a colorless oil]-6-azaspiro [3.4]]Octane-7-carboxylic acid methyl ester (300 mg,784.33umol,67.22% yield, assuming 100% purity).

Step 3:6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butyryl ] -6-azaspiro [3.4] octane-7-carboxylic acid

To 6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butanoyl]-6-azaspiro [3.4]]Octane-7-carboxylic acid methyl ester (300 mg, 784.33. Mu. Mol,1 eq.) in THF (1.5 mL) and H ₂ LiOH.H was added to a solution in O (0.5 mL) ₂ O (98.74 mg,2.35mmol,3 eq.). The reaction was then stirred at 20℃for 16h. After completion, the reaction mixture was quenched with an additional 5mL EtOAc at 20 ℃The material was then treated with 20mL H ₂ O dilution. Next, the desired phase was separated, adjusted to ph=1 with 1M HCl and extracted with 15mL EtOAc (5 mL x 3). The combined organic layers were washed with 5mL saturated NaHCO ₃ Washing with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 6- [ (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butyryl as a colorless gum residue ]-6-azaspiro [3.4]]Octane-7-carboxylic acid (270 mg, crude material). The crude product was used directly in the next step.

Step 4: n- [ (1S) -1- [7- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carbonyl ] -2, 2-dimethyl-propyl ] carbamic acid tert-butyl ester

To 6- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butanoyl at-20 ℃C]-6-azaspiro [3.4]]Octane-7-carboxylic acid (210 mg,569.93umol,1 eq.) and (2S) -2-amino-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]To a solution of propionamide (112.41 mg,569.93umol,1 eq.) in DMF (2 mL) was added DMF (1 mL) containing PyBOP (296.59 mg,569.93umol,1 eq.) and TEA (115.34 mg,1.14mmol,158.66uL,2 eq.). Subsequently, the reaction was stirred at 20℃for 2h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 30mL EtOAc (10 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=50/1 to 0/1, 10% MeOH) to give the compound N- [ (1S) -1- [7- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] as a white oil ]Hept-6-yl]Methyl group]Ethyl group]Carbamoyl group]-6-azaspiro [3.4]]Octane-6-carbonyl group]-2, 2-dimethyl-propyl]Tert-butyl carbamate (175 mg,319.53umol,56.06% yield, assuming 100% purity).

Step 5:6- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] -6-azaspiro [3.4] octane-7-carboxamide

A solution of tert-butyl N- [ (1S) -1- [7- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carbonyl ] -2, 2-dimethyl-propyl ] carbamate (140 mg,255.62umol,1 eq.) in 4MHCl/EtOAc (5 mL). Subsequently, the reaction was stirred at 20℃for 2h. After completion, the reaction was concentrated to dryness in vacuo to give the compound 6- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] -6-azaspiro [3.4] octane-7-carboxamide (133 mg, crude material, 2 HCl) as a white solid. The crude product was used directly in the next step.

Step 6: n- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] -6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -6-azaspiro [3.4] octane-7-carboxamide

To 6- [ (2S) -2-amino-3, 3-dimethyl-butyryl]-N- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]Methyl group]Ethyl group]-6-azaspiro [3.4]]To a solution of octane-7-carboxamide (133 mg,255.53umol,1 eq, 2 HCl) in DCM (3 mL) was added DIEA (132.10 mg,1.02mmol,178.03ul,4 eq) followed by DCM (0.5 mL) containing TFAA (134.17 mg,638.82umol,88.86ul,2.5 eq). Subsequently, the reaction was stirred at 20℃for 2h. After completion, by adding 10mL of H at 20 ℃ ₂ O quench the reaction mixture and then dilute with 10mL EtOAc and extract with 20mL EtOAc (10 mL x 2). The combined organic layers were washed with 10mL saturated NaCl (10 mL x 1), dried over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives the compound N- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] as a brown solid]Hept-6-yl]Methyl group]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4]]Octane-7-carboxamide (110 mg, crude). The crude product was used directly in the next step.

Step 7: n- [ (1S) -1-cyano-2- [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] ethyl ] -6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4 ]]Hept-6-yl]Methyl group]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4 ]]To a solution of octane-7-carboxamide (110 mg,202.36umol,1 eq.) in DCM (5 mL) was added the Buerger's reagent (106.09 mg,445.20umol,2.2 eq.). Subsequently, the reaction was stirred at 25℃for 16h. After completion, use N ₂ The reaction was blow-dried. By preparative HPLC (column: waters Xbridge Prep OBD C18.times.40 mm.times.10 um; mobile phase: [ water (NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -55%,8 min) and preparative HPLC (column: phenomenex Luna C18, 75 x 30mm x 3um; mobile phase: [ Water (FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,8 min) to give N- [ (1S) -1-cyano-2- [ (6R) -5-oxo-4-azaspiro [2.4 ] as a white solid]Hept-6-yl]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide (isomer 1, 10mg,18.63umol,9.21% yield, 97.9% purity). MS (ESI) m/z 526.1[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.41(br d,J＝9.0Hz,1H),8.56(d,J＝8.3Hz,1H),7.83(s,1H),5.03-4.80(m,1H),4.67(d,J＝8.8Hz,1H),4.19(t,J＝7.3Hz,1H),3.83(d,J＝10.3Hz,1H),3.61(d,J＝10.3Hz,1H),2.58-2.52(m,1H),2.25(dd,J＝8.3,12.5Hz,1H),2.11-1.92(m,5H),1.91-1.75(m,6H),0.97(s,9H),0.79-0.69(m,1H),0.63-0.48(m,3H)。

Obtaining N- [ (1S) -1-cyano-2- [ (6R) -5-oxo-4-azaspiro [2.4 ] as a white solid]Hept-6-yl]Ethyl group]-6- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] ]Butyryl group]-6-azaspiro [3.4 ]]Octane-7-carboxamide (isomer 2, 20mg,34.40umol,17.00% yield, 90.4% purity). MS (ESI) m/z 526.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.35(br d,J＝7.1Hz,1H),8.89(d,J＝8.7Hz,1H),7.73(s,1H),5.04-4.82(m,1H),4.52(br d,J＝5.9Hz,1H),4.18(t,J＝8.1Hz,1H),3.88(d,J＝9.9Hz,1H),3.49(d,J＝10.0Hz,1H),2.76-2.68(m,1H),2.28-2.15(m,2H),2.05-1.92(m,4H),1.86(br d,J＝2.8Hz,4H),1.80-1.65(m,2H),1.00(s,9H),0.79-0.67(m,1H),0.63-0.47(m,3H)。

EXAMPLE 238 Synthesis of viral protease inhibitor Compound 3133

Step 1: (S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamic acid tert-butyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (5 g,16.65mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 50mL,21.02 eq.) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in DCM (10 mL) and concentrated twice under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Tert-butyl carbamate (10.1 g, crude) and used directly in the next step. MS (ESI) m/z 286.1[ M+H ]] ⁺

Step 2: (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propanamide

N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]A mixture of tert-butyl carbamate (10.1 g,35.40mmol,1 eq.) in HCl/EtOAc (4M, 151.50mL,17.12 eq.) was stirred at 20deg.C for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in toluene (10 mL) and concentrated twice under reduced pressure to give methyl (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl as a pale yellow solid ]Propionamide (10.1 g, crude) which was used directly in the next step. MS (ESI) m/z 186.2[ M+H ]] ⁺

Step 3: ((4- (tert-butyl) phenyl) (2- ((4, 4-difluorocyclohexyl) amino) -1- (5-fluoropyridin-3-yl) -2-oxoethyl) carbamoyl) -L-proline methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Propionamide (0.5 g,2.26mmol,1 eq, HCl) and (1 r,2S, 5S) -3- [ (2S) -2- (tert-butoxycarbonylamino) -3, 3-di-n-Methyl-butyryl]-6, 6-dimethyl-3-azabicyclo [3.1.0]To a solution of hexane-2-carboxylic acid (831.06 mg,2.26mmol,1 eq.) in DMF (10 mL) was added PyBOP (1.29 g,2.48mmol,1.1 eq.) and then Et ₃ N (456.46 mg,4.51mmol,627.86uL,2 eq.). The mixture was stirred at-30℃for 2h. After completion, the combined reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (40 mL x 2). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate/meoh=20/1/0 to 0/0/1) to give N- [ (1S) -1- [ (1 r,2S, 5S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a pale yellow solid]Methyl group ]Ethyl group]Carbamoyl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-3-carbonyl]-2, 2-dimethyl-propyl]Tert-butyl carbamate (290 mg,541.37umol,24.00% yield). MS (ESI) m/z 436.2[ M-100+H] ⁺

Step 4: (1R, 2S, 5S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- ((S) -2-amino-3, 3-dimethylbutyryl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

A mixture of tert-butyl N- [ (1S) -1- [ (1R, 2S, 5S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-3-carbonyl ] -2, 2-dimethyl-propyl ] carbamate (290 mg,541.37umol,1 eq.) in HCl/EtOAc (4M, 50mL,369.43 eq) was stirred at 20℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in toluene (10 mL) and concentrated twice under reduced pressure to give (1 r,2S, 5S) -3- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide (300 mg, crude material, HCl) as a yellow solid and used directly in the next step.

Step 5: (1R, 2S, 5S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

To (1R, 2S, 5S) -3- [ (2S) -2-amino-3, 3-dimethyl-butyryl at 0deg.C]-N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]To a solution of hexane-2-carboxamide (0.2 g,423.71umol,1 eq, HCl) in DCM (10 mL) was added DIEA (164.29 mg,1.27mmol,221.41uL,3 eq) and TFAA (102.34 mg,487.27umol,67.78uL,1.15 eq). The mixture was stirred at 0℃for 1h. After completion, the combined reaction mixture was poured into NaHCO ₃ Aqueous solution (20 mL) and extracted with DCM (10 mL x 2). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (1R, 2S, 5S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide (294 mg, crude) which was used directly in the next step. MS (ESI) m/z 532.2[ M+H ] ] ⁺

Step 6: (1R, 2S, 5S) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

To (1R, 2S, 5S) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]To a solution of hexane-2-carboxamide (194 mg,364.96umol,1 eq.) in DCM (10 mL) was added methoxycarbonyl- (triethylammonium) sulfonyl-imide (260.91 mg,1.09mmol,3 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was quenched with water (0.5 mL) at 20 ℃ and with N ₂ The system was blow-dried to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,10 min) to give (1R, 2S, 5S) -N- [ (1S) -1-cyanogen as a white solid1-methyl-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0 ]Hexane-2-carboxamide (60.22 mg,117.26umol,32.13% yield, 100% purity). MS (ESI) m/z 514.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.41(br d,J＝7.2Hz,1H),8.99(d,J＝8.3Hz,1H),7.51(s,1H),5.01(ddd,J＝5.9,8.4,10.2Hz,1H),4.44-4.38(m,1H),4.16(s,1H),3.90(dd,J＝5.5,10.3Hz,1H),3.68(d,J＝10.5Hz,1H),3.12-3.05(m,2H),2.37-2.19(m,2H),1.85(br dd,J＝3.7,12.7Hz,1H),1.79-1.66(m,2H),1.60-1.52(m,2H),1.43-1.33(m,1H),1.29(d,J＝7.7Hz,1H),1.02(s,3H),0.99(s,9H),0.84(s,3H)。

EXAMPLE 239 Synthesis of (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propionitrile

Step 1: (S) -2- (((benzyloxy) carbonyl) amino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 0deg.C]To a solution of methyl propionate (10 g,38.02mmol,90% purity, 1 eq., HCl) in IPA (100 mL) was added NaOH (4M, 10.00mL,1.05 eq.) and NaHCO ₃ (194.40 g,2.31mol,90.00ml,60.86 eq.) in solution (ph=11). Next, benzyl (2, 5-dioxopyrrolidin-1-yl) carbonate (10.42 g,41.83mmol,1.1 eq.) was added. The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was poured into 100mL of H at 20 ℃ ₂ O, and then extracted with EtOAc (100 ml x 3). The combined organic layers were washed with brine (100 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 30/1 to give (2S) -2- (benzyloxycarbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl as a yellow oil ]Methyl propionate (10.5 g,31.40mmol,82.59% yield).

Step 2: (S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamic acid benzyl ester

(2S) -2- (Benzylmethoxycarbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (10 g,14.95mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 50mL,23.41 eq.) was stirred at 65℃for 48h. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Benzyl carbamate (9.55 g, crude material).

Step 3: ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) carbamic acid benzyl ester

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of benzyl carbamate (9.55 g,29.90mmol,1 eq.) in DCM (100 mL) was added the Buerger reagent (14.25 g,59.81mmol,2 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was poured into 120mL of H at 20 ℃ ₂ O, and then extracted with DCM (120 ml×3). The combined organic layers were washed with brine (100 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 0/1) the residue was purified to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Ethyl group]Benzyl carbamate (7.4 g,24.56mmol,82.12% yield).

Step 4: (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propionitrile

To N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]To a solution of benzyl carbamate (300 mg, 995.55. Mu. Mol,1 eq.) in IPA (6 mL) was added Pd (OH) 2 (699.05 mg, 995.55. Mu. Mol,20% purity, 1 eq.). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM/meoh=1/0 to 80/1 to give (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl as a white solid]Propionitrile (147 mg,820.24umol,82.39% yield, 93.3% pure)Degree). MS (ESI) m/z 168.1[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝4.11(t,J＝7.9Hz,1H),3.30-3.23(m,2H),2.56-2.45(m,1H),2.27-2.15(m,1H),2.03(d,J＝3.0,6.3,12.9Hz,1H),1.92-1.84(m,1H),1.81-1.70(m,2H),1.64-1.51(m,1H)。

EXAMPLE 240 Synthesis of viral protease inhibitor Compounds 247

Step 1: (S) -2-amino-3- (1H-indol-3-yl) propanamide

Methyl (2S) -2-amino-3- (1H-indol-3-yl) propionate (20 g,78.52mmol,1 eq. HCl) in NH ₃ A solution in MeOH (7M, 200.00mL,17.83 eq.) was stirred at 80℃for 38h. After completion, the reaction mixture was concentrated under reduced pressure to give (S) -2-amino-3- (1H-indol-3-yl) propionamide (17 g, crude material) as a yellow solid. MS (ESI) m/z 204.1[ M+H ]] ⁺ 。

Step 2: (2S) -2-amino-3- (2-oxoindolin-3-yl) propanamide

To a mixture of (2S) -2-amino-3- (1H-indol-3-yl) propionamide (17 g,83.64mmol,1 eq.) in AcOH (170 mL) was slowly added a mixture of HCl (12M, 27.88mL,4 eq.) and DMSO (9.80 g,125.47mmol,9.80mL,1.5 eq.) and the mixture was then stirred at 20℃for 14H. After completion, the reaction mixture was concentrated under reduced pressure to remove HCl and AcOH, followed by addition of NH at 20 ℃ ₃ .H ₂ O to quench until adjusted to pH>8. By preparative HPLC (column: xtime C18 u 250mm 80mm; mobile phase: [ water (0.05% NH.) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified 0% -20%,30 min) to give (2S) -2-amino-3- (2-oxoindolin-3-yl) propanamide (3.0 g,10.95mmol,13.09% yield, 80% purity) as a yellow solid. MS (ESI) m/z 220.2[ M+H ]] ⁺ 。

Step 3: 4-methoxy-1H-indole-2-carbonyl chloride

To 4-methoxy-1H-indole-2-carboxylic acid (10) g,52.31mmol,1 eq.) in DCM (100 mL) ₂ (26.56 g,209.22mmol,18.31mL,4 eq.) and DMF (191.16 mg,2.62mmol, 201.22. Mu.L, 0.05 eq.) followed by stirring at 40℃for 5h. After completion, the reaction mixture was concentrated under reduced pressure to give 4-methoxy-1H-indole-2-carbonyl chloride (11 g, crude material) as a brown solid.

Step 4: (S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propionic acid

To (2S) -2-amino-3-cyclopropyl-propionic acid (9.49 g,73.46mmol,1.4 eq.) in DCM (100 mL) and saturated Na at 25 ℃ ₂ CO ₃ To the mixture in (50 mL) was added dropwise a solution of 4-methoxy-1H-indole-2-carbonyl chloride (11 g,52.47mmol,1 eq.) in DCM (100 mL). Next, the mixture was stirred at 25℃for 0.5h. The reaction mixture was adjusted to ph=1 with 1N HCl, followed by extraction with DCM (50 ml x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered, concentrated under reduced pressure and purified by preparative HPLC (column: phenomenex luna C18 (250X 70mm,15 um): mobile phase [ water (0.05% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:23% -53%,27 min) to give (S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propionic acid (8.0 g,23.82mmol,45.39% yield, 90% purity) as a yellow solid. MS (ESI) m/z 303.1[ M+H ] ] ⁺ 。

Step 5: n- (1- (((2S) -1-amino-1-oxo-3- (2-oxoindolin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of (2S) -2-amino-3- (2-oxoindol-3-yl) propionamide (2 g,7.30mmol,80% purity, 1 eq.) in DCM (20 mL) and DMF (5 mL) was added (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]Propionic acid (2.75 g,7.30mmol,90% purity, 1 eq., HCl), DIEA (1.89 g,14.60mmol,2.54mL,2 eq.), HOBt (1.97 g,14.60mmol,2 eq.) and EDCI (2.80 g,14.60mmol,2 eq.) was then added at 0deg.C. The mixture was then stirred at 20℃for 2h. By adding H at 20 ℃ ₂ O (1 mL) to quench the reaction mixture, followed by concentration under reduced pressure and purification by preparative HPLC (column: phe)nomnex luna C18 (250 x 70mm,15 um); mobile phase: [ Water (0.05% HCl) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:22% -52%,27 min) to give N- (1- (((2S) -1-amino-1-oxo-3- (2-oxoindol-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (2.5 g,4.87mmol,66.67% yield, 98% purity) as a yellow solid. MS (ESI) m/z 504.2[ M+H ] ] ⁺ 。

Step 6: n- (1- (((1S) -1-cyano-2- (2-oxoindolin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a mixture of N- (1- (((2S) -1-amino-1-oxo-3- (2-oxoindol-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (2.5 g,4.87mmol,98% purity, 1 eq.) in DCM (25 mL) was added the bergs reagent (3.48 g,14.60mmol,3 eq.) and stirred for 2H at 20 ℃. Then the Bolus reagent (3.48 g,14.60mmol,3 eq.) was added and stirred at 20℃for 2h. By adding H at 20 ℃ ₂ O (2.5 mL) to quench the reaction mixture, followed by depressurization [. Sub.<Concentrated at 30 ℃ and purified by preparative HPLC (column: agela DuraShell C18, 250 x 70mm x 10um; mobile phase: [ Water (0.225% FA) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,20 min) to give N- (1- (((1S) -1-cyano-2- (2-oxoindol-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (800 mg,1.59mmol,32.65% yield, 96.4% purity) as a white solid. MS (ESI) m/z 486.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(br d,J＝11.0Hz,1H),10.52(br d,J＝18.5Hz,1H),9.13-8.94(m,1H),8.51(br d,J＝6.9Hz,1H),7.36(br s,1H),7.33-7.24(m,1H),7.22-7.15(m,1H),7.13-7.05(m,1H),7.02-6.91(m,2H),6.84(br t,J＝6.1Hz,1H),6.50(br d,J＝7.0Hz,1H),5.32-5.02(m,1H),4.59-4.40(m,1H),3.88(br s,3H),3.52-3.46(m,1H),2.41-2.15(m,2H),1.90-1.67(m,1H),1.64-1.35(m,1H),0.91-0.70(m,1H),0.53-0.31(m,2H),0.27-0.00(m,2H)

EXAMPLE 241 Synthesis of viral protease inhibitor Compound 689

Step 1: (S) -2- ((S) -3-cyclopropyl-2- (4-methoxy-1H-indole-2-carboxamido) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (150 mg,460.97umol,1 eq), 4-methoxy-1H-indole-2-carboxylic acid (105.76 mg,553.16umol,1.2 eq) in DCM (5 mL) was added DMAP (112.63 mg,921.94umol,2 eq) and EDCI (132.55 mg,691.45umol,1.5 eq). The mixture was stirred at 20℃for 2h. After completion, by adding 10mL H ₂ O quench the reaction mixture and then extract with DCM (10 ml x 3). The combined organic layers were washed with 15mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow oil]Propionyl radical]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (125 mg,248.21umol,53.85% yield, 99% purity). MS (ESI) m/z 499.2[ M+H ]] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (110 mg,220.63umol,1 eq.) in NH ₃ A solution in MeOH (220.63. Mu. Mol,10mL,1 eq.) was stirred at 65℃for 14h. After completion, the mixture was concentrated under reduced pressure to remove NH ₃ MeOH. DCM (10 mL) was added (three times) and the resulting solution was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (105 mg, crude). MS (ESI) m/z 484.2[ M+H ]] ⁺ 。

Step 3: n- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (105 mg,217.14umol,1 eq.) in DCM (5 mL) was added the Buerger's reagent (113.84 mg,477.71umol,2.2 eq.). The mixture was stirred at 25℃for 2h. After completion, at N ₂ DCM was removed. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the 35% -65%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (26 mg,55.85umol,25.72% yield, 100% purity). MS (ESI) m/z 466.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(s,1H),8.91(d,J＝8.2Hz,1H),8.51(br d,J＝7.6Hz,1H),7.82(s,1H),7.36(d,J＝1.3Hz,1H),7.13-7.06(m,1H),7.03-6.98(m,1H),6.50(d,J＝7.7Hz,1H),5.00-4.92(m,1H),4.49-4.41(m,1H),3.88(s,3H),2.64-2.55(m,1H),2.22-2.13(m,1H),1.99(dd,J＝8.5,12.3Hz,1H),1.89-1.70(m,2H),1.57-1.42(m,2H),1.19-1.03(m,6H),0.80(br dd,J＝6.0,7.3Hz,1H),0.47-0.33(m,2H),0.25-0.03(m,2H)

EXAMPLE 242 Synthesis of viral protease inhibitor Compound 731

At 0 ℃, to 2-tert-butoxycarbonyl-2-azaspiro [4.5]]Decane-3-carboxylic acid (200 mg,564.65umol,80% purity, 1 eq.) was added to a mixture of (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl in DCM (3 mL)]Methyl propionate (133.65 mg,564.65umol,1 eq., HCl), DMAP (206.95 mg,1.69mmol,3 eq.) and EDCI (216.49 mg,1.13mmol,2 eq.) were then stirred for 1h at 20 ℃. After completion, the reaction mixture was quenched by addition of water (3 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give 3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] as a yellow solid]Decane-2-carboxylic acid tert-butyl ester (200 mg,429.57umol,76.08% yield). MS (ESI) m/z 466.2[ M+H ]] ⁺ 。

3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] at 0deg.C]A mixture of tert-butyl decane-2-carboxylate (200 mg,386.61umol,90% purity, 1 eq.) in HCl/MeOH (4M, 2.67mL,27.59 eq.) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [ 4.5) as a yellow solid]Decane-3-carboxamido) propionic acid methyl ester (170 mg, crude material, HCl). MS (ESI) m/z 366.1[ M+H ]] ⁺ 。

Step 3: (2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [4.5] at 0 ]]To a mixture of methyl decane-3-carboxamido) propionate (172 mg,427.94umol,1 eq, HCl) in DCM (3 mL) was added 7-chloro-1H-indole-2-carboxylic acid (83.71 mg,427.94umol,1 eq)) DMAP (156.84 mg,1.28mmol,3 eq.) and EDCI (164.07 mg,855.88umol,2 eq.) and then stirring the resulting mixture at 20℃for 1h. After completion, the reaction mixture was quenched by addition of water (3 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give (2S) -2- (2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [ 4.5) as a yellow solid]Decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (100 mg,178.80umol,41.78% yield, 97.1% purity). MS (ESI) m/z 543.2[ M+H ]] ⁺ 。

Step 4: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH was added to a mixture of methyl propionate (100 mg,184.14umol,1 eq) ₃ MeOH (7 m,3ml,114.04 eq.) and then stirring the resulting mixture at 65 ℃ for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (90 mg, crude). MS (ESI) m/z 528.2[ M+H ]] ⁺

Step 5:2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (80 mg,151.50umol,1 eq.) in DCM (3 mL) was added the Bungeus reagent (72.21 mg,303.01umol,2 eq.) and stirred at 25℃for 2h. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ]<30 ℃ C.). By making Preparative HPLC (column: waters Xbridge BEH C100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide (4 mg,7.47 mol,4.93% yield, 95.2% purity). MS (ESI) m/z 510.1[ M+H ]] ⁺ .1HNMR (400 MHz, methanol-d 4) δ=7.63 (br d, J=7.9 Hz, 1H), 7.33-7.21 (m, 1H), 7.20-6.75 (m, 2H), 5.16-5.05 (m, 1H), 4.73-4.55 (m, 1H), 3.99-3.83 (m, 1H), 3.72 (br d, J=10.5 Hz, 1H), 3.28-2.93 (m, 2H), 2.73-2.39 (m, 2H), 2.39-2.23 (m, 1H), 2.03 (br s, 1H), 1.98-1.89 (m, 1H), 1.85-1.37 (m, 14H).

EXAMPLE 243 Synthesis of viral protease inhibitor Compound 818

To (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,3.80mmol,90% purity, 1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (932.77 mg,3.80mmol,1 eq) in DCM (20 mL) was added DMAP (1.16 g,9.51mmol,2.5 eq) and EDCI (1.46 g,7.60mmol,2 eq) and the mixture was then stirred at 20 ℃ for 1h. After completion, the reaction mixture was quenched with 50mL H ₂ O was diluted and extracted with DCM (30 ml x 3). The combined organic layers were washed with 40mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.2 g,2.81mmol,73.82% yield). MS (ESI) m/z 428.3[ M+H ]] ⁺ 。

A mixture of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (1.2 g,2.81mmol,1 eq.) in HCl/MeOH (4M, 20mL,28.50 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (1 g, crude material, HCl) as a white solid.

Step 3: 7-fluoro-1H-indole-2-carboxylic acid

To 7-fluoro-1H-indole-2-carboxylic acid ethyl ester (900 mg,4.34mmol,1 eq.) in THF (10 mL) and H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (546.77 mg,13.03mmol,3 eq.) and then the mixture was stirred at 60℃for 3h. After completion, 60mL of H was added by adding at 0deg.C ₂ O quench the reaction mixture and add 1MHCl drop wise to ph=5 and then extract with ethyl acetate (40 ml x 3). The combined organic layers were washed with 30mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 7-fluoro-1H-indole-2-carboxylic acid (700 mg, crude material) as a yellow solid.

Step 4: (S) -2- ((S) -2- (7-fluoro-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 7-fluoro-1H-indole-2-carboxylic acid (443.09 mg,2.47mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (0.9 g,2.47mmol,1 eq, HCl) in DCM (30 mL) was added DMAP (755.41 mg,6.18mmol,2.5 eq) and EDCI (948.29 mg,4.95mmol,2 eq) and the mixture was then stirred at 20℃for 2h. After completion, the reaction mixture was quenched with 60mL H ₂ O was diluted and extracted with DCM (40 ml x 3). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) to give the residue as white(2S) -2- [ [ (2S) -2- [ (7-fluoro-1H-indole-2-carbonyl) amino ] as a color solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g,1.64mmol,66.21% yield). MS (ESI) m/z 489.3[ M+H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-fluoro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g,1.64mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,85.50 eq.) was stirred at 30℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-7-fluoro-1H-indole-2-carboxamide (0.7 g, crude). MS (ESI) m/z 474.3[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 7-fluoro-1H-indole-2-carboxamide (0.6 g,1.27mmol,1 eq.) in DCM (15 mL) was added the berg reagent (452.92 mg,1.90mmol,1.5 eq.) and the mixture was then stirred at 40 ℃ for 1.5H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-7-fluoro-1H-indole-2-carboxamide (230 mg,495.32umol,39.09% yield, 98.1% purity). MS (ESI) m/z 456.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.03(s,1H),8.97(d,J＝7.7Hz,1H),8.59(d,J＝7.9Hz,1H),7.54(br s,1H),7.48-7.44(m,1H),7.29(d,J＝3.1Hz,1H),7.07-6.97(m,2H),5.09-5.01(m,1H),4.59-4.51(m,1H),3.11-3.02(m,2H),2.30-2.18(m,2H),1.87-1.77(m,2H),1.76-1.64(m,3H),1.59-1.48(m,1H),1.44-1.34(m,1H),0.94(s,9H)。

EXAMPLE 244 Synthesis of viral protease inhibitor Compound 826

Step 1: (Z) -2-azido-3- (2-chloro-5-fluoro-phenyl) prop-2-enoic acid methyl ester

A mixture of NaOMe (3.41 g,63.07mmol,2 eq.) in MeOH (30 mL) was cooled to-10deg.C, a mixture of 2-chloro-5-fluoro-benzaldehyde (5 g,31.53mmol,1 eq.) and ethyl 2-azidoacetate (8.14 g,63.07mmol,7.21mL,2 eq.) in MeOH (100 mL) was added dropwise, and the mixture was then stirred at 25deg.C for 18h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue, which was taken up in 60mL of H ₂ O was diluted and extracted with 90mL EA (30 mL x 3). The combined organic layers were washed with 45mL brine (45 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=1/0) to give methyl (Z) -2-azido-3- (2-chloro-5-fluoro-phenyl) prop-2-enoate (2.6 g,10.17mmol,32.25% yield) as a yellow solid.

Step 2: 4-chloro-7-fluoro-1H-indole-2-carboxylic acid methyl ester

A mixture of (Z) -2-azido-3- (2-chloro-5-fluoro-phenyl) prop-2-enoic acid methyl ester (2.4 g,9.39mmol,1 eq.) in xylene (25 mL) was stirred at 170℃for 1h. After completion, the reaction mixture was filtered to give methyl 4-chloro-7-fluoro-1H-indole-2-carboxylate (700 mg,3.08mmol,32.76% yield) as a white solid.

Step 3: 4-chloro-7-fluoro-1H-indole-2-carboxylic acid

To 4-chloro-7-fluoro-1H-indole-2-carboxylic acid methyl ester (700 mg,3.08mmol,1 eq.) in THF (4 mL) and H at 25 ℃ ₂ LiOH.H was added at once to the mixture in O (4 mL) ₂ O (258.08 mg,6.15mmol,2 eq.). The mixture was stirred at 60℃for 1 hour. After completion, the reaction mixture was made acidic with 1M HCl and extracted with 90mL EA (30 mL x 3). The combined organic layers were washed with 45mL brine (45 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 4-chloro-7-fluoro-1H-indole-2-carboxylic acid (600 mg,2.81mmol,91.34% yield) as a white solid. (ESI) M/z211.9[ M-H] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (4-chloro-7-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 4-chloro-7-fluoro-1H-indole-2-carboxylic acid (500 mg,2.34mmol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl [ (2S) -2 at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (895.68 mg,2.57mmol,1.1 eq., HCl) in DCM (10 mL) and DMF (3 mL) was added DMAP (857.96 mg,7.02mmol,3 eq.) and EDCI (897.50 mg,4.68mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was quenched with 30mL H ₂ O was diluted and extracted with 60mL EA (20 mL x 3). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (4-chloro-7-fluoro-1H-indole-2-carbonyl) amino ] as a white solid ]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,1.97mmol,84.27% yield). MS (ESI) m/z 505.0[ M-H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-chloro-7-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (4-chloro-7-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.21 g,2.17mmol,91% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,64.52 eq.) was stirred at 60℃for 16h. The reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-chloro-7-fluoro-1H-indole-2-carboxamide (850 mg,1.38mmol,63.70% yield, 80% purity). MS (ESI) M/z492.2[ M+H ]] ⁺

Step 6: 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃ ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-chloro-7-fluoro-1H-indole-2-carboxamide (780 mg,1.27mmol,80% purity, 1 eq.) in DCM (15 mL) was added the Bogis reagent (604.57 mg,2.54mmol,2 eq.) in one portion. The mixture was stirred at 40℃for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: waters Xbridge C18 150X 50mM 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give 4-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-fluoro-1H-indole-2-carboxamide (250 mg,527.51umol,41.59% yield). MS (ESI) m/z 474.1[ M+H ]]. ¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.48(br s,1H),8.96(d,J＝7.9Hz,1H),8.79(d,J＝7.5Hz,1H),7.54(br s,1H),7.44(d,J＝2.6Hz,1H),7.14-7.02(m,2H),5.07(q,J＝7.8Hz,1H),4.55-4.44(m,1H),3.17-3.00(m,2H),2.31-2.20(m,2H),1.91-1.65(m,4H),1.57(br d,J＝3.7Hz,1H),1.52-1.34(m,2H),0.89-0.75(m,1H),0.49-0.35(m,2H),0.26-0.05(m,2H)

Example 245 Synthesis of viral protease inhibitor Compounds 834 and 836

Step 1: (Z) -2-azido-3- (4-chloro-3-fluoro-phenyl) prop-2-enoic acid ethyl ester

To a solution of NaOMe (6.81 g,126.14mmol,2 eq.) in MeOH (70 mL) was added MeOH (100 mL) containing 4-chloro-3-fluoro-benzaldehyde (10 g,63.07mmol,1 eq.) and ethyl 2-azidoacetate (17.10 g,132.44mmol,15.13mL,2.1 eq.) at-10 ℃. After completion, the mixture was stirred at 20 ℃ for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH. By adding 100mL of H ₂ O quench the reaction mixture and extract with 100mL ethyl acetate (50 mL x 2). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 10/1, petroleum ether: ethyl acetate=5:1, (I2)) to give ethyl (Z) -2-azido-3- (4-chloro-3-fluoro-phenyl) prop-2-enoate (8.9 g,29.70mmol,47.10% yield, 90% purity) as a yellow solid. MS (ESI) m/z 256.02[ M+H ]] ⁺

Step 2: 6-chloro-7-fluoro-1H-indole-2-carboxylic acid methyl ester and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (4-chloro-3-fluoro-phenyl) prop-2-enoic acid ethyl ester (4 g,14.83mmol,1 eq.) in xylene (40 mL). The mixture was stirred at 170℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 5/1, petroleum ether: ethyl acetate=5:1, (UV 254 nm)) to give a mixture of methyl 6-chloro-7-fluoro-1H-indole-2-carboxylate (2.85 g,5.61mmol,37.82% yield, 44.8% purity) and methyl 6-chloro-5-fluoro-1H-indole-2-carboxylate (2.85 g,6.41mmol,43.22% yield, 51.2% purity) as a yellow solid. MS (ESI) m/z 228.01[ M+H ] ] ⁺

Step 3: 6-chloro-7-fluoro-1H-indole-2-carboxylic acid and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid

To 6-chloro-7-fluoro-1H-indole-2-carboxylic acid methyl ester (1 g,1.97mmol,44.8% purity, 1 eq.) and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid methyl ester (1 g,2.25mmol,51.2% purity)1.14 eq.) in THF (10 mL) and H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (247.76 mg,5.90mmol,3 eq.). The mixture was stirred at 60℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove THF, then HCl (1M) was added to the reaction mixture until ph=3, and then extracted with 10mL EA. The combined organic layers were washed with 10mL brine, filtered and concentrated under reduced pressure to give 6-chloro-7-fluoro-1H-indole-2-carboxylic acid (680 mg, crude) and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid (680 mg, crude) as yellow solids. MS (ESI) m/z 214.00[ M+H ]] ⁺

Step 4: (2S) -2- [ [ (2S) -2- [ (6-chloro-7-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester and (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.1 g,3.16mmol,1 eq, HCl) and 6-chloro-7-fluoro-1H-indole-2-carboxylic acid (405.28 mg,758.98 mol,63.67ul,40% purity, 0.24 eq) and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid (270.19 mg,758.98 mol,60% purity, 0.24 eq) in DCM (45 mL) were added followed by DMAP (1.16 g,9.49mmol,3 eq) and then EDCI (1.82 g,9.49mmol,3 eq). The mixture was stirred at 20℃for 2h. After completion, by adding 30mL H ₂ O quench the reaction mixture and then extract with 40mL DCM (20 mL x 2). The combined organic layers were washed with 30mL HCl (1M) (15 mL x 2), the combined organic layers were washed with 30mL brine, and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=3/1 to 0/1, dichloromethane: methanol=10:1, (UV 254 nm)) the residue was purified to give (2S) -2- [ [ (2S) -2- [ (6-chloro-7-fluoro-1H-indole-2-carbonyl) amino as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,685.76umol,21.68% yield, 40.9% purity) and (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino) ]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (850 mg,989.24umol,31.28% yield, 59% purity). MS (ESI) m/z 507.17[ M+H ]] ⁺ 。

Step 5: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-7-fluoro-1H-indole-2-carboxamide and N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-5-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-7-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (830 mg,669.63umol,40.9% purity, 1 eq.) and (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (830.00 mg,965.97umol,59% purity, 1.44 eq.) in NH ₃ A solution in MeOH (7M, 4.08mL,42.68 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent and to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-7-fluoro-1H-indole-2-carboxamide (800 mg, crude material) and N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-5-fluoro-1H-indole-2-carboxamide (800 mg, crude). MS (ESI) m/z 492.17[ M+H ]] ⁺ 。

Step 6: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-fluoro-1H-indole-2-carboxamide and 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl group]-6-chloro-7-fluoro-1H-indole-2-carboxamide (780 mg,635.81umol,40.1% purity, 1 eq.) and N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl)]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 6-chloro-5-fluoro-1H-indole-2-carboxamide (780.00 mg,948.16 mol,59.8% purity, 1.49 eq.) in DCM (18 mL) was added the bergs reagent (303.04 mg,1.27mmol,2 eq.). The mixture was stirred at 30℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -65%,10 min) to give 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-fluoro-1H-indole-2-carboxamide (500 mg,422.01umol,66.37% yield, 40% purity) and 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-fluoro-1H-indole-2-carboxamide (500 mg,633.01umol,99.56% yield, 60% purity). MS (ESI) m/z 474.16[ M+H ]] ⁺ 。

Step 7: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-fluoro-1H-indole-2-carboxamide and 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALCEL OD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH3H2O MEOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:38% -38%,6 min) isolation of 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-5-fluoro-1H-indole-2-carboxamide (500 mg) to give 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-fluoro-1H-indole-2-carboxamide (109.43 mg,230.21umol,54.55% yield, 99.7% purity). MS (ESI) m/z 474.16[ M+H ]] ⁺ 。1H NMR(400MHz,DMSO-d ₆ )δ＝0.04-0.29(m,2H),0.30-0.52(m,2H),0.71-0.90(m,1H),1.32-1.63(m,3H),1.65-1.90(m,4H),2.16-2.37(m,2H),2.97-3.18(m,2H),4.41-4.55(m,1H),4.97-5.13(m,1H),7.13(d,J＝8.56Hz,1H),7.34(d,J＝3.06Hz,1H),7.43-7.58(m,2H),8.65(d,J＝7.46Hz,1H),8.97(d,J＝7.95Hz,1H),12.31(s,1H)

EXAMPLE 246 Synthesis of viral protease inhibitor Compounds 844

Step 1: (Z) -2-azido-3- (4-bromo-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (1.00 g,18.60mmol,2 eq.) in MeOH (10 mL) was added MeOH (50 mL) containing 4-bromo-2-methoxy-benzaldehyde (2 g,9.30mmol,1 eq.) and ethyl 2-azidoacetate (2.40 g,18.60mmol,2.13mL,2 eq.) at 0deg.C. The mixture was stirred at 20℃for 18h. After the reaction was complete, the mixture was concentrated in vacuo, water (150 mL) was added and then extracted with EtOAc (50 mL x 3). The resulting mixture was concentrated in vacuo and passed through a column (SiO ₂ PE: EA=1:0 to 10:1) to give methyl (Z) -2-azido-3- (4-bromo-2-methoxyphenyl) acrylate (1.6 g,5.13mmol,55.12% yield) as a yellow solid.

Step 2: 6-bromo-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of methyl (Z) -2-azido-3- (4-bromo-2-methoxyphenyl) acrylate (1.6 g,5.13mmol,1 eq.) in xylene (10 mL) was stirred at 170℃for 1h. After the reaction was complete, the mixture was concentrated in vacuo and wet-triturated with petroleum ether (10 mL) and filtered to give 6-bromo-4-methoxy-1H-indole-2-carboxylic acid methyl ester (1.2 g,4.22mmol,82.40% yield) as a white solid. MS (ESI) m/z 283.8[ M+H ]] ⁺

Step 3: 6-bromo-4-methoxy-1H-indole-2-carboxylic acid

To 6-bromo-4-methoxy-1H-indole-2-carboxylic acid methyl ester (1.2 g,4.22mmol,1 eq.) in THF (12 mL) and H ₂ LiOH.H was added to the solution in O (6 mL) ₂ O (531.69 mg,12.67mmol,3 eq.) and then the mixture was stirred at 50℃for 5h. After the reaction was complete, the mixture was concentrated in vacuo, adjusted to pH-1 with 1M HCl (15 mL) and then extracted with EtOAc (5 mL x 3), followed by concentration in vacuo to give 6-bromo-4-methoxy-1H-indole-2-carboxylic acid (1 g, crude material) as a white solid. MS (ESI) m/z 268.0[ M-H ]] ⁺

Step 4: (S) -2- ((S) -2- (6-bromo-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 6-bromo-4-methoxy-1H-indole-2-carboxylic acid (480 mg,1.78mmol,1.2 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (461.16 mg,1.48mmol,1 eq.) in DMF (20 mL) was added DMF (1 mL) containing TEA (449.60 mg,4.44mmol,618.43uL,3 eq.) and PyBop (770.73 mg,1.48mmol,1 eq.) and the mixture was then stirred at-40℃for 2h. After completion of the reaction, the mixture was quenched with water (30 mL) and extracted with DCM (10 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ PE: ea=1:1 to 0:1 to DCM: meoh=10:1) to give methyl (S) -2- ((S) -2- (6-bromo-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionate (800 mg,993.90umol,67.11% yield, 70% purity) as a yellow gum. MS (ESI) m/z 563.2[ M-H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-bromo-4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (6-bromo-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (200 mg, 354.96. Mu. Mol,1 eq.) was reacted with NH ₃ A solution in MeOH (7M, 20 mL) was stirred at 50℃for 8h. After completion of the reaction, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-bromo-4-methoxy-1H-indole-2-carboxylic acid as a white solid Amine (600 mg, crude material).

Step 6: 6-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-bromo-4-methoxy-1H-indole-2-carboxamide (580 mg,1.06mmol,1 eq.) in DCM (8 mL) was added the bergs reagent (756.07 mg,3.17mmol,3 eq.) and the resulting mixture was then stirred at 30 ℃ for 1H. After the reaction was completed, the mixture was quenched with water (1 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give 6-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-methoxy-1H-indole-2-carboxamide (130 mg,227.94umol,21.55% yield, 93% purity) as a white solid. MS (ESI) m/z 530.0[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm11.72(d,J＝1.9Hz,1H),8.90(d,J＝8.1Hz,1H),8.57(d,J＝7.5Hz,1H),7.52(s,1H),7.38(d,J＝1.5Hz,1H),7.19(s,1H),6.66(d,J＝1.3Hz,1H),5.06(q,J＝8.1Hz,1H),4.49-4.39(m,1H),3.91(s,3H),3.14-3.02(m,2H),2.30-2.21(m,2H),1.88-1.75(m,3H),1.74-1.66(m,1H),1.62-1.51(m,1H),1.49-1.32(m,2H),0.86-0.74(m,1H),0.46-0.35(m,2H),0.24-0.05(m,2H)。

EXAMPLE 247 Synthesis of viral protease inhibitor Compound 846

Step 1: (Z) -2-azido-3- (5-bromo-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (2.51 g,46.50mmol,2 eq.) in MeOH (25 mL) was added 5-bromo-2-methoxy-benzaldehyde (5 g,23.25mmol,1 eq.) and ethyl 2-azidoacetate (6.30 g,48.83mmol,5.58mL,2.1 eq.) of MeOH (25 mL). The mixture was stirred at 20℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was taken up in 100mL H ₂ O was diluted and extracted with EtOAc (100 ml x 3). The combined organic layers were washed with solvent brine (100 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give methyl (Z) -2-azido-3- (5-bromo-2-methoxy-phenyl) prop-2-enoate (2.1 g, crude material) as a yellow solid.

Step 2: 7-bromo-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (5-bromo-2-methoxy-phenyl) prop-2-enoic acid methyl ester (2.1 g,6.73mmol,1 eq.) in xylene (43 mL) was stirred at 170℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give methyl 7-bromo-4-methoxy-1H-indole-2-carboxylate (100 mg, crude material) as a yellow solid.

Step 3: 7-bromo-4-methoxy-1H-indole-2-carboxylic acid

To 7-bromo-4-methoxy-1H-indole-2-carboxylic acid methyl ester (100 mg, 351.98. Mu. Mol,1 eq.) in THF (7 mL) and H ₂ LiOH.H was added to a solution in O (3.5 mL) ₂ O (44.31 mg,1.06mmol,3 eq.). The mixture was stirred at 50℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. 1M HCl was added to adjust the pH to 3, followed by filtration and concentration under reduced pressure to give 7-bromo-4-methoxy-1H-indole-2-carboxylic acid (50 mg, crude material) as a yellow solid.

Step 4: (S) -2- ((S) -2- (7-bromo-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 7-bromo-4-methoxy-1H-indole-2-carboxylic acid (500 mg,1.85mmol,1 eq.) (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (772.74 mg,2.22mmol,1.2 eq.) and DMAP (678.51 mg,5.55mmol,3 eq.) in DCM (10 mL) was added EDCI (709.80 mg,3.70mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ In OAnd then extracted with DCM (35 ml x 3). The combined organic layers were washed with brine (35 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=50/1 to 10/1) to give (2S) -2- [ [ (2S) -2- [ (7-bromo-4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (460 mg,816.41umol,44.10% yield).

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-bromo-4-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (450 mg,798.67umol,1 eq.) in NH ₃ Solution in MeOH (7M, 18mL,157.76 eq.). The mixture was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH to give N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-bromo-4-methoxy-1H-indole-2-carboxamide (430 mg, crude). MS (ESI) m/z 548.1[ M+H ]] ⁺ 。

Step 6: 7-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-bromo-4-methoxy-1H-indole-2-carboxamide (420 mg,765.82umol,1 eq.) in DCM (5 mL) was added the Bungeus reagent (365.00 mg,1.53mmol,2 eq.). The mixture was stirred at 30℃for 3h. After completion, the reaction mixture was poured into 20mL of H at 20 ℃ ₂ O, and then extracted with DCM (25 ml x 3). The combined organic layers were washed with brine (20 ml x 2), dried over Na ₂ SO ₄ Drying, filtering and reducingConcentration under reduced pressure gave a residue. By preparative HPLC (column Phenomenex Luna C, 18, 200X 40mm X10 um; mobile phase: [ water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -70%,8 min) to give 7-bromo-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (95.7 mg,180.43umol,23.56% yield, 100% purity). MS (ESI) m/z 530.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.46(d,J＝1.5Hz,1H),8.98(d,J＝7.9Hz,1H),8.72(d,J＝7.6Hz,1H),7.52(s,1H),7.35(d,J＝8.2Hz,1H),7.28(d,J＝2.1Hz,1H),6.54(d,J＝8.3Hz,1H),5.07(d,J＝7.9Hz,1H),4.51(d,J＝6.2Hz,1H),3.89(s,3H),3.17-3.00(m,2H),2.26(t,J＝8.7Hz,2H),1.81(dd,J＝8.5,14.2Hz,4H),1.49(s,3H),0.89-0.73(m,1H),0.52-0.36(m,2H),0.25-0.04(m,2H)。

EXAMPLE 248 Synthesis of viral protease inhibitor Compound 850

Step 1: 7-bromo-6-fluoro-1H-indole

At-40 ℃ under N ₂ To a solution of 2-bromo-1-fluoro-3-nitro-benzene (8 g,36.36mmol,1 eq.) in THF (110 mL) was added dropwise magnesium bromide (1 m,127.28mL,3.5 eq.). The reaction mixture was stirred at-40℃for a further 1.5hr. After completion, at N ₂ Pouring the residue into NH ₄ Aqueous Cl (200 mL) and stirred for 10min. The aqueous phase was extracted with ethyl acetate (200 ml x 4). The combined organic phases were washed with brine (300 ml x 2), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 10/1) to give 7-bromo-6-fluoro-1H-indole (2.4 g,11.21mmol,30.84% yield, N/A purity) as a yellow oil. MS (ESI) M/z213.0[ M+H ]] ⁺ 。

Step 2: 7-bromo-6-fluoro-1H-indole-1-carboxylic acid tert-butyl ester

To 7-bromo-6-fluoro-1H-indole (2.4 g,11.21 m)Boc was added to a solution of mol,1 eq.) and TEA (1.36 g,13.46mmol,1.87mL,1.2 eq.) in DCM (25 mL) ₂ O (2.69 g,12.33mmol,2.83mL,1.1 eq.) and DMAP (273.98 mg,2.24mmol,0.2 eq.). The mixture was stirred at 25℃for 2hr. After completion, by adding 40mL H ₂ O quench the reaction mixture and extract with DCM (20 ml x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/0 to 10/1) to give 7-bromo-6-fluoro-indole-1-carboxylic acid tert-butyl ester (3 g,8.88mmol,79.20% yield, 93% purity) as a yellow oil. MS (ESI) m/z 313.0[ M+H ]] ⁺ 。

Step 3: 7-bromo-1- (tert-butoxycarbonyl) -6-fluoro-1H-indole-2-carboxylic acid

At-60 ℃ under N ₂ To a solution of 7-bromo-6-fluoro-indole-1-carboxylic acid tert-butyl ester (3 g,9.55mmol,1 eq.) in THF (30 mL) was added LDA (2 m,5.25mL,1.1 eq.) dropwise. The reaction mixture was stirred at-60 ℃ for 0.5h, then the solution was added to solid carbon dioxide (21.01 g,1.00 eq). The reaction mixture was stirred at 25℃for an additional 1hr. After completion, at N ₂ The reaction mixture was poured into H ₂ O (80 mL) and stirred for 10min. To the aqueous phase was added 1M HCl at 0 ℃ to adjust to pH-2 and extracted with ethyl acetate (40 ml x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column Phenomenex luna C (250X 70mm,15 um); mobile phase: [ water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,20 min) to give 7-bromo-1-tert-butoxycarbonyl-6-fluoro-indole-2-carboxylic acid (900 mg,2.51mmol,26.31% yield, N/a purity) as a white solid. MS (ESI) m/z 357.0[ M+H ] ] ⁺ 。

Step 4: 7-bromo-6-fluoro-1H-indole-2-carboxylic acid

To a solution of 7-bromo-1-tert-butoxycarbonyl-6-fluoro-indole-2-carboxylic acid (900 mg,2.51mmol,1 eq.) in THF (10 mL) was added HBr (14.90 g,73.66mmol,10mL,40% purity, 29.31 eq.). At 25 DEG CThe mixture was stirred for 8hr. After completion, by adding H ₂ O (40 mL) quenched the reaction mixture and extracted with EtOAc (25 mL x 4). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The compound 7-bromo-6-fluoro-1H-indole-2-carboxylic acid (650 mg, crude material) was obtained as a yellow solid. MS (ESI) m/z 256.9[ M+H ]] ⁺ 。

Step 5: (S) -2- ((S) -2- (7-bromo-6-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 7-bromo-6-fluoro-1H-indole-2-carboxylic acid (650 mg,2.52mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (876.18 mg,2.52mmol,1 eq., HCl) in DCM (25 mL) was added DMAP (615.47 mg,5.04mmol,2 eq.) and EDCI (724.33 mg,3.78mmol,1.5 eq.). The mixture was stirred at 25℃for 1h. After completion, the reaction was slowly quenched with 60mL H ₂ O quenched and then extracted with DCM (30 ml x 3). The combined organic phases were washed with brine (45 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=3/1 to 0/1) to give (2S) -2- [ [ (2S) -2- [ (7-bromo-6-fluoro-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.75 g,1.29mmol,51.30% yield, 95% purity). MS (ESI) m/z 550.1[ M+H ]] ⁺ 。

Step 6: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-6-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-bromo-6-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (800 mg,1.45mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 40mL,192.99 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to remove DCM to give a brown solidN- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group ]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-bromo-6-fluoro-1H-indole-2-carboxamide (730 mg, crude). MS (ESI) m/z 535.1[ M+H ]] ⁺ 。

Step 7: 7-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-bromo-6-fluoro-1H-indole-2-carboxamide (730 mg,1.36mmol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (648.64 mg,2.72mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion, at N ₂ The reaction mixture was concentrated at 25 ℃. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mm X10 um; mobile phase: [ water (0.05% NH) ₃ H ₂ O+10mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -55%,8 min) to give 7-bromo-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-fluoro-1H-indole-2-carboxamide (235 mg,453.34umol,33.31% yield, 100% purity). MS (ESI) m/z 517.1[ M+H ] ] ⁺ 。1H NMR(400MHz,DMSO-d ₆ )δ＝11.72-11.49(m,1H),9.11-8.90(m,1H),8.55-8.37(m,1H),7.67-7.49(m,2H),7.06-6.93(m,1H),6.68-6.60(m,1H),5.17-5.04(m,1H),4.65-4.55(m,1H),3.17-3.00(m,2H),2.37-2.19(m,2H),1.94-1.66(m,4H),1.62-1.32(m,3H),0.84-0.72(m,1H),0.52-0.37(m,2H),0.21-0.05(m,2H)。

Example 249 Synthesis of viral protease inhibitor Compounds 854

Step 1: (Z) -2-azido-3- (5-bromo-2-fluorophenyl) acrylic acid ethyl ester and (Z) -2-azido-3- (5-bromo-2-fluorophenyl) acrylic acid methyl ester

NaO is processed byA mixture of Me (2.66 g,49.26mmol,2 eq.) in MeOH (30 mL) was cooled to-10deg.C, and then a mixture of 5-bromo-2-fluoro-benzaldehyde (5 g,24.63mmol,1 eq.) and ethyl 2-azidoacetate (6.36 g,49.26mmol,5.63mL,2 eq.) in MeOH (70 mL) was added dropwise to the foregoing solution. The mixture was stirred at 20℃for 18h. After completion, the reaction mixture was concentrated under reduced pressure to remove 60mL of MeOH. The residue was taken up in 100mL H ₂ O was diluted and extracted with EtOAc (100 ml x 3). The combined organic layers were washed with 100mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether Ethyl acetate=1:0 to 100:1) the residue was purified to give the product (Z) -2-azido-3- (5-bromo-2-fluoro-phenyl) prop-2-enoic acid ethyl ester (1.6 g,5.09mmol,20.68% yield) as a yellow solid and (Z) -2-azido-3- (5-bromo-2-fluoro-phenyl) prop-2-enoic acid methyl ester (1.6 g,5.33mmol,21.65% yield) as a yellow solid.

Step 2: 7-bromo-4-fluoro-1H-indole-2-carboxylic acid ethyl ester and 7-bromo-4-fluoro-1H-indole-2-carboxylic acid methyl ester

A mixture of (Z) -2-azido-3- (5-bromo-2-fluoro-phenyl) prop-2-enoic acid ethyl ester (1.6 g,5.09mmol,1 eq.) and (Z) -2-azido-3- (5-bromo-2-fluoro-phenyl) prop-2-enoic acid methyl ester (1.6 g,5.33mmol,1.05 eq.) in xylene (30 mL) was stirred at 170℃for 1.5h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 100:1) to give ethyl 7-bromo-4-fluoro-1H-indole-2-carboxylate (0.35 g,1.22mmol,24.02% yield) as a white solid and methyl 7-bromo-4-fluoro-1H-indole-2-carboxylate (0.35 g,1.29mmol,25.26% yield) as a white solid.

Step 3: 7-bromo-4-fluoro-1H-indole-2-carboxylic acid

To 7-bromo-4-fluoro-1H-indole-2-carboxylic acid ethyl ester (250 mg,873.83umol,1 eq.) in THF (6 mL) and H ₂ LiOH.H was added to the solution in O (3 mL) ₂ O (110.00 mg,2.62mmol,3 eq.) and then the mixture was stirred at 60℃for 3.5h. After completion, 60mL of H was added by adding at 0deg.C ₂ O to quench the reactionThe mixture was added dropwise to adjust the pH to about 5, and then extracted with EtOAc (40 ml x 3). The combined organic layers were washed with 30mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 7-bromo-4-fluoro-1H-indole-2-carboxylic acid (200 mg, crude) as a white solid. MS (ESI) m/z 255.9[ M-H ]] ⁺ 。

7-bromo-4-fluoro-1H-indole-2-carboxylic acid methyl ester (350 mg,1.29mmol,1 eq.) in THF (6 mL) and H ₂ A solution in O (3 mL) was then added LiOH.H ₂ O (161.94 mg,3.86mmol,3 eq.) and the mixture stirred at 60℃for 3.5h. After completion, 60mL of H was added by adding at 0deg.C ₂ O quench the reaction mixture and add 1M HCl dropwise to ph=5 and then extract with EtOAc (40 ml x 3). The combined organic layers were washed with 30mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 7-bromo-4-fluoro-1H-indole-2-carboxylic acid (300 mg, crude) as a white solid.

Step 4: (S) -2- ((S) -2- (7-bromo-4-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

7-bromo-4-fluoro-1H-indole-2-carboxylic acid (500 mg,1.94mmol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A solution of methyl propionate (876.18 mg,2.52mmol,1.30 eq., HCl) in DCM (10 mL) was then added DMAP (710.16 mg,5.81mmol,3 eq.) and EDCI (742.91 mg,3.88mmol,2 eq.) and the mixture stirred at 20deg.C for 2h. After completion, 60mL of H was added by adding at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (40 ml x 3). The combined organic layers were washed with 60mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) the residue was purified to give the product (2S) -2- [ [ (2S) -2- [ (7-bromo-4-fluoro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (600 mg,1.09mmol,56.16% yield). MS (ESI) m/z 551.1[ M+H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-bromo-4-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-bromo-4-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.6 g,1.09mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 30.00mL,192.99 eq.) was stirred at 60℃for 12h. The reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Amino group ]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-bromo-4-fluoro-1H-indole-2-carboxamide (550 mg, crude). MS (ESI) m/z 536.1[ M+H ]] ⁺ 。

Step 6: 7-bromo-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-bromo-4-fluoro-1H-indole-2-carboxamide (550 mg,1.03mmol,1 eq.) in DCM (20 mL) was added the berg reagent (488.70 mg,2.05mmol,2 eq.) and the mixture was then stirred at 40 ℃ for 12H. The reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product 7-bromo-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-fluoro-1H-indole-2-carboxamide (230 mg,443.69umol,43.27% yield, 100% purity). MS (ESI) m/z 518.0[ M+H ] ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.76(br s,1H),9.01(d,J＝7.9Hz,1H),8.84(d,J＝7.5Hz,1H),7.53(br s,1H),7.48-7.40(m,1H),7.34(s,1H),6.94-6.79(m,1H),5.15-5.02(m,1H),4.60-4.46(m,1H),3.17-3.00(m,2H),2.31-2.17(m,2H),1.90-1.66(m,4H),1.63-1.35(m,3H),0.91-0.75(m,1H),0.53-0.38(m,2H),0.24-0.17(m,1H),0.15-0.07(m,1H)。

Example 250 Synthesis of viral protease inhibitor Compounds 858

Step 1: (S) -2- ((S) -2- (4-cyano-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpiperidin-3-yl) propanoate (1.05 g,2.40mmol,80% purity, 1.1 eq, HCl) in DCM (4 mL) and DMF (1 mL) was added 4-cyano-1H-indole-2-carboxylic acid (500 mg,2.69mmol,1 eq). After EDCI (1.03 g,5.37mmol,2 eq.) and DMAP (984.36 mg,8.06mmol,3 eq.) were added at 0deg.C, the mixture was stirred at 20deg.C for 2h. After completion, the reaction mixture was quenched by addition of water (3 mL) and then extracted with DCM (3 mL x 2). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with DCM: meoh=100:1 to 10:1 to give methyl (S) -2- ((S) -2- (4-cyano-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (750 mg,1.49mmol,55.32% yield, 95% purity) as a yellow solid. MS (ESI) m/z 480.1[ M+H ] ] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-cyano-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (4-cyano-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (700 mg,1 eq) in NH ₃ The mixture in MeOH (7M, 5mL,745.93 eq.) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (N- ((S) -1- (((S) -1-amino) as a yellow solid)-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-cyano-1H-indole-2-carboxamide (670 mg, crude material). MS (ESI) m/z 465.2[ M+H ]] ⁺ 。

Step 3: 4-cyano-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-cyano-1H-indole-2-carboxamide (640 mg,1.37mmol,95% purity, 1 eq.) in DCM (10 mL) was added the Buerger's reagent (979.63 mg,4.11mmol,3 eq.) and stirred at 20deg.C for 14H. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ] <30 ℃ C.). By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -55%,10 min) to give 4-cyano-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-indole-2-carboxamide (40.4 mg,90.48umol,6.60% yield, 100% purity) as an off-white solid. MS (ESI) m/z 447.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.84-7.68(m,1H),7.60-7.46(m,1H),7.46-7.39(m,1H),7.39-7.29(m,1H),5.18-5.01(m,1H),4.66-4.48(m,1H),3.29-3.15(m,2H),2.57-2.27(m,2H),2.05-1.46(m,7H),1.01-0.73(m,1H),0.61-0.45(m,2H),0.28-0.11(m,2H)

EXAMPLE 245 Synthesis of viral protease inhibitor Compounds 864

Step 1: 5-chloro-1H-pyrrole-2-carboxylic acid

To a mixture of methyl 5-chloro-1H-pyrrole-2-carboxylate (500 mg,3.13mmol,1 eq.) in MeOH (2 mL) was added NaOH (250.66 mg,6.27mmol,2 eq.) in H ₂ O (2 mL), and then at 80 ℃The resulting mixture was stirred for 14h. After completion, the mixture was concentrated under reduced pressure to give 5-chloro-1H-pyrrole-2-carboxylic acid (500 mg, crude material) as a yellow oil. MS (ESI) m/z 146.0[ M+H ]] ⁺ 。

Step 2: (S) -2- ((S) -2- (5-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]Methyl propionate (1.49 g,3.44mmol,80% purity, 1 eq, HCl) was added to a mixture of DMF (1 mL) and DCM (4 mL), and then DMAP (1.26 g,10.31mmol,3 eq) and EDCI (1.32 g,6.87mmol,2 eq) were added at 0 ℃, followed by stirring the mixture at 25 ℃ for 2H. After completion, the reaction mixture was quenched by addition of water (10 mL) and then extracted with DCM (10 mL x 3). The combined organic layers were washed with HCl (1M, 10 mL), then brine (10 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=100:1 to 10:1 to give methyl (S) -2- ((S) -2- (5-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (300 mg,683.52umol,19.90% yield) as a yellow solid. MS (ESI) m/z 439.1[ M+H ]] ⁺ 。

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-chloro-1H-pyrrole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (5-chloro-1H-pyrrole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group ]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (300 mg,683.52umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,102.41 eq.) was stirred at 80℃for 30h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-chloro-1H-pyrrole-2-carboxamide (280 mg, crude material) as a yellow solid. MS (ESI) m/z 424.1[ M+H ]] ⁺ 。

Step 4: 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 5-chloro-1H-pyrrole-2-carboxamide (220 mg,467.10umol,90% purity, 1 eq.) in DCM (3 mL) was added the berg reagent (333.94 mg,1.40mmol,3 eq.) and the resulting mixture was then stirred at 25 ℃ for 4H. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ]<30 ℃ C.). By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide (30.49 mg,75.12umol,16.08% yield, 100% purity) as a white solid. MS (ESI) m/z 406.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.36(s,1H),9.10-8.81(m,1H),8.13-8.03(m,1H),7.52(br s,1H),6.96-6.50(m,1H),6.45-6.02(m,1H),5.17-4.88(m,1H),4.49-4.31(m,1H),3.15-3.01(m,2H),2.29-2.14(m,2H),1.88-1.66(m,4H),1.61-1.48(m,1H),1.47-1.25(m,2H),0.84-0.63(m,1H),0.53-0.24(m,2H),0.20-0.01(m,2H)

Example 251 Synthesis of viral protease inhibitor Compounds 868

Step 1: 5-chloro-1H-imidazole-2-carboxylic acid ethyl ester

To a solution of ethyl 1H-imidazole-2-carboxylate (5 g,35.68mmol,1 eq.) in DMF (150 mL) was added acetic acid (1 mL) dropwise, and then NCS (3.00 g,22.47mmol,0.63 eq.) containing DMF (30 mL) was added at 0deg.C, the mixture was stirred at 20deg.C for 20H, and then 45℃for 24H, and then 80 ℃And stirring for 2 hours. After completion, the reaction mixture was diluted with water (100 mL) and extracted with EA (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE: ea=1:0 to 65:35 to give the product 5-chloro-1H-imidazole-2-carboxylic acid ethyl ester (2 g,8.02mmol,22.48% yield, 70% purity) as a pale yellow solid. MS (ESI) m/z 175.1[ M+H ] ] ⁺ 。

Step 2: 5-chloro-1H-imidazole-2-carboxylic acid

To 5-chloro-1H-imidazole-2-carboxylic acid ethyl ester (2.5 g,10.02mmol,70% purity, 1 eq.) in THF (30 mL) and H ₂ To a mixture of O (10 mL) and MeOH (3 mL) was added LiOH.H ₂ O (1.05 g,25.06mmol,2.5 eq.). The mixture was stirred at 40℃for 24h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (TFA column: waters X bridge C18:18:150.50 mm.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -15%,10 min) to give the product 5-chloro-1H-imidazole-2-carboxylic acid (1.0 g,6.82mmol,68.08% yield) as a white solid. MS (ESI) m/z 145.1[ M+H ]] ⁺ 。

Step 3: (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -3-cyclopropyl-propionamide

To N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of tert-butyl carbamate (2.1 g,5.55mmol,1 eq.) in EA (12 mL) was added HCl/EA (4M, 12mL,8.65 eq.). The mixture was stirred at 20℃for 1h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with ethyl acetate (20 ml x 3) and concentrated under reduced pressure to give the product (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]3-cyclopropyl-propionamide (1.7 g, crude material, HCl). MS (ESI) M/z279.1[ M+H] ⁺ 。

Step 4: 5-chloro-N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-imidazole-2-carboxamide

To a solution of 5-chloro-1H-imidazole-2-carboxylic acid (800 mg,5.46mmol,2.02 eq.) in DMF (20 mL) was added HOBt (729.67 mg,5.40mmol,2 eq.), EDCI (1.04 g,5.40mmol,2 eq.) and (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]3-cyclopropyl-propionamide (1.7 g,2.70mmol,50% purity, 1 eq, HCl). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with EA (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters X bridge C, 150X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -45%,10 min) to give the product 5-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (500 mg,1.23mmol,45.51% yield). MS (ESI) m/z 407.1[ M+H ] ] ⁺ 。 ¹ HNMR(400MHz,DMSO-d ₆ )δ＝11.59(br s,1H),8.96-8.79(m,1H),8.42-8.28(m,1H),7.52(br s,1H),7.40(s,1H),5.11-4.88(m,1H),4.53-4.35(m,1H),3.07(br s,2H),2.32-2.10(m,2H),1.81(br s,4H),1.62-1.34(m,3H),0.78-0.63(m,1H),0.39(br d,J＝7.8Hz,2H),0.21-0.05(m,2H)。

By SFC (column: DAICEL CHIRALPAK IF (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:27% -27%,9 min) isolation of 5-chloro-N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (300 mg,737.34umol,1 eq.) gives 5-chloro-N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide isomer 1 (197.02 mg,484.24umol,65.67% yield). MS (ESI) M/z407.1[ M+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝13.47(br s,1H),8.85(br d,J＝8.0Hz,1H),8.40(br d,J＝8.0Hz,1H),7.52(br s,1H),7.43(s,1H),5.17-4.92(m,1H),4.54-4.25(m,1H),3.09(br s,2H),2.34-2.18(m,2H),1.88-1.65(m,4H),1.62-1.31(m,3H),0.70(br s,1H),0.39(br d,J＝7.6Hz,2H),0.19-0.00(m,2H)。

Obtaining 5-chloro-N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (10.42 mg,25.61umol,3.47% yield). MS (ESI) m/z407.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝13.44(br s,1H),8.86(br d,J＝7.6Hz,1H),8.35(br d,J＝8.0Hz,1H),7.52(br s,1H),7.42(s,1H),5.11-4.95(m,1H),4.51-4.38(m,1H),3.09(br s,2H),2.26-2.08(m,2H),1.89-1.66(m,4H),1.63-1.34(m,3H),0.69(br s,1H),0.38(br s,2H),0.09(br d,J＝13.7Hz,2H)。

Obtaining 5-chloro-N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (39.82 mg,97.87umol,13.27% yield). MS (ESI) m/z407.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝13.45(br s,1H),8.92(br d,J＝7.2Hz,1H),8.41-8.32(m,1H),7.53(br s,1H),7.43(s,1H),5.07-4.83(m,1H),4.45(br d,J＝5.4Hz,1H),3.09(br s,2H),2.35-2.12(m,2H),1.92-1.67(m,4H),1.65-1.35(m,3H),0.70(br s,1H),0.38(br s,2H),0.09(br d,J＝16.8Hz,2H)。

Example 252 Synthesis of viral protease inhibitor Compound 870

Step 1: (S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid tert-butyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (5 g,12.15mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 50mL,28.80 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give N- [ (1S) -2-amino ] as a yellow solid-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Tert-butyl carbamate (4.8 g, crude material). MS (ESI) m/z 397.2[ M+H ]] ⁺ 。

Step 2: (S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid tert-butyl ester

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of tert-butyl carbamate (4.8 g,12.11mmol,1 eq.) in DCM (50 mL) was added the Buerger reagent (5.77 g,24.21mmol,2 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was poured into 60mL of H at 20 ℃ ₂ O, and then extracted with DCM (60 ml x 3). The combined organic layers were washed with brine (60 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 0/1) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]Tert-butyl carbamate (4.3 g,11.36mmol,93.85% yield). MS (ESI) m/z 379.2[ M+H ]] ⁺ 。

Step 3: (S) -2-amino-N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -3-cyclopropylpropionamide

N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]A solution of tert-butyl carbamate (2.5 g,6.61mmol,1 eq.) in EA (10 mL) and HCl/EtOAc (4M, 10mL,6.06 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue, which gave (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]3-cyclopropyl-propionamide (2 g, crude material, HCl). MS (ESI) m/z 279.1[ M+H ]] ⁺ 。

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-imidazole-2-carboxamide

To 1H-imidazole-2-carboxylic acid (267.03 mg,2.38mmol,1 eq), (2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]To a solution of 3-cyclopropyl-propionamide (1.5 g,2.38mmol,50% purity, 1 eq., HCl), DMAP (873.15 mg,7.15mmol,3 eq.) in DCM (10 mL) was added EDCI (913.42 mg,4.76mmol,2 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:10% -40%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (130 mg,293.22umol,12.31% yield, 84% purity). MS (ESI) m/z 373.1[ M+H ]] ⁺ 。

Step 5: n- (1- (((1S) -1-cyano-2- (2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-imidazole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK IG (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-MeOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%,12 min) isolation of N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (130 mg,293.22umol,12.31% yield, 84% purity) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (14.9 mg,40.01umol,11.46% yield, 100% purity). MS (ESI) m/z 373.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.40-7.00(m,2H),5.23-5.05(m,1H),4.51(t,J＝7.0Hz,1H),3.27-3.17(m,2H),2.55-2.35(m,2H),2.06-1.62(m,6H),1.60-1.44(m,1H),0.90-0.76(m,1H),0.60-0.44(m,2H),0.26-0.07(m,2H)

Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-)Ethyl group]-1H-imidazole-2-carboxamide (27.3 mg,72.20umol,20.69% yield, 98.5% purity). MS (ESI) m/z 373.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.43-7.01(m,2H),5.06(t,J＝7.3Hz,1H),4.56(dd,J＝6.2,7.4Hz,1H),3.29-3.19(m,2H),2.48-2.39(m,1H),2.34(td,J＝6.7,13.8Hz,1H),2.01-1.66(m,6H),1.62-1.50(m,1H),0.89-0.75(m,1H),0.56-0.42(m,2H),0.24-0.10(m,2H)。

Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-imidazole-2-carboxamide (12.5 mg,32.59umol,9.34% yield, 97.1% purity). MS (ESI) m/z 373.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.33-7.09(m,2H),5.15-5.03(m,1H),4.53(d,J＝6.0,7.3Hz,1H),3.26-3.21(m,2H),2.44-2.27(m,2H),2.04-1.96(m,1H),1.93-1.77(m,3H),1.76-1.68(m,2H),1.59-1.51(m,1H),0.89-0.76(m,1H),0.56-0.45(m,2H),0.23-0.12(m,2H)

Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-1H-imidazole-2-carboxamide (5.5 mg,13.32umol,3.82% yield, 90.2% purity). MS (ESI) m/z 373.2[ M+H ]] ⁺ 。 ¹ H NMR (400 MHz, methanol-d) ₄ )δ＝7.35-7.05(m,2H),5.11(t,J＝7.2Hz,1H),4.55(t,J＝6.9Hz,1H),3.27-3.21(m,2H),2.50-2.32(m,2H),2.06-1.98(m,1H),1.97-1.81(m,3H),1.79-1.68(m,2H),1.63-1.56(m,1H),1.65-1.53(m,1H),0.88-0.77(m,1H),0.52(d,J＝7.8Hz,2H),0.23-0.09(m,2H)。

EXAMPLE 253 Synthesis of viral protease inhibitor Compound 896

To (2S) -2-amino-3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (2 g,8.98mmol,1 eq, HCl) in DCM (20 mL) and DMF (2 mL) was added 2-tert-butoxycarbonyl-2-azaspiro [4.5]]Decane-3-carboxylic acid (2.80 g,9.88mmol,1.1 eq.), T3P (11.43 g,17.96mmol,10.68mL,50% purity, 2 eq.) and TEA (5.45 g,53.89mmol,7.50mL,6 eq.) were stirred at 20℃for 3h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE: ea=4/1 to 0/1 to give the product 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (2.5 g,4.43mmol,49.31% yield, 80% purity). MS (ESI) m/z 452.3[ M+H ]] ⁺ 。

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl]Methyl group]Ethyl group]Carbamoyl group]-2-azaspiro [4.5]]A mixture of tert-butyl decane-2-carboxylate (2.1 g,3.72mmol,80% purity, 1 eq.) in HCl/MeOH (4M, 25mL,26.88 eq.) was stirred at 20℃for 3h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product (2S) -2- (2-azaspiro [ 4.5) as a white oil]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate(1.4 g, crude material, HCl). MS (ESI) m/z 352.2[ M+H ]] ⁺ 。

To (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxopyrrolidin-3-yl]To a mixture of methyl propionate (1.4 g,3.61mmol,1 eq, HCl) in DCM (20 mL) was added 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (1.06 g,4.69mmol,1.3 eq), DMAP (1.10 g,9.02mmol,2.5 eq) and EDCI (1.38 g,7.22mmol,2 eq) and the resulting mixture was then stirred at 20 ℃ for 1H. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ PE: EA=2/1-0/1) purification of the residue to give (2S) -2-azaspiro [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.5 g,2.68mmol,74.34% yield). MS (ESI) m/z 559.2[ M+H ]] ⁺ 。

(2S) -2- [ [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] ]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (1.46 g,2.61mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,53.61 eq.) was stirred at 30℃for 20h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl as a yellow oil]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.35 g, crude). MS (ESI) m/z 544.2[ M+H ]] ⁺ 。

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxopyrrolidin-3-yl at 30 ℃C]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.35 g,2.11mmol,85% purity, 1 eq.) in DCM (15 mL) was added the Bungeus reagent (1.51 g,6.33mmol,3 eq.) for 1h. After completion, the mixture was quenched with water (1 mL) and with N ₂ And (5) blow-drying. By preparative HPLC (column: waters X bridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) was purified and passed through SFC (column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,12 min) to give the product 2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxopyrrolidin-3-yl as a white solid]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (322.82 mg,613.70umol,29.10% yield). MS (ESI) M/z526.2[ M+H ]] ⁺ 。

EXAMPLE 254 Synthesis of viral protease inhibitor Compound 910

Step 1:2- ((diphenylmethylene) amino) -4-methylpent-4-enoate ethyl ester

at-78deg.C, at N ₂ To a solution of ethyl 2- (benzhydryleneamino) acetate (10 g,37.41mmol,1 eq.) in THF (40 mL) was added LiHMDS (1 m,41.15mL,1.1 eq.) (-0.5 h) dropwise and the mixture was then stirred at-78 ℃ for 0.5h. After dropwise addition of 3-bromo-2-methyl-prop-1-ene (5.05 g,37.41mmol,3.77ml,1 eq.) to the mixture, the mixture was stirred at 0 ℃ for 0.5h, and then the mixture was warmed to 20 ℃ and stirred at 20 ℃ for 1h. After completion, the mixture was quenched with water (200 mL) and concentrated in vacuo. The mixture was then extracted with EA (70 mL x 3), washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, concentrated in vacuo and passed through a column (SiO ₂ PE: ea=1:0 to 40:1) to give ethyl 2- ((diphenylmethylene) amino) -4-methylpent-4-enoate (9.5 g,26.60mmol,71.11% yield, 90% purity) as a yellow solid. MS (ESI) m/z 322.2[ M+H ]] ⁺

Step 2: 2-amino-4-methylpent-4-enoic acid ethyl ester

2- ((diphenylmethylene) amino) -4-methylpent-4-enoate ethyl ester (9 g,28.00mmol,1 eq) in aqueous HCl (2M, 1)40.01mL,10 equivalents) was stirred at 20℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give ethyl 2-amino-4-methylpent-4-enoate (9 g, crude material) as a yellow gum. MS (ESI) M/z158.2[ M+H] ⁺

Step 3:2- (((Benzylmethoxy) carbonyl) amino) -4-methylpent-4-enoic acid ethyl ester

To a solution of ethyl 2-amino-4-methylpent-4-enoate (9 g,28.62mmol,1 eq.) in THF (100 mL) was added saturated Na ₂ CO ₃ (7.21 g,68.06mmol,14.84ul,2.38 eq.) and then benzyl chloroformate (9.77 g,57.25mmol,8.14ml,2 eq.) was added. The resulting mixture was stirred at 20℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo, followed by addition of H ₂ O (500 mL) and extracted with EA (150 mL x 3). The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ Drying, passing through a column (SiO ₂ Pe:ea=60:1 to 40:1) and purified by preparative HPLC (column: xtime C18 u250mm 80mm; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,30 min) to give ethyl 2- (((benzyloxy) carbonyl) amino) -4-methylpent-4-enoate (3.4 g,10.50mmol,36.69% yield, 90% purity) as a yellow oil. MS (ESI) m/z 292.1[ M+H ]] ⁺

Step 4:2- (((Benzylmethoxy) carbonyl) amino) -3- (1-methylcyclopropyl) propanoic acid ethyl ester

At 0℃under N ₂ Downward ZnEt ₂ To a solution of (1M, 14.27mL,4.16 eq.) in DCM (50 mL) was added dropwise DCM (25 mL) containing diiodomethane (3.79 g,14.17mmol,1.14mL,4.13 eq.) followed by N at 0deg.C ₂ The mixture was stirred for 0.5h, followed by the addition of DCM (25 mL) containing ethyl 2- (((benzyloxy) carbonyl) amino) -4-methylpent-4-enoate (1 g,3.43mmol,1 eq.) at 0deg.C. The mixture was stirred at 20℃for 48h. After completion of the reaction, the mixture was poured into aqueous HCl (30 ml,1 m) at 0 ℃ followed by extraction with DCM (10 ml x 3) and the organic phase was taken over Na ₂ SO ₄ Dried, concentrated in vacuo and passed through a column (SiO ₂ PE: EA=60:1 to 30:1) to give 2-fold as a yellow oil((Benzylmethoxy) carbonyl) amino) -3- (1-methylcyclopropyl) propanoic acid ethyl ester (800 mg,2.36mmol,68.69% yield, 90% purity). MS (ESI) m/z 306.1[ M+H ] ] ⁺

Step 5:2- (((benzyloxy) carbonyl) amino) -3- (1-methylcyclopropyl) propanoic acid

To ethyl 2- (((benzyloxy) carbonyl) amino) -3- (1-methylcyclopropyl) propionate (800 mg,2.62mmol,1 eq.) in THF (5 mL) and H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (329.81 mg,7.86mmol,3 eq.) and the mixture was stirred at 30℃for 16h. After the reaction was completed, the mixture was adjusted to ph=1 with aqueous HCl (1 m,10 ml) and extracted with ethyl acetate (5 ml x 2). The resulting mixture was concentrated in vacuo to give 2- (((benzyloxy) carbonyl) amino) -3- (1-methylcyclopropyl) propanoic acid (700 mg, crude material) as a pale yellow oil. MS (ESI) m/z 276.1[ M-H ]] ⁺

Step 6: (2S) -2- (2- (((phenylmethoxy) carbonyl) amino) -3- (1-methylcyclopropyl) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To a solution of 2- (((benzyloxy) carbonyl) amino) -3- (1-methylcyclopropyl) propanoic acid (600 mg,2.16mmol,1 eq.) in ACN (1 mL) was added (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (614.54 mg,2.60mmol,1.2 eq. HCl), hexafluorophosphoric acid [ chloro (dimethylamino) methylene ]]Dimethyl-ammonium (910.59 mg,3.25mmol,1.5 eq.) followed by 1-methylimidazole (532.90 mg,6.49mmol,517.37ul,3 eq.) and the mixture stirred at 20℃for 1h. After completion, the reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with 1N HCl (10 mL), then brine (20 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=0:1 to 10:1 to give methyl (2S) -2- (2- (((phenylmethoxy) carbonyl) amino) -3- (1-methylcyclopropyl) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionate (1.1 g,1.80mmol,82.98% yield, 75% purity) as a yellow solid. MS (ESI) m/z 460.2[ M+H ]] ⁺ 。

Step 7: (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) carbamic acid benzyl ester

Methyl (2S) -2- (2- (((phenylmethoxy) carbonyl) amino) -3- (1-methylcyclopropyl) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1 g,1.63mmol,75% purity, 1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 10.71mL,45.95 eq.) was stirred at 30℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give benzyl (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) carbamate (1 g, crude material) as a yellow solid. MS (ESI) m/z 445.2[ M+H ]] ⁺ 。

Step 8: 2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- (1-methylcyclopropyl) propanamide

At N ₂ To a solution of benzyl (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) carbamate (1 g,2.25mmol,1 eq.) in IPA (20 mL) was added HCl (12 m,243.71ul,1.3 eq.) and Pd/C (1 g,833.33umol,10% purity, 0.37 eq.). The suspension was degassed in vacuo and purified by H ₂ Purging 3 times. The mixture is put in H ₂ (15 psi) and stirred at 25℃for 1h. After completion, the reaction mixture was filtered through celite, followed by concentration under reduced pressure to give 2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- (1-methylcyclopropyl) propanamide (680 mg, crude material) as a yellow solid. MS (ESI) m/z 311.2[ M+H ]] ⁺ 。

Step 9: n- (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of 2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- (1-methylcyclopropyl) propanamide (680 mg,1.96mmol,1 eq, HCl) in DCM (7 mL) was added 4-methoxy-1H-indole-2-carboxylic acid (449.78 mg, 2.35mmol,1.2 eq.) DMAP (718.54 mg,5.88mmol,3 eq.) followed by EDCI (751.65 mg,3.92mmol,2 eq.) at 0deg.C followed by stirring the mixture at 25deg.C for 2h. After completion, the mixture was quenched with water (10 mL) and extracted with DCM (10 mL x 3). The organic layer was washed sequentially with 1N HCl (10 mL) and brine (20 mL), and then with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=0:1 to 10:1 to give N- (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (550 mg,1.02mmol,52.21% yield, 90% purity) as a yellow solid. MS (ESI) m/z 484.2[ M+H ]] ⁺ 。

Step 10: n- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a mixture of N- (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (550 mg,1.02mmol,90% purity, 1 eq.) in DCM (1 mL) was added the berges reagent (731.85 mg,3.07mmol,3 eq.) and stirred for 3H at 25 ℃. After completion, by adding H at 20℃ ₂ O (0.5 mL) to quench the reaction mixture, and then under reduced pressure<Concentrated at 30 ℃ to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -55%,10 min) to give N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxoprop-2-yl) -4-methoxy-1H-indole-2-carboxamide (250 mg,531.64umol,51.93% yield, 99% purity) as a white solid. MS (ESI) M/z466.2[ M+H] ⁺ 。

Step 11: n- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,15 min) purification of N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (250 mg,531.64umol,99% purity, 1 eq) gave N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (63.08 mg,135.50umol,25.49% yield, 100% purity) as a white solid. MS (ESI) m/z466.2[ M+H ] ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.58(br s,1H),8.96(br d,J＝8.1Hz,1H),8.43(br d,J＝7.8Hz,1H),7.52(br s,1H),7.33(s,1H),7.09(t,J＝7.9Hz,1H),7.01(d,J＝8.3Hz,1H),6.50(d,J＝7.6Hz,1H),5.10-5.01(m,1H),4.66-4.57(m,1H),3.88(s,3H),3.13-3.02(m,2H),2.32-2.20(m,2H),1.86-1.72(m,3H),1.72-1.51(m,3H),1.39(br d,J＝11.0Hz,1H),1.07(s,3H),0.56-0.49(m,1H),0.29-0.23(m,1H),0.23-0.12(m,2H)

N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (1-methylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (85.13 mg,182.86umol,34.40% yield, 100% purity) was obtained as a white solid. MS (ESI) m/z 466.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.60(br s,1H),8.95(br d,J＝8.1Hz,1H),8.45(br d,J＝8.4Hz,1H),7.53(br s,1H),7.34(s,1H),7.10(t,J＝7.9Hz,1H),7.01(d,J＝8.3Hz,1H),6.51(d,J＝7.7Hz,1H),5.09-5.00(m,1H),4.70-4.62(m,1H),3.89(s,3H),3.13-3.03(m,2H),2.25-2.14(m,2H),1.89-1.69(m,4H),1.65-1.50(m,2H),1.48-1.36(m,1H),1.05(s,3H),0.59-0.50(m,1H),0.27-0.20(m,1H),0.19-0.11(m,2H)

EXAMPLE 255 Synthesis of viral protease inhibitor Compound 912

Step 1:2- ((diphenylmethylene) amino) -5-methyl hex-4-enoic acid ethyl ester

To a solution of ethyl 2- (benzhydryleneamino) acetate (5 g,18.70mmol,1 eq.) in THF (50 mL) at-78 ℃ was added LiHMDS (1 m,20.57mL,1.1 eq.), stirred for 30min and then 1-bromo-3-methyl-but-2-ene (2.79 g,18.70mmol,2.16mL,1 eq.) was added. The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 50mL of H at 20 ℃ ₂ O, and then extracted with EtOAc (60 ml x 3). The combined organic layers were washed with brine (50 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 30/1 to give ethyl 2- (benzhydryleneamino) -5-methyl-hex-4-enoate (3 g,8.94mmol,47.82% yield) as a yellow oil. MS (ESI) M/z336.2[ M+H ] ⁺ 。

Step 2: 2-amino-5-methylhex-4-enoic acid ethyl ester

A solution of ethyl 2- (benzhydrylamino) -5-methyl-hex-4-enoate (2.7 g,8.05mmol,1 eq.) in HCl (2M, 27mL,6.71 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove H ₂ O, 2-amino-5-methyl-hex-4-enoic acid ethyl ester (1.3 g, crude material, HCl) was obtained as a white solid.

Step 3:2- (((Benzylmethoxy) carbonyl) amino) -5-methylhex-4-enoic acid ethyl ester

Na was added to a solution of ethyl 2-amino-5-methyl-hex-4-enoate (900 mg,5.26mmol,1 eq.) in IPA (10 mL) at 0deg.C ₂ CO ₃ (1.39 g,13.14mmol,9mL,2.5 eq.) and NaOH (210.22 mg,5.26mmol,1mL,1 eq.) to adjust the pH to 11, and benzyl (2, 5-dioxopyrrolidin-1-yl) carbonate (1.44 g,5.78mmol,1.1 eq.) was then added. The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ O, and then extracted with EtOAc (35 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give ethyl 2- (benzyloxycarbonylamino) -5-methyl-hex-4-enoate as a yellow solid (1.3 g, Crude material). MS (ESI) m/z 306.1[ M+H ]] ⁺ 。

Step 4:2- (((Benzylmethoxy) carbonyl) amino) -3- (2, 2-dimethylcyclopropyl) propanoic acid ethyl ester

2- (Benzylmethoxycarbonylamino) -5-methyl-hex-4-enoic acid ethyl ester (1.2 g,2.75mmol,70% purity, 1 eq.) in DCM (20 mL) and ZnEt ₂ The solution in (1M, 6.88mL,2.5 eq.) was stirred at-40℃for 10min, followed by addition of CH ₂ I ₂ (1.47 g,5.50mmol,443.83uL,2 eq.). The mixture was stirred for 15h 50min at 20 ℃. After completion, by adding 30mL NH at 20 ℃ ₄ The reaction mixture was quenched with Cl and then extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 40/1) to give ethyl 2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propionate (1 g, crude material) as a yellow oil. MS (ESI) m/z 320.2[ M+H ]] ⁺ 。

Step 5:2- (((benzyloxy) carbonyl) amino) -3- (2, 2-dimethylcyclopropyl) propanoic acid

To ethyl 2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propionate (1 g,3.13mmol,1 eq.) in THF (9 mL) and H ₂ LiOH.H was added to the solution in O (3 mL) ₂ O (394.15 mg,9.39mmol,3 eq.). The mixture was stirred at 50℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. 1M HCl was added to adjust pH to 3 followed by extraction with DCM (35 mL. Times.3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propanoic acid (1.1 g, crude material) as a yellow oil.

Step 6: (2S) -2- (2- (((benzyloxy) carbonyl) amino) -3- (2, 2-dimethylcyclopropyl) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To 2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propanoic acid (1.08 g,3.71mmol,1 eq), (2S)-2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.75 g,7.41mmol,2 equivalents, HCl) in DCM (10 mL) was added DMAP (1.13 g,9.27mmol,2.5 equivalents), followed by EDCI (1.42 g,7.41mmol,2 equivalents). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 50mL of H at 20 ℃ ₂ O, and then extracted with DCM (50 ml×3). The combined organic layers were washed with brine (50 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM meoh=1/0 to 80/1 to give (2S) -2- [ [2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propionyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.5 g,3.17mmol,85.45% yield). MS (ESI) m/z 474.2[ M+H ]] ⁺ 。

Step 7: (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (2, 2-dimethylcyclopropyl) -1-oxopropan-2-yl) carbamic acid benzyl ester

(2S) -2- [ [2- (benzyloxycarbonylamino) -3- (2, 2-dimethylcyclopropyl) propanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.5 g,3.17mmol,1 eq.) in NH ₃ A solution in MEOH (7M, 15.00mL,33.15 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH to give N- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]Benzyl carbamate (1.45 g, crude material). MS (ESI) m/z 459.2[ M+H ]] ⁺ 。

Step 8: 2-amino-N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- (2, 2-dimethylcyclopropyl) propanamide

To N- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]To a solution of benzyl carbamate (1.45 g,3.16mmol,1 eq.) in i-PrOH (15 mL) was added HCl (12M, 263.51uL,1 eq.) followed by Pd/C (1.45g,3.16mmol,20% purity, 1 eq). At H ₂ The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give (2S) -2- [ [ 2-amino-3- (2, 2-dimethylcyclopropyl) propionyl ] as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Propionamide (1 g, crude). MS (ESI) m/z 325.2[ M+H ]] ⁺ 。

Step 9: n- (1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3- (2, 2-dimethylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To (2S) -2- [ [ 2-amino-3- (2, 2-dimethylcyclopropyl) propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of propionamide (990 mg,3.05mmol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (700.10 mg,3.66mmol,1.2 eq.) in DCM (10 mL) was added DMAP (932.03 mg,7.63mmol,2.5 eq.) followed by EDCI (1.17 g,6.10mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 25mL of H at 20 ℃ ₂ O, and then extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (25 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with DCM: meoh=100/0 to 95/5 to give N- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (1.01 g,2.03mmol,66.52% yield). MS (ESI) m/z 498.2[ M+H ]] ⁺ 。

Step 10: n- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-dimethylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (1 g,2.01mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (957.85 mg,4.02mmol,2 equivalents). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ O, and then extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (280 mg,583.86umol,29.05% yield, 100% purity). MS (ESI) m/z 480.2[ M+H ]] ⁺ 。

Step 11: n- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-dimethylcyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-IPA)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -50%,18 min) isolation of N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (280 mg,583.86umol,29.05% yield, 100% purity) to give N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group ]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (55 mg,113.31umol,19.41% yield, 98.8% purity). MS (ESI) m/z 480.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.26(s,1H),7.19-7.10(m,1H),7.07-6.98(m,1H),6.51(d,J＝7.2Hz,1H),5.17-5.08(m,1H),4.58-4.46(m,1H),3.93(s,3H),3.27-3.15(m,2H),2.55-2.39(m,2H),2.08-1.89(m,3H),1.86-1.66(m,3H),1.56-1.45(m,1H),1.26-0.96(m,6H),0.71-0.58(m,1H),0.50(d,J＝2.5,4.5Hz,1H),0.14-0.01(m,1H)。

Obtaining N- [2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] as a white solid]Ethyl group]Amino group]-1- [ (2, 2-dimethylcyclopropyl) methyl]-2-oxoRadical-ethyl radical]-4-methoxy-1H-indole-2-carboxamide (155 mg,322.56umol,55.25% yield, 99.8% purity). MS (ESI) m/z 480.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27(s,1H),7.19-7.11(m,1H),7.04(d,J＝8.3Hz,1H),6.52(d,J＝7.7Hz,1H),5.06(d,J＝6.2,10.0Hz,1H),4.59(d,J＝5.8,8.4Hz,1H),3.93(s,3H),3.22-3.11(m,2H),2.40(d,J＝6.2,10.2,13.8Hz,1H),2.33-2.22(m,1H),2.12-2.02(m,1H),2.00-1.85(m,2H),1.84-1.73(m,2H),1.61(d,J＝2.4Hz,1H),1.49(d,J＝11.0Hz,1H),1.14-1.07(m,3H),1.07-0.99(m,3H),0.69-0.58(m,1H),0.48(d,J＝4.3,8.7Hz,1H),0.14-0.03(m,1H)。

EXAMPLE 256 Synthesis of viral protease inhibitor Compound 918

Step 1: (1S, 3aR,7 aS) -1- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) hexahydro-1H-isoindole-2 (3H) -carboxylic acid tert-butyl ester

To (1S, 3aR,7 aS) -2-tert-butoxycarbonyl-1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxylic acid (450 mg,1.67mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (571.23 mg,2.17mmol,90% purity, 1.3 eq., HCl) in DCM (5 mL) and DMF (1.5 mL) was added DMAP (612.36 mg,5.01mmol,3 eq.) and EDCI (640.58 mg,3.34mmol,2 eq.) followed by stirring the mixture at 20℃for 2h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (50 ml x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (1S, 3aR,7 aS) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Carbamoyl group]-tert-butyl 1, 3a,4,5,6,7 a-octahydroisoindole-2-carboxylate (680 mg,1.36mmol,81.12% yield, 90% purity). MS (ESI) m/z 452.2[ M+H ]] ⁺ 。

Step 2: (S) -2- ((1S, 3aR,7 aS) -octahydro-1H-isoindole-1-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester hydrochloride

A solution of (1S, 3aR,7 aS) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -1, 3a,4,5,6,7 a-octahydroisoindole-2-carboxylic acid tert-butyl ester (680 mg,1.51mmol,1 eq.) in HCl/MeOH (4M, 10mL,26.56 eq.) and the mixture stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (1S, 3ar,7 as) -2, 3a,4,5,6,7 a-octahydro-1H-isoindole-1-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (580 mg, crude, HCl) as a white solid.

Step 3: (S) -2- ((1S, 3aR,7 aS) -2- (4-methoxy-1H-indole-2-carbonyl) octahydro-1H-isoindole-1-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (371.62 mg,1.94mmol,1.3 eq.) and (2S) -2- [ [ (1S, 3aR,7 aS) -2, 3a,4,5,6,7 a-octahydro-1H-isoindole-1-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (580 mg,1.50mmol,1 eq., HCl) in DCM (15 mL) and DMF (3 mL) was added DMAP (548.02 mg,4.49mmol,3 eq.) and EDCI (573.27 mg,2.99mmol,2 eq.) followed by stirring the mixture at 20℃for 2h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (50 ml x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) to give the product (2S) -2- [ [ (1S, 3ar,7 as) -2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carbonyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (710 mg,1.26mmol,84.18% yield, 93% purity). MS (ESI) m/z 525.2[ M+H ] ] ⁺ 。

Step 4: (1S, 3ar,7 as) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) octahydro-1H-isoindole-1-carboxamide

(2S) -2- [ [ (1S, 3aR,7 aS) -2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (710 mg,1.35mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 10mL,51.72 eq.) and then the mixture was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (1S, 3ar,7 as) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (640 mg, crude). MS (ESI) m/z 510.2[ M+H ]] ⁺ 。

Step 5: (1S, 3aR,7 aS) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) octahydro-1H-isoindole-1-carboxamide

(1S, 3aR,7 aS) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-solution of 2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (640 mg,1.26mmol,1 eq.) and a bergiut reagent (598.57 mg,2.51mmol,2 eq.) in DCM (10 mL) and the mixture stirred at 30 ℃ for 3H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge C18. Times.50 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give the product (1S, 3aR,7 aS) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (450 mg,906.26umol,72.16% yield, 99% purity). MS (ESI) m/z 492.2[ M+H ]] ⁺ 。

Step 6:2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O MEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,4 min) purification of 2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5]Decane-3-carboxamide gives the product (3 aR,7 aS) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide isomer 1 (136.17 mg,277.00umol,30.26% yield, 100% purity). MS (ESI) m/z 492.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.56(s,1H),9.30-8.85(m,1H),7.54(br s,1H),7.15-7.07(m,1H),7.07-6.95(m,2H),6.52(d,J＝7.6Hz,1H),5.06(br d,J＝7.7Hz,1H),4.58-4.20(m,1H),4.06-3.94(m,1H),3.91-3.81(m,3H),3.78(br dd,J＝5.8,9.8Hz,1H),3.15-2.89(m,2H),2.45-2.37(m,1H),2.31-2.11(m,3H),1.46(br d,J＝3.5Hz,13H)。

The product (3 aR,7 aS) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid ]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide isomer 2 (161.76 mg,329.06umol,35.95% yield, 100% purity). MS (ESI) m/z 492.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.62-11.55(m,1H),9.15-8.84(m,1H),7.55-7.39(m,1H),7.15-7.09(m,1H),7.06-6.97(m,2H),6.61-6.50(m,1H),5.05(br d,J＝8.1Hz,1H),4.47-4.24(m,1H),4.00(dd,J＝7.1,9.7Hz,1H),3.90-3.83(m,3H),3.82-3.75(m,1H),3.08(br s,2H),2.44-2.37(m,1H),2.24(br d,J＝7.1Hz,3H),1.83-1.32(m,13H)。

Example 257 Synthesis of viral protease inhibitor Compounds 930

Step 1:3- [ [ (1S) -1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

At 25 ℃, to (2S) -2-amino-3- [ (3R)-5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (750.00 mg,2.99mmol,1 eq. HCl) and 2-tert-butoxycarbonyl-2-azaspiro [4.5]]Decane-3-carboxylic acid (932.40 mg,3.29mmol,1.1 eq.) DMAP (1.10 g,8.97mmol,3 eq.) and EDCI (1.15 g,5.98mmol,2 eq.) were added in one portion to a mixture of DCM (10 mL) and DMF (3 mL). The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was quenched with 30mL H ₂ O was diluted and extracted with 60mL EA (20 mL x 3). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give 3- [ [ (1S) -1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl as a yellow oil ]-2-methoxy-2-oxo-ethyl]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carboxylic acid tert-butyl ester (1.2 g,2.50mmol,83.64% yield). MS (ESI) m/z 480.3[ M+H ]] ⁺

Step 2: (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

A mixture of 3- [ [ (1S) -1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester (1.2 g,2.50mmol,1 eq.) in HCl/MeOH (20 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (1 g,2.40mmol,96.09% yield, HCl) as a yellow solid.

Step 3: (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester

To 7-chloro-1H-indole-2-carboxylic acid (427.50 mg,2.19mmol,1 eq.) and (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionic acid methyl ester (1 g,2.40mmol,1.1 eq. HCl) to a mixture of DCM (12 mL) and DMF (3 mL) was added DMAP (801.02 mg,6.56mmol,3 eq.) and EDC I (837.95 mg,4.37mmol,2 eq.) the mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 30mL H ₂ O was diluted and extracted with 60mL EA (20 mL x 3). The combined organic layers were washed with 30mL brine (30 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a white solid]Decane-3-carbonyl]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (700 mg,1.26mmol,57.49% yield). MS (ESI) m/z 557.3[ M+H ]] ⁺

Step 4: n- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl ] -2-oxo-ethyl ] -2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-methyl 3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (700 mg,1.26mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 105.00mL,334.25 eq.) was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl as a white solid ]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (660 mg,1.22mmol,96.90% yield). MS (ESI) m/z 542.3[ M+H ]] ⁺

Step 5:2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-1- [ (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) methyl at 25 ℃]-2-oxo-ethyl]-2- (7-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (660 mg,1.03mmol,85% purity, 1 eq.) in DCM (10 mL) was added at once the Bunges reagent (863.22 mg,3.62mmol,3.5 eq.). The mixture was stirred at 25℃for 8 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product.By preparative HPLC (neutral conditions; column: waters Xbridge Prep OBD C: 150 x 40mM x 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -60%,8 min) to give the desired compound as a white solid (270 mg,49% yield, 100% purity), which was purified by SFC (condition: column: DAICEL CHIRALPAK IC (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:43% -43%,7 min) to give 2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl as a white solid]-2-azaspiro [4.5 ]]Decane-3-carboxamide (100 mg,190.82umol,18.44% yield). MS (ESI) m/z 524.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.66-11.45(m,1H),8.95(d,J＝8.2Hz,1H),7.81(s,1H),7.64(d,J＝7.9Hz,1H),7.29(d,J＝7.3Hz,1H),7.14(s,1H),7.11-6.97(m,1H),4.99-4.75(m,1H),4.50(t,J＝8.6Hz,1H),3.83(br d,J＝10.1Hz,1H),3.66(d,J＝10.4Hz,1H),2.76-2.64(m,1H),2.29-2.13(m,2H),1.99(dd,J＝8.6,11.9Hz,1H),1.82-1.66(m,1H),1.65-1.28(m,12H),1.18-1.07(m,3H),1.02(s,3H)

EXAMPLE 258 Synthesis of viral protease inhibitor Compounds 934

Step 1: benzyl N- [1- (hydroxymethyl) cyclopropyl ] carbamate

350mL of buffer (pH=11) (saturated NaHCO adjusted to pH=11 with 4M NaOH) ₃ ) To a solution of (1-aminocyclopropyl) methanol (20 g,229.57mmol,1 eq.) in IPA (350 mL). The reaction mixture was cooled to 0deg.C and benzyl 2, 5-dioxopyrrolidine-1-carboxylate (53.54 g,229.57mmol,1 eq.) was added. The reaction mixture was stirred at 20℃for 16h. After completion, the reaction mixture was filtered and then concentrated under reduced pressure to remove IPA. The residue was taken up in 100mL H ₂ O was diluted and extracted with 200mL EA (100 mL x 2). The combined organic layers were washed with 100mL brine (100 mL x 1), dried over Na ₂ SO ₄ Drying, filtering and depressurizingConcentration under reduced pressure gave a residue. The crude product was wet-milled with DCM at 20 ℃ for 20min. Obtaining the compound N- [1- (hydroxymethyl) cyclopropyl ] as a white solid ]Benzyl carbamate (35 g,142.37mmol,62.02% yield, 90% purity).

Step 2: benzyl N- (1-formyl cyclopropyl) carbamate

To N- [1- (hydroxymethyl) cyclopropyl ]]To a mixture of toluene (130 mL), DMSO (130.00 g,1.66mol,130.00mL,28.32 eq.) and TFA (3.35 g,29.38mmol,2.18mL,0.5 eq.) and pyridine (4.65 g,58.76mmol,4.74mL,1 eq.) and DCC (36.37 g,176.27mmol,35.66mL,3 eq.) were added. The mixture was stirred at 25℃for 16h. After completion, the reaction mixture was filtered and diluted with 100mL EA and 300mL H ₂ O (100 ml x 3) washes. The combined organic layers were washed with 100mL brine (100 mL x 1) and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=8/1 to 5/1) to give benzyl N- (1-formylcyclopropyl) carbamate (9 g,39.00mmol,66.37% yield, 95% purity) as a white solid.

Step 3: n- [1- [ (E) -2-methoxyvinyl ] cyclopropyl ] carbamic acid phenylmethyl ester

To a solution of methoxymethyl (triphenylphosphonium chloride (25.02 g,72.98mmol,4 eq.) in THF (80 mL) at-10deg.C was added dropwise a solution of t-BuOK (1M, 72.80mL,3.99 eq.). The reaction mixture was warmed to 20 ℃ and stirred at 20 ℃ for 1h. Benzyl N- (1-formylcyclopropyl) carbamate (4 g,18.25mmol,1 eq.) in THF (40 mL) at 0deg.C is added at N ₂ The solution was stirred at 20℃for a further 1h. After completion, the reaction mixture was quenched with 100mL H ₂ O was diluted and extracted with 200mL ethyl acetate (100 mL x 2). The combined organic layers were washed with 100mL brine (100 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=30/1 to 10/1) the residue was purified to give N- [1- [ (E) -2-methoxyvinyl as a yellow oil]Cyclopropyl group]Benzyl carbamate (2.1 g,764mmol,41.89% yield, 90% purity). MS (ESI) m/z 246.1[ M-H ]] ⁺

Step 4: benzyl N- [1- (2-oxoethyl) cyclopropyl ] carbamate

To N- [1- [ (E) -2-methoxyvinyl]Cyclopropyl group]To a mixture of benzyl carbamate (1.9 g,7.68mmol,1 eq.) in THF (20 mL) was added HCl (19.38 g,53.15mmol,19.00mL,10% purity, 6.92 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 100mL H ₂ O was diluted and extracted with 300mL EA (150 mL x 2). The combined organic layers were washed with 300mL brine (300 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=20/1 to 5/1) the residue was purified. Obtaining the compound N- [1- (2-oxoethyl) cyclopropyl as a white solid ]Benzyl carbamate (1.3 g,5.02mmol,65.28% yield, 90% purity).

Step 5: (2S) -4- [2- [1- (Benzylmethoxycarbonylamino) cyclopropyl ] -1-hydroxy-ethyl ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

At-60 ℃ under N ₂ To a mixture of (2S) -5-oxopyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (450 mg,1.85mmol,1 eq.) in THF (8 mL) was added LiHMDS (1M, 2.40mL,1.3 eq.) in one portion. The mixture was stirred at-60℃for 30min and then N- [1- (2-oxoethyl) cyclopropyl-containing was added at-60 ℃]Benzyl carbamate (431.51 mg,1.85mmol,1 eq.) in THF (4 mL) and stirred at-60℃for 2h. After completion, the reaction mixture was quenched by adding 8mL THF containing 4mL AcOH at-60 ℃ and concentrated under reduced pressure to give a residue which was used directly in the next step. The compound (2S) -4- [2- [1- (benzyloxycarbonylamino) cyclopropyl was obtained as a yellow oil]-1-hydroxy-ethyl]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (900 mg, crude material). MS (ESI) m/z 377.1[ M+H-100 ]] ⁺

Step 6: (2S, 4E) -4- [2- [1- (benzyloxycarbonylamino) cyclopropyl ] ethylene ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

At 25 ℃, to (2S) -4- [2- [1- (benzyloxycarbonylamino) cyclopropyl]-1-hydroxy-ethyl]To a mixture of O1-tert-butyl O2-methyl 5-oxo-pyrrolidine-1, 2-dicarboxylic acid (900 mg,1.89mmol,1 eq.) in DCM (20 mL) was added the Bolus reagent (1.35 g,5.67mmol,3 eq.). The mixture was stirred at 40℃for 16h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 3/1) the residue was purified. Obtaining (2S, 4E) -4- [2- [1- (benzyloxycarbonylamino) cyclopropyl ] as a colorless oil]Ethylene group]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (420 mg,824.42umol,43.65% yield, 90% purity). MS (ESI) m/z 459.2[ M+H ]] ⁺

Step 7: (2S) -4- [2- (1-Aminocyclopropyl) ethyl ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

To (2S, 4E) -4- [2- [1- (benzyloxycarbonylamino) cyclopropyl]Ethylene group]To a mixture of O1-tert-butyl O2-methyl 5-oxo-pyrrolidine-1, 2-dicarboxylic acid (700 mg,1.53mmol,1 eq.) in i-PrOH (10 mL) was added Pd/C (300 mg,1.53mmol,10% purity, 1.00 eq.). At 25℃at H ₂ (3.08 mg,1.53mmol,1 eq.) the mixture was stirred at 15Psi for 1h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue which was used directly in the next step. The (2S) -4- [2- (1-aminocyclopropyl) ethyl compound was obtained as a colorless oil]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (400 mg, crude material).

Step 8: (2S) -2- (tert-Butoxycarbonylamino) -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propionic acid methyl ester

At 80℃to (2S) -4- [2- (1-aminocyclopropyl) ethyl]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (350 mg,1.07mmol,1 eq.) in MeOH (5 mL) and CHCl ₃ KOAc (210.48 mg,2.14mmol,2 eq.) was added to the mixture in (0.5 mL). The mixture was stirred at 80℃for 48h. After completion, the residue was taken up with 5mL H ₂ O was diluted and extracted with 10mL EA (5 mL x 2). The combined organic layers were taken up in 10mLBrine (10 ml x 1), washed with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue which is used directly in the next step. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5:1 to 1:1) purification residue. The compound (2S) -2- (tert-butoxycarbonylamino) -3- (5-oxo-4-azaspiro [ 2.5) was obtained as a colorless oil ]Methyl oct-6-yl propionate (200 mg,586.42umol,54.69% yield, 95.7% purity).

Step 9: (2S) -2-amino-3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propionic acid methyl ester

To methyl (2S) -2- (tert-butoxycarbonylamino) -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propionate (170 mg,520.85umol,1 eq.) was added HCl/MeOH (4M, 42.50mL,326.39 eq.). The mixture was stirred at 25℃for 60min. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound methyl (2S) -2-amino-3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propionate (136 mg, crude material, HCl) was obtained as a colorless oil.

Step 10: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propanoic acid methyl ester

To (2S) -2-amino-3- (5-oxo-4-azaspiro [2.5]]To a mixture of methyl oct-6-yl propionate (136 mg,517.64umol,1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (118.68 mg,517.64umol,1 eq) in DCM (9 mL) was added DMAP (126.48 mg,1.04mmol,2 eq) and EDCI (198.46 mg,1.04mmol,2 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 40mL EA (20 mL x 2). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5:1/1 to 1/1) the residue was purified. The compound (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl was obtained as a colorless oil]Amino group]-3- (5-oxo-4-azaspiro [2.5]]Methyl oct-6-yl propionate (186 mg,403.86umol,78.02% yield,95% purity). MS (ESI) M/z438.3[ M+H ]] ⁺

Step 11: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propanoic acid methyl ester

To methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propanoate (162 mg,370.26umol,1 eq) was added HCl/MeOH (4M, 12.21mL,131.86 eq). The mixture was stirred at 25℃for 60min. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propanoic acid methyl ester (138 mg, crude material, HCl) was obtained as a white solid.

Step 12: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.5] oct-6-yl) propionic acid methyl ester

To 7-chloro-1H-indole-2-carboxylic acid (72.20 mg,369.11umol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.5]]To a mixture of methyl oct-6-yl propionate (138 mg, 369.11. Mu. Mol,1 eq, HCl) in DCM (5 mL) were added EDCI (141.52 mg, 738.22. Mu. Mol,2 eq) and DMAP (90.19 mg, 738.22. Mu. Mol,2 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 40mL EA (20 mL x 2). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=3/1 to 1/1) the residue was purified. The compound (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] was obtained as a colorless oil]-3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.5]]Methyl oct-6-yl propionate (150 mg,282.52umol,76.54% yield, 97% purity). MS (ESI) m/z 515.2[ M+H ]] ⁺

Step 13: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.5] oct-6-yl) methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]Propionyl radical]Amino group]-3- (5-oxo-4-azaspiro [2.5]]Methyl oct-6-yl propionate (130 mg,254.61 mol,1 eq.) in NH ₃ A solution in MeOH (7M, 13.16mL,361.91 eq.) was stirred at 60℃for 48h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.5] as a white solid]Oct-6-yl) methyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (120 mg,217.93umol,85.59% yield, 90% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺

Step 14: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.5] oct-6-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.5]]Oct-6-yl) methyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 7-chloro-1H-indole-2-carboxamide (120 mg, 240.01. Mu. Mol,1 eq.) in DCM (6 mL) was added the Buerger's reagent (114.39 mg, 480.01. Mu. Mol,2 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was quenched with 5mL H ₂ O was diluted and extracted with 10mL DCM (5 mL x 2). The combined organic layers were concentrated by blow-drying to give a residue. By neutral prep HPLC (column: waters Xbridge BEH C18 100. Times.30 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -55%,10 min) to give 50mg of a mixture. By SFC (column: DAICEL CHIRALPAK IF (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%,12 min) to purify 50mg of the mixture. The compound 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.5 ] as a white solid was obtained]Oct-6-yl) ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (12 mg,24.65umol,10.27% yield)Yield, 99% purity). MS (ESI) m/z 482.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.73(br s,1H),9.02(d,J＝8.1Hz,1H),8.72(d,J＝7.7Hz,1H),7.67-7.57(m,2H),7.35-7.29(m,1H),7.26(s,1H),7.07(t,J＝7.8Hz,1H),5.09(q,J＝8.0Hz,1H),4.59-4.47(m,1H),2.40-2.21(m,2H),1.98-1.71(m,4H),1.63-1.33(m,3H),0.88-0.65(m,2H),0.61-0.37(m,5H),0.26-0.03(m,2H)。

Obtaining 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.5 ] as a white solid]Oct-6-yl) ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (4 mg,8.30umol,3.46% yield). MS (ESI) m/z 482.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.73(br d,J＝1.8Hz,1H),9.04(br d,J＝7.5Hz,1H),8.75(br d,J＝7.9Hz,1H),7.71-7.54(m,2H),7.35-7.23(m,2H),7.07(t,J＝7.8Hz,1H),5.02(q,J＝7.2Hz,1H),4.61-4.51(m,1H),2.35-2.26(m,2H),2.01-1.91(m,1H),1.87-1.71(m,3H),1.67-1.40(m,3H),0.88-0.65(m,2H),0.62-0.37(m,5H),0.26-0.06(m,2H)。

EXAMPLE 259 Synthesis of viral protease inhibitor Compound 936

Step 1: (2S) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl ] -hydroxy-methyl ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

To a solution of benzyl N- (1-formylcyclopropyl) carbamate (1.80 g,8.22mmol,1 eq.) in THF (30 mL) at-60℃was added LiHMDS (1M, 10.69mL,1.3 eq.). The solution was stirred at-60℃for 1h. (2S) -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (2000 mg,8.22mmol,1 eq.) was added. The solution was stirred at-60℃for 2.5h. After completion, the solution is treated with H ₂ O (60 mL) was quenched and extracted with EA (50 mL. Times.3) and concentrated to give crude (2S) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl) as a yellow oil]-hydroxy-methyl]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (3.3 g, crude material). The crude material was used directly in the next stepIs a kind of medium. MS (ESI) m/z 463.2[ M+H ]] ⁺

Step 2: (2S, 4E) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl ] methylene ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

At 20 ℃, to (2S) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl ]]-hydroxy-methyl]To a solution of O1-tert-butyl O2-methyl-5-oxo-pyrrolidine-1, 2-dicarboxylate (3300 mg,7.14mmol,1 eq.) in DCM (50 mL) was added the Bolus reagent (5.10 g,21.41mmol,3 eq.). The solution was stirred at 40℃for 10h. After completion, the solution was concentrated to give a crude material. Through the column (SiO) ₂ The crude material was purified by pe:ea=10:1 to 0:1 to give the product (2 s,4 e) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl) as a yellow material]Methylene group]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (1.9 g,4.27mmol,59.91% yield). MS (ESI) m/z 445.1[ M+H ]] ⁺

Step 3: (2S) -4- [ (1-Aminocyclopropyl) methyl ] -5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl O2-methyl ester

At N ₂ In the atmosphere, to (2S, 4E) -4- [ [1- (benzyloxycarbonylamino) cyclopropyl ]]Methylene group]To a solution of O1-tert-butyl O2-methyl 5-oxo-pyrrolidine-1, 2-dicarboxylate (1400 mg,3.15mmol,1 eq.) in IPA (25 mL) was added Pd/C (261.12 mg,220.48umol,10% purity, 0.07 eq.) (10%). The suspension was degassed and used with H ₂ Purging 3 times. The mixture is put in H ₂ (15 psi) was stirred at 25℃for 2h. After completion, the mixture was filtered and concentrated to give crude (2S) -4- [ (1-aminocyclopropyl) methyl as a yellow oil]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (950 mg, crude material). The crude material was used directly in the next step. MS (ESI) m/z 313.1[ M+H ]] ⁺

Step 4: (2S) -2- (tert-Butoxycarbonylamino) -3- (5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

To (2S) -4- [ (1-aminocyclopropyl) methyl]-5-oxo-pyrrolidine-1, 2-dicarboxylic acid O1-tert-butyl ester O2-methyl ester (950 mg,3.04mmol,1 eq.) in MeOH (15 mL) and CHCl ₃ KOAc (895.46 mg,9.12mmol,3 eq.) was added to a solution in (1.5 mL). The solution was stirred at 60℃for 3h. After completion, the solution was concentrated and taken up with H ₂ O (50 mL) was diluted and extracted with EA (50 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ The crude material was purified with PE: ea=10:1 to 0:1 to give the product (2S) -2- (tert-butoxycarbonylamino) -3- (5-oxo-4-azaspiro [ 2.4) as a white solid]Methyl hept-6-yl) propionate (430 mg,1.38mmol,45.26% yield). MS (ESI) m/z 313.1[ M+H ]] ⁺

Step 5: (2S) -2-amino-3- (5-oxo-4-azaspiro [2.4] hept-6-yl) propionic acid methyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- (5-oxo-4-azaspiro [ 2.4)]A solution of methyl hept-6-yl) propionate (260 mg, 832.37. Mu. Mol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 1h. After completion, the solution was concentrated to dryness to give crude (2S) -2-amino-3- (5-oxo-4-azaspiro [2.4] as a white solid]Methyl hept-6-yl) propionate (207 mg, crude, HCl). The crude material was used directly in the next step. MS (ESI) m/z 213.2[ M+H ] ] ⁺

Step 6: (2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

To (2S) -2-amino-3- (5-oxo-4-azaspiro [2.4]]To a solution of methyl hept-6-yl) propionate (207 mg,832.31 mol,1 eq, HCl) in DCM (7 mL) was added DMAP (203.37 mg,1.66mmol,2 eq) and (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionic acid (200.37 mg,873.92 mol,1.05 eq) EDCI (319.11 mg,1.66mmol,2 eq). The solution was stirred at 20℃for 2h. After completion, the solution is treated with H ₂ O (40 mL) was diluted and extracted with EA (50 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ The crude material was purified with PE: ea=10:1 to 0:1 to give the product (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -3-cyclopropyl-propionyl as a white solid]Amino group]-3- (5-oxo-4-azaspiro [2.4]]Methyl hept-6-yl) propionate (290 mg,684.77umol,82.27% yield). MS (ESI) m/z 424.2[ M+H ]] ⁺

Step 7: (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.4] ]A solution of methyl hept-6-yl) propionate (284 mg, 672.96. Mu. Mol,1 eq.) in HCl/MeOH (10 mL) was stirred at 25℃for 1h. After completion, the solution was concentrated to dryness to give crude (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl as a white solid]Amino group]-3- (5-oxo-4-azaspiro [2.4]]Methyl hept-6-yl) propionate (245 mg, crude, HCl). The crude material was used directly in the next step. MS (ESI) m/z 324.2[ M+H ]] ⁺

Step 8: (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- (5-oxo-4-azaspiro [2.4] hept-6-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.4]]To a solution of methyl hept-6-yl) propionate (240 mg,666.95umol,1 eq, HCl) in DCM (6 mL) was added DMAP (162.96 mg,1.33mmol,2 eq) and 7-chloro-1H-indole-2-carboxylic acid (130.46 mg,666.95umol,1 eq) and EDCI (255.71 mg,1.33mmol,2 eq). The solution was stirred at 25℃for 1h. After completion, the solution is treated with H ₂ O (60 mL) was diluted and extracted with EA (50 mL x 3) and concentrated to give the crude material. Through the column (SiO) ₂ PE: ea=10:1 to 0:1) to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino ] as a pale yellow solid ]-3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.4]]Methyl hept-6-yl) propionate (185 mg,369.28umol,55.37% yield). MS (ESI) m/z 501.2[ M+H ]]+

Step 9: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.4] hept-6-yl) methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- (5-oxo-4-azaspiro [2.4]]Methyl hept-6-yl) propionate (185 mg,369.28umol,1 eq.) in NH ₃ A solution in MeOH (7M, 10.55mL,200 eq.) was stirred at 60℃for 20h. After completion, the solution was concentrated to dryness to give a white colorCrude N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.4] as a color solid]Hept-6-yl) methyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-1H-indole-2-carboxamide (184 mg, crude). The crude material was used directly in the next step. MS (ESI) m/z 486.2[ M+H ]]+

Step 10: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.4] hept-6-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -1H-indole-2-carboxamide

At 20℃to N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ (5-oxo-4-azaspiro [2.4 ]]Hept-6-yl) methyl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-1H-indole-2-carboxamide (155 mg,318.95umol,1 eq.) in DCM (15 mL) was added the Buerger's reagent (228.03 mg,956.86umol,3 eq.). The solution was stirred at 20℃for 4h. After completion, the solution was concentrated to give a crude material. By preparative TLC (SiO) ₂ PE: EA=0:1) to give the product (70 mg), and purification was continued by SFC to give the product

7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.4 ] as a white solid]Hept-6-yl) ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (27 mg,57.70umol,18.09% yield, 100% purity) and 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-oxo-4-azaspiro [2.4 ] as a white solid]Hept-6-yl) ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-1H-indole-2-carboxamide (5 mg,10.36umol,3.25% yield, 97% purity). SFC method (neutral), column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-IPA ]；B％：45％-45％，6min。MS(ESI)m/z 468.2[M+H] ⁺ Isomer 1: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.72(br s,1H),9.02(d,J＝8.1Hz,1H),8.72(d,J＝7.5Hz,1H),7.80(s,1H),7.63(dd,J＝0.8,8.0Hz,1H),7.33-7.24(m,2H),7.07(t,J＝7.8Hz,1H),5.01-4.93(m,1H),4.55-4.47(m,1H),3.50-3.37(m,1H),3.33-3.27(m,1H),2.68-2.59(m,1H),2.56-2.51(m,1H),2.20(ddd,J＝5.7,9.1,13.7Hz,1H),2.01-1.76(m,4H),1.50(ddd,J＝6.2,7.6,14.0Hz,1H),1.03(d,J＝6.1Hz,1H),0.86-0.76(m,1H),0.76-0.67(m,1H),0.58-0.48(m,3H),0.48-0.38(m,2H),0.23-0.15(m,1H),0.15-0.07(m,1H)

isomer 2: ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.74(br s,1H),9.09(d,J＝7.7Hz,1H),8.76(d,J＝7.7Hz,1H),7.85(s,1H),7.63(d,J＝7.5Hz,1H),7.32(d,J＝7.4Hz,1H),7.26(s,1H),7.07(t,J＝7.8Hz,1H),4.96(d,J＝7.2Hz,1H),4.58(br d,J＝6.1Hz,1H),3.56-3.38(m,2H),2.58(br s,1H),2.55-2.52(m,1H),2.32-2.23(m,1H),2.12-2.05(m,1H),2.03-1.93(m,1H),1.88-1.73(m,2H),1.54(s,1H),1.23(br s,1H),1.03(d,J＝6.1Hz,1H),0.80(br s,1H),0.77-0.71(m,1H),0.61-0.48(m,3H),0.48-0.39(m,2H),0.19(br d,J＝2.4Hz,1H),0.12(br d,J＝2.4Hz,1H)

EXAMPLE 260 Synthesis of viral protease inhibitor Compound 1059

Step 1: n- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK IE (250 mm. Times.30 mm,10 um); mobile phase: [ heptane-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -80%,19 min) isolation of N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (210 mg) to give N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 1 (50 mg,102.56umol,100% purity) as a white solid. MS (ESI) M/z488.1[ M+H ]] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27(s,1H),7.20-7.10(m,1H),7.04-7.02(m,1H),6.52-6.50(m,1H),5.17-5.08(m,1H),4.60-4.56(m,1H),3.93(s,3H),3.26-3.18(m,2H),2.53-2.37(m,2H),2.49-2.40(m,1H),2.04-1.88(m,3H),1.87-1.64(m,3H),1.58-1.43(m,2H),1.18-1.05(m,1H)。

By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -55%,8 min) to give N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 2 (25 mg,47.03umol,91.7% purity) as a white solid. MS (ESI) m/z488.1[ M+H ] ] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27(s,1H),7.20-7.11(m,1H),7.04-7.02(m,1H),6.52-6.50(m,1H),5.18-5.06(m,1H),4.62-4.59(m,1H),3.93(s,3H),3.26-3.17(m,2H),2.52-2.37(m,2H),2.23-2.10(m,1H),2.02-1.88(m,3H),1.86-1.76(m,1H),1.75-1.62(m,2H),1.59-1.44(m,2H),1.21-1.09(m,1H)。

EXAMPLE 261 Synthesis of viral protease inhibitor Compound 1059

By SFC (column: DAICEL CHIRALPAK IE (250 mm. Times.30 mm,10 um); mobile phase: [ heptane-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,20 min) isolation of N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]Amino group]-1- [ (2, 2-difluorocyclopropyl) methyl]-2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (210 mg) to give N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 2_2 (50 mg,102.15umol,99.6% purity) as a white solid. MS (ESI) m/z 488.1[ M+H ]] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27(s,1H),7.18-7.16(m,1H),7.04-7.02(m,1H),6.53-6.51(m,1H),5.08-5.04(m,1H),4.64-4.60(m,1H),3.98(s,3H),3.20-3.19(m,2H),2.39-2.33(m,2H),2.31-2.05(m,5H),2.01-1.96(m,2H),1.72-1.43(m,2H),1.18-1.09(m,1H)。

By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:15% -55%,8 min) to give N- (1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3- (2, 2-difluorocyclopropyl) -1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide isomer 2_1 (50 mg,102.56umol,100% purity) as a white solid. MS (ESI) m/z 488.1[ M+H ] ] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.27(s,1H),7.20-7.11(m,1H),7.04-7.02(m,1H),6.52-6.50(m,1H),5.18-5.06(m,1H),4.62-4.59(m,1H),3.93(s,3H),3.26-3.17(m,2H),2.52-2.37(m,2H),2.23-2.10(m,1H),2.02-1.88(m,3H),1.86-1.76(m,1H),1.75-1.62(m,2H),1.59-1.44(m,2H),1.16-1.12(m,1H)。

EXAMPLE 262 Synthesis of (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -3-trimethylsilyl-propionic acid

Step 1: (R) -2- (4-methoxy-1H-indole-2-carboxamide) -3- (trimethylsilyl) propionic acid

(2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]A solution of tert-butyl-3-trimethylsilyl-propionate (30 mg, 76.82. Mu. Mol,1 eq.) in DCM (1.2 mL) was cooled to 0deg.C and then TFA/H was added dropwise at 0deg.C ₂ O10:1 (0.8 mL). Subsequently, the reaction was stirred at 25℃for 2h. After completion, the reaction was concentrated to dryness in vacuo at below 30 ℃. The residue was poured into water (20 mL). The aqueous phase was extracted with ethyl acetate (12 ml x 2). The combined organic phases were dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo. By preparative HPLC (column Phenomenex luna C, 18, 80X 40mm X3 um; mobile phase: [ water (0.04% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:32% -58%,7 min) to give (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a white solid]-3-trimethylsilyl-propionic acid (2.74 mg,8.19umol,10.67% yield, 100% purity). MS (ESI) m/z 335.1[ M+H ]] ⁺¹ H NMR(400MHz,CDCl ₃ -d)δ＝9.81-9.58(s,1H),7.21(t,J＝8.0Hz,1H),7.09-7.02(m,2H),6.60(br d,J＝7.9Hz,1H),6.51(d,J＝7.8Hz,1H),4.94-4.74(m,1H),3.96(s,3H),1.37(dd,J＝5.6,14.7Hz,1H),1.16(br dd,J＝9.7,14.7Hz,1H),0.11(s,9H)

EXAMPLE 263 Synthesis of viral protease inhibitor Compound 1083

Step 1: (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propionic acid methyl ester

At 20℃under N ₂ To (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propionic acid (500 mg,2.29mmol,1 eq.) was added HCl/MeOH (4M, 25.00mL,43.65 eq.) in one portion. The mixture was stirred at 20℃for 12h. After completion, the reaction mixture was concentrated to give the product. Methyl (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propionate (550 mg,2.05mmol,89.33% yield, HCl) was obtained as a purple solid and used directly in the next step. MS (ESI) m/z 233.1[ M+H ]] ⁺ 1H NMR(400MHz,DMSO-d ₆ )δppm 11.03(br s,1H),8.49(br s,4H),7.09-7.22(m,2H),6.92(t,J＝7.61Hz,1H),6.71(d,J＝7.06Hz,1H),4.11(br t,J＝7.28Hz,1H),3.65(s,3H),3.39-3.48(m,1H),3.27(br d,J＝8.16Hz,1H),2.59(s,3H)

Step 2: (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propanamide

At 20℃under N ₂ To methyl (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propionate (550 mg,2.05mmol,1 eq., HCl) was added NH in one portion ₃ MeOH (7M, 20.00mL,68.41 eq.). The mixture was stirred at 80℃for 12h. After completion, the reaction mixture was cooled to 25 ℃ and concentrated to give the product. (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propionamide (520 mg, crude material) was obtained as a pale yellow solid and used directly in the next step. MS (ESI) m/z 218.1[ M+H ]] ⁺

Step 3: (2S) -2-amino-3- (4-methyl-2-oxo-indolin-3-yl) propanamide

At 20℃under N ₂ Next (2S) -2-amino-3- (4-methyl-1H-indol-3-yl) propanamide (490 mg,2.26mmol,1 eq.) was added in one portion to a mixture in AcOH (10 mL)A solution of DMSO (264.32 mg,3.38mmol,264.32uL,1.5 eq.) and HCl (12M, 751.77uL,4 eq.) was added. The mixture was stirred at 20℃for 2h. After completion, water (10 mL) was added to the reaction mixture and concentrated to 10mL. By preparative HPLC (column Phenomenex luna C, 250X 50mm X10 um; mobile phase: [ water (0.04% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -30%,10 min) to purify the crude product. (2S) -2-amino-3- (4-methyl-2-oxo-indolin-3-yl) propanamide (124 mg, crude material, HCl) was obtained as a pale green solid. MS (ESI) m/z 234.1[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ (4-methyl-2-oxo-indolin-3-yl) methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

at-30deg.C, at N ₂ Downward (2S) -2-amino-3- (4-methyl-2-oxo-indolin-3-yl) propanamide (120 mg,514.43umol,1 eq.) and (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a mixture of propionic acid (194.41 mg,514.43umol,80% purity, 1 eq.) in DMF (1.2 mL) was added PyBop (267.71 mg,514.43umol,1 eq.) and TEA (156.17 mg,1.54mmol,214.81uL,3 eq.). The mixture was stirred at-30℃for 2h. After completion, the reaction mixture was quenched by addition of water (10 mL) at 20 ℃ and then diluted with DCM (10 mL) and extracted with DCM (5 mL x 2). The combined organic layers were concentrated under reduced pressure to give a residue. The crude product was purified by preparative TLC. Obtaining N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (4-methyl-2-oxo-indolin-3-yl) methyl ] as a pale yellow solid ]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (130 mg, crude). MS (ESI) m/z 518.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -1-cyano-2- (4-methyl-2-oxo-indolin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

At 20℃under N ₂ Downward N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (4-methyl-2-oxo-indolin-3-yl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxoRadical-ethyl radical]To a mixture of 4-methoxy-1H-indole-2-carboxamide (130 mg,251.17umol,1 eq.) in DCM (20 mL) was added the Burgess reaction (179.57 mg,753.51umol,3 eq.) in one portion. The mixture was stirred at 20℃for 12h. After completion, water (5 mL) was added to the reaction mixture and stirred for 20min. The reaction mixture was then concentrated to give the crude product. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to purify the crude product. Obtaining N- [ (1S) -2- [ [ (1S) -1-cyano-2- (4-methyl-2-oxo-indolin-3-yl) ethyl as a white solid ]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (55 mg,110.10umol,43.83% yield). MS (ESI) m/z 500.2[ M+H ]] ⁺

Step 6: n- [2- [ [ (1S) -1-cyano-2- (4-methyl-2-oxo-indolin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ .H ₂ O EtOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:55% -55%,20 min). Isomer 1: obtaining N- [2- [ [ (1S) -1-cyano-2- (4-methyl-2-oxo-indolin-3-yl) ethyl as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (10.8 mg,21.34umol,19.38% yield, 98.7% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δppm11.57(s,1H)10.41-10.59(m,1H)8.95(m,1H)8.42-8.59(m,1H)7.36(m,1H)7.05-7.14(m,2H)6.99(br d,J＝8.33Hz,1H)6.73-6.82(m,1H)6.66(m,1H)6.50(d,J＝7.89Hz,1H)4.99-5.14(m,1H)4.38-4.54(m,1H)3.88(s,3H)3.50-3.65(m,1H)3.50-3.65(m,1H)2.62-2.72(m,1H)2.28(d,J＝12.93Hz,2H)2.17(m,1H)1.92-2.04(m,1H)1.71-1.87(m,1H)1.43-1.61(m,2H)0.73-0.88(m,1H)0.33-0.49(m,2H)0.02-0.25(m,2H)

Isomer 2: obtaining N- [2- [ [ (1S) -1-cyano-2- (4-methyl-2-oxo-indolin-3-yl) ethyl as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxylic acid methyl esterAmide (17.2 mg,34.29umol,31.15% yield, 99.6% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δppm11.55(br d,J＝1.53Hz,1H)10.40-10.54(m,1H)8.93(m,1H)8.48(m,1H)7.34(m,1H)7.03-7.14(m,2H)6.98(br d,J＝8.11Hz,1H)6.71-6.79(m,1H)6.64(m,1H)6.45-6.52(m,1H)4.97-5.10(m,1H)4.36-4.53(m,1H)3.86(s,3H)3.49-3.64(m,1H)2.57-2.64(m,1H)2.26(d,J＝12.94Hz,3H)2.16(m,1H)1.96(m,1H)1.70-1.84(m,1H)1.44-1.57(m,1H)0.77(m,1H)0.34-0.44(m,2H)0.04-0.22(m,2H)

EXAMPLE 264 Synthesis of viral protease inhibitor Compound 1085

Step 1: (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionic acid methyl ester

A mixture of (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionic acid (1 g,4.58mmol,1 eq.) in HCl/MeOH (4M, 20 mL) was stirred at 25℃for 16H. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionate (1.1 g,4.09mmol,89.33% yield, HCl) as a yellow solid.

Step 2: (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propanamide

Methyl (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionate (1.1 g,4.74mmol,1 eq.) was reacted with NH ₃ The mixture in MeOH (7M, 34.71mL,51.30 eq.) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionamide (1 g,4.60mmol,97.19% yield) as a yellow solid. MS (ESI) m/z 218.1[ M+H ]] ⁺

Step 3: (2S) -2-amino-3- (5-methyl-2-oxo-indolin-3-yl) propanamide

A mixture of DMSO (539.43 mg,6.90mmol,539.43uL,1.5 eq.) and HCl (12M, 1.53mL,4 eq.) was added to a mixture of (2S) -2-amino-3- (5-methyl-1H-indol-3-yl) propionamide (1 g,4.60mmol,1 eq.) in AcOH (10 mL),the mixture was stirred at 25℃for 16h. After completion, the mixture was purified by preparative HPLC (HCl conditions; column: welch Xtime C18X 25mm X3 um; mobile phase: [ water (0.04% HCl) -ACN) ]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -10%,8 min) to give (2S) -2-amino-3- (5-methyl-2-oxo-indolin-3-yl) propanamide (370 mg,682.05umol,14.82% yield, 43% purity) as a yellow solid. MS (ESI) m/z 234.0[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5-methyl-2-oxo-indolin-3-yl) methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To (2S) -2-amino-3- (5-methyl-2-oxo-indolin-3-yl) propanamide (370 mg,682.05umol,43% purity, 1 eq.) and (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino]To a solution of propionic acid (226.82 mg,750.26 mmol, 1.1 eq.) in DCM (5 mL) and DMF (2 mL) was added DMAP (249.98 mg,2.05mmol,3 eq.) and EDCI (261.50 mg,1.36mmol,2 eq.). The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 10mL H ₂ O was diluted and extracted with 18mL EA (6 mL x 3). The combined organic layers were washed with 9mL brine (9 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By preparative TLC (SiO) ₂ Purification of the crude material to give N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5-methyl-2-oxo-indolin-3-yl) methyl) as a white solid was performed by EA: meoh=10:1) ]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (150 mg,289.81umol,42.49% yield). MS (ESI) m/z 518.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-methyl-2-oxo-indolin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (5-methyl-2-oxo-indolin-3-yl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (120 mg,185.48umol,80% purity, 1 eq.) in DCM (2 mL)To the mixture of (2) was added at once a primary (88.40 mg,370.96umol,2 equivalents). The mixture was stirred at 40℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: waters Xbridge Prep OBD C: 150 x 40mM x 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- (5-methyl-2-oxo-indolin-3-yl) ethyl as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-4-methoxy-1H-indole-2-carboxamide (30 mg,60.05umol,32.38% yield). MS (ESI) m/z 500.1[ M+H ]] ⁺¹ HNMR(400MHz,DMSO-d ₆ )δ＝11.62-11.48(m,1H),10.47-10.30(m,1H),9.12-8.89(m,1H),8.58-8.45(m,1H),7.37(br d,J＝6.7Hz,1H),7.19-7.04(m,2H),7.02-6.95(m,2H),6.77-6.63(m,1H),6.55-6.15(m,1H),5.25-5.02(m,1H),4.58-4.40(m,1H),3.89(d,J＝3.3Hz,3H),3.51-3.38(m,1H),2.32-2.13(m,5H),1.87-1.70(m,1H),1.59-1.39(m,1H),0.80(br s,1H),0.50-0.28(m,2H),0.27--0.03(m,2H)

EXAMPLE 265 Synthesis of viral protease inhibitor Compound 1087

Step 1: (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionic acid methyl ester

At 20℃under N ₂ To (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionic acid (1.00 g,4.58mmol,1 eq.) was added HCl/MeOH (4M, 50.00mL,43.65 eq.) in one portion. The mixture was stirred at 20℃for 12h. After completion, the reaction mixture was concentrated to give methyl (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionate (1.15 g, crude material, HCl) as a pale yellow solid and was used directly in the next step. MS (ESI) M/z233.1[ M+H] ⁺

Step 2: (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propanamide

(2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionic acid methyl ester (1.15 g,4.28mmol,1 eq. HCl) was dissolved at 20 ℃By dissolving in NH ₃ In MeOH (7M, 20mL,32.72 eq.). The mixture was stirred at 80℃for 12h. After completion, the reaction mixture was cooled to 20 ℃ and concentrated to give the product. (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionamide (1.1 g, crude material) was obtained as a pale yellow solid and used directly in the next step. MS (ESI) m/z 218.1[ M+H ] ] ⁺

Step 3: (2S) -2-amino-3- (6-methyl-2-oxo-indolin-3-yl) propanamide

At 20℃under N ₂ To a solution of (2S) -2-amino-3- (6-methyl-1H-indol-3-yl) propionamide (1.08 g,4.97mmol,1 eq.) in AcOH (10 mL) was added dropwise a solution of DMSO (582.58 mg,7.46mmol,582.58uL,1.5 eq.) and HCl (12M, 1.66mL,4 eq.). The mixture was stirred at 20℃for 2h. After completion, saturated aqueous sodium bicarbonate solution was added to the reaction mixture to pH-6. By preparative HPLC (column Phenomenex luna C, 250X 50mm X10 um; mobile phase: [ water (0.04% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -30%,10 min) to give (2S) -2-amino-3- (6-methyl-2-oxo-indolin-3-yl) propanamide (0.22 g,754.50umol,15.18% yield, 80% purity) as a green solid. MS (ESI) m/z 234.1[ M+H ]] ⁺

Step 4: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ (6-methyl-2-oxo-indolin-3-yl) methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

at-30deg.C, at N ₂ Downward (2S) -2-amino-3- (6-methyl-2-oxo-indolin-3-yl) propanamide (200.00 mg,857.39umol,1 eq.) and (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino group ]To a mixture of propionic acid (324.01 mg,857.39umol,80% purity, 1 eq.) in DMF (0.5 mL) was added PyBop (446.18 mg,857.39umol,1 eq.) and TEA (260.28 mg,2.57mmol,358.01uL,3 eq.) in one portion. The mixture was stirred at-30℃for 2h. After completion, the reaction mixture was quenched by addition of water (20 mL) at 20 ℃ and then diluted with DCM (20 mL) and extracted with DCM (10 mL x 2). The combined organic layers were concentrated under reduced pressure to give N- [ (1S) -2- [ [ (1S) -2-ammonia as a pale yellow oil1- [ (6-methyl-2-oxo-indolin-3-yl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (200 mg, crude). MS (ESI) m/z 518.2[ M+H ]] ⁺

Step 5: n- [ (1S) -2- [ [ (1S) -1-cyano-2- (6-methyl-2-oxo-indolin-3-yl) ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4-methoxy-1H-indole-2-carboxamide

At 25℃under N ₂ Downward N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (6-methyl-2-oxo-indolin-3-yl) methyl]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 4-methoxy-1H-indole-2-carboxamide (100 mg,193.21umol,1 eq.) in DCM (20 mL) was added the Bungeus reagent (138.13 mg,579.63umol,3 eq.) in one portion. The mixture was stirred at 25℃for 2h. After completion, water (5 mL) was added to the reaction mixture and stirred for 20min. The reaction mixture was then concentrated to give the crude product. By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- (6-methyl-2-oxo-indolin-3-yl) ethyl as a white solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (2.11 mg,4.20umol,2.17% yield, 99.4% purity). MS (ESI) m/z 500.2[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.67-11.47(m,1H),10.52-10.34(m,1H),9.11-8.93(m,1H),8.59-8.44(m,1H),7.40-7.31(m,1H),7.18-7.03(m,2H),7.01-6.94(m,1H),6.82-6.70(m,1H),6.64(d,J＝7.2Hz,1H),6.54-6.45(m,1H),5.19-5.01(m,1H),4.53-4.41(m,1H),3.91-3.83(m,3H),3.49-3.36(m,1H),2.31(br d,J＝2.0Hz,5H),1.85-1.68(m,1H),1.59-1.38(m,1H),0.86-0.70(m,1H),0.44-0.28(m,2H),0.24--0.01(m,2H)。

EXAMPLE 266 Synthesis of viral protease inhibitor Compound 1091

Step 1: (S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionic acid methyl ester

A mixture of (2S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionic acid (1 g,4.19mmol,1 eq.) in HCl/MeOH (4M, 20mL,19.09 eq.) was stirred at 25℃for 14H. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionate (1 g, crude material) as a yellow solid. MS (ESI) m/z 251.1[ M-H ]] ⁺ .

Step 2: (S) -2-amino-3- (4-chloro-1H-indol-3-yl) propanamide

Methyl (2S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionate (1 g,3.96mmol,1 eq.) is reacted with NH ₃ A solution in MeOH (7M, 20.00mL,35.38 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give (2S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionamide (0.9 g, crude material) as a yellow solid. MS (ESI) m/z 238.1[ M+H ] ] ⁺ 。

Step 3: (2S) -2-amino-3- (4-chloro-2-oxoindolin-3-yl) propanamide

A solution of (2S) -2-amino-3- (4-chloro-1H-indol-3-yl) propionamide (500 mg,2.10mmol,1 eq.) in t-BuOH (6 mL), etOH (4 mL) and AcOH (2 mL) was then added BLAH; pyridin-1-ium (672.78 mg,2.10mmol,1 eq.) was stirred at 20℃for 3h, followed by the addition of AcOH (2 mL) and Zn (1.05 g,15.99mmol,7.6 eq.) and the mixture was stirred at 20℃for 15h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column Phenomenex luna C, 18, 80X 40mm X3 um; mobile phase: [ water (0.04% HCl) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:1% -25%,7 min) to give (2S) -2-amino-3- (4-chloro-2-oxo-indolin-3-yl) propanamide (35 mg,137.97umol,6.56% yield) as a white solid. MS (ESI) m/z 254.1[ M+H ]] ⁺ 。

Step 4: n- ((2S) -1- (((2S) -1-amino-3- (4-chloro-2-oxoindol-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To (2S) -2-amino-3- (4-chloro-2-oxo-indoline-3-yl) propanamide (30 mg,103.40umol,1 eq, HCl) and (2S) -3-cyclopropyl-2- [ (4-methoxy-1H-indole-2-carbonyl) amino ]To a solution of propionic acid (40.64 mg,134.42umol,1.3 eq.) in DMF (1 mL) was added Py-Bop (53.81 mg,103.40umol,1 eq.) and cooled to-30℃followed by dropwise addition of an Et-containing solution ₃ N (31.39 mg,310.19umol,43.17uL,3 eq.) in DMF (0.5 mL) and the mixture was stirred at-30℃for 2h. After completion, by adding 20mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (10 ml x 3). The combined organic layers were washed with 15mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with EtOAc: meoh=20:1) to give N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (4-chloro-2-oxo-indolin-3-yl) methyl as a white solid]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (35 mg,65.06umol,62.92% yield). MS (ESI) m/z 538.2[ M+H ]] ⁺ 。

Step 5: n- ((2S) -1- (((1S) -2- (4-chloro-2-oxoindol-3-yl) -1-cyanoethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ (4-chloro-2-oxo-indolin-3-yl) methyl]-2-oxo-ethyl ]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (35 mg,65.06umol,1 eq.) in DCM (2 mL) was added the Buerger's reagent (31.01 mg,130.11umol,2 eq.) followed by stirring the mixture at 20℃for 12H. After completion, by adding 0.5mL H ₂ O to quench the reaction mixture, and concentrate under reduced pressure to give a residue. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give N- [ (1S) -2- [ [ (1S) -2- (4-chloro-2-oxo-indolin-3-yl) -1-cyano-ethyl ] as a yellow solid]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4-methoxy-1H-indole-2-carboxamide (4 mg,7.69umol,11.82% yield, 100% purity). MS (ESI) m/z 520.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.29-7.23(m,1H),7.21-7.10(m,2H),7.07-6.95(m,2H),6.88-6.65(m,1H),6.55-6.49(m,1H),5.28-4.53(m,1H),3.93(d,J＝2.6Hz,3H),3.85-3.72(m,1H),3.01-2.85(m,1H),2.62-2.28(m,1H),1.88-1.78(m,1H),1.70-1.55(m,1H),1.41-1.26(m,1H),0.95-0.76(m,1H),0.58-0.39(m,2H),0.27-0.09(m,2H)。

EXAMPLE 267 Synthesis of viral protease inhibitor Compound 1101

Step 1: (S) -2-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- (tert-butoxycarbonylamino) -3- (5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) propionate (500.00 mg,1.59mmol,1 eq.) in HCl/MeOH (4M, 10.00mL,25.15 eq.) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionate (390 mg, crude material, HCl) as a white solid.

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (390 mg,1.82mmol,1 eq.) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (580.48 mg,2.37mmol,1.3 eq.) were added DMAP (667.11 mg,5.46mmol,3 eq.) and EDCI (697.88 mg,3.64mmol,2 eq.) in DCM (10 mL) and DMF (3 mL), and the mixture was then stirred at 20℃for 2h. After completion, by adding 30mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (30 ml x 3). The combined organic layers were washed with brine (30 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) to give the residue as a white solid(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (510 mg,924.00umol,50.76% yield, 80% purity). MS (ESI) m/z 442.2[ M+H ] ] ⁺ 。

Step 3: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (250 mg,566.17 mol,1 eq) in HCl/MeOH (4M, 5mL,35.32 eq) was stirred at 25℃for 1h. The reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (190 mg, crude material, HCl) as a white solid.

Step 4: (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2- (4-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (190 mg, 502.77. Mu. Mol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (124.96 mg, 653.60. Mu. Mol,1.3 eq) in DCM (4 mL) and DMF (1 mL) and then DMAP (184.27 mg,1.51mmol,3 eq) and EDCI (192.76 mg,1.01mmol,2 eq) were added and the mixture was stirred at 20℃for 2H. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (50 ml x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give the product (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]Methyl propionate (200 mg,349.78umol,69.57% yield, 9)0% purity). MS (ESI) m/z 515.2[ M+H ]] ⁺ 。

Step 5: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

(2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]-2- [ [ (2S) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]Methyl propionate (200 mg,388.64umol,1 eq.) in NH ₃ A solution in MeOH (7M, 5mL,90.06 eq.) was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid ]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (190 mg, crude). MS (ESI) M/z500.2[ M+H] ⁺ 。

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 4-methoxy-1H-indole-2-carboxamide (160 mg,320.26umol,1 eq.) in DCM (3 mL) was added the berg reagent (91.58 mg,384.31umol,1.2 eq.) and the mixture was then stirred at 25 ℃ for 3H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: xtimate C18 u 250mM 80mM; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,35 min) to give the product N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (67.35 mg,139.85umol,43.67% yield, 100% purity). MS (ESI) m/z 482.2[ M+H ] ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.54(d,J＝1.6Hz,1H),8.87(d,J＝8.2Hz,1H),8.46(d,J＝8.1Hz,1H),7.81(s,1H),7.33(d,J＝1.5Hz,1H),7.12-7.06(m,1H),7.03-6.98(m,1H),6.50(d,J＝7.6Hz,1H),4.98-4.89(m,1H),4.51(dt,J＝3.6,8.5Hz,1H),3.88(s,3H),2.60-2.54(m,1H),2.17(dt,J＝4.8,9.0Hz,1H),1.95(dd,J＝8.6,12.2Hz,1H),1.83-1.72(m,2H),1.71-1.63(m,1H),1.49(t,J＝11.6Hz,1H),1.14(s,3H),1.02(s,3H),0.93(s,9H)

EXAMPLE 268 Synthesis of viral protease inhibitor Compounds 1103

Step 1: (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester hydrochloride

A solution of (3S) -3- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester (400 mg,834.01umol,1 eq) in HCl/MeOH (4M, 5mL,23.98 eq) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (3S) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (340 mg, crude material, HCl) as a white solid.

Step 2: (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (170.98 mg,894.33umol,1.2 eq.) and (2S) -2- [ [ (3S) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (310 mg,745.28 mmol, 1 eq., HCl) in DCM (15 mL) and DMF (3 mL) was added DMAP (273.15 mg,2.24mmol,3 eq.) and EDCI (285.74 mg,1.49mmol,2 eq.) followed by stirring the mixture at 20℃for 2h. After completion, by adding H at 0deg.C ₂ O (50 mL) to quench the reaction mixture and then extract with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL1) washing with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give the product (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]-2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carbonyl]Amino group]Methyl propionate (320 mg,561.65umol,75.36% yield, 97% purity). MS (ESI) m/z 553.2[ M+H ]] ⁺ 。

Step 3: (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]-2- [ [ (3S) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carbonyl]Amino group]Methyl propionate (320 mg,579.02umol,1 eq.) in NH ₃ A solution in MeOH (7M, 5.00mL,60.45 eq.) was stirred at 60℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid ]Methyl group]-2-oxo-ethyl]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (310 mg, crude). MS (ESI) m/z 538.3[ M+H ]] ⁺ 。

Step 4: (S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]A solution of decane-3-carboxamide (310 mg,501.63umol,87% purity, 1 eq.) and Prague reagent (239.09 mg,1.00mmol,2 eq.) in DCM (5 mL) was stirred at 30℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]；B％：35％-65％，8min) purification of the residue to give the product (3S) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carboxamide (115.37 mg,222.02umol,44.26% yield, 100% purity). MS (ESI) m/z 520.2[ M+H ] ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.53(s,1H),8.89(d,J＝8.1Hz,1H),7.81(s,1H),7.15-7.08(m,1H),7.02(d,J＝8.3Hz,1H),6.94(d,J＝1.2Hz,1H),6.52(d,J＝7.7Hz,1H),4.96-4.89(m,1H),4.50(t,J＝8.5Hz,1H),3.89(s,3H),3.87-3.79(m,1H),3.67(d,J＝10.4Hz,1H),2.26-2.11(m,2H),2.00(dd,J＝8.4,12.0Hz,1H),1.78-1.70(m,1H),1.63-1.31(m,13H),1.14(s,3H),1.08-1.00(m,3H)。

EXAMPLE 269 Synthesis of viral protease inhibitor Compound 1105

Step 1:7- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester

To (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (450 mg,2.10mmol,1 eq.) and 6-tert-butoxycarbonyl-6-azaspiro [3.4]]To a mixture of octane-7-carboxylic acid (589.83 mg,2.31mmol,1.1 eq.) in DCM (10 mL) was added DMAP (769.75 mg,6.30mmol,3 eq.) and EDCI (805.24 mg,4.20mmol,2 eq.) and the mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 25mL H ₂ O was diluted and extracted with 45mL DCM (15 mL x 3). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) purification of the crude material to give 7- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-methoxy-2-oxo-ethyl]Carbamoyl group]-6-azaspiro [3.4]]Tert-butyl octane-6-carboxylate (0.9 g,1.79mmol,85.41% yield, 90% purity) 。MS(ESI)m/z 452.3[M+H] ⁺

Step 2: (2S) -2- (6-azaspiro [3.4] octane-7-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

A mixture of 7- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -6-azaspiro [3.4] octane-6-carboxylic acid tert-butyl ester (0.9 g,1.79mmol,90% purity, 1 eq.) in HCl/MeOH (4M, 15 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- (6-azaspiro [3.4] octane-7-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionate (800 mg, crude material, HCl) as a white solid.

Step 3: (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] -2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carbonyl ] amino ] propionic acid methyl ester

To (2S) -2- (6-azaspiro [3.4]]Octane-7-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (386.00 mg,995.10umol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (199.76 mg,1.04mmol,1.05 eq) in DCM (10 mL) was added DMAP (364.71 mg,2.99mmol,3 eq) and EDCI (381.53 mg,1.99mmol,2 eq) and then N ₂ The mixture was stirred for 2h at 25℃under an atmosphere. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 45mL EA (15 mL x 3). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]-2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]Methyl propionate (390 mg,691.37umol,69.48% yield, 93% purity). MS (ESI) m/z 525.3[ M+H ]] ⁺

Step 4: n- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

(2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]-2- [ [6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carbonyl group]Amino group]Methyl propionate (370 mg,705.29umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 15mL,148.88 eq.) was stirred at 50deg.C for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid ]Methyl group]-2-oxo-ethyl]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]Octane-7-carboxamide (320 mg,571.43umol,81.02% yield, 91% purity). MS (ESI) m/z 510.3[ M+H ]] ⁺

Step 5: n- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4] octane-7-carboxamide

To N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4]]To a mixture of octane-7-carboxamide (320 mg,571.43umol,91% purity, 1 eq.) in DCM (10 mL) was added the Buerger's reagent (272.36 mg,1.14mmol,2 eq.) and the resulting mixture was stirred at 20℃for 5h. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (neutral conditions; column: phenomenex Gemini-NX C18 75X 30mM 3um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give 200mg of the desired compound as a white solid, which was purified by SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ₃ H ₂ OIPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:52% -52%,12 min) to give N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 1 (90 mg,181.62umol,31.78% yield, 99.2% purity). MS (ESI) m/z 492.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(s,1H),8.83(d,J＝8.2Hz,1H),7.82(s,1H),7.16-7.07(m,1H),7.05-6.94(m,2H),6.52(d,J＝7.7Hz,1H),4.93(q,J＝8.1Hz,1H),4.45(t,J＝7.4Hz,1H),4.04-3.69(m,5H),2.70-2.56(m,1H),2.35-2.25(m,1H),2.21-1.69(m,10H),1.50(br t,J＝11.5Hz,1H),1.20-0.85(m,6H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.39-11.15(m,1H),8.83-8.51(m,1H),7.66-7.53(m,1H),7.17-7.02(m,2H),7.01-6.88(m,1H),6.53(d,J＝7.7Hz,1H),4.99-4.86(m,1H),4.64-4.44(m,1H),4.05-3.82(m,5H),2.54(br s,1H),2.39-2.25(m,1H),2.20-1.73(m,10H),1.61-1.42(m,1H),1.23-1.03(m,6H)。

Obtaining N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-6- (4-methoxy-1H-indole-2-carbonyl) -6-azaspiro [3.4 ]]Octane-7-carboxamide isomer 2 (100 mg,203.43umol,35.60% yield, 100% purity). MS (ESI) m/z 492.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69-11.46(s,1H),8.74(d,J＝8.2Hz,1H),7.87-7.66(m,1H),7.17-7.09(m,1H),7.06-6.95(m,2H),6.60-6.40(m,1H),5.00-4.74(m,1H),4.45(t,J＝7.2Hz,1H),3.98(q,J＝10.1Hz,2H),3.93-3.77(m,3H),2.49-2.41(m,1H),2.28(br dd,J＝8.0,12.2Hz,1H),2.14(br dd,J＝4.0,9.3Hz,2H),2.05-1.80(m,7H),1.80-1.67(m,1H),1.50(br t,J＝11.5Hz,1H),1.20-1.02(m,6H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.35-11.25(m,1H),8.59(br d,J＝5.5Hz,1H),7.56(br s,1H),7.18-7.03(m,2H),7.02-6.89(m,1H),6.53(d,J＝7.7Hz,1H),4.98-4.88(m,1H),4.63-4.49(m,1H),4.02-3.88(m,5H),2.49-2.42(m,1H),2.36-2.26(m,1H),2.21-2.10(m,2H),2.08-1.85(m,8H),1.58-1.46(m,1H),1.25-1.01(m,6H)。

Example 270 Synthesis of viral protease inhibitor Compound 1115

Step 1: (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (150 mg,414.52 mol,1 eq, HCl) and 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (140.29 mg,621.78 mol,1.5 eq) in DCM (4 mL) was added DMAP (101.28 mg,829.04 mol,2 eq) and EDCI (119.20 mg,621.78 mol,1.5 eq). The mixture was stirred at 20℃for 2h. After completion, by adding 40mL H ₂ O quench the reaction mixture and then extract with DCM (20 ml x 3). The combined organic layers were washed with brine (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give (2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (198mg, 360.33umol,86.93% yield, 97% purity). MS (ESI) m/z 532.2[ M+H ]] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-5-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (160 mg,300.18umol,1 eq.) in NH ₃ A solution in MeOH (7M, 10mL,233.19 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a yellow solid ]Methyl group]-2-oxo-ethyl]Amino group]-1 (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-5-methoxy-1H-indole-2-carboxamide (166.1 mg, crude material). MS (ESI) m/z 517.2[ M+H ]] ⁺ 。

Step 3: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-5-methoxy-1H-indole-2-carboxamide (140 mg,270.27umol,1 eq.) in DCM (4 mL) was added the berg reagent (141.69 mg,594.59umol,2.2 eq.). The mixture was stirred at 25℃for 2h. After completion, at N ₂ DCM in the reaction mixture was removed. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl ]-5-methoxy-1H-indole-2-carboxamide (45 mg,90.00umol,33.30% yield, 100% purity). MS (ESI) m/z 499.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.67-11.34(m,1H),9.06-8.96(m,1H),8.69-8.62(m,1H),7.90-7.79(m,1H),7.19-7.11(m,2H),7.02-6.96(m,1H),5.02-4.92(m,1H),4.57-4.45(m,1H),3.82-3.73(m,3H),2.63-2.54(m,1H),2.26-2.12(m,1H),2.03-1.95(m,1H),1.86-1.73(m,2H),1.58-1.44(m,2H),1.16-1.12(m,3H),1.09-1.05(m,3H),0.85-0.77(m,1H),0.51-0.38(m,2H),0.26-0.16(m,1H),0.14-0.06(m,1H)。

EXAMPLE 271 Synthesis of viral protease inhibitor Compound 1119

A solution of tert-butyl (3S) -3- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylate (400 mg,834.01 mol,1 eq) in HCl/MeOH (4M, 5.00mL,23.98 eq) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (3S) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (340 mg, crude material, HCl) as a white solid.

Step 2: (S) -2- ((S) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

(2S) -2- [ [ (3S) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ]Methyl propionate (310 mg,745.28umol,1 eq, HCl) and 6-chloro-1H-indole-2-carboxylic acid (174.93 mg,894.33umol,1.2 eq) in DCM (10 mL) and DMF (3 mL) and then DMAP (273.15 mg,2.24mmol,3 eq) and EDCI (285.74 mg,1.49mmol,2 eq) were added and the mixture was stirred at 20℃for 2H. After completion, the reaction mixture was quenched by adding 30ml h2o at 0 ℃ and then extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (30 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (2S) -2- [ [ (3S) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Propionate (420 mg,467.44umol,62.72% yield, 62% purity). MS (ESI) m/z 557.2[ M+H ]] ⁺ 。

Step 3: (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [ (3S) -2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5 ]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (420 mg,753.93umol,1 eq.) in NH ₃ Solution in MeOH (7M, 5mL,46.42 eq.) and stirred at 60 ℃The mixture was stirred for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-oxo-ethyl]-2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (400 mg, crude). MS (ESI) M/z542.2[ M+H] ⁺ 。

Step 4: (S) -2- (6-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (6-chloro-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (400 mg,553.44umol,75% purity, 1 eq.) in DCM (5 mL) was added the Bungeus reagent (791.32 mg,3.32mmol,6 eq.) and the mixture was then stirred at 30℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,8 min) to give the product (3S) -2- (6-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (80.38 mg,153.38umol,27.71% yield, 99.7% purity). MS (ESI) m/z 524.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.62(d,J＝8.5Hz,1H),7.45(s,1H),7.09-6.99(m,2H),5.06-4.94(m,1H),4.61(dd,J＝7.7,9.8Hz,1H),3.99(br d,J＝10.2Hz,1H),3.72(d,J＝10.3Hz,1H),2.97(br dd,J＝5.0,8.7Hz,1H),2.51-2.34(m,1H),2.30(br dd,J＝7.7,12.3Hz,1H),2.16(dd,J＝8.5,12.3Hz,1H),1.91-1.81(m,1H),1.74(dd,J＝10.2,12.4Hz,1H),1.67-1.37(m,11H),1.22(s,3H),1.13-0.80(m,3H)。

EXAMPLE 272 Synthesis of viral protease inhibitor Compounds 1121

Step 1: (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

(S) -3- (((S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-methoxy-1-oxopropan-2-yl) carbamoyl) -2-azaspiro [4.5]A solution of tert-butyl decane-2-carboxylate (350 mg,729.76umol,1 eq.) in HCl/MeOH (4M, 4 mL) was stirred at 20℃for 1h. After completion of the reaction, the reaction mixture was concentrated in vacuo to give (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2-azaspiro [4.5] as a white solid]Decane-3-carboxamido) propionic acid methyl ester (280 mg, crude material). MS (ESI) M/z380.2[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((S) -2-azaspiro [4.5]To a solution of methyl decane-3-carboxamido) propionate (280 mg,673.15umol,1 eq, HCl) in DCM (2 mL) was added 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (182.26 mg,807.78umol,1.2 eq), DMAP (205.60 mg,1.68mmol,2.5 eq) and EDCI (258.09 mg,1.35mmol,2 eq) and the mixture was stirred at 20 ℃ for 1H. After the reaction was complete, the reaction mixture was quenched by the addition of water (5 mL) and then extracted with DCM (2 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL) followed by saturated NaHCO ₃ (2 mL) is adjusted to pH-7, and the mixture is treated by Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester (300 mg,510.98umol,75.91% yield). MS (ESI) M/z587.3[ M+H ]] ⁺

Step 3: (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [ 4.5)]Decane-3-formylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester (300 mg,510.98umol,1 eq.) in NH ₃ A solution in MeOH (7M, 5 mL) was stirred at 50℃for 20h. After completion of the reaction, the mixture was concentrated in vacuo to give (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (260 mg, crude). MS (ESI) m/z 572.3[ M+H ]] ⁺

Step 4: (S) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To (S) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (260 mg,461.46umol,1 eq.) in DCM (2 mL) was added the Bungeus reagent (329.91 mg,1.38mmol,3 eq.) and stirred at 30℃for 1.5h. After the reaction was completed, the mixture was quenched with water (1 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge BEH C, 100 x 25mM x 5um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,10 min) to give (S) -2- (7-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -2-azaspiro [4.5 ] as a white solid]Decane-3-carboxamide (61.15 mg,110.36umol,23.92% yield, 100% purity). MS (ESI) m/z 554.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.12-10.96(m,1H),8.79-8.63(m,1H),7.63-7.51(m,1H),7.22(s,1H),7.05-6.91(m,1H),6.61-6.53(m,1H),4.96-4.86(m,1H),4.67-4.51(m,1H),3.95-3.88(m,3H),3.87-3.78(m,1H),3.70-3.54(m,1H),2.65-2.55(m,1H),2.30-2.09(m,2H),1.85-1.74(m,1H),1.71-1.62(m,1H),1.60-1.33(m,12H),1.21-1.14(m,3H),1.12-1.03(m,3H)。

Example 273 Synthesis of viral protease inhibitor Compound 1123

Step 1: (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate hydrochloride (130 mg,359.25 mol,1.1 eq, HCl) in DCM (5 mL) was added 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (73.69 mg,326.59 mol,1 eq), DMAP (119.70 mg,979.78 mol,3 eq) and EDCI (125.22 mg,653.19 mol,2 eq). The mixture was stirred at 25℃for 2h. After completion, the reaction was quenched with H ₂ O (20 mL) was quenched and extracted with DCM (10 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered, concentrated in vacuo and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give the product methyl (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (150 mg,281.42umol,86.17% yield) as a yellow solid. MS (ESI) m/z 533.2[ M+H ]] ⁺

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide

A solution of methyl (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (150 mg,281.42umol,1 eq.) in ammonia (in MeOH solution, 7M,20mL,497.48 eq.) was stirred at 30℃for 10H. After completion, the reaction was concentrated in vacuo to give the product N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclo as a yellow solidPropyl-1-oxopropan-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide (130 mg, crude material). MS (ESI) M/z518.2[ M+H ] ⁺

Step 3: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide (130 mg,250.96umol,1 eq) in DCM (10 mL) was added the bergs reagent (179.42 mg,752.89umol,3 eq) and the mixture stirred at 25 ℃ for 1H. After completion, the mixture was concentrated in vacuo and purified by preparative HPLC (column Phenomenex Gemini-NX 80 x 40mM x 3um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the product 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-1H-indole-2-carboxamide (40 mg,80.00umol,31.88% yield, 100% purity) as a white solid. MS (ESI) m/z 500.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝8.99-8.97(m,1H),8.67-8.65(m,1H),7.82(s,1H),7.29-7.16(m,2H),6.57-6.55(m,1H),4.99-4.93(m,1H),4.56-4.37(m,1H),3.89(s,3H),2.61(br s,1H),2.23-2.12(m,1H),2.01-1.97(m,1H),1.87-1.72(m,2H),1.55-1.43(m,2H),1.15(s,3H),1.07(s,3H),0.83-0.81(m,1H),0.44-0.42(m,2H),0.25-0.05(m,2H)

EXAMPLE 274 Synthesis of viral protease inhibitor Compounds 1131

A solution of methyl (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (260 mg,588.82umol,1 eq) in HCl/MeOH (4M, 5mL,33.97 eq) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (210 mg, crude material, HCl) as a white solid.

Step 2: (S) -2- ((S) -2- (7-chloro-4-methoxy-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

(2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (210 mg,555.69umol,1 eq, HCl) and 7-chloro-4-methoxy-1H-indole-2-carboxylic acid (162.99 mg,722.40umol,1.3 eq) in DCM (5 mL) and DMF (1.5 mL) were added DMAP (203.67 mg,1.67mmol,3 eq) and EDCI (213.05 mg,1.11mmol,2 eq) and the resulting mixture was then stirred at 20 ℃ for 2H. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] as a yellow solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (270 mg,393.40umol,70.79% yield, 80% purity). MS (ESI) m/z 549.2[ M+H ]] ⁺ 。

Step 3: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (270.00 mg,491.75umol,1 eq.) inNH ₃ A solution in MeOH (7M, 5mL,71.17 eq.) was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid ]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]-7-chloro-4-methoxy-1H-indole-2-carboxamide (240 mg, crude). MS (ESI) m/z 534.2[ M+H ]] ⁺ 。

Step 4: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 7-chloro-4-methoxy-1H-indole-2-carboxamide (240 mg,373.00umol,83% purity, 1 eq.) in DCM (5 mL) was added the berg reagent (222.22 mg,932.50umol,2.5 eq.) and the mixture was then stirred at 25 ℃ for 3H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: xtimate C18 u 250mM 80mM; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,35 min) to give the product 7-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl ]-4-methoxy-1H-indole-2-carboxamide (107.95 mg,207.10umol,55.52% yield, 99% purity). MS (ESI) m/z 516.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.67(br s,1H),8.99(d,J＝7.9Hz,1H),8.64(d,J＝7.9Hz,1H),7.82(s,1H),7.27(s,1H),7.20(d,J＝8.1Hz,1H),6.55(d,J＝8.3Hz,1H),4.94(br d,J＝7.5Hz,1H),4.54(br d,J＝6.8Hz,1H),3.88(s,3H),2.58-2.53(m,1H),2.21-2.14(m,1H),1.94(dd,J＝8.4,12.2Hz,1H),1.77(td,J＝3.3,6.4Hz,1H),1.72(br d,J＝6.4Hz,2H),1.49(t,J＝11.4Hz,1H),1.13(s,3H),1.01(s,3H),0.94(s,9H)

EXAMPLE 275 Synthesis of viral protease inhibitor Compounds 1133

Step 1:2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-norpinan-3-ylidene ] amino ] acetic acid ester

To a solution of tert-butyl 2-aminoacetate (5.75 g,43.84mmol,1.47 eq.) and (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-one (5 g,29.72mmol,1 eq.) in toluene (135 mL) at 20deg.C was added BF ₃ .Et ₂ O(513.39mg，3.62mmol，446.43uL，1.22e ^-1 Equivalent weight). The reaction was then stirred at 120℃for 12h in the presence of a dean-Stark trap. After completion, the reaction mixture was concentrated in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=50/1 to 5/1,1% TEA to give 2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-ylidene as a yellow oil]Amino group]Tert-butyl acetate (6 g,21.32mmol,71.74% yield).

Step 2: (2R) -2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-norpinan-3-ylidene ] amino ] -3-trimethylsilyl-propionic acid tert-butyl ester

To a solution of N-isopropyl-2-amine (4.67 g,46.20mmol,6.53mL,2.5 eq.) in anhydrous THF (100 mL) at-10deg.C was added a solution of N-BuLi (2.5M, 18.48mL,2.5 eq.) very slowly. After 30min, the mixture was cooled to-60 ℃ and 2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-ylidene dissolved in THF (10 mL) was added ]Amino group]Tert-butyl acetate (5.2 g,18.48mmol,1 eq.). After 30min, iodomethyl (trimethyl) silane (7.12 g,33.26mmol,4.95mL,1.8 eq.) was added. The mixture was stirred at-60℃for 1h. The mixture was then cooled to 0 ℃ and held for 12h. After completion, the reaction was quenched with saturated ammonium chloride solution (40 mL). The aqueous phase was then extracted with ethyl acetate (20 ml x 3). The organic phase was purified by Na ₂ SO ₄ Dried, filtered and concentrated to dryness in vacuo. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=50/1 to 10/1,1% tea to give (2R) -2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-ylidene as a yellow oil]Amino group]-3-trimethylsilyl-propionic acid tert-butyl ester (3.4 g,8.32mmol,45.05% yield, 90% purity).

Step 3: (2R) -2-amino-3-trimethylsilyl-propionic acid tert-butyl ester

To (2R) -2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-ylidene]Amino group]To a solution of tert-butyl 3-trimethylsilyl-propionate (0.65 g,1.77mmol,1 eq.) in THF (3.5 mL) was added a solution of citric acid (10 mL,15% purity). The mixture was stirred at 50℃for 16h. After completion, THF was removed in vacuo, and the aqueous layer was extracted with EtOAc (15 mL) to remove chiral inducer. Then, the pH was increased to 8-9 by adding potassium carbonate. The free amine was then extracted with EtOAc (30 ml x 3). The organic layers were combined, taken up over Na ₂ SO ₄ Drying and concentration at room temperature due to amine volatility afforded (2R) -2-amino-3-trimethylsilyl-propionic acid tert-butyl ester (380 mg, crude material) as a yellow oil.

Step 4: (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -3-trimethylsilyl-propionic acid tert-butyl ester

To a solution of 4-methoxy-1H-indole-2-carboxylic acid (380 mg,1.99mmol,1 eq.) in DMF (5 mL) was added (2R) -2-amino-3-trimethylsilyl-propionic acid tert-butyl ester (380 mg,1.75mmol,8.79e-1 eq.), EDCI (495.34 mg,2.58mmol,1.3 eq.), TEA (603.38 mg,5.96mmol,829.96uL,3 eq.). Next, HOBt (349.14 mg,2.58mmol,1.3 eq.) was added to the reaction at-10-0deg.C for 10min. Subsequently, the reaction was stirred at 20℃for 1h. After completion, the reaction mixture was taken up with H ₂ O (100 mL) was diluted and extracted with 150mL EA (50 mL. Times.3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE/ea=10/1 to 1/1) to give (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a white solid]-3-trimethylsilyl-propionic acid tert-butyl ester (600 mg,1.54mmol,77.29% yield, assuming 100% purity).

Step 5: (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino ] -3-trimethylsilyl-propionic acid

At 0 DEG CTo (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]To a solution of tert-butyl-3-trimethylsilyl-propionate (300 mg,768.15umol,1 eq.) in DCM (3 mL) was added TFA/H ₂ O (2 mL, 10/1). Subsequently, the reaction was stirred at 20℃for 3h. After completion, the reaction was concentrated to dryness in vacuo at below 30 ℃. Quench the reaction mixture by adding EA (30 mL) at 20deg.C, and then with H ₂ O (20 mL) was diluted and extracted with 20mL EA (10 mL. Times.2). The combined organic layers were washed with 20mL saturated NaCl (20 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino as a yellow oily residue]-3-trimethylsilyl-propionic acid (300 mg, crude material).

Step 6: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -2-oxo-1- (trimethylsilylmethyl) ethyl ] -4-methoxy-1H-indole-2-carboxamide isomer 1 and N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] amino ] -2-oxo-1- (trimethylsilylmethyl) ethyl ] -4-methoxy-1H-indole-2-carboxamide isomer 2

at-20deg.C, to (2R) -2- [ (4-methoxy-1H-indole-2-carbonyl) amino group]-3-trimethylsilyl-propionic acid (230 mg,687.71umol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of propionitrile (114.99 mg, 687.71. Mu.l, 1 eq.) in DMF (3 mL) was added DMF (1 mL) containing PyBop (357.88 mg, 687.71. Mu.l, 1 eq.) and TEA (139.18 mg,1.38mmol, 191.44. Mu.L, 2 eq.). Next, the reaction was stirred at-20℃for 2h. After completion, the reaction was diluted with MeCN (2 mL) and filtered. By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,8 min) to give-70 mg of epimer. Next, the mixture was subjected to SFC (column: DAICEL CHIRALCEL OD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ OMEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -30%,15 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-2-oxo-1- (trimethylsilylmethyl) ethyl]-4-methyloxy-1H-indole-2-carboxamide isomer 1 (20 mg,40.53umol,5.89% yield, 98% purity). MS (ESI) m/z 484.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23(s,1H),7.19-7.10(m,1H),7.03(d,J＝8.2Hz,1H),6.52(d,J＝7.7Hz,1H),5.04(dd,J＝6.3,9.9Hz,1H),4.60(t,J＝8.0Hz,1H),3.94(s,3H),3.21-3.11(m,2H),2.45-2.23(m,2H),2.05-1.95(m,1H),1.90(td,J＝6.8,13.6Hz,1H),1.82(dt,J＝3.9,9.0Hz,1H),1.73-1.60(m,1H),1.57-1.43(m,1H),1.25-1.15(m,2H),0.10(s,9H)。

Obtaining N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid ]Ethyl group]Amino group]-2-oxo-1- (trimethylsilylmethyl) ethyl]-4-methoxy-1H-indole-2-carboxamide isomer 2 (70 mg,144.74umol,21.05% yield, 100% purity). MS (ESI) m/z 484.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23(s,1H),7.19-7.10(m,1H),7.02(d,J＝8.3Hz,1H),6.51(d,J＝7.6Hz,1H),5.09(dd,J＝6.1,9.8Hz,1H),4.61(t,J＝7.9Hz,1H),3.93(s,3H),3.22-3.17(m,2H),2.52-2.37(m,2H),2.02-1.88(m,2H),1.83-1.74(m,1H),1.72-1.60(m,1H),1.57-1.42(m,1H),1.21(d,J＝8.0Hz,2H),0.16-0.05(m,9H)。

EXAMPLE 276 Synthesis of viral protease inhibitor Compounds 1135

Step 1: (S) -2- ((tert-Butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionic acid

A solution of (2S) -2- (tert-Butoxycarbonylamino) -3-cyclopropyl-propionic acid (6 g,26.17mmol,1 eq.) in THF (60 mL) was cooled at 0deg.C and NaH (2.30 g,57.57mmol,60% purity, 2.2 eq.) was added and the mixture was warmed and stirred at 25deg.C for 1.5h, followed by CH addition ₃ I (8.17 g,57.57mmol,3.58mL,2.2 eq.) and stirred for 2.5h. After completion, the mixture was treated with H ₂ O (200 mL) was quenched and adjusted to ph=1 with HCl (1M) and extracted with ethyl acetate (150 mL x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ Drying, filtering andconcentrated in vacuo to give (S) -2- ((tert-butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionic acid (6.17 g, crude material) as a yellow oil. MS (ESI) m/z 244.1[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To a solution of (S) -2- ((tert-butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionic acid (6.17 g,25.36mmol,1 eq.) in DCM (100 mL) was added methyl (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propionate (6.60 g,27.90mmol,1.1 eq., HCl), DMAP (9.29 g,76.08mmol,3 eq.) and EDCI (9.72 g,50.72mmol,2 eq.) and then the mixture was stirred at 25 ℃ for 2h. After completion, the mixture was treated with H ₂ O (200 mL) was quenched and extracted with DCM (200 mL x 3). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/10 to 0/1) to yield the product methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (7 g,16.45mmol,64.87% yield) as a yellow oil. MS (ESI) m/z 426.3[ M+H ]] ⁺

Step 3: ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) (methyl) carbamic acid tert-butyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1.02 g,2.40mmol,1 eq.) in ammonia (7M, 40mL,116.81 eq.) was stirred at 30℃for 10h. After completion, the reaction was concentrated in vacuo to give tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) (methyl) carbamate (990 mg, crude material) as a yellow solid. MS (ESI) m/z 411.3[ M+H ] ] ⁺

Step 4: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropyl-2- (methylamino) propanamide; (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

A solution of tert-butyl ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) (methyl) carbamate (990 mg,2.41mmol,1 eq.) in HCl/MeOH (30 mL) was stirred at 25℃for 1h. After completion. The reaction was concentrated in vacuo to give (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropyl-2- (methylamino) propanamide (1.1 g, crude material, 30% purity) and (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (1.1 g, crude material, 60% purity) as a yellow solid. MS (ESI) m/z 311.2[ M+H ]] ⁺ ,MS(ESI)m/z 326.2[M+H] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-N-methyl-1H-indole-2-carboxamide

To a solution of (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3-cyclopropyl-2- (methylamino) propanamide (1 g,966.52umol,30% purity, 1 eq), 4-methoxy-1H-indole-2-carboxylic acid (184.78 mg,966.52umol,1 eq) in DCM (30 mL) was added DMAP (354.24 mg,2.90mmol,3 eq), EDCI (370.56 mg,1.93mmol,2 eq) and the mixture stirred at 25 ℃ for 2H. After completion, the reaction was quenched with H ₂ O (200 mL) was quenched and extracted with DCM (100 mL x 3). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give the product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-N-methyl-1H-indole-2-carboxamide as a yellow solid (90 mg,186.12umol,19.26% yield). MS (ESI) m/z 484.2[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-N-methyl-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-N-methyl-1H-indole-2-carboxamide (86 mg,177.85umol,1 eq) in DCM (3 mL) was added the bergs reagent (127.15 mg,533.54umol,3 eq) and the mixture stirred at 25 ℃ for 2H. After completion, the reaction was concentrated in vacuo and purified by preparative HPLC (column Phenomenex Luna C18 75 x 30mm x 3um; mobile phase: [ water (0.2% fa) -ACN ]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give the product N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-methoxy-N-methyl-1H-indole-2-carboxamide (35 mg,75.18umol,42.27% yield, 100% purity) as a white solid. MS (ESI) m/z 466.1[ M+H ]] ⁺¹ HNMR(400MHz,DMSO-d ₆ )δ＝11.60(br s,1H),8.87(br s,1H),7.56(br s,1H),7.15-7.07(m,1H),7.05-6.99(m,1H),6.91(br s,1H),6.52-6.50(m,1H),5.13-4.96(m,2H),3.87(s,3H),3.09(br s,2H),2.23(br s,2H),1.96-1.77(m,3H),1.74-1.71(m,1H),1.66-1.34(m,3H),0.80-0.02(m,5H)

EXAMPLE 277 Synthesis of viral protease inhibitor Compounds 1137

Step 1: (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) (methyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (2 g,4.70mmol,1 eq.) in HCl/MeOH (50 mL) was stirred at 25℃for 3h. After completion, the reaction was concentrated in vacuo to give the crude product methyl (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (2 g, crude material) as a yellow solid. MS (ESI) m/z 326.2[ M+H ]] ⁺

Step 2: (S) -2- ((S) -2- (7-chloro-N-methyl-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (1 g,2.76mmol,1 eq, HCl) in DCM (30 mL) was added 7-chloro-1H-indole-2-carboxylic acid (540.54 mg,2.76mmol,1 eq), DMAP (1.01 g,8.29mmol,3 eq), EDCI (1.06 g,5.53mmol,2 eq) and the mixture was stirred at 25 ℃ for 2H. After completion, the reaction was quenched with H ₂ O (200 mL) was quenched and extracted with DCM (100 mL x 3). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/9 to 0/1) to give the product methyl (S) -2- ((S) -2- (7-chloro-N-methyl-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (530 mg,1.05mmol,38.13% yield) as a yellow solid. MS (ESI) m/z 503.2[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-N-methyl-1H-indole-2-carboxamide

A solution of methyl (S) -2- ((S) -2- (7-chloro-N-methyl-1H-indole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (530 mg,1.05mmol,1 eq.) in ammonia (7M, 30mL,199.30 eq.) was stirred at 30℃for 10H. After completion, the reaction was concentrated in vacuo to give the crude product N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-N-methyl-1H-indole-2-carboxamide (440 mg, crude material) as a yellow solid. MS (ESI) m/z 488.2[ M+H ] ] ⁺

To N- ((S) -1- ((. About.S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-N-methyl-1H-indole-2-carboxamide (440 mg,901.68umol,1 eq) in DCM (15 mL) was added with the Bogis reagent (644.62 mg,2.71mmol,3 eq) and the mixture stirred at 25℃for 4H. After completion, the reaction was concentrated in vacuo and purified by preparative HPLC (column Waters Xbridge Prep OBD C18 150 x 40mm x 10um; mobile phase: [ water (10 mm nh4hco 3) -ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -N-methyl-1H-indole-2-carboxamide (220 mg,468.12umol,51.92% yield, 100% purity) as a white solid. MS (ESI) m/z 470.1[ M+H ]] ⁺¹ HNMR(400MHz,DMSO-d ₆ )δ＝11.93-11.60(m,1H),8.90(br s,1H),7.57(br s,2H),7.28-7.26(m,1H),7.08-7.04(m,1H),7.02-6.57(m,1H),5.12-5.02(m,1H),5.00-4.71(m,1H),3.30-3.16(m,2H),3.13-2.93(m,3H),2.30-2.16(m,2H),1.95-1.39(m,7H),0.84-0.18(m,5H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.34(br s,1H),8.73-8.71(m,1H),7.61-7.59(m,1H),7.35-7.23(m,2H),7.10-7.02(m,1H),6.87(br s,1H),5.07-5.01(m,1H),4.93(br s,1H),3.14(br s,5H),2.34-2.19(m,2H),1.98-1.81(m,3H),1.80-1.56(m,3H),1.51-1.44(m,1H),0.71(br s,1H),0.53-0.37(m,1H),0.53-0.37(m,1H),0.19-0.04(m,2H)

EXAMPLE 278 Synthesis of viral protease inhibitor Compounds 1141

Step 1: 7-chloro-2- (trichloromethyl) -1H-benzo [ d ] imidazole

To a solution of 3-chlorobenzene-1, 2-diamine (2 g,14.03mmol,1 eq.) in AcOH (20 mL) was added methyl 2, 2-trichloroethyliminoate (2.97 g,16.83mmol,2.08mL,1.2 eq.). The mixture was stirred at 25℃for 2h. After completion, by addition of saturated NaHCO ₃ (50 mL) to adjust to ph=7-8, and then quenching the reaction mixture with ethylEthyl acetate (30 ml x 3) extraction. The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 7-chloro-2- (trichloromethyl) -1H-benzimidazole (3 g, crude) as a yellow solid. MS (ESI) m/z 270.9[ M+H ]] ⁺

Step 2: 7-chloro-1H-benzo [ d ] imidazole-2-carboxylic acid methyl ester

To a solution of 7-chloro-2- (trichloromethyl) -1H-benzimidazole (3 g,11.11mmol,1 eq.) in MeOH (40 mL) was added Na ₂ CO ₃ (1.18 g,11.11mmol,1 eq.). The mixture was stirred at 70℃for 14h. After completion, the mixture was concentrated under reduced pressure to give the product methyl 7-chloro-1H-benzimidazole-2-carboxylate (3 g, crude material) as a yellow solid. MS (ESI) m/z 210.9[ M+H ]] ⁺

Step 3: 7-chloro-1H-benzo [ d ] imidazole-2-carboxylic acid

To 7-chloro-1H-benzimidazole-2-carboxylic acid methyl ester (3 g,14.24mmol,1 eq.) in THF (20 mL) and H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (1.79 g,42.73mmol,3 eq.). The mixture was stirred at 60℃for 2h. After completion, the reaction mixture was quenched by addition of 1N HCl (20 mL) to adjust to ph=3-5, and then extracted with EA (30 ml×3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the product 7-chloro-1H-benzimidazole-2-carboxylic acid (2 g, crude material) as a yellow solid. MS (ESI) m/z 197.0[ M+H ]] ⁺

Step 4: (S) -2- ((S) -3-cyclopropyl-2- (methylamino) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

(2S) -2- [ [ (2S) -2- [ tert-Butoxycarbonyl (methyl) amino ]]-3-cyclopropyl-propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1 g,2.35mmol,1 eq.) in HCl/MeOH (4M, 20mL,59.57 eq.) was stirred at 20deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give the product (2S) -2- [ [ (2S) -3-cyclopropyl-2- (methylamino) propionyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g, crude material, HCl). MS (ESI) m/z 326.2[ M+H] ⁺

Step 5: (S) -2- ((S) -2- (7-chloro-N-methyl-1H-benzo [ d ] imidazole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- [ [ (2S) -3-cyclopropyl-2- (methylamino) propanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (700 mg,1.93mmol,1 eq, HCl) and 7-chloro-1H-benzimidazole-2-carboxylic acid (456.35 mg,2.32mmol,1.2 eq) in DMF (20 mL) was added DMAP (472.65 mg,3.87mmol,2 eq) and EDCI (741.67 mg,3.87mmol,2 eq). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=5/1 to 0/1) to give the product (2S) -2- [ [ (2S) -2- [ (7-chloro-1H-benzimidazole-2-carbonyl) -methyl-amino ] as a yellow solid]-3-cyclopropyl propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.5 g,992.11umol,51.29% yield). MS (ESI) m/z 504.2[ M+H ]] ⁺

Step 6: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-chloro-N-methyl-1H-benzo [ d ] imidazole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-1H-benzimidazole-2-carbonyl) -methyl-amino]-3-cyclopropyl propionyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (450 mg,892.90umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,78.40 eq.) was stirred at 30℃for 16h. After completion, the mixture was concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid ]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-N-methyl-1H-benzimidazole-2-carboxamide (400 mg, crude). MS (ESI) m/z 489.2[ M+H ]] ⁺

Step 7: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -N-methyl-1H-benzo [ d ] imidazole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-N-methyl-1H-benzimidazole-2-carboxamide (200 mg,409.03umol,1 eq.) in DCM (10 mL) was added the berg reagent (194.95 mg,818.05umol,2 eq.). The mixture was stirred at 20℃for 4h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product 7-chloro-N- [ (1S) -2- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxoethyl]-N-methyl-1H-benzimidazole-2-carboxamide (50 mg,106.17umol,12.98% yield, 100% purity). MS (ESI) m/z 471.2[ M+H ] ] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.72-7.48(s,1H),7.45-7.25(m,3H),5.92(dd,J＝5.8,9.2Hz,1H),5.32-5.17(m,1H),5.14-4.94(m,1H),3.62-3.55(m,1H),3.20(dd,J＝4.6,8.8Hz,3H),3.07(s,3H),2.57-2.33(m,3H),2.12-1.92(m,4H),1.88-1.77(m,2H),1.74-1.46(m,4H),0.62-0.39(m,3H),0.35-0.10(m,3H),0.009-0.037(m,1H)。

Example 279 Synthesis of viral protease inhibitor Compounds 1143

Step 1: (S) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester

To (S) -2- (tert-Butoxycarbonyl) -2-azaspiro [4.5]]Decane-3-carboxylic acid (300 mg,846.97umol,80% purity, 1 eq.) methyl (169.59 mg,846.97umol,1 eq.) of (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propanoate was added to a solution of (S) -2-amino-3- ((S) -2-oxopiperidin-3-yl) propanoate in DCM (8 mL), and then DMAP (310.42 mg,2.54mmol,3 asAmount) and EDCI (324.73 mg,1.69mmol,2 eq.) followed by stirring the mixture at 20 ℃ for 1h. After the reaction was complete, the reaction mixture was quenched by the addition of water (10 mL) and extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 4 mL) followed by saturated NaHCO ₃ (4 mL) is adjusted to pH 7, and the mixture is treated by Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (S) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [ 4.5) as a yellow solid]Decane-2-carboxylic acid tert-butyl ester (268 mg,575.62 mol,67.96% yield). MS (ESI) m/z 466.3[ M+H ] ] ⁺

Step 2: (S) -3- ((S) -2-oxopiperidin-3-yl) -2- ((S) -2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester

(S) -3- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -2-azaspiro [4.5] at 0deg.C]A solution of tert-butyl decane-2-carboxylate (240 mg,515.48umol,1 eq.) in HCl/MeOH (4M, 4 mL) was stirred at 20℃for 1h. After completion of the reaction, the mixture was concentrated in vacuo to give (S) -3- ((S) -2-oxopiperidin-3-yl) -2- ((S) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamido) propionic acid methyl ester (180 mg, crude material, HCl). MS (ESI) m/z 366.2[ M+H ]] ⁺

Step 3: (S) -2- ((S) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (S) -3- ((S) -2-oxopiperidin-3-yl) -2- ((S) -2-azaspiro [4.5]Decane-3-carboxamido) methyl propionate (180 mg,447.84umol,1 eq, HCl) to a mixture of DCM (4 mL) was added 5-methoxy-1H-indole-2-carboxylic acid (85.62 mg,447.84umol,1 eq) and then DMAP (109.43 mg,895.69umol,2 eq) and EDCI (128.78 mg,671.76umol,1.5 eq) were added at 20℃and the mixture was stirred for 1H at 20 ℃. After the reaction was complete, the reaction mixture was quenched by the addition of water (10 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 4 mL) followed by saturated NaHCO ₃ (4 mL) is adjusted to pH 7, and the mixture is treated by Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (S) -2- ((S) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester (220 mg,408.44umol,91.20% yield). MS (ESI) m/z 539.3[ M+H ]] ⁺

Step 4: (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To (S) -2- ((S) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]NH was added to a solution of methyl decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (200 mg,371.31umol,1 eq) ₃ MeOH (7 m,5 ml) and then the mixture was stirred at 30 ℃ for 18h. After the reaction was completed, the reaction mixture was concentrated in vacuo to give (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carboxamide (150 mg, crude). MS (ESI) m/z 524.3[ M+H ]] ⁺

Step 5: (S) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To (S) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (150 mg,286.47umol,1 eq.) in DCM (3 mL) was added the Bungeus reagent (204.80 mg,859.40umol,3 eq.) and stirred at 30℃for 1.5h. After the reaction was completed, the reaction mixture was quenched with water (1 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge BEH C, 100 x 25mM x 5um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give (S) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2- (5-methoxy-1H-indole-2-carbonyl) -2-azaspiro as a white solid[4.5]Decane-3-carboxamide (10.54 mg,20.85umol,7.28% yield, 100% purity). MS (ESI) m/z 506.2[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.44-11.33(m,1H),9.11-8.84(m,1H),7.58-7.41(m,1H),7.36-7.24(m,1H),7.15(d,J＝1.5Hz,1H),6.98-6.92(m,1H),6.89-6.76(m,1H),4.89(s,1H),4.54-4.45(m,1H),3.94-3.84(m,1H),3.75(s,3H),3.72(s,1H),2.88(s,2H),2.30-2.23(m,1H),2.22-2.14(m,1H),2.03-1.82(m,1H),1.81-1.63(m,2H),1.59-1.12(m,14H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δppm 11.14(s,1H),8.72(s,1H),7.35(d,J＝9.0Hz,1H),7.27(s,1H),7.09(s,1H),6.86(dd,J＝1.9,8.9Hz,2H),5.03-4.91(m,1H),4.69-4.54(m,1H),3.89(d,J＝11.0Hz,1H),3.78(s,3H),3.66-3.51(m,1H),2.30-2.10(m,3H),1.86-1.62(m,4H),1.59-1.36(m,14H)。

EXAMPLE 280 Synthesis of viral protease inhibitor Compound 1145

Step 1: (2S) -3- ((S) -2-oxopiperidin-3-yl) -2- (2-azaspiro [4.5] decane-3-carboxamido) propionic acid methyl ester hydrochloride

A solution of tert-butyl 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carboxylate (3.5 g,7.52mmol,1 eq.) in HCl/MeOH (4M, 50mL,26.60 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give the product methyl (2S) -2- (2-azaspiro [4.5] decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (3 g, crude material, HCl) as a white solid.

Step 2: (2S) -2- (2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

At 0 ℃, to (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.59 g,3.96mmol,9.92e-1 eq, HCl) and 5-chloro-4-methoxy-1H-indole-2-carboxylic acid (900 m)g,3.99mmol,1 eq.) DMAP (1.46 g,11.97mmol,3 eq.) and EDCI (1.53 g,7.98mmol,2 eq.) are added to a solution in DCM (15 mL) and DMF (5 mL), and the resulting mixture is then stirred at 20℃for 2h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=5:1 to 0:1) the residue was purified to give the product (2S) -2- [ [2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (2 g,3.35mmol,83.99% yield, 96% purity). MS (ESI) m/z 573.2[ M+H ] ] ⁺ 。

Step 3: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.95 g,3.40mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 50mL,102.86 eq.) was stirred at 40℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]-2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.8 g, crude). MS (ESI) m/z 558.2[ M+H ]] ⁺ 。

Step 4:2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (1.80 g,3.23mmol,1 eq.) in DCM (25 mL) was added the Bungeus reagent (2.31 g,9.68mmol,3 eq.) and then at 25 ℃ The resulting mixture was stirred for 3h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: xtimate C18 u 250mM 80mM; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,35 min) to give the product 2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (1.5 g,2.75mmol,85.25% yield, 99% purity). MS (ESI) m/z 540.2[ M+H ]] ⁺ 。

Step 5:2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -2-azaspiro [4.5] decane-3-carboxamide

By SFC (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm,10 um); mobile phase: [0.1% NH) ₃ H ₂ O MEOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,4 min) isolation of 2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (1.5 g) gave the product 2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide isomer 1 (308.55 mg,569.62umol,20.51% yield, 99.7% purity). MS (ESI) m/z 540.2[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23-6.86(m,3H),5.09-5.00(m,1H),4.63(dd,J＝8.0,9.5Hz,1H),4.10-4.02(m,3H),3.94(br s,1H),3.80(d,J＝10.3Hz,1H),3.23-3.01(m,2H),2.42-2.05(m,3H),2.01-1.38(m,16H)

The product 2- (5-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide isomer 2 (269.63 mg,499.27umol,17.98% yield, 100% purity). MS (ESI) m/z 540.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.25-6.82(m,3H),5.11(dd,J＝5.7,10.5Hz,1H),4.62(dd,J＝7.9,9.6Hz,1H),4.01(s,3H),4.00-3.82(m,1H),3.75(d,J＝10.1Hz,1H),3.28-3.05(m,2H),2.62-1.69(m,7H),1.68-1.33(m,12H)

EXAMPLE 281 Synthesis of viral protease inhibitor Compounds 1147

Step 1: (2S) -2- [ [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

(2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]A mixture of methyl propionate (1.4 g,3.48mmol,1 eq, HCl) in DCM (20 mL) was added followed by 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (1.2 g,5.32mmol,1.53 eq), DMAP (1.06 g,8.71mmol,2.5 eq) and EDCI (1.34 g,6.97mmol,2 eq) and stirred at 20deg.C for 1H. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ The residue was purified with PE: ea=2:1 to 0:1 to give the product (2S) -2-azaspiro [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid ]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.3 g,2.27mmol,65.13% yield). MS (ESI) M/z573.2[ M+H ]] ⁺ 。

Step 2: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1.26 g,2.20mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 20mL,63.68 eq.) was stirred at 30℃for 20h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (30 ml x 3) and concentrated under reduced pressure to give the product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperaquine as a yellow solidBoydo group]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (1.2 g, crude). MS (ESI) m/z 558.3[ M+H ]] ⁺ 。

Step 3:2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (1.20 g,2.15mmol,1 eq.) in DCM (20 mL) was added the Buerger's reagent (1.49 g,6.24mmol,2.9 eq.) and stirred for 1h at 30 ℃. After completion, the mixture was quenched with water (1 mL) and with N ₂ And (5) blow-drying. By preparative HPLC (column: waters X bridge C, 150X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) was purified and passed through SFC (column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-MeOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:60% -60%,7 min) to give the product 2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white oil]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide isomer 1 (251.52 mg,465.73umol,21.66% yield). MS (ESI) m/z 540.3[ M+H ]] ⁺ 。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.12(d,J＝1.8Hz,1H),7.02(s,1H),6.98-6.91(m,1H),5.17-4.94(m,1H),4.61(s,1H),3.95-3.85(m,1H),3.85-3.76(m,3H),3.70(br d,J＝10.4Hz,1H),3.29-3.13(m,2H),2.64-2.23(m,3H),2.09-1.87(m,2H),1.82-1.68(m,2H),1.64-1.39(m,12H)。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.05(br s,1H),8.72(br d,J＝7.5Hz,1H),7.28(br s,1H),7.12(br s,1H),6.97(s,2H),4.97(br s,1H),4.60(br s,1H),3.80(s,4H),3.61(br s,1H),3.08-3.03(m,1H),2.49-2.47(m,1H),2.45-2.08(m,3H),2.00-1.62(m,4H),1.59-1.32(m,12H)。

The product 2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidine was obtained as a white solidBase group]Ethyl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide isomer 2 (366.62 mg,678.86umol,31.57% yield). MS (ESI) m/z 540.3[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.01(br s,1H),8.67(br s,1H),7.25(br s,1H),7.20-7.09(m,1H),6.98(s,2H),4.98(br d,J＝7.1Hz,1H),4.59(br s,1H),3.80(s,4H),3.62(br s,1H),3.12-3.10(m,1H),3.08-3.06(m,1H),2.50-2.47(m,2H),2.20(br s,3H),1.83(br s,2H),1.67(br d,J＝11.5Hz,2H),1.57-1.33(m,12H)。

¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.13(d,J＝2.1Hz,1H),7.04(s,1H),7.00-6.93(m,1H),5.01(dd,J＝6.2,10.2Hz,1H),4.62(dd,J＝7.8,9.8Hz,1H),3.94(br d,J＝10.4Hz,1H),3.84-3.72(m,4H),3.23-3.02(m,2H),2.45-2.22(m,3H),2.04-1.85(m,2H),1.84-1.68(m,2H),1.65-1.50(m,7H),1.44(br d,J＝10.1Hz,5H)。

EXAMPLE 282 Synthesis of viral protease inhibitor Compound 1149

Step 1: (Z) -2-azido-3- (4-chloro-2-methoxyphenyl) acrylic acid methyl ester

A mixture of NaOMe (6.33 g,117.24mmol,2 eq.) in MeOH (150 mL) was cooled to-10deg.C, and then a mixture of 4-chloro-2-methoxy-benzaldehyde (10 g,58.62mmol,1 eq.) and ethyl 2-azidoacetate (15.14 g,117.24mmol,13.40mL,2 eq.) in MeOH (150 mL) was added dropwise to the foregoing solution. The mixture was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and then purified by adding H ₂ O (100 mL) and then extracted with EA (30 mL x 3). The combined organic layers were washed with brine (100 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with PE: ea=1:0 to 5:1 to give methyl (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoate (8 g,29.89mmol,50.99% yield) as a yellow solid.

Step 2: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

Xylene (80 mL) containing methyl (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoate (7.60 g,28.40mmol,1 eq.) was stirred at 170℃for 2h. After completion, the mixture was concentrated under reduced pressure to give a residue. The crude product was wet-milled with PE (10 mL) at 0deg.C for 20min to give methyl 6-chloro-4-methoxy-1H-indole-2-carboxylate (4 g,16.69mmol,58.77% yield) as a white solid. MS (ESI) m/z 240.1[ M+H ] ] ⁺

Step 3: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (1 g,4.08mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (1.16 g,4.89mmol,1.2 eq., HCl) in DCM (20 mL) was added DMAP (996.01 mg,8.15mmol,2 eq.) and EDCI (1.56 g,8.15mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g, crude material). MS (ESI) M/z428.3[ M+H ]] ⁺

Step 4: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To methyl propionate (1 g,2.34mmol,1 eq.) was added HCl/MeOH (4M, 10mL,17.10 eq.) and the mixture was then stirred at 20deg.C for 1h. After completion, the mixture was concentrated under reduced pressure to give (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl as a yellow solid ]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.8 g, crude material, HCl). MS (ESI) m/z 328.2[ M+H ]] ⁺

Step 5: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid

To 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (4 g,16.69mmol,1 eq.) in THF (30 mL) and H ₂ LiOH.H was added to the mixture in O (10 mL) ₂ O (2.10 g,50.07mmol,3 eq.). The mixture was stirred at 60℃for 2h. After completion, by adding H ₂ O (50 mL) to quench the mixture, and then aqueous HCl (1M) was added to adjust to ph=3-4 and extracted with EA (50 ml×3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (4 g, crude material) as a yellow solid. MS (ESI) m/z 226.0[ M+H ]] ⁺

Step 6: (S) -2- ((S) -2- (6-chloro-4-methoxy-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (0.8 g,2.20mmol,1 eq, HCl) and 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (744.08 mg,3.30mmol,1.5 eq) in DCM (20 mL) was added DMAP (537.18 mg,4.40mmol,2 eq) and EDCI (842.93 mg,4.40mmol,2 eq). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with PE: ea=10:1 to 0:1 to give (2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (1 g,1.87mmol,85.01% yield). MS (ESI) m/z 535.2[ M+H ]] ⁺

Step 7: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -6-chloro-4-methoxy-1H-indole-2-carboxamide

Will contain (2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]NH of methyl propionate (1 g,1.87mmol,1 eq) ₃ MeOH (7M, 15mL,56.18 eq.) was stirred at 80℃for 16h. After completion, the mixture was concentrated under reduced pressure to give N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]Carbamoyl group ]-3, 3-dimethyl-butyl]-6-chloro-4-methoxy-1H-indole-2-carboxamide (0.8 g, crude material). MS (ESI) m/z 520.2[ M+H ]] ⁺

Step 8: 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 6-chloro-4-methoxy-1H-indole-2-carboxamide (700 mg,1.35mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (962.35 mg,4.04mmol,3 eq.). The mixture was stirred at 20℃for 8h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give 6-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (230 mg,458.16umol,34.04% yield, 100% purity). MS (ESI) m/z 502.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.72(br s,1H),8.88(d,J＝8.2Hz,1H),8.53(br d,J＝7.8Hz,1H),7.51(br s,1H),7.35(s,1H),7.04(s,1H),6.55(d,J＝1.6Hz,1H),5.14-4.93(m,1H),4.51-4.48(m,1H),3.91(s,3H),3.16-2.99(m,2H),2.32-2.17(m,2H),1.88-1.73(m,3H),1.71-1.62(m,2H),1.59-1.46(m,1H),1.44-1.31(m,1H),0.93(s,9H)

Example 283 Synthesis of viral protease inhibitor Compound 1151

Step 1: (5R) -5- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-1, 3-azasilacyclopentane (azasilolidine) -1-carboxylic acid tert-butyl ester

To (5R) -1-tert-butoxycarbonyl-3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxylic acid (300 mg,1.16mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl at 25 ℃]To a mixture of methyl propionate (301.15 mg,1.27mmol,1.1 eq., HCl) in DCM (6 mL) and DMF (2 mL) was added DMAP (423.91 mg,3.47mmol,3 eq.) and EDCI (443.46 mg,2.31mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2 hours. After completion, the reaction mixture was quenched with 22mL H ₂ O was diluted and extracted with 45mL EA (15 mL x 3). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ The crude material was purified with petroleum ether/ethyl acetate=5/1 to 0/1 to give (5R) -5- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-3, 3-dimethyl-1, 3-azasilacyclopentane-1-carboxylic acid tert-butyl ester (500 mg,1.13mmol,97.89% yield). (ESI) m/z 442.3[ M+H ] ] ⁺

Step 2: (2S) -2- [ [ (5R) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

A mixture of (5R) -5- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -3, 3-dimethyl-1, 3-azasilacyclopentane-1-carboxylic acid tert-butyl ester (500 mg,1.13mmol,1 eq.) in HCl/MeOH (5 mL) was stirred at 25℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ [ (5R) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (420 mg,1.11mmol,98.15% yield, HCl) as a white solid.

Step 3: (2S) -2- [ [ (5R) -1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To 4-methoxy-1H-indole-2-carboxylic acid (193.15 mg,1.01mmol,1 eq.) and (2S) -2- [ [ (5R) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl at 25 ℃]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a mixture of methyl propionate (0.42 g,1.11mmol,1.1 eq., HCl) in DCM (4 mL) and DMF (1 mL) was added DMAP (370.27 mg,3.03mmol,3 eq.) and EDCI (387.34 mg,2.02mmol,2 eq.) in one portion. The mixture was stirred at 25℃for 2h. After completion, the reaction mixture was quenched with 25mL H ₂ O was diluted and extracted with 45mL EA (15 mL x 3). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=5/1 to 0/1) to give (2S) -2- [ [ (5R) -1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (450 mg,874.39umol,86.55% yield). (ESI) m/z 515.2[ M+H ]] ⁺

Step 4: (5R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxamide

(2S) -2- [ [ (5R) -1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (750 mg,1.46mmol,1 eq.) in NH ₃ The mixture in MeOH (7M, 40mL,192.13 eq.) was stirred at 25℃for 48h. After completion, the reaction mixture was concentrated under reduced pressure to give (5R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid ]Methyl group]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxamide (720 mg,1.44mmol,98.88% yield). MS (ESI) m/z 500.2[ M+H ]] ⁺

Step 5: (5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxamide

To (5R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl at 25 ℃]Methyl group]Ethyl group]To a mixture of 1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxamide (640 mg,922.28umol,72% purity, 1 eq.) in DCM (10 mL) was added the berges reagent (549.47 mg,2.31mmol,2.5 eq.) in one portion. The mixture was stirred at 25℃for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product. By preparative HPLC (FA conditions; column: phenomenex Luna C, 200X 40mm X10 um; mobile phase: [ water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -75%,8 min) to give (5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Ethyl group]-1- (4-methoxy-1H-indole-2-carbonyl) -3, 3-dimethyl-1, 3-azasilacyclopentane-5-carboxamide (190 mg,394.50umol,42.77% yield). MS (ESI) m/z 482.1[ M+H ] ] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.47(br s,1H),8.78(br d,J＝6.7Hz,1H),7.52(br s,1H),7.19-6.97(m,3H),6.50(d,J＝7.6Hz,1H),5.01(q,J＝7.9Hz,2H),3.88(br s,3H),3.35(br s,1H),3.28-3.17(m,1H),3.08(br s,2H),2.31-2.10(m,2H),1.99-1.47(m,4H),1.44-1.16(m,2H),0.98(br d,J＝14.8Hz,1H),0.33-0.15(m,6H)

Example 284 Synthesis of viral protease inhibitor Compound 1153

Step 1:3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -2-aza-5-silaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester

To 2-tert-Butoxycarbonyl-2-aza-5-silaspiro [4.4]]Nonane-3-carboxylic acid (0.7 g,2.45mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (580.52 mg,2.45mmol,1 eq., HCl) in DCM (8 mL) was added DMAP (898.90 mg,7.36mmol,3 eq.) and EDCI (940.33 mg,4.91mmol,2 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and extract with DCM (10 mL x 4). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ Petroleum ether ethyl acetate=2:1 to 1:1 to 0:1) to give the product 3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a yellow oil]Methyl group]Ethyl group]Carbamoyl group]-2-aza-5-silaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester (0.8 g,1.37mmol,55.80% yield, 80% purity). MS (ESI) m/z 468.3[ M+H ]] ⁺

Step 2: (2S) -2- (2-aza-5-silaspiro [4.4] nonane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

3- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-2-aza-5-silaspiro [4.4]]A solution of tert-butyl nonane-2-carboxylate (0.75 g,1.28mmol,80% purity, 1 eq.) in HCl/MeOH (4M, 24.00mL,74.82 eq.) was stirred at 20℃for 2h. After completion, the reaction was concentrated under reduced pressure to give the crude product (2S) -2- (2-aza-5-silaspiro [ 4.4) as a yellow oil]Nonane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.5 g, crude material, HCl). MS (ESI) m/z 368.1[ M+H ]] ⁺

Step 3: (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4] nonane-3-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propionic acid methyl ester

To (2S) -2- (2-aza-5-silaspiro [4.4]]Nonane-3-carbonylamino) -3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (0.5 g,1237.71 mol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (236.63 mg,1237.71 mol,1 eq) in DCM (10 mL) was added DMAP (453.63 mg,1.49mmol,3 eq) and EDCI (474.53 mg,1.49mmol,2 eq). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (80 mL) to quench the reaction and extract with DCM (15 mL x 6). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by column chromatographySpectrum (SiO) ₂ Petroleum ether/ethyl acetate=9/1 to 4/1 to 1/1 to 0/1, and then dichloromethane/methanol=10/1) to give the product (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [ 4.4) as a yellow oil]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.227 g,274.89umol,89.59% yield, 85% purity). MS (ESI) m/z 541.3[ M+H ]] ⁺

Step 4: n- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4] nonane-3-carboxamide

(2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4]]Nonane-3-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.47 g,869.27umol,1 eq.) in NH ₃ A solution in MeOH (1 mL, 7M) was stirred at 20℃for 42h. After completion, the reaction was concentrated under reduced pressure to give the crude product N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a yellow solid]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4]Nonane-3-carboxamide (0.45 g, crude). MS (ESI) m/z 526.3[ M+H ] ] ⁺

Step 5: n- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl ] ethyl ] -2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4] nonane-3-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4]To a solution of nonane-3-carboxamide (0.46 g,875.07umol,1 eq.) in DCM (5 mL) was added the Bunges reagent (625.62 mg,2.63mmol,3 eq.). The mixture was stirred at 30℃for 6h. After completion, the mixture was quenched with water (3 mL) and with N ₂ Blow dried and purified by preparative HPLC (column: kromasil C18 (250X 50mM X10 um); mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -55%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4]Nonane-3-carboxylic acid methyl esterAmide isomer 1 (57 mg,111.16umol,12.70% yield, 99% purity). MS (ESI) m/z 508.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.52(s,1H),8.86(br d,J＝7.2Hz,1H),7.51(br s,1H),7.17-7.00(m,3H),6.51(d,J＝7.7Hz,1H),5.00-4.99(m,1H),5.09-4.94(m,1H),3.88(s,3H),3.51-3.42(m,1H),3.08(br s,2H),2.31-2.10(m,2H),1.89-1.70(m,3H),1.70-1.46(m,6H),1.46-1.33(m,2H),1.01(br d,J＝15.2Hz,1H),0.92-0.64(m,4H)。

The product N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl was obtained as a white solid]Ethyl group]-2- (4-methoxy-1H-indole-2-carbonyl) -2-aza-5-silaspiro [4.4 ]Nonane-3-carboxamide isomer 2 (47 mg,91.66umol,10.47% yield, 99% purity). ¹ HNMR(400MHz,DMSO-d ₆ )δ＝11.48(br s,1H),8.84(br d,J＝7.3Hz,1H),7.52(br s,1H),7.19-7.00(m,3H),6.50(d,J＝7.7Hz,1H),5.16-4.91(m,2H),3.88(s,3H),3.52-3.41(m,1H),3.07(br s,2H),2.31-2.19(m,2H),1.88-1.73(m,2H),1.72-1.46(m,7H),1.45-1.32(m,2H),1.11-0.99(m,1H),0.91-0.70(m,4H)。

EXAMPLE 285 Synthesis of viral protease inhibitor Compounds 1163

Step 1: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (180.00 mg,717.93umol,1 eq, HCl) in DMF (1 mL) and DCM (3 mL) was added DMAP (263.12 mg,2.15mmol,3 eq), and then (S) -2- ((t-butoxycarbonyl) amino) -4, 4-dimethylpentanoic acid (211.34 mg,865.51umol,1.2 eq) and EDCI (275.26 mg,1.44mmol,2 eq). The resulting solution was stirred at 15 ℃ for 2h, and then diluted with water (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (5 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was used directly in the next step. Obtaining the obtainedThe compound (S) -methyl 2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (0.3 g, crude material) was obtained as a yellow solid. MS (ESI) m/z 441.2[ M+H ] ] ⁺ 。

Step 2: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

A mixture of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (0.28 g,634.12umol,1 eq.) in HCl/MeOH (4 mL) was stirred at 15℃for 3h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (0.18 g, crude material, HCl) as a yellow solid.

Step 3: (2S) -2- (2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5] decane-3-carboxamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (0.13 g,380.74umol,1 eq, HCl) in DMF (0.7 mL) and DCM (1.3 mL) was added PyBop (198.13 mg,380.71umol,1 eq), and then 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (85.90 mg,380.71umol,1 eq) and NEt ₃ (115.58 mg,1.14mmol,3 eq.) the solution was stirred at 15℃for 2h. After completion, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=10:1 to 0:1) to give (2S) -2- [ [2- (4-methoxy-1H-indole-2-carbonyl) -8-oxa-2-azaspiro [4.5 ] as a white solid]Decane-3-carbonyl]Amino group]-3- [ (3S) -2-oxopyrrolidin-3-yl]Methyl propionate (0.16 g,291.41umol,76.54% yield, 90% purity). MS (ESI) m/z 549.2[ M+H ]] ⁺ 。

Step 4: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (7-chloro-5-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (0.13 g,246.88umol,1 eq.) was reacted in NH ₃ The mixture in MeOH (7M, 3mL,85.06 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (0.12 g, crude material) as a yellow oil. MS (ESI) m/z 534.2[ M+H ] ] ⁺ 。

Step 5: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -5-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (0.11 g,205.97umol,1 eq) in DCM (1 mL) was added the berg reagent (147.26 mg,617.92umol,3 eq) and the solution stirred at 25 ℃ for 2H. After completion, the reaction mixture was quenched with water (1 mL) and air dried. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]) The residue was purified to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -5-methoxy-1H-indole-2-carboxamide (30.00 mg,58.14umol,28.22% yield, 99% purity) as a white solid. MS (ESI) m/z 516.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.23-7.08(m,2H),7.08-6.98(m,1H),6.53(br d,J＝7.6Hz,1H),5.02(br dd,J＝5.7,10.1Hz,1H),4.72-4.62(m,2H),4.19-4.03(m,1H),3.98-3.81(m,4H),3.77-3.62(m,4H),3.29-3.17(m,1H),2.52-2.20(m,3H),2.02-1.42(m,8H)。

Example 286 Synthesis of viral protease inhibitor Compounds 1167

Step 1: 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid ethyl ester (200 mg,843.08umol,1 eq.) in THF (4 mL) and H ₂ LiOH.H was added to the solution in O (2 mL) ₂ O (106.14 mg,2.53mmol,3 eq.) and the mixture stirred at 60℃for 3h. After the reaction was complete, the mixture was concentrated in vacuo and adjusted to pH-1 with 1M HCl (3 mL) and extracted with EA (10 mL x 3), followed by concentration in vacuo to give 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (170 mg, crude material) as a white solid. MS (ESI) m/z 208.1[ M-H] ⁺

Step 2: (S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamide) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (150 mg,414.52umol,1 eq, HCl) and 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (104.05 mg,497.43umol,1.2 eq) in DCM (8 mL) was added DMAP (101.28 mg,829.04umol,2 eq), followed by EDCI (158.93 mg,829.04umol,2 eq) to the mixture which was stirred at 20 ℃ for 2H. After completion of the reaction, the mixture was filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ Ea=1) to give methyl (S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamido) propionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,252.05umol,60.81% yield, 93% purity) as a yellow solid. MS (ESI) m/z 517.1[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

(S) -2- ((S) -3-cyclopropyl-2- (7-fluoro-4-methoxy-1H-indole-2-carboxamido) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester (140 mg,271.02umol,1 eq.) in NH ₃ A solution in MeOH (4 mL, 7M) was stirred at 30deg.C for 16h. After the reaction was complete, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (130 mg, crude material) as a yellow solid. MS (ESI) m/z 502.2[ M+H ]] ⁺

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (120 mg,239.26umol,1 eq) in DCM (6 mL) was added a berg reagent (285.09 mg,1.20mmol,5 eq) at 30 ℃ and the resulting mixture was then stirred for 1H at 30 ℃. After the reaction was completed, the reaction mixture was quenched with water (1 mL) and was purged with N ₂ Dried and purified by preparative HPLC (column: waters Xbridge C18 150 x 50mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (25 mg,51.70umol,21.61% yield, 100% purity) as a white solid. MS (ESI) M/z484.2[ M+H] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.14-11.90(m,1H),9.03-8.85(m,1H),8.54(d,J＝7.4Hz,1H),7.88-7.73(m,1H),7.38-7.29(m,1H),6.97-6.86(m,1H),6.46-6.37(m,1H),5.01-4.90(m,1H),4.51-4.40(m,1H),3.87(s,3H),2.63-2.54(m,1H),2.22-2.11(m,1H),2.07(s,1H),1.87-1.72(m,2H),1.56-1.44(m,2H),1.16(s,3H),1.07(s,3H),0.81(s,1H),0.42(d,J＝6.4Hz,2H),0.24-0.16(m,1H),0.10(s,1H)。

EXAMPLE 287 Synthesis of viral protease inhibitor Compounds 1173

Step 1: (S) -2- ((S) -3-cyclopropyl-2- (4, 5-dimethyl-1H-pyrrole-2-carboxamido) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,340.46 mol,88% purity, 1 eq, HCl) in DCM (3 mL) was added 4, 5-dimethyl-1H-pyrrole-2-carboxylic acid (56.85 mg,408.55 mol,1.2 eq) and then DMAP (124.78 mg,1.02mmol,3 eq) and EDCI (130.53 mg,680.92 mol,2 eq) at 0 ℃. The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was quenched by addition to water (3 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1 to give methyl (S) -2- ((S) -3-cyclopropyl-2- (4, 5-dimethyl-1H-pyrrole-2-carboxamido) propionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (105 mg,216.33umol,63.54% yield, 92% purity) as a yellow solid. MS (ESI) m/z 447.1[ M+H ]] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4, 5-dimethyl-1H-pyrrole-2-carboxamide

Methyl (S) -2- ((S) -3-cyclopropyl-2- (4, 5-dimethyl-1H-pyrrole-2-carboxamido) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (100 mg,206.03umol,92% purity, 1 eq.) was reacted in NH ₃ The mixture in MeOH (7M, 5.48mL,186.06 eq.) was stirred at 20℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4, 5-dimethyl-1H-pyrrole-2-carboxamide (100 mg, crude material) as a white solid. MS (ESI) M/z430.1[ M-H] ⁺ 。

Step 3: n- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 5-dimethyl-1H-pyrrole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 5-dimethyl-1H-pyrrole-2-carboxamide (90 mg,177.28umol,85% purity, 1 eq) in DCM (1 mL) was added the bergs reagent (92.94 mg,390.01umol,2.2 eq) and stirred for 14H at 20 ℃. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ] <30 ℃ C.). By preparative HPLC (column Phenomenex Luna C, 75X 30mm X3 um; mobile phase: [ Water (0.2% FA) -ACN)]The method comprises the steps of carrying out a first treatment on the surface of the B%: the residue was purified from 35% -65%,8 min) to give N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 5-dimethyl-1H-pyrrole-2-carboxamide (4.27 mg,10.12umol,5.71% yield, 98.0% purity) as a white solid. MS (ESI) m/z 414.0[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝10.94(br d,J＝1.5Hz,1H),8.80(d,J＝8.2Hz,1H),7.81(s,1H),7.77-7.71(m,1H),6.61(d,J＝2.6Hz,1H),4.98-4.87(m,1H),4.40-4.32(m,1H),2.60-2.53(m,1H),2.18-2.10(m,1H),2.10-2.04(m,3H),1.97(dd,J＝8.6,12.3Hz,1H),1.90(s,3H),1.80-1.69(m,2H),1.53-1.44(m,1H),1.43-1.35(m,1H),1.19-1.12(m,3H),1.06(s,3H),0.80-0.67(m,1H),0.45-0.31(m,2H),0.18-0.00(m,2H)。

EXAMPLE 288 Synthesis of viral protease inhibitor Compound 1175

Step 1: (S) -2- ((S) -2- (4-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (150 mg,364.78umol,88% purity, 1 eq, HCl) in DCM (3 mL) was added 4-chloro-1H-pyrrole-2-carboxylic acid (63.71 mg,437.73umol,1.2 eq). Next, HOBT (98.58 mg,729.56umol,2 eq.) and DIEA (94.29 mg,729.56umol,127.08uL,2 eq.) and EDCI (139.86 mg,729.56umol,2 eq.) were added at 0deg.C. The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was quenched by addition of water (3 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give methyl (S) -2- ((S) -2- (4-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (110 mg,242.86umol,66.58% yield) as a yellow solid. MS (ESI) m/z 451.0[ M-H] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-chloro-1H-pyrrole-2-carboxamide

Methyl (S) -2- ((S) -2- (4-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (100 mg,220.78umol,100% purity, 1 eq.) was reacted in NH ₃ The mixture in MeOH (7M, 3mL,95.12 eq.) was stirred at 50deg.C for 20h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4-chloro-1H-pyrrole-2-carboxamide (95 mg, crude) as a white solidA substance). MS (ESI) m/z 438.2[ M+H ] ] ⁺ 。

Step 3: 4-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4-chloro-1H-pyrrole-2-carboxamide (95 mg,190.90umol,88% purity, 1 eq) in DCM (1 mL) was added the bergs reagent (136.48 mg,572.71umol,3 eq) and stirred for 1.5H at 20 ℃. After completion, the reaction mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ (]<30 ℃ C.). By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,10 min) to give 4-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide (28.63 mg,68.18umol,35.72% yield, 100% purity) as a white solid. MS (ESI) m/z 420.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.84-11.68(m,1H),8.88(d,J＝8.2Hz,1H),8.18(d,J＝7.5Hz,1H),7.83(s,1H),6.95(br d,J＝7.3Hz,2H),5.01-4.86(m,1H),4.46-4.29(m,1H),2.62-2.53(m,1H),2.19-2.09(m,1H),2.03-1.92(m,1H),1.81-1.71(m,2H),1.54-1.36(m,2H),1.16(s,3H),1.10-0.99(m,3H),0.83-0.69(m,1H),0.47-0.33(m,2H),0.21-0.02(m,2H)。

EXAMPLE 289 Synthesis of viral protease inhibitor Compounds 1177

/>

Step 1: (S) -2- ((S) -2- (5-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidino-To a solution of methyl 3-yl) propionate (150.00 mg,460.97umol,1 equivalent, HCl) in ACN (5 mL) was added NMI (113.54 mg,1.38mmol,3 equivalent), and then 5-chloro-1H-pyrrole-2-methylpropanoic acid (carboxpuriylic acid) (67.09 mg,460.97umol,1.0 equivalent) and TCFH (129.34 mg,460.97umol,1 equivalent) were added, and the solution was stirred at 20℃for 2H. After completion, the reaction mixture was diluted with water (10 mL), extracted with EtOAc (5 mL x 3) and taken up in Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (Phenomnex Luna 80X 30mm X3 um; mobile phase: [ water (0.1% TFA) -ACN)]) The residue was purified. The compound (S) -methyl 2- ((S) -2- (5-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (60.0 mg,132.37umol,28.74% yield, 90% purity) was obtained as a white solid. MS (ESI) m/z 453.2[ M+H ]] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-chloro-1H-pyrrole-2-carboxamide

A mixture of methyl (S) -2- ((S) -2- (5-chloro-1H-pyrrole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (60.00 mg,132.47 mol,1 eq.) in HCl/MeOH (4M, 4 mL) was stirred at 60℃for 18H. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-chloro-1H-pyrrole-2-carboxamide (50.00 mg, crude material) as a white solid.

Step 3: 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-chloro-1H-pyrrole-2-carboxamide (50.00 mg,114.18umol,1 eq) in DCM (1 mL) was added the berg reagent (81.63)mg,342.53umol,3 equivalents), and then the solution was stirred at 25 ℃ for 4h. After completion, the reaction mixture was quenched with water (1 mL) and air dried. By preparative HPLC (Waters Xbridge BEH C18 x 30mM x 10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]) The residue was purified to give 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-pyrrole-2-carboxamide (5.00 mg,11.91umol,10.43% yield) as a white solid. MS (ESI) m/z 420.1[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δppm 6.82(d,J＝3.81Hz,1H)6.03(d,J＝3.93Hz,1H)5.01(dd,J＝10.37,5.72Hz,1H)4.45(t,J＝7.39Hz,1H)2.87-2.71(m,1H)2.34(ddd,J＝13.77,10.43,5.13Hz,1H)2.14(dd,J＝12.40,8.46Hz,1H)1.94-1.76(m,2H)1.65-1.53(m,2H)1.25(s,3H)1.16(s,3H)0.87-0.72(m,1H)0.51(d,J＝8.11Hz,2H)0.22-0.10(m,2H)

Example 290 Synthesis of viral protease inhibitor Compounds 1181

Step 1: (S) -2- ((S) -3-cyclopropyl-2- (4, 6-dichloro-1H-indole-2-carboxamide) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,386.89 mol,1 eq, HCl) in DCM (3 mL) was added 4, 6-dichloro-1H-indole-2-carboxylic acid (89.00 mg,386.89 mol,1 eq), DMAP (141.80 mg,1.16mmol,3 eq) and EDCI (148.33 mg,773.77 mol,2 eq) followed by stirring at 20 ℃ for 2H. After completion, the reaction mixture was quenched by addition of water (3 mL) and then extracted with DCM (5 mL x 3). The combined organic layers were washed with HCl (1M, 5 mL), then brine (5 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. To give (S) -2- ((S) -3-cyclopropyl-2- (4, 6-) -as a yellow soliddichloro-1H-indole-2-carboxamido) propionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester (130 mg,241.89umol,62.52% yield). MS (ESI) m/z 537.2[ M+H] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -3-cyclopropyl-2- (4, 6-dichloro-1H-indole-2-carboxamido) propanamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (110 mg,204.68umol,1 eq.) in HN ₃ The mixture in MeOH (7M, 2mL,68.40 eq.) was stirred at 80℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide (110 mg, crude material) as a white solid. MS (ESI) m/z 522.2[ M+H ]] ⁺ 。

Step 3:4, 6-dichloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-indole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -4, 6-dichloro-1H-indole-2-carboxamide (105 mg,180.89umol,90% purity, 1 eq) in DCM (2 mL) was added the bergs reagent (258.64 mg,1.09mmol,6 eq) followed by stirring at 20 ℃ for 2H. After completion, the mixture was quenched with water (0.1 mL) and concentrated under reduced pressure to give residue [ ]<30 ℃ C.). By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give 4, 6-dichloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -1H-indole-2-carboxamide as a white solid. MS (ESI) M/z504.0[ M+H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.99-11.77(m,1H),8.84-8.66(m,1H),8.60-8.49(m,1H),7.55-7.35(m,2H),7.40-7.32(m,1H),7.19(d,J＝1.5Hz,1H),5.03-4.88(m,1H),4.64-4.45(m,1H),2.60-2.54(m,1H),2.28-2.14(m,1H),2.12-1.99(m,1H),1.90-1.75(m,2H),1.66-1.51(m,2H),1.22-1.09(m,6H),0.91-0.73(m,1H),0.48-0.39(m,2H),0.28-0.03(m,2H)

EXAMPLE 291 Synthesis of viral protease inhibitor Compounds 1191

Step 1: (2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To 7-chloro-6-fluoro-1H-indole-2-carboxylic acid (0.1 g,374.54umol,80% purity, 1.2 eq.) and (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl ]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (112.94 mg,312.12umol,1 eq, HCl) in DCM (3 mL) were added EDCI (119.67 mg,624.24umol,2 eq) and DMAP (114.40 mg,936.36umol,3 eq), and the resulting mixture was then stirred at 20 ℃ for 1h. After completion, by adding H ₂ O (20 mL) to quench the reaction mixture and extract with DCM (6 mL x 5). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (0.14 g,241.85umol,77.49% yield, 90% purity). MS (ESI) m/z 521.2[ M+H ]] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -7-chloro-6-fluoro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (7-chloro-6-fluoro-1H-indole)Indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ]Methyl propionate (0.12 g,230.34umol,1 eq.) in NH ₃ A solution in MeOH (3 mL) was stirred at 30℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-7-chloro-6-fluoro-1H-indole-2-carboxamide (0.11 g, crude material). MS (ESI) m/z 506.2[ M+H ]] ⁺

Step 3: 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 7-chloro-6-fluoro-1H-indole-2-carboxamide (0.11 g,173.92umol,80% purity, 1 eq.) in DCM (3 mL) was added the berg reagent (82.90 mg,347.85umol,2 eq.) and the mixture stirred at 30 ℃ for 2H. After completion, the mixture was quenched with water (1 mL) and with N ₂ Blow dried and purified by preparative HPLC (column Waters Xbridge Prep OBD C18.40.5.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,8 min) to give the product 7-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-fluoro-1H-indole-2-carboxamide (0.058 g,118.86umol,68.34% yield, 100% purity). MS (ESI) m/z 488.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.89(br s,1H),9.01(d,J＝7.9Hz,1H),8.69(d,J＝7.5Hz,1H),7.83(s,1H),7.66(dd,J＝4.8,8.7Hz,1H),7.27(s,1H),7.13(dd,J＝8.9,10.0Hz,1H),4.97(br d,J＝7.9Hz,1H),4.50(br d,J＝6.6Hz,1H),2.63-2.55(m,1H),2.26-2.09(m,1H),2.00(dd,J＝8.4,12.2Hz,1H),1.90-1.72(m,2H),1.59-1.44(m,2H),1.15(s,3H),1.07(s,3H),0.90-0.75(m,1H),0.55-0.35(m,2H),0.25-0.16(m,1H),0.15-0.07(m,1H)。

EXAMPLE 292 Synthesis of viral protease inhibitor Compounds 1193

Step 1: (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (0.1 g,276.35 mol,1 eq, HCl) and 6-chloro-5-fluoro-1H-indole-2-carboxylic acid (70.83 mg,331.62 mol,1.2 eq) in DCM (3 mL), DMF (0.5 mL) was added EDCI (105.95 mg,552.70 mol,2 eq), DMAP (101.28 mg,829.04 mol,3 eq). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and extract with DCM (8 mL x 5). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give the product (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino ] as a white solid]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (0.08 g,141.27umol,51.12% yield, 92% purity). MS (ESI) M/z521.2[ M+H] ⁺

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-5-fluoro-1H-indole-2-carboxamide

Will contain (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]NH of methyl propionate (0.07 g,134.36umol,1 eq) ₃ MeOH (1 mL) was stirred at 50deg.C for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo as a white solidSubstituted-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-5-fluoro-1H-indole-2-carboxamide (0.05 g, crude material). MS (ESI) m/z 506.2[ M+H ]] ⁺

Step 3: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 6-chloro-5-fluoro-1H-indole-2-carboxamide (0.03 g,56.33umol,95% purity, 1 eq.) in DCM (1 mL) was added the berg reagent (53.69 mg,225.31umol,4 eq.). The mixture was stirred at 30℃for 1h. After completion, the mixture was quenched with water (1 mL) and with N ₂ Blow dried and purified by preparative HPLC (column Waters Xbridge Prep OBD C18, 150 x 40mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give the product 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-fluoro-1H-indole-2-carboxamide (0.017 g,34.84umol,61.85% yield, 100% purity). MS (ESI) m/z 488.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.52(d,J＝6.4Hz,1H),7.46-7.42(m,1H),7.17(d,J＝0.7Hz,1H),5.04(dd,J＝5.7,10.5Hz,1H),4.54(t,J＝7.5Hz,1H),2.92-2.77(m,1H),2.40-2.31(m,1H),2.21-2.11(m,1H),1.88(br s,2H),1.71-1.55(m,2H),1.26-1.23(m,3H),1.14(s,3H),0.91-0.77(m,1H),0.59-0.46(m,2H),0.19(dd,J＝5.0,10.6Hz,2H)

EXAMPLE 293 Synthesis of viral protease inhibitor Compounds 1195

Step 1: (S) -2- ((S) -2- (6-chloro-7-fluoro-1H-indole-2-carboxamide) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a mixture of methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,344.19umol,80% purity, 1 eq) in DCM (3 mL) was added 6-chloro-7-fluoro-1H-indole-2-carboxylic acid (88.22 mg,413.03umol,1.2 eq), DMAP (126.15 mg,1.03mmol,3 eq) and EDCI (131.96 mg,688.38umol,2 eq) followed by stirring the resulting mixture at 20 ℃ for 2H. After completion, the reaction mixture was quenched by addition of water (1 mL) and then extracted with DCM (3 mL x 3). The combined organic layers were washed with HCl (1M, 3 mL), then brine (3 mL), over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give methyl (S) -2- ((S) -2- (6-chloro-7-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,241.85umol,70.27% yield, 90% purity) as a yellow solid. MS (ESI) m/z 519.0[ M-H ]] ⁺ .

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -6-chloro-7-fluoro-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (6-chloro-7-fluoro-1H-indole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (140 mg,241.85umol,90% purity, 1 eq.) was reacted at NH ₃ The mixture in MeOH (7M, 3mL,86.83 eq.) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -6-chloro-7-fluoro-1H-indole-2-carboxamide (140 mg, crude material) as a white solid. MS (ESI) m/z 504.0[ M-H ]] ⁺ 。

Step 3: 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-fluoro-1H-indole-2-carboxamide

To a mixture of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-chloro-7-fluoro-1H-indole-2-carboxamide (130 mg,205.55umol,80% purity, 1 eq) in DCM (2 mL) was added the berg reagent (146.95 mg,616.64umol,3 eq) followed by stirring at 20 ℃ for 2H. After completion, the mixture was quenched with water (1 mL) and concentrated under reduced pressure to give a residue [ ] <30 ℃ C.). By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,10 min) to give 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-1H-indole-2-carboxamide (29.53 mg,60.09umol,29.24% yield, 99.3% purity) as a white solid. MS (ESI) m/z 488.0[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.35-12.19(m,1H),8.99(d,J＝7.9Hz,1H),8.66(d,J＝7.5Hz,1H),7.83(s,1H),7.50(d,J＝8.6Hz,1H),7.32(d,J＝3.1Hz,1H),7.13(dd,J＝6.4,8.6Hz,1H),5.02-4.90(m,1H),4.53-4.42(m,1H),2.61-2.54(m,1H),2.22-2.11(m,1H),1.99(dd,J＝8.4,12.1Hz,1H),1.89-1.72(m,2H),1.57-1.42(m,2H),1.18-1.03(m,6H),0.88-0.73(m,1H),0.50-0.34(m,2H),0.26-0.03(m,2H)

Example 294 Synthesis of viral protease inhibitor Compounds 1201

Step 1: 3-chloro-5-fluoro-2-hydroxybenzaldehyde

To 2-chloro-4-fluoro-phenol (5 g,34.12mmol,1 eq.) and MgCl ₂ To a solution of (9.80 g,102.93mmol,4.22mL,3.02 eq.) in ACN (300 mL) was added TEA (9.45 g,93.40mmol,13.00mL,2.74 eq), (HCHO) n (3.5 g,34.12mmol,1.00 eq.) and the mixture stirred at 80℃for 60h. After the reaction was complete, the mixture was concentrated in vacuo and acidified to pH 1 with aqueous HCl (100 ml,3 m) and extracted with EtOAc (50 ml x 3), then in vacuoThe organic phase was concentrated and passed through a column (SiO ₂ Pe=1) to give 3-chloro-5-fluoro-2-hydroxybenzaldehyde (5 g,25.21mmol,73.88% yield, 88% purity) as a yellow solid. MS (ESI) m/z 173.1[ M-H ] ⁺

Step 2: 3-chloro-5-fluoro-2-methoxybenzaldehyde

To a solution of 3-chloro-5-fluoro-2-hydroxybenzaldehyde (1 g,5.73mmol,1 eq.) in ACN (20 mL) was added K ₂ CO ₃ (2.38 g,17.19mmol,3 eq.) and CH ₃ I (1.30 g,9.17mmol,570.62uL,1.6 eq.) the mixture was stirred at 80℃for 3h. After completion of the reaction, the mixture was concentrated in vacuo and water (60 mL) was added and extracted with DCM (20 mL x 3), followed by Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give 3-chloro-5-fluoro-2-methoxybenzaldehyde (1 g, crude material) as a yellow oil. MS (ESI) m/z 189.1[ M+H ]] ⁺

Step 3: (Z) -2-azido-3- (3-chloro-5-fluoro-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (572.94 mg,10.61mmol,2 eq.) in MeOH (20 mL) was added a solution of 3-chloro-5-fluoro-2-methoxybenzaldehyde (1 g,5.30mmol,1 eq.) and ethyl 2-azidoacetate (1.37 g,10.61mmol,1.21mL,2 eq.) in MeOH (10 mL) at 0deg.C. The mixture was stirred at 20℃for 16h. After completion of the reaction, the mixture was concentrated in vacuo and water (60 mL) was added and extracted with EtOAc (30 mL x 3), then concentrated in vacuo and passed through a column (SiO ₂ PE: EA=1:0 to 50:1) to give methyl (Z) -2-azido-3- (3-chloro-5-fluoro-2-methoxyphenyl) acrylate (0.35 g,1.05mmol,19.81% yield) as a yellow solid.

Step 4: 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of methyl (Z) -2-azido-3- (3-chloro-5-fluoro-2-methoxyphenyl) acrylate (350.00 mg,1.23mmol,1 eq.) in xylene (5 mL) was stirred at 170℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (300 mg, crude material) as a yellow solid.

Step 5: 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (300 mg,1.16mmol,1 eq.) in THF (2 mL) and H ₂ LiOH.H was added to the solution in O (2 mL) ₂ O (146.59 mg,3.49mmol,3 eq.) and the mixture was stirred at 30℃for 16h. After completion of the reaction, the mixture was concentrated in vacuo and adjusted to pH-1 with aqueous HCl (15 ml,1 m), followed by extraction with EtOAc (5 ml x 3), followed by concentration in vacuo to give 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (250 mg, crude material) as a yellow solid. MS (ESI) m/z 189.1[ M+H ]] ⁺

Step 6: (S) -2- ((S) -2- (5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (250 mg,690.87umol,1 eq, HCl) and 5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (168.31 mg,690.87umol,1 eq) in DCM (5 mL) was added DMAP (253.21 mg,2.07mmol,3 eq) and EDCI (264.88 mg,1.38mmol,2 eq) and the resulting mixture was then stirred at 20 ℃ for 1H. After completion of the reaction, the mixture was adjusted to pH-1 with 1M HCl (3 mL) and water (6 mL) was added, followed by extraction with DCM (3 mL x 3), then the organic phase was Na ₂ SO ₄ The mixture was dried, concentrated in vacuo and purified by preparative TLC (SiO ₂ Etoac=1) to give methyl (S) -2- ((S) -2- (5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (180 mg,294.01umol,42.56% yield, 90% purity) as a white solid. MS (ESI) m/z 551.2[ M+H ]] ⁺

Step 7: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide

(S) -2- ((S) -2- (5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cycloPropyl propionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester (160 mg,290.38umol,1 eq.) in NH ₃ A solution in MeOH (8 mL, 7M) was stirred at 30℃for 16h. After the reaction was complete, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide (100 mg, crude material) as a yellow solid. MS (ESI) m/z 536.1[ M+H ]] ⁺

Step 8: 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide (90 mg,167.91umol,1 eq) in DCM (2 mL) was added a berg reagent (120.05 mg,503.74umol,3 eq) at 30 ℃ and the resulting mixture was then stirred for 3H at 30 ℃. After the reaction was completed, the mixture was quenched with water (0.5 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge BEH C18 100 x 25mM x 5um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -70%,10 min) to give 5-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (15 mg,28.88umol,17.20% yield, 99.74% purity) as a white solid. MS (ESI) M/z518.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.36(s,1H),9.07-8.91(m,1H),8.76-8.63(m,1H),7.84-7.79(m,1H),7.70-7.32(m,1H),7.17(d,J＝10.4Hz,1H),4.97(q,J＝8.2Hz,1H),4.53-4.43(m,1H),4.03-3.77(m,3H),2.61-2.54(m,1H),2.21-2.12(m,1H),2.05-1.96(m,1H),1.89-1.73(m,2H),1.57-1.43(m,2H),1.19-1.13(m,3H),1.10-1.04(m,3H),0.87-0.76(m,1H),0.48-0.38(m,2H),0.24-0.07(m,2H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δppm 12.05(s,1H),8.71(d,J＝7.8Hz,1H),8.44-8.33(m,1H),7.56-7.49(m,1H),7.44-7.35(m,1H),7.09(d,J＝10.4Hz,1H),5.00-4.93(m,1H),4.60-4.53(m,1H),4.04-3.81(m,3H),2.63-2.55(m,1H),2.25-2.16(m,1H),2.14-2.06(m,1H),1.90-1.75(m,2H),1.69-1.52(m,2H),1.23-1.17(m,3H),1.16-1.11(m,3H),0.88-0.79(m,1H),0.50-0.42(m,2H),0.24-0.09(m,2H)。

Example 295 Synthesis of viral protease inhibitor Compounds 1203

Step 1: 4-chloro-3-fluoro-2-methoxy-benzaldehyde

At-70 ℃ under N ₂ To a mixture of 1-chloro-2-fluoro-3-methoxy-benzene (5 g,31.14mmol,1 eq.) in THF (100 mL) was added n-BuLi (2.5 m,13.70mL,1.1 eq.) in one portion. The mixture was stirred at-70 ℃ for 1h, then DMF (18.66 g,255.35mmol,19.65mL,8.2 eq.) in THF (35 mL) was added at-70 ℃ and then the resulting mixture was stirred at-70 ℃ for 1h, then HCl (1 m,75mL,2.41 eq.) was added and heated to 25 ℃ and stirred for 16h. Residual 20% of the reactants, 100mL of H was used for the reaction mixture ₂ O was diluted and extracted with 200mL EA (100 mL x 2). The combined organic layers were washed with 100mL brine (100 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue.

Neutral prep HPLC (column: welch Xtime C18X 70mM #10um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,20 min) of the purified residue. The compound 4-chloro-3-fluoro-2-methoxy-benzaldehyde (1 g,4.77mmol,15.33% yield, 90% purity) was obtained as a white solid. MS (ESI) m/z 189.0[ M+H ]] ⁺

Step 2: (Z) -2-azido-3- (4-chloro-3-fluoro-2-methoxy-phenyl) prop-2-enoic acid methyl ester

A mixture of NaOMe (229.18 mg,4.24mmol,2 eq.) in MeOH (8 mL) was cooled to-10deg.C, to this solution was added dropwise a mixture of 4-chloro-3-fluoro-2-methoxy-benzaldehyde (400 mg,2.12mmol,1 eq.) and ethyl 2-azidoacetate (547.73 mg,4.24mmol,484.72uL,2 eq.) in MeOH (4 mL). The mixture was stirred at 25 ℃ for 16h and a yellow solid was observed. After completion, the reaction mixture was filtered to give a residue. The residue was purified by preparative TLC (petroleum ether: ethyl acetate=20:1). The compound (Z) -2-azido-3- (4-chloro-3-fluoro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (200 mg,630.12umol,29.71% yield, 90% purity) was obtained as a white solid.

Step 3: 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (4-chloro-3-fluoro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (140 mg,490.10umol,1 eq.) in xylene (10 mL) was stirred at 170℃for 4h. Residual reactant by 10%. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (petroleum ether: ethyl acetate=2:1). The compound 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (96 mg,353.98umol,72.23% yield, 95% purity) was obtained as a white solid.

Step 4: 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (96 mg,372.61 mol,1 eq.) in THF (2 mL) and H ₂ LiOH.H was added to the mixture in O (1 mL) ₂ O (31.27 mg,745.21umol,2 eq.). The mixture was stirred at 60℃for 1h. After completion, the reaction mixture was adjusted to ph=3 by addition of HCl, and then with 30mL of H ₂ O was diluted and extracted with 100mL EA (50 mL x 2). The combined organic layers were washed with 50mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid (70 mg, crude material) was obtained as a yellow solid. MS (ESI) m/z 241.9[ M-H ] ] ⁺

Step 5: (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-4-methoxy-1H-indole-2-carbonyl) amino ] -3-cyclopropyl-propionyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxylic acid (70 mg,287.33umol,1 eq.) and (2S) -2-amino-3-cyclopropyl-propionyl [ (2S) -2]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (103.98 mg, 287.33. Mu. Mol,1 eq., HCl) in DCM (6 mL) and DMF (3 mL) was added DMAP (70.21 mg, 574.67. Mu. Mol,2 eq.) and EDCI (110.17 mg, 574.67. Mu. Mol,2 eq.). The mixture was stirred at 20℃for 1h. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 40mL EA (20 mL x 2). The combined organic layers were washed with 20mL brine (20 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ DCM: meoh=10:1) purification residue. The compound (2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-4-methoxy-1H-indole-2-carbonyl) amino was obtained as a colorless oil]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (120 mg,206.89umol,72.00% yield, 95% purity). MS (ESI) M/z551.1[ M+H ] ⁺

Step 6: n- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-5-fluoro-4-methoxy-1H-indole-2-carbonyl) amino group]-3-cyclopropyl-propionyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (100 mg,181.49umol,1 eq.) in NH ₃ A solution in MeOH (7M, 15.00mL,578.56 eq.) was stirred at 60℃for a further 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue and used directly in the next step. Obtaining the compound N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxamide (90 mg, crude material). MS (ESI) m/z 536.2[ M+H ]] ⁺

Step 7: 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -5-fluoro-4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a mixture of 6-chloro-5-fluoro-4-methoxy-1H-indole-2-carboxamide (90 mg,167.91umol,1 eq.) in DCM (5 mL) was added the berg reagent (80.03 mg,335.82umol,2 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was quenched with 5mL H ₂ O was diluted and extracted with 10mL DCM (5 mL x 2). The combined organic layers were concentrated by blow-drying to give a residue. By neutral prep HPLC (column: waters Xbridge BEH C18100. Times.30 mM. Times.10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,10 min) of the purified residue. Obtaining the compound 6-chloro-N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-5-fluoro-4-methoxy-1H-indole-2-carboxamide (52 mg,97.18umol,57.87% yield, 96.8% purity). MS (ESI) M/z518.1[ M+H ]] ⁺¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.81(br d,J＝1.1Hz,1H),8.96(d,J＝8.1Hz,1H),8.69(d,J＝7.6Hz,1H),7.82(s,1H),7.57(d,J＝0.9Hz,1H),7.26-7.16(m,1H),5.05-4.88(m,1H),4.54-4.40(m,1H),4.12(d,J＝1.3Hz,3H),2.55(br s,1H),2.23-2.11(m,1H),1.98(dd,J＝8.6,12.3Hz,1H),1.89-1.71(m,2H),1.57-1.40(m,2H),1.15(s,3H),1.05(s,3H),0.86-0.74(m,1H),0.49-0.34(m,2H),0.25-0.16(m,1H),0.14-0.04(m,1H)。

EXAMPLE 296 Synthesis of viral protease inhibitor Compounds 1205

HMTA (5.00 g, 35)67mmol,1.05 eq.) was added in small portions to TFA (80 mL) and the resulting mixture was heated to reflux at 78 ℃. A solution of 3-chloro-4-fluorophenol (5 g,34.12mmol,1 eq.) in TFA (30 mL) was then added dropwise and the mixture stirred for an additional 1h. After completion, the mixture was cooled to room temperature and concentrated in vacuo. The residue was poured into ice water (50 mL) and stirred overnight. The mixture was filtered and the filter cake was dissolved in EA (50 mL), over Na ₂ SO ₄ Dried and concentrated in vacuo to give 4-chloro-5-fluoro-2-hydroxybenzaldehyde (5.5 g, crude material) as a yellow oil. MS (ESI) M/z175.0[ M+H] ⁺ 。

Step 2: 4-chloro-5-fluoro-2-methoxybenzaldehyde

To a solution of 4-chloro-5-fluoro-2-hydroxybenzaldehyde (5.35 g,15.32mmol,50% purity, 1 eq.) in DMF (1 mL) was added K ₂ CO ₃ (4.24 g,30.65mmol,2 eq.) MeI (4.35 g,30.65mmol,1.91mL,2 eq.) is then added dropwise at 0deg.C and the mixture stirred at 25deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure and purified by adding 50mL of H at 0 ℃ ₂ O to quench the residue and then extract with DCM (50 ml x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 5:1) to give 4-chloro-5-fluoro-2-methoxybenzaldehyde (920 mg,4.63mmol,30.24% yield, 95% purity) as a yellow solid. MS (ESI) m/z 189.0[ M+H ]] ⁺ 。

Step 3: (Z) -2-azido-3- (4-chloro-5-fluoro-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (515.61 mg,9.54mmol,2 eq.) in MeOH (10 mL) was added dropwise, at-10deg.C, meOH (10 mL) containing ethyl 2-azidoacetate (1.23 g,9.54mmol,1.09mL,2 eq.) and 4-chloro-5-fluoro-2-methoxybenzaldehyde (900 mg,4.77mmol,1 eq.). The mixture was stirred at 25℃for 18h. After completion, the reaction mixture was concentrated under reduced pressure and purified by adding 50mL of H at 0 ℃ ₂ O to quench the residue and then extract with 150mL DCM (50 mL x 3). The combined organic layers were washed with brine (50 ml x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 20:1) the residue was purified to give methyl (Z) -2-azido-3- (4-chloro-5-fluoro-2-methoxyphenyl) acrylate (390 mg,1.30mmol,27.18% yield, 95% purity) as a yellow solid. MS (ESI) m/z 286.0[ M+H ] ] ⁺ 。

Step 4: 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of methyl (Z) -2-azido-3- (4-chloro-5-fluoro-2-methoxyphenyl) acrylate (390 mg,1.37mmol,1 eq.) in xylene (10 mL) was stirred at 170℃for 1.5h. After completion, the reaction mixture was cooled to 25 ℃, solid precipitated, followed by filtration, and the crude product was washed with 100mL PE to give methyl 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylate (220 mg,811.19umol,59.42% yield, 95% purity) as a white solid. MS (ESI) m/z 258.0[ M+H ]] ⁺ 。

Step 5: 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (200 mg, 776.26. Mu. Mol,1 eq.) in THF (2 mL) and H ₂ LiOH.H was added to a solution in O (0.5 mL) ₂ O (97.72 mg,2.33mmol,3 eq.) and then stirring the resulting mixture at 60℃for 2h. After completion, the resulting solution was adjusted to pH-5 with 1M HCl and then extracted with EtOAc (5 ml x 2). The combined organic phases were taken up in Na ₂ SO ₄ Dried, filtered and concentrated to give 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (190 mg, crude material) as a white solid. MS (ESI) m/z 244.0[ M+H ]] ⁺ 。

Step 6: (S) -2- ((S) -2- (6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To a solution of 6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (190 mg,779.91umol,1 eq.) in DCM (2 mL) and DMF (2 mL) was added (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxoMethyl alkylpyrrolidin-3-yl) propionate (416.92 mg,1.01mmol,88% purity, 1.3 eq, HCl), DMAP (285.84 mg,2.34mmol,3 eq) and EDCI (299.02 mg,1.56mmol,2 eq) were added and the mixture was stirred at 25 ℃ for 1h. After completion, the resulting solution was poured into brine (10 mL) and then extracted with EtOAc (10 mL x 2), the combined organic layers were washed with citric acid (20 mL x 2), then NaHCO ₃ (10 mL), brine (10 mL. Times.3), washed with Na ₂ SO ₄ Drying, filtration and concentration gave the crude product. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=1/1 to 0/1) the residue was purified to give methyl (S) -2- ((S) -2- (6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionate (210 mg,381.12umol,48.87% yield) as a white solid. MS (ESI) m/z 551.2[ M+H ]] ⁺ 。

Step 7: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide

Methyl (S) -2- ((S) -2- (6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamido) -3-cyclopropylpropionylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanoate (190 mg,344.82umol,1 eq.) was reacted in NH ₃ A solution in MeOH (7M, 3.83mL,77.66 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent to give N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -3-cyclopropyl-1-oxoprop-2-yl) -6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide (186 mg, crude material) as a white solid. MS (ESI) m/z 536.2[ M+H ]] ⁺ 。

Step 8: 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide

To N- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) amino) -3-cycloTo a solution of propyl-1-oxopropan-2-yl) -6-chloro-7-fluoro-4-methoxy-1H-indole-2-carboxamide (186 mg,347.02umol,1 eq) in DCM (4 mL) was added a bergiut reagent (165.40 mg,694.04umol,2 eq) and the mixture was then stirred at 25 ℃ for 3H. LCMS showed most starting material remaining, followed by addition of the berg reagent (82.70 mg,347.02umol,1 eq.) and stirring for 12h. LCMS showed little starting material remaining, followed by addition of the berg reagent (82.70 mg,347.02umol,1 eq.) and stirring for a further 6h. After completion, the resulting solution was treated with H ₂ O (0.5 mL) was quenched and then concentrated in vacuo (25 ℃ C.). By preparative HPLC (column: waters Xbridge BEH C, 100X 25mM X5 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,10 min) to give 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -7-fluoro-4-methoxy-1H-indole-2-carboxamide (50.52 mg,97.53umol,28.11% yield, 100% purity) as a white solid. MS (ESI) m/z 518.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )8.94(d,J＝8.2Hz,1H),8.60(d,J＝7.6Hz,1H),7.82(s,1H),7.37(d,J＝2.8Hz,1H),6.59(d,J＝4.6Hz,1H),5.04-4.88(m,1H),4.54-4.35(m,1H),3.89(s,3H),2.61-2.53(m,1H),2.21-2.12(m,1H),1.99(dd,J＝8.4,12.3Hz,1H),1.84-1.73(m,2H),1.53-1.44(m,2H),1.16(s,3H),1.06(s,3H),0.87-0.74(m,1H),0.44-0.34(m,2H),0.21-0.05(m,2H)

Example 297 Synthesis of viral protease inhibitor Compound 1215

Step 1: (Z) -2-azido-3- (4-chloro-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (9.50 g,175.86mmol,2 eq.) in MeOH (100 mL) was added MeOH (100 mL) containing 4-chloro-2-methoxy-benzaldehyde (15 g,87.93mmol,1 eq.) and ethyl 2-azidoacetate (23.84 g,184.65mmol,21.10mL,2.1 eq.) at-10deg.C. The mixture was stirred at 20℃for 18h. After completion ofBy adding 50mL of H at 20 DEG C ₂ O to quench the reaction mixture and then concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 80/1 to give methyl (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoate (15.5 g,57.91mmol,65.86% yield) as a yellow solid.

Step 2: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (4-chloro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (10 g,37.36mmol,1 eq.) in xylene (100 mL) was stirred at 170℃for 2h. After completion, the reaction mixture was filtered and concentrated under reduced pressure to give methyl 6-chloro-4-methoxy-1H-indole-2-carboxylate (4 g, crude material) as a white solid.

Step 3: 6-chloro-4-methoxy-1H-indole-2-carboxylic acid

To 6-chloro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (4 g,16.69mmol,1 eq.) in THF (30 mL) and H ₂ LiOH.H was added to the solution in O (10 mL) ₂ O (2.10 g,50.07mmol,3 eq.). The mixture was stirred at 50℃for 5h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. 1M HCl was added to adjust the pH to 3, followed by filtration and concentration under reduced pressure to give 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (3.5 g, crude material) as a white solid.

Step 4: hexahydrocyclopenta [ c ] pyrrole-1, 2 (1H) -dicarboxylic acid (1S, 3aR,6 aS) -2-tert-butyl 1-ethyl ester

To (3S, 3aS,6 aR) -1,2, 3a,4,5,6 a-octahydrocyclopenta [ c ]]To a solution of pyrrole-3-carboxylic acid ethyl ester (1.5 g,8.19mmol,1 eq.) and TEA (993.96 mg,9.82mmol,1.37mL,1.2 eq.) in DCM (15 mL) was added (Boc) ₂ O (2.14 g,9.82mmol,2.26mL,1.2 eq.) and then DMAP (200.01 mg,1.64mmol,0.2 eq.) were added. The mixture was stirred at 20℃for 16h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ O, and then extracted with DCM (35 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By passing throughColumn chromatography (SiO) ₂ The residue was purified with petroleum ether/ethyl acetate=1/0 to 80/1) to give (3 s,3as,6 ar) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ] as a colorless oil]Pyrrole-2, 3-dicarboxylic acid O2-tert-butyl ester O3-ethyl ester (2 g,7.06mmol,86.22% yield).

Step 5: (1S, 3aR,6 aS) -2- (tert-Butoxycarbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxylic acid

To (3S, 3aS,6 aR) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ]]Pyrrole-2, 3-dicarboxylic acid O2-tert-butyl ester O3-ethyl ester (2 g,7.06mmol,1 eq.) in THF (15 mL) and H ₂ LiOH.H was added to the solution in O (5 mL) ₂ O (888.55 mg,21.17mmol,3 eq.). The mixture was stirred at 50℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to remove the solvent. The pH was adjusted to 3 by addition of 1M HCl, and then filtered and concentrated under reduced pressure to give (3S, 3aS,6 aR) -2-t-butoxycarbonyl-3, 3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ] as a white solid ]Pyrrole-3-carboxylic acid (1.7 g, crude material).

Step 6: (1S, 3aR,6 aS) -1- (((S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-methoxy-1-oxopropan-2-yl) carbamoyl) hexahydrocyclopenta [ c ] pyrrole-2 (1H) -carboxylic acid tert-butyl ester

To (3S, 3aS,6 aR) -2-tert-Butoxycarbonyl-3, 3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ]]Pyrrole-3-carboxylic acid (534.61 mg,2.09mmol,1.5 eq), (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (350 mg,1.40mmol,1 eq., HCl) in DCM (7 mL) was added DMAP (426.36 mg,3.49mmol,2.5 eq.) and EDCI (535.22 mg,2.79mmol,2 eq.) was then added. The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 15mL of H at 20 ℃ ₂ O, and then extracted with DCM (20 ml x 3). The combined organic layers were washed with brine (15 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=60/1 to 40/1) to give (3S, 3as,6 ar) -3- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-methoxy-2-oxo-ethyl]Carbamoyl group ]-3,3a,4,5,6 a-hexahydro-1H-cyclopenta [c]Pyrrole-2-carboxylic acid tert-butyl ester (600 mg,1.20mmol,85.67% yield, 90% purity).

Step 7: (S) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -2- ((1S, 3aR,6 aS) -octahydrocyclopenta [ c ] pyrrole-1-carboxamido) propanoic acid methyl ester

A solution of (3S, 3aS,6 aR) -3- [ [ (1S) -1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-methoxy-2-oxo-ethyl ] carbamoyl ] -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ] pyrrole-2-carboxylic acid tert-butyl ester (550 mg,1.22mmol,1 eq.) in HCl/MeOH (10 mL) was stirred at 20deg.C for 2H. After completion, the reaction mixture was concentrated under reduced pressure to remove MeOH to give methyl (2S) -2- [ [ (3S, 3as,6 ar) -1,2, 3a,4,5,6 a-octahydrocyclopenta [ c ] pyrrole-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propanoate (472 mg, crude, HCl) as a yellow solid.

Step 8: (S) -2- ((1S, 3aR,6 aS) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxamido) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (363.64 mg,1.61mmol,1.2 eq.) 2S-2- [ [ (3S, 3aS,6 aR) -1,2, 3a,4,5,6 a-octahydrocyclopenta [ c ] ]Pyrrole-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (470 mg,1.34mmol,1 eq.) in DCM (10 mL) was added DMAP (410.19 mg,3.36mmol,2.5 eq.) and EDCI (514.93 mg,2.69mmol,2 eq.) was then added. The resulting mixture was stirred at 20℃for 2h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ O, and then extracted with DCM (35 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=80/1 to 30/1) to give (2S) -2- [ [ (3S, 3as,6 ar) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ] as a yellow solid]Pyrrole-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (650 mg,1.16mmol,86.57% yield).

Step 9: (1S, 3ar,6 as) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide

(2S) -2- [ [ (3S, 3aS,6 aR) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ] ]Pyrrole-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (650 mg,1.16mmol,1 eq.) in NH ₃ A solution in MEOH (7M, 13.00mL,78.27 eq.) was stirred at 65℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give (3S, 3as,6 ar) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-oxo-ethyl]-2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ]]Pyrrole-3-carboxamide (630 mg, crude).

Step 10: (1S, 3ar,6 as) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) octahydrocyclopenta [ c ] pyrrole-1-carboxamide

To (3S, 3as,6 ar) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ]]To a solution of pyrrole-3-carboxamide (630 mg,1.16mmol,1 eq.) in DCM (12 mL) was added the Buerger's reagent (551.91 mg,2.32mmol,2 eq.). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was poured into 30mL of H at 20 ℃ ₂ O, and then extracted with DCM (30 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -75%,10 min) to give (3S, 3as,6 ar) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-3,3a,4,5,6 a-hexahydro-1H-cyclopenta [ c ]]Pyrrole compounds3-carboxamide (215 mg,408.73umol,35.30% yield, 100% purity).

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.69(s,1H),8.92(d,J＝7.9Hz,1H),7.83(s,1H),7.18-6.85(m,2H),6.57(s,1H),4.95(d,J＝7.1Hz,1H),4.74-4.30(m,1H),4.16(d,J＝8.9Hz,1H),3.97-3.62(m,4H),3.33-3.26(m,1H),2.95-2.75(m,1H),2.69-2.57(m,1H),2.23-1.32(m,10H),1.24-0.68(m,6H)

¹ H NMR(400MHz,DMSO-d ₆ ,273+80k)δ＝11.37(s,1H),8.85-8.58(m,1H),7.51(s,1H),7.10(s,1H),6.88(s,1H),6.55(s,1H),5.01-4.88(m,1H),4.49(s,1H),4.08(s,1H),3.92(s,3H),3.74(d,J＝4.5,10.8Hz,1H),3.00(s,4H),2.88-2.75(m,1H),2.65-2.54(m,2H),2.22-2.10(m,1H),2.08-1.92(m,2H),1.90-1.71(m,3H),1.68-1.58(m,2H),1.57-1.47(m,2H),1.19(s,3H),1.08(s,3H)。

EXAMPLE 298 Synthesis of viral protease inhibitor Compound 1219

Step 1: (1S, 2S, 5R) -2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) -3-azabicyclo [3.1.0] hexane-3-carboxylic acid tert-butyl ester

To (1S, 2S, 5R) -3-tert-Butoxycarbonyl-3-azabicyclo [3.1.0]Hexane-2-carboxylic acid (500 mg,2.20mmol,1 eq), (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (755.22 mg,2.64mmol,70% purity, 1.2 eq.) in DCM (15 mL) was added DMAP (537.58 mg,4.40mmol,2 eq.) and EDCI (632.66 mg,3.30mmol,1.5 eq.). The mixture was stirred at 20℃for 3h. After completion, by adding H ₂ O (40 mL) to quench the reaction mixture and then extract with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (SiO ₂ Methanol: dichloromethane=10:1) the residue was purified to give (1S, 2S,5 r) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-methylphenidate as a white solidBoydo group]Methyl group]Ethyl group]Carbamoyl group]-3-azabicyclo [3.1.0]Hexane-3-carboxylic acid tert-butyl ester (811 mg,1.47mmol,66.61% yield, 74% purity). MS (ESI) m/z 410.2[ M+H ]] ⁺ 。

Step 2: (S) -2- ((1S, 2S, 5R) -3-azabicyclo [3.1.0] hexane-2-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

(1S, 2S, 5R) -2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]Carbamoyl group]-3-azabicyclo [3.1.0]A solution of tert-butyl hexane-3-carboxylate (750 mg,1.83mmol,1 eq.) in HCl/MeOH (4M, 14 mL) was stirred at 20℃for 1h. After completion, the reaction mixture was concentrated under reduced pressure to remove HCl/MeOH, and DCM (20 mL) was added and concentrated under reduced pressure (repeated three times) to give (2S) -2- [ [ (1S, 2S,5 r) -3-azabicyclo [3.1.0] as a white solid ]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (500 mg, crude material, HCl). MS (ESI) m/z 310.2[ M+H ]] ⁺ 。

Step 3: (S) -2- ((1S, 2S, 5R) -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0] hexane-2-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (1S, 2S, 5R) -3-azabicyclo [3.1.0]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (800 mg,2.59mmol,1 eq.) and 4-methoxy-1H-indole-2-carboxylic acid (494.40 mg,2.59mmol,1 eq.) in DCM (20 mL) were added DMAP (631.86 mg,5.17mmol,2 eq.) and EDCI (743.60 mg,3.88mmol,1.5 eq.). The mixture was stirred at 20℃for 2h. After completion, by adding 50mL H ₂ O quench the reaction mixture and then extract with DCM (30 ml x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with ethyl acetate/meoh=5/1) to give (2S) -2- [ [ (1S, 2S,5 r) -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0] as a white solid]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl ]Methyl propionate (810 mg,1.59mmol,61.67% yield, 95% purity). MS (ESI) m/z 483.2[M+H] ⁺ 。

Step 4: (1S, 2S, 5R) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0] hexane-2-carboxamide

(2S) -2- [ [ (1S, 2S, 5R) -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0]Hexane-2-carbonyl group]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (810 mg,1.76mmol,1 eq.) in NH ₃ A solution in MeOH (7M, 10mL,39.74 eq.) was stirred at 65℃for 14h. After completion, the reaction mixture was concentrated under reduced pressure to remove HN ₃ MeOH, and DCM (30 mL) was added and concentrated under reduced pressure (repeated three times) to give (1S, 2S, 5R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0]Hexane-2-carboxamide (800 mg, crude material). MS (ESI) M/z468.2[ M+H ]] ⁺ 。

Step 5: (1S, 2S, 5R) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0] hexane-2-carboxamide

To (1S, 2S, 5R) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ]Methyl group]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0]To a solution of hexane-2-carboxamide (720 mg,1.73mmol,1 eq.) in DCM (10 mL) was added the Buerger's reagent (908.33 mg,3.81mmol,2.2 eq.). The mixture was stirred at 25℃for 2h. After completion, at N ₂ DCM was removed. By preparative TLC (SiO) ₂ The residue was purified with EA: meoh=20:1 to give the desired compound (460 mg,98% purity) as a white solid, which was purified by SFC (column: REGIS (S, S) WHELK-O1 (250 mm 25mm,10 um); mobile phase: [ Neu-ETOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,7 min) to give (1S, 2S,5 r) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0]Hexane-2-carboxamide isomer 1 (120 mg,266.96umol,15.41% yield, 100% purity). MS (ESI) m/z 450.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.55(s,1H),9.43-8.98(m,1H),7.60-7.46(m,1H),7.11(q,J＝8.3Hz,1H),7.06-6.97(m,1H),6.96-6.59(m,1H),6.55-6.43(m,1H),5.07(q,J＝7.7Hz,1H),4.92-4.63(m,1H),4.18-3.95(m,2H),3.93-3.78(m,3H),3.14-2.86(m,2H),2.32-2.14(m,2H),1.83-1.36(m,6H),1.30-1.01(m,1H),0.89-0.75(m,1H),0.21(br d,J＝3.8Hz,1H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.29(br s,1H),9.10-8.56(m,1H),7.27(br d,J＝1.3Hz,1H),7.16-6.74(m,3H),6.52(d,J＝7.5Hz,1H),5.11-5.00(m,1H),4.83-4.72(m,1H),4.06(br d,J＝9.9Hz,2H),3.90(s,3H),3.18-3.08(m,2H),2.31-2.21(m,2H),1.92-1.40(m,7H),0.88-0.78(m,1H),0.21(q,J＝4.1Hz,1H)。

To give (1S, 2S, 5R) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-3- (4-methoxy-1H-indole-2-carbonyl) -3-azabicyclo [3.1.0]Hexane-2-carboxamide isomer 2 (182 mg,392.75umol,22.67% yield, 97% purity). MS (ESI) m/z 450.2[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.61-11.48(m,1H),9.26-8.95(m,1H),7.58-7.43(m,1H),7.17-6.97(m,2H),6.96-6.56(m,1H),6.55-6.46(m,1H),5.12-5.03(m,1H),4.82-4.66(m,1H),4.16-4.00(m,2H),3.92-3.82(m,3H),3.13-2.96(m,2H),2.33(br s,2H),1.88-1.38(m,7H),0.86-0.74(m,1H),0.25-0.11(m,1H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.37-11.24(m,1H),8.99-8.67(m,1H),7.30-7.22(m,1H),7.15-7.09(m,1H),7.07-7.02(m,1H),6.96-6.80(m,1H),6.52(d,J＝7.7Hz,1H),5.13-5.00(m,1H),4.79-4.71(m,1H),4.13-3.94(m,2H),3.90(s,3H),3.18-3.09(m,2H),2.31-2.08(m,2H),1.94-1.57(m,6H),1.50-1.38(m,1H),0.87-0.77(m,1H),0.24-0.15(m,1H)。

Example 299 Synthesis of viral protease inhibitor Compounds 1221

Step 1: (R) -2-amino-3- (trimethylsilyl) propionic acid tert-butyl ester

To (2R) -2- [ (Z) - [ (2R) -2-hydroxy-2, 6-trimethyl-pinan-3-ylidene]Amino group]To a solution of tert-butyl 3-trimethylsilyl-propionate (1.2 g,3.26mmol,1 eq.) in THF (6 mL) was added a solution of citric acid (18 mL,15% purity). The mixture was stirred at 50℃for 3h. LCMS showed reaction incomplete, followed by stirring for a further 12h. After completion, THF was removed in vacuo and the aqueous layer was extracted with EtOAc (15 ml x 2) to remove chiral inducer. Then, the pH was increased to 8-9 with potassium carbonate. The free amine was then extracted with EtOAc (3 x 30 ml). The organic layers were combined, taken up over Na ₂ SO ₄ Drying and concentration at room temperature due to amine volatility afforded (2R) -2-amino-3-trimethylsilyl-propionic acid tert-butyl ester (510 mg,2.35mmol,71.87% yield) as a pale yellow oil. MS (ESI) m/z 218.1[ M+H ]] ⁺

Step 2: (R) -2- (6-chloro-4-methoxy-1H-indole-2-carboxamido) -3- (trimethylsilyl) propionic acid tert-butyl ester

To a solution of 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (477.45 mg,2.12mmol,1 eq.) in DMF (5 mL) was added tert-butyl (2R) -2-amino-3-trimethylsilyl-propionate (460 mg,2.12mmol,1 eq.), EDCI (527.36 mg,2.75mmol,1.3 eq.), TEA (642.38 mg,6.35mmol,883.61uL,3 eq.) and HOBt (371.72 mg,2.75mmol,1.3 eq.) at 0deg.C. The resulting reaction was stirred at 25℃for 1h. LCMS showed the reaction was incomplete, then EDCI (527.36 mg,2.75mmol,1.3 eq.) HOBt (371.72 mg,2.75mmol,1.3 eq.) and TEA (363.50 mg,3.59mmol,0.5ml,1.70 eq.) were added and stirred for a further 14h. After completion, the reaction mixture was taken up with H ₂ O (20 mL) was diluted and extracted with EA (20 mL x 2). The combined organic layers were washed with brine (10 ml x 5), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through a silica gel column (SiO) ₂ PE/EA=10:1) to give (2R) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]-3-trimethylsilyl-propionic acid tert-butyl ester (770 mg,1.79mmol,84.76% yield, 99% purity). MS (ESI) m/z 369.1[ M+H-56 ]] ⁺

Step 3: (R) -2- (6-chloro-4-methoxy-1H-indole-2-carboxamido) -3- (trimethylsilyl) propionic acid

At 0℃to (2R) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]To a solution of tert-butyl-3-trimethylsilyl-propionate (750 mg,1.76mmol,1 eq.) in DCM (9 mL) was added TFA/H ₂ O10:1 (6 mL). Subsequently, the reaction was stirred at 25℃for 2h. After completion, the reaction was concentrated in vacuo to dryness (below 30 ℃). The residue was poured into water (20 mL). The aqueous phase was extracted with ethyl acetate (12 ml x 2). The combined organic phases were dried over anhydrous Na ₂ SO ₄ Dried, filtered and concentrated in vacuo to give (2R) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino as a pale yellow solid]-3-trimethylsilyl-propionic acid (740 mg, crude material). MS (ESI) m/z 369.1[ M+H ] ] ⁺

Step 4: 6-chloro-N- ((R) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -1-oxo-3- (trimethylsilyl) propan-2-yl) -4-methoxy-1H-indole-2-carboxamide

at-30deg.C, to (2R) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-3-trimethylsilyl-propionic acid (100 mg,271.09umol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of propionitrile (67.99 mg, 406.63. Mu.L, 1.5 eq.) in DMF (2 mL) was added a solution of PyBOP (211.61 mg, 406.63. Mu.L, 1.5 eq.) and TEA (82.29 mg, 813.26. Mu.L, 113.20. Mu.L, 3 eq.) in DMF (2 mL). Next, the reaction was stirred at-30℃for 1h. After completion, the reaction mixture was quenched with water (10 mL) at-30 ℃, followed by extraction with EA (10 mL x 2) and the combined organic phases were taken over Na ₂ SO ₄ Drying and concentrating. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,8 min) to obtain the product. By SFC (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm,10 um); mobile phase: [0.1% NH ] ₃ H ₂ O IPA]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -40%,12 min) to give isomer 1 (rt=1.344 min) 6-chloro-N- [ (1R) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3 ] Piperidinyl group]Ethyl group]Amino group]-2-oxo-1- (trimethylsilylmethyl) ethyl]-4-methoxy-1H-indole-2-carboxamide (4.39 mg,8.40umol,3.10% yield, 99.1% purity). MS (ESI) m/z 518.1[ M+H ]] ⁺

1H NMR(400MHz,MeOD-d ₄ )δ＝7.21(s,1H),7.05(s,1H),6.52(d,J＝1.4Hz,1H),5.08(br dd,J＝6.1,9.8Hz,1H),4.59(t,J＝7.9Hz,1H),3.93(s,3H),3.23-3.16(m,2H),2.53-2.36(m,2H),2.03-1.84(m,2H),1.78(dt,J＝4.6,9.0Hz,1H),1.72-1.60(m,1H),1.55-1.43(m,1H),1.20(d,J＝7.9Hz,2H),0.10(s,9H)

Example 300 Synthesis of viral protease inhibitor Compound 1227

Step 1: (2S) -2- [ [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- (2-azaspiro [4.5]]Decane-3-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (0.1 g,240.41 mol,1 eq, HCl) and 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (81.37 mg,360.62 mol,1.5 eq) in DCM (2 mL) was added DMAP (88.11 mg,721.23 mol,3 eq), EDCI (92.17 mg,480.82 mol,2 eq). The mixture was stirred at 20℃for 2h. After completion, by adding H ₂ O (10 mL) was used to quench the reaction mixture and extracted with DCM (5 mL x 4). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give (2S) -2- [ [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow oil ]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (0.089 g,128.85umol,53.60% yield, 85% purity). MS (ESI) m/z 587.2[ M+H ]] ⁺

Step 2: n- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

Will contain (2S) -2- [ [2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]NH methyl propionate (0.079 g,134.56umol,1 eq) ₃ MeOH (3 mL) was stirred at 60℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a yellow solid]Methyl group]-2-oxo-ethyl]-2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (75 mg, crude). MS (ESI) m/z 572.2[ M+H ]] ⁺

Step 3: (3S) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (0.075 g,131.10umol,1 eq.) in DCM (1 mL) was added the Bunges reagent (93.73 mg,393.29umol,3 eq.). The mixture was stirred at 30℃for 3h. After completion, the mixture was quenched with water (0.5 mL) and with N ₂ Blow dried and purified by preparative HPLC (column Waters Xbridge BEH C18 100 x 30mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,8 min) to give the product (3S) -2- (6-chloro-4-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (19 mg,34.29umol,26.16% yield, 100% purity). MS (ESI) m/z 554.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.76-11.58(m,1H),8.90(d,J＝7.9Hz,1H),7.81(s,1H),7.15-6.85(m,2H),6.57(s,1H),4.92(br d,J＝6.6Hz,1H),4.49(t,J＝8.4Hz,1H),3.92(s,3H),3.84(br s,1H),3.67(br d,J＝10.4Hz,1H),2.66-2.58(m,1H),2.28-2.04(m,2H),1.99(dd,J＝8.3,11.8Hz,1H),1.82-1.68(m,1H),1.61-1.31(m,12H),1.19-1.06(m,3H),1.06-0.80(m,3H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.51-11.34(m,1H),8.85-8.58(m,1H),7.62-7.53(m,1H),7.13-6.82(m,2H),6.60-6.50(m,1H),5.04-4.81(m,1H),4.73-4.42(m,1H),3.96-3.90(m,3H),3.88-3.83(m,1H),3.76-3.51(m,1H),2.65-2.55(m,1H),2.27-2.01(m,2H),1.82-1.36(m,14H),1.20-1.14(m,3H),1.12-1.00(m,3H)。

Example 301 Synthesis of viral protease inhibitor Compound 1229

Step 1: (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] -2- [ [2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] propionic acid methyl ester

To (2S) -2- (2-azaspiro [4.5] ]Decane-3-carbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (82.86 mg,199.20umol,1 eq, HCl), 7-fluoro-4-methoxy-1H-indole-2-carboxylic acid (0.05 g,239.04umol,1.2 eq) in DCM (1 mL) was added DMAP (73.01 mg,597.59umol,3 eq) and EDCI (76.37 mg,398.39umol,2 eq). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (15 mL) was used to quench the reaction mixture and extracted with DCM (5 mL). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure and purified by preparative TLC (SiO ₂ DCM: meoh=10:1) to give the product (2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]-2- [ [2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]Methyl propionate (0.08 g,141.27umol,57.11% yield, 97% purity). MS (ESI) m/z 571.3[ M+H ]] ⁺

Step 2: n- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ]-2- [ [2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]Methyl propionate (0.07 g,122.67umol,1 eq.) in NH ₃ A solution in MeOH (7M, 3 mL) was stirred at 50℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Methyl group]-2-oxo-ethyl]-2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (0.065 g, crude). MS (ESI) m/z 556.3[ M+H ]] ⁺

Step 3: n- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

To N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a solution of decane-3-carboxamide (0.06 g,102.58umol,95% purity, 1 eq.) in DCM (1 mL) was added the berg reagent (97.79 mg,410.34umol,4 eq.) and the resulting mixture was stirred at 30 ℃ for 1h. After completion, the mixture was quenched with water (0.5 mL) and with N ₂ Blow dried and purified by preparative HPLC (column Waters Xbridge BEH C18100 x 25mM x 5um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -65%,10 min) to give the product N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-2- (7-fluoro-4-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (0.015 g,26.67umol,26.00% yield, 95.6% purity). MS (ESI) m/z 538.3[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.13-11.88(m,1H),9.12-8.84(m,1H),7.88-7.69(m,1H),7.07-6.63(m,2H),6.48-6.31(m,1H),4.99-4.83(m,1H),4.50(t,J＝8.7Hz,1H),3.90-3.77(m,4H),3.68(d,J＝10.3Hz,1H),2.73-2.59(m,1H),2.27-2.12(m,2H),2.06-1.96(m,1H),1.82-1.69(m,1H),1.62-1.29(m,12H),1.19-1.06(m,3H),1.05-0.87(m,3H)。

¹ H NMR(400MHz,DMSO-d ₆ ,273+80K)δ＝11.73-11.58(m,1H),8.80-8.61(m,1H),7.66-7.51(m,1H),7.05-6.80(m,2H),6.52-6.28(m,1H),5.00-4.84(m,1H),4.73-4.48(m,1H),3.92-3.80(m,4H),3.75-3.50(m,1H),2.65-2.54(m,1H),2.29-2.18(m,1H),2.11-1.93(m,1H),1.87-1.61(m,2H),1.58-1.28(m,12H),1.20-1.14(m,3H),1.10-1.00(m,3H)。

EXAMPLE 302 Synthesis of viral protease inhibitor Compounds 1231

Step 1: (2S) -2- [ [ (3S) -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

To (2S) -2- [ [ (3S) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (600 mg,1.44mmol,1 eq, HCl) in DCM (10 mL) was added 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (650.92 mg,2.88mmol,2 eq), DMAP (440.56 mg,3.61mmol,2.5 eq) and EDCI (553.05 mg,2.88mmol,2 eq). The mixture was stirred at 20℃for 3h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ Petroleum ether ethyl acetate=5/1 to 0/1) purification of the residue to give the product (2S) -2- [ [ (3S) -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] as a yellow solid]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (620 mg,844.82umol,58.57% yield, 80% purity). MS (ESI) m/z 587.3[ M+H ]] ⁺ 。

Step 2: (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5] decane-3-carboxamide

(2S) -2- [ [ (3S) -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (620 mg,844.82umol,80% purity, 1 eq.) in NH ₃ The mixture in MeOH (7M, 10mL,82.86 eq.) was stirred at 50deg.C for 16h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid ]Methyl group]-2-oxo-ethyl]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]Decane-3-carboxamide (480 mg, crude). MS (ESI) m/z 572.3[ M+H ]] ⁺ 。

Step 3: (3S) -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -2-azaspiro [4.5]To a mixture of decane-3-carboxamide (480 mg,839.02umol,1 eq.) in DCM (6 mL) was added the Bungeus reagent (599.83 mg,2.52mmol,3 eq.). The mixture was stirred at 30℃for 1h. After completion, the mixture was quenched with water (1 mL) and quenched with N ₂ And (5) blow-drying. By preparative HPLC (column: waters X bridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%: from 35% to 70%,8 min) to give the product (3S) -2- (7-chloro-5-methoxy-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]-2-azaspiro [4.5]]Decane-3-carboxamide (215.9 mg,386.54umol,46.07% yield, 99.2% purity). MS (ESI) m/z 554.3[ M+H ] ] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.51-11.25(m,1H),9.03-8.86(m,1H),7.87-7.73(m,1H),7.17-7.10(m,1H),7.03(s,1H),7.00-6.96(m,1H),4.97-4.76(m,1H),4.49(t,J＝8.6Hz,1H),3.70-3.86(m,4H),3.63(d,J＝10.4Hz,1H),2.78-2.63(m,1H),2.30-2.11(m,2H),2.04-1.95(m,1H),1.85-1.68(m,1H),1.64-1.28(m,12H),1.18-1.08(m,3H),1.05-0.86(m,3H)。

EXAMPLE 303 Synthesis of viral protease inhibitor Compound 1237

Step 1: (S) -2- ((S) -2- (6, 7-dichloro-1H-indole-2-carboxamide) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To 6, 7-dichloro-1H-indole-2-carboxylic acid (91.31 mg,396.92umol,1 eq.) and (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (150 mg,396.92 mol,1 eq, HCl) in DCM (5 mL) was added DMAP (96.98 mg,793.85 mol,2 eq) and EDCI (152.18 mg,793.85 mol,2 eq). The mixture was stirred at 20℃for 1h. After completion, by adding H ₂ O (30 mL) to quench the reaction mixture and then extract with EA (20 mL x 3). The combined organic layers were washed with brine (30 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with PE: ea=0:1) to give the product (2S) -2- [ [ (2S) -2- [ (6, 7-dichloro-1H-indole-2-carbonyl) amino ] as a yellow solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (110 mg,198.74umol,50.07% yield). MS (ESI) m/z 553.2[ M+H ] ] ⁺

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -6, 7-dichloro-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6, 7-dichloro-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (110 mg,198.74umol,1 eq.) in NH ₃ The mixture in MeOH (7M, 7.86mL,276.74 eq.) was stirred at 80℃for 16h. After completion, atThe mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]-6, 7-dichloro-1H-indole-2-carboxamide (100 mg, crude). MS (ESI) M/z538.2[ M+H ]] ⁺

Step 3:6, 7-dichloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl ]To a solution of 6, 7-dichloro-1H-indole-2-carboxamide (90 mg,167.14umol,1 eq.) in DCM (3 mL) was added the bergs reagent (79.66 mg,334.28umol,2 eq.). The mixture was stirred at 20℃for 5h. After completion, the mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,8 min) to give the product 6, 7-dichloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-1H-indole-2-carboxamide (20 mg,38.24umol,22.88% yield, 99.5% purity). MS (ESI) m/z 520.2[ M+H ]] ⁺¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.56(d,J＝8.4Hz,1H),7.27-7.15(m,2H),5.00(dd,J＝5.6,10.4Hz,1H),4.65(dd,J＝4.6,8.2Hz,1H),2.78(dd,J＝5.6,8.2Hz,1H),2.36-2.07(m,1H),2.10(dd,J＝8.6,12.4Hz,1H),1.95-1.84(m,2H),1.83-1.74(m,1H),1.65-1.52(m,1H),1.21(s,3H),1.08(s,3H),1.05-1.01(m,9H)。

EXAMPLE 304 Synthesis of viral protease inhibitor Compound 1239

Step 1: (S) -2- ((S) -2- (6-chloro-4-methoxy-1H-indole-2-carboxamido) -4, 4-dimethylpentanoylamino) -3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of methyl propionate (200 mg,529.23 mol,1 eq, HCl) and 6-chloro-4-methoxy-1H-indole-2-carboxylic acid (143.29 mg,635.08 mol,1.2 eq) in DCM (10 mL) were added EDCI (202.91 mg,1.06mmol,2 eq) and DMAP (193.97 mg,1.59mmol,3 eq) and the mixture was stirred at 20 ℃ for 1H. After completion, the reaction mixture was taken up with H ₂ O (60 mL) was diluted and extracted with DCM (40 mL x 3). The combined organic layers were washed with brine (50 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative TLC (SiO) ₂ The residue was purified with DCM: meoh=10:1) to give the product (2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino ] as a white solid]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (180 mg,327.83umol,61.95% yield). MS (ESI) m/z 549.3[ M+H ]] ⁺ 。

Step 2: n- ((S) -1- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxoprop-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -6-chloro-4-methoxy-1H-indole-2-carboxamide

(2S) -2- [ [ (2S) -2- [ (6-chloro-4-methoxy-1H-indole-2-carbonyl) amino group]-4, 4-dimethyl-pentanoyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (180.00 mg,327.83umol,1 eq.) in NH ₃ A solution in MeOH (7M, 5mL,106.76 eq.) was stirred at 60℃for 12h. After completion, the reaction mixture was concentrated under reduced pressure to give the product N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a yellow solid ]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]-6-chloro-4-methoxy-1H-indole-2-carboxamide (150 mg, crude). MS (ESI) m/z 534.3[ M+H ]] ⁺ 。

Step 3: 6-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -4-methoxy-1H-indole-2-carboxamide

To N- [ (1S) -1- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Carbamoyl group]-3, 3-dimethyl-butyl]To a solution of 6-chloro-4-methoxy-1H-indole-2-carboxamide (130.00 mg,243.42umol,1 eq.) in DCM (6 mL) was added the Bogis reagent (232.04 mg,973.70umol,4 eq.) and the mixture stirred at 25℃for 4H. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge Prep OBD C, 150X 40mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -65%,8 min) to give the product 6-chloro-N- [ (1S) -1- [ [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Ethyl group]Carbamoyl group]-3, 3-dimethyl-butyl]-4-methoxy-1H-indole-2-carboxamide (30 mg,58.14umol,23.88% yield, 100% purity). MS (ESI) m/z 516.2[ M+H ] ] ⁺ 。 ¹ H NMR(400MHz,MeOD-d ₄ )δ＝7.23(s,1H),7.04(s,1H),6.52(d,J＝1.4Hz,1H),4.99(dd,J＝5.8,10.4Hz,1H),4.63(dd,J＝4.6,8.3Hz,1H),3.93(s,3H),2.78-2.68(m,1H),2.41-2.29(m,1H),2.09(dd,J＝8.6,12.4Hz,1H),1.92-1.75(m,3H),1.62-1.54(m,1H),1.21(s,3H),1.13-0.96(m,12H)。

EXAMPLE 305 Synthesis of viral protease inhibitor Compound 1249

Step 1: (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

A solution of methyl (S) -2- ((S) -2- ((tert-butoxycarbonyl) amino) -3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (5 g,12.15mmol,1 eq.) in HCl/MeOH (4M, 60 mL) was stirred at 20deg.C for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (4 g, crude material) as a white solid. MS (ESI) M/z312.2[ M+H] ⁺

Step 2: (S) -2- ((S) -3-cyclopropyl-2- (5- (trifluoromethyl) -1H-pyrrole-2-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoic acid methyl ester

To methyl (S) -2- ((S) -2-amino-3-cyclopropylpropionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (680 mg,1.95mmol,1 eq. HCl) and 5- (trifluoromethyl) -1H-pyrrole-2-carboxylic acid (350.12 mg,1.95mmol,1 eq.) in CH ₃ To a solution of CN (8 mL) was added NMI (481.50 mg,5.86mmol,467.48uL,3 eq.) and TCFH (548.52 mg,1.95mmol,1 eq.) and the mixture was stirred at 20℃for 1h. After completion of the reaction, the mixture was diluted with water (20 mL) and then extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL), and dried over Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by column chromatography (SiO ₂ PE/ea=1/0 to 0/1) followed by preparative TLC (SiO ₂ EA: meoh=10:1) to give methyl (S) -2- ((S) -3-cyclopropyl-2- (5- (trifluoromethyl) -1H-pyrrole-2-carboxamido) propionylamino) -3- ((S) -2-oxopiperidin-3-yl) propanoate (350 mg,740.81umol,37.89% yield) as a yellow solid. MS (ESI) m/z 473.2[ M+H ]] ⁺

Step 3: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5- (trifluoromethyl) -1H-pyrrole-2-carboxamide

Methyl (S) -2- ((S) -3-cyclopropyl-2- (5- (trifluoromethyl) -1H-pyrrole-2-carboxamido) propanamido) -3- ((S) -2-oxopiperidin-3-yl) propanoate (300 mg, 634.98. Mu. Mol,1 eq.) was reacted in NH ₃ A solution in MeOH (10 mL, 7M) was stirred at 25℃for 12h, followed by 30℃for 24h. After the reaction was completed, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5- (trifluoromethyl) -1H-pyrrole-2-carboxamide (330 mg, crude material) as a yellow solid. MS (ESI) m/z 458.2[ M+H ] ] ⁺

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5- (trifluoromethyl) -1H-pyrrole-2-carboxamide

To a solution of N- ((S) -1- (((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5- (trifluoromethyl) -1H-pyrrole-2-carboxamide (300 mg,459.07umol,1 eq) in DCM (5 mL) was added the bergs reagent (328.20 mg,1.38mmol,3 eq) and the mixture stirred at 30 ℃ for 4H. After the reaction was completed, the mixture was quenched with water (1 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge Prep OBD C, 18, 150 x 40mM x 10um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -50%,8 min) to give N- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -5- (trifluoromethyl) -1H-pyrrole-2-carboxamide (4.36 mg,9.92umol,2.16% yield, 100% purity) as a white solid. MS (ESI) M/z440.1[ M+H] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.73(s,1H),8.96-8.85(m,1H),8.45-8.32(m,1H),7.52(s,1H),6.95(d,J＝3.2Hz,1H),6.66-6.57(m,1H),5.05(q,J＝8.0Hz,1H),4.48-4.39(m,1H),3.13-3.03(m,2H),2.29-2.18(m,2H),1.88-1.65(m,4H),1.62-1.50(m,1H),1.49-1.34(m,2H),0.83-0.71(m,1H),0.47-0.35(m,2H),0.22-0.04(m,2H)。

Example 306 Synthesis of viral protease inhibitor Compounds 1251

Step 1: (Z) -2-azido-3- (4-fluoro-2-methoxyphenyl) acrylic acid methyl ester

To a solution of NaOMe (700.98 mg,12.98mmol,2 eq.) at-10℃was added 4-fluoro-2-methoxy-benzaldehyde (1 g,6.49mmol,1 eq.) and ethyl 2-azidoacetate (1.68 g,12.98mmol,1.48mL,2 eq.) in MeOH (30 mL) and stirred at 25℃for 16h. After completion, by adding 30mL H ₂ O quench the reaction mixture and then extract with EA (30 ml x 3). Combined organic layer 3Washed with 0mL of brine, dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=100/1 to 50/1 to give methyl (Z) -2-azido-3- (4-fluoro-2-methoxy-phenyl) prop-2-enoate (685 mg,2.45mmol,37.83% yield, 90% purity) as a yellow solid.

Step 2: 6-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester

A solution of (Z) -2-azido-3- (4-fluoro-2-methoxy-phenyl) prop-2-enoic acid methyl ester (685 mg,2.73mmol,1 eq.) in xylene (10 mL) was stirred at 170℃for 2h. After completion, the reaction mixture was cooled to 25 ℃, and then a solid was obtained by filtration and washed with 10mL PE to give methyl 6-fluoro-4-methoxy-1H-indole-2-carboxylate (400 mg, crude material) as a white solid. MS (ESI) m/z 224.1[ M+H ] ] ⁺ 。

Step 3: 6-fluoro-4-methoxy-1H-indole-2-carboxylic acid

To 6-fluoro-4-methoxy-1H-indole-2-carboxylic acid methyl ester (400 mg,1.79mmol,1 eq.) in THF (8 mL) and H ₂ LiOH.H was added to the solution in O (2 mL) ₂ O (150.41 mg,3.58mmol,2 eq.). The mixture was stirred at 40℃for 3h. After completion, by adding 20mL H ₂ O to quench the reaction mixture, HCl (1M) was added to the aqueous phase to ph=3 and extracted with DCM (15 ml x 3). The combined organic layers were washed with 20mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 6-fluoro-4-methoxy-1H-indole-2-carboxylic acid (460 mg, crude material) as a white solid. MS (ESI) m/z 210.0[ M+H ]] ⁺ 。

Step 4: n- ((S) -1- (((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) amino) -3-cyclopropyl-1-oxopropan-2-yl) -6-fluoro-4-methoxy-1H-indole-2-carboxamide

To 6-fluoro-4-methoxy-1H-indole-2-carboxylic acid (81.01 mg,387.28umol,1.1 eq.) 2S) -2-amino-N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl]Ethyl group]To a solution of 3-cyclopropyl-propionamide (140 mg,352.08umol,70% purity, 1 eq.) in DCM (4 mL) was added DMAP (86.02 mg,704.15umol,2 equivalents) and EDCI (101.24 mg,528.11umol,1.5 equivalents). The mixture was stirred at 25℃for 2h. After completion, by adding 15mL H ₂ O quench the reaction mixture and then extract with EA (15 ml x 3). The combined organic layers were washed with 20mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) to give N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-6-fluoro-4-methoxy-1H-indole-2-carboxamide (20 mg,41.75umol,11.86% yield, 98% purity). MS (ESI) m/z 470.2[ M+H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.64(s,1H),8.89(d,J＝8.2Hz,1H),8.49(d,J＝7.5Hz,1H),7.52(br s,1H),7.36(d,J＝1.5Hz,1H),6.73(dd,J＝1.3,9.5Hz,1H),6.46(dd,J＝2.0,12.1Hz,1H),5.10-5.01(m,1H),4.48-4.40(m,1H),3.90(s,3H),3.15-3.03(m,2H),2.29-2.22(m,2H),1.87-1.68(m,4H),1.61-1.35(m,3H),0.85-0.73(m,1H),0.48-0.34(m,2H),0.25-0.04(m,2H)

EXAMPLE 307 Synthesis of viral protease inhibitor Compounds 1253

Step 1:1- (tert-Butoxycarbonyl) -4-cyclopentylpyrrolidine-2-carboxylic acid

To a solution of 4-cyclopentylpyrrolidine-2-carboxylic acid (900 mg,4.10mmol,1 eq, HCl) in DCM (20 mL) was added TEA (497.41 mg,4.92mmol,684.19uL,1.2 eq), DMAP (100.09 mg, 819.27. Mu. Mol,0.2 eq) and (Boc) ₂ O (983.42 mg,4.51mmol,1.04mL,1.1 eq.) and then stirring the resulting mixture at 20℃for 12h. After completion, by adding 50mL of H at 0deg.C ₂ O quench the reaction mixture and then extract with 30mL DCM. 1M HCl was added to the aqueous layer to pH=4, followed by extraction with DCM (30 mL. Times.3), washing with 40mL brine, washing with Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give 1-tert-butoxycarbonyl-4-cyclopentyl-pyrrolidine-2-carboxylic acid (750 mg, crude material) as a yellow solid.

Step 2: 4-cyclopentyl-2- (((S) -1-methoxy-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) carbamoyl) pyrrolidine-1-carboxylic acid tert-butyl ester

To 1-tert-butoxycarbonyl-4-cyclopentyl-pyrrolidine-2-carboxylic acid (750 mg,2.65mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (904.92 mg,3.44mmol,90% purity, 1.3 eq, HCl) in DCM (20 mL) was added DMAP (808.38 mg,6.62mmol,2.5 eq) and EDCI (1.01 g,5.29mmol,2 eq) and the mixture was then stirred at 20 ℃ for 1h. After completion, 60mL of H was added by adding at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (30 ml x 3). The combined organic layers were washed with 40mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:0 to 0:1) to give 4-cyclopentyl-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group ]Carbamoyl group]Pyrrolidine-1-carboxylic acid tert-butyl ester (1.0 g,2.15mmol,81.15% yield). MS (ESI) m/z 466.2[ M+H ]] ⁺

Step 3: (2S) -2- (4-Cyclopentylpyrrolidine-2-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

A mixture of tert-butyl 4-cyclopentyl-2- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] pyrrolidine-1-carboxylate (1 g,2.15mmol,1 eq.) in HCl/MeOH (4M, 20mL,37.25 eq.) was stirred at 20deg.C for 1h. After completion, the reaction mixture was concentrated under reduced pressure to give methyl (2S) -2- [ (4-cyclopentylpyrrolidine-2-carbonyl) amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (0.8 g, crude material, HCl) as a white solid.

Step 4: (2S) -2- (4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ (4-cyclopentylpyrrolidine-2-carbonyl) amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (0.8 g,1.99mmol,1 eq, HCl) and 4-methoxy-1H-indole-2-carboxylic acid (456.64 mg,2.39mmol,1.2 eq) in DCM (20 mL) was added DMAP (607.91 mg,4.98mmol,2.5 eq) and EDCI (763.13 mg,3.98mmol,2 eq) and the resulting mixture was then stirred at 20 ℃ for 2H. After completion, 60mL of H was added by adding at 0deg.C ₂ O quench the reaction mixture and then extract with DCM (30 ml x 3). The combined organic layers were washed with 40mL brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether ethyl acetate=1:4 to 0:1) to give (2S) -2- [ [ 4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl as a yellow solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.9 g,1.60mmol,80.59% yield, 96% purity). MS (ESI) m/z 539.3[ M+H ]] ⁺

Step 5: n- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

(2S) -2- [ [ 4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (0.9 g,1.67mmol,1 eq.) in NH ₃ The mixture was stirred at 50℃for 12h with MeOH (4M, 20mL,47.88 eq). After completion, the reaction mixture was concentrated under reduced pressure to give N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as a white solid]Methyl group]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (0.9 g, crude). MS (ESI) m/z 524.3[ M+H ] ] ⁺

Step 6: n- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) -4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide

To N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide (8)50mg,1.62mmol,1 eq.) of the Buerger's reagent (773.68 mg,3.25mmol,2 eq.) are added to a solution of the reagent in DCM (8 mL) and the mixture is stirred for 1h at 25 ℃. LCMS showed-50% of reactant 1 remained, followed by addition of the berg reagent (386.84 mg,1.62mmol,1.00 eq.) and stirring for an additional 2h. After completion, the reaction was quenched with water (0.8 mL), stirred for 10min and concentrated in vacuo (below 30 ℃). By preparative HPLC (column: waters Xbridge BEH C, 250X 50mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -75%,10 min) followed by SFC (column: DAICEL CHIRALCEL OD (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-MeOH]The method comprises the steps of carrying out a first treatment on the surface of the B%:45% -45%,15 min) to give isomer 1 (rt=1.409 min), N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl, as a white solid]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide isomer 1 (75.72 mg,149.76umol,9.23% yield, 100% purity). MS (ESI) m/z 506.2[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.21(br s,1H),8.64(br s,1H),7.23-7.04(m,3H),6.89(br s,1H),6.53(d,J＝7.3Hz,1H),4.98(br d,J＝7.2Hz,1H),4.58(br s,1H),4.12(dd,J＝7.8,10.0Hz,1H),3.91(s,3H),3.50(br s,1H),3.10(br s,2H),2.47-2.06(m,5H),1.90-1.74(m,4H),1.71-1.46(m,7H),1.45-1.13(m,3H)

Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide isomer 2 (98.03 mg,193.89umol,11.94% yield, 100% purity). MS (ESI) M/z506.2[ M+H] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.20(br s,1H),8.53(br s,1H),7.09(br dd,J＝7.8,16.0Hz,3H),6.91(br s,1H),6.61-6.43(m,1H),4.97(br s,1H),4.57(br s,1H),4.12(br d,J＝8.8Hz,1H),3.91(br d,J＝7.7Hz,3H),3.53(br s,1H),3.09(br s,2H),2.28-2.02(m,4H),1.90-1.54(m,12H),1.45-1.11(m,3H)

Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole)-2-carbonyl) pyrrolidine-2-carboxamide isomer 3 (250 mg,494.46umol,23.04% yield, 100% purity). By SFC (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm,10 um); mobile phase: [ Neu-EtOH)]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -50%,15 min) to give N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl) as a white solid]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide isomer 3_1 (90.63 mg,179.25umol,11.04% yield, 100% purity). MS (ESI) m/z506.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.25(br s,1H),8.62(br s,1H),7.19-7.04(m,3H),6.88(br s,1H),6.52(d,J＝7.3Hz,1H),5.00(br d,J＝6.4Hz,1H),4.68(br s,1H),4.07(br s,1H),3.91(s,3H),3.52(br s,1H),3.10(br s,2H),2.36-2.13(m,3H),2.05-1.68(m,8H),1.67-1.37(m,6H),1.20(br d,J＝10.6Hz,2H)

Obtaining N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-4-cyclopentyl-1- (4-methoxy-1H-indole-2-carbonyl) pyrrolidine-2-carboxamide isomer 3_2 (89.82 mg,177.65umol,10.94% yield, 100% purity). MS (ESI) M/z506.2[ M+H ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.24(br s,1H),8.66(br s,1H),7.37-7.01(m,3H),6.89(br s,1H),6.53(br d,J＝6.7Hz,1H),5.00(br s,1H),4.69(br s,1H),4.05(br s,1H),3.90(br d,J＝4.5Hz,3H),3.51(br s,1H),3.11(br s,2H),2.31-2.19(m,3H),2.07-1.68(m,8H),1.65-1.38(m,6H),1.20(br s,2H)。

Example 308 Synthesis of viral protease inhibitor Compounds 1268

/>

Step 1: (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-difluoro-1H-indole-2-carbonyl) amino ] propionyl ] amino ] -3- [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] propionic acid methyl ester

To (2S) -2- [ [ (2S) -2-amino-3-cyclopropyl-propionyl]Amino group]-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]Methyl propionate (200)To a solution of mg,555.79 mol,1 eq, HCl) in DCM (5 mL) was added 4, 7-difluoro-1H-indole-2-carboxylic acid (164.35 mg,833.69 mol,1.5 eq), DMAP (169.75 mg,1.39mmol,2.5 eq) and EDCI (213.09 mg,1.11mmol,2 eq). The mixture was stirred at 25℃for 1h. After completion, the reaction mixture was adjusted to acidic (pH 4-5) with 1M HCl (50 mL) and extracted with DCM (30 mL x 3). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through the column (SiO) ₂ Petroleum ether ethyl acetate=5/1-0/1) the residue was purified to give the product (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-difluoro-1H-indole-2-carbonyl) amino ] as a white solid]Propionyl radical]Amino group]-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]Methyl propionate (100 mg,199.00umol,35.81% yield). MS (ESI) m/z 503.2[ M+H ] ] ⁺ 。

Step 2: n- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] methyl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 7-difluoro-1H-indole-2-carboxamide

To (2S) -2- [ [ (2S) -3-cyclopropyl-2- [ (4, 7-difluoro-1H-indole-2-carbonyl) amino group]Propionyl radical]Amino group]-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]NH was added to a solution of methyl propionate (100 mg,199.00umol,1 eq) ₃ MeOH (7M, 2mL,70.35 eq.). The mixture was stirred at 65℃for 20h. After completion, the mixture was concentrated under reduced pressure to give a residue, which was then dissolved with DCM (10 ml x 3) and concentrated under reduced pressure to give the product N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] as a white solid]Hept-6-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4, 7-difluoro-1H-indole-2-carboxamide (90 mg, crude material). MS (ESI) m/z 488.2[ M+H ]] ⁺ 。

Step 3: n- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (6R) -5-oxo-4-azaspiro [2.4] hept-6-yl ] ethyl ] amino ] -1- (cyclopropylmethyl) -2-oxo-ethyl ] -4, 7-difluoro-1H-indole-2-carboxamide

To N- [ (1S) -2- [ [ (1S) -2-amino-2-oxo-1- [ [ (6R) -5-oxo-4-azaspiro [2.4] ]Hept-6-yl]Methyl group]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]To a solution of 4, 7-difluoro-1H-indole-2-carboxamide (90 mg,184.62umol,1 eq.) in DCM (1 mL) was added the berg reagent (131.98 mg,553.85umol,3 eq.). The mixture was stirred at 30℃for 1h. After completion, the mixture was quenched with water (1 mL) and with N ₂ And (5) blow-drying. By preparative HPLC (column: waters X bridge BEH C18 100 x 25mm x 5um; mobile phase: [ water (NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:20% -55%,10 min) to give the product N- [ (1S) -2- [ [ (1S) -1-cyano-2- [ (6R) -5-oxo-4-azaspiro [2.4 ] as a white solid]Hept-6-yl]Ethyl group]Amino group]-1- (cyclopropylmethyl) -2-oxo-ethyl]-4, 7-difluoro-1H-indole-2-carboxamide (10.52 mg,21.96umol,11.89% yield, 98% purity). MS (ESI) m/z 470.2[ M+H ]] ⁺ 。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.45(s,1H),8.98(d,1H),8.70(d,1H),7.80(s,1H),7.40(d,J＝2.4Hz,1H),7.06-6.96(m,1H),6.85-6.76(m,1H),5.04-4.89(m,1H),4.58-4.43(m,1H),2.70-2.57(m,1H),2.29-2.15(m,1H),2.01-1.75(m,4H),1.55-1.38(m,1H),0.90-0.77(m,1H),0.73-0.68(m,1H),0.62-0.48(s,3H),0.47-0.38(m,2H),0.26-0.01(m,2H)。

EXAMPLE 309 Synthesis of viral protease inhibitor Compound 1282

To (1S, 3aR,7 aS) -2-tert-butoxycarbonyl-1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxylic acid (450 mg,1.67mmol,1 eq.) and (2S) -2-amino-3- [ (3S) -2-oxo-3-piperidinyl ]To a solution of methyl propionate (571.23 mg,2.17mmol,90% purity, 1.3 eq, HCl) in DMF (5 mL) and DCM (1.5 mL) was added EDCI (640.58 mg,3.34mmol,2 eq) and DMAP (612.34 mg,5.01mmol,3 eq) and the mixture was then stirred at 25 ℃ for 2h. After completion, the reaction mixture was quenched with 20mL of H at 0deg.C ₂ O was diluted and then extracted with EA (10 ml x 2). The combined organic layers were washed with brine (10 ml x 5), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. Through a silica gel column (SiO) ₂ PE/ea=1:1 to EA). Next, diluted with EA (20 mL) and washed with 15% citric acid (10 mL x 2), the combined organic layers were washed with NaHCO ₃ (10 mL), brine (10 mL), washed over Na ₂ SO ₄ Drying, filtration and concentration under reduced pressure gives (1S, 3aR,7 aS) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl) as a white solid]Methyl group]Ethyl group]Carbamoyl group]-tert-butyl 1, 3a,4,5,6,7 a-octahydroisoindole-2-carboxylate (600 mg,1.33mmol,79.53% yield). MS (ESI) m/z 452.3[ M+H ]] ⁺

A solution of (1S, 3aR,7 aS) -1- [ [ (1S) -2-methoxy-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl ] methyl ] ethyl ] carbamoyl ] -1, 3a,4,5,6,7 a-octahydroisoindole-2-carboxylic acid tert-butyl ester (500 mg,1.11mmol,1 eq.) in HCl/MeOH (4M, 10mL,36.12 eq.) was stirred at 25℃for 1h. After completion, the reaction was concentrated in vacuo to give methyl (2S) -2- [ [ (1S, 3ar,7 as) -2, 3a,4,5,6,7 a-octahydro-1H-isoindole-1-carbonyl ] amino ] -3- [ (3S) -2-oxo-3-piperidinyl ] propanoate (430 mg, crude material, HCl) as a white solid.

Step 3: (S) -2- ((1S, 3aR,7 aS) -2- (7-chloro-1H-indole-2-carbonyl) octahydro-1H-isoindole-1-carboxamido) -3- ((S) -2-oxopiperidin-3-yl) propionic acid methyl ester

To (2S) -2- [ [ (1S, 3aR,7 aS) -2, 3a,4,5,6,7 a-octahydro-1H-isoindole-1-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]To a solution of methyl propionate (430 mg,1.11mmol,1 eq, HCl) and 7-chloro-1H-indole-2-carboxylic acid (281.88 mg,1.44mmol,1.3 eq) in DMF (6 mL) and DCM (2 mL) was added EDCI (425.01 mg,2.22mmol,2 eq) and DMAP (406.29 mg,3.33mmol,3 eq) followed by stirring the mixture at 25 ℃ for 2H. After completion, the reaction mixture was quenched with H at 0deg.C ₂ Dilution with O (20 mL)And then extracted with EA (20 ml x 2). The combined organic layers were washed with 15% citric acid (20 ml x 2) and the combined organic layers were washed with NaHCO ₃ (20 mL), brine (20 mL), washed over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By chromatography on silica gel (SiO ₂ The residue was purified with PE/ea=0:1) to give (2S) -2- [ [ (1S, 3ar,7 as) -2- (7-chloro-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carbonyl as an off-white solid]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (440 mg,798.45umol,72.03% yield, 96% purity). MS (ESI) m/z 529.2[ M+H ]] ⁺

Step 4: (1S, 3ar,7 as) -N- ((S) -1-amino-1-oxo-3- ((S) -2-oxopiperidin-3-yl) propan-2-yl) -2- (7-chloro-1H-indole-2-carbonyl) octahydro-1H-isoindole-1-carboxamide

(2S) -2- [ [ (1S, 3aR,7 aS) -2- (7-chloro-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carbonyl]Amino group]-3- [ (3S) -2-oxo-3-piperidinyl]Methyl propionate (440 mg, 831.72. Mu. Mol,1 eq.) in NH ₃ A solution in MeOH (7M, 12mL,101.00 eq.) and the mixture stirred at 40℃for 24h. After completion, the reaction mixture was concentrated under reduced pressure to give (1S, 3ar,7 as) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl as an off-white solid ]Methyl group]Ethyl group]-2- (7-chloro-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (430 mg, crude material). MS (ESI) m/z 514.2[ M+H ]] ⁺

Step 5: (1S, 3aR,7 aS) -2- (7-chloro-1H-indole-2-carbonyl) -N- ((S) -1-cyano-2- ((S) -2-oxopiperidin-3-yl) ethyl) octahydro-1H-isoindole-1-carboxamide

To (1S, 3aR,7 aS) -N- [ (1S) -2-amino-2-oxo-1- [ [ (3S) -2-oxo-3-piperidinyl]Methyl group]Ethyl group]To a solution of 2- (7-chloro-1H-indole-2-carbonyl) -1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (430 mg,836.55umol,1 eq.) in DCM (5 mL) was added a bergiut reagent (598.08 mg,2.51mmol,3 eq.) and the mixture was then stirred at 25 ℃ for 3H. After completion, the residue was quenched with water (0.5 mL) and stirred for 10min, followed by concentration below 30 ℃. By preparative HPLC (HPLC column: phenomnex Gem)ini-NX 80 x 40mm x 3um; mobile phase: [ Water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:25% -55%,8 min) of the purified residue. Subsequently, purification by SFC gave isomer 1 (rt=0.878 min) (1S, 3ar,7 as) -2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (103.07 mg,207.80umol,24.84% yield, 100% purity). MS (ESI) m/z 496.1[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )(273+80K)δ＝11.13(br s,1H),8.74(br s,1H),7.62(br s,1H),7.31-7.23(m,2H),7.08(br t,J＝7.8Hz,2H),5.00(br s,1H),4.31(br d,J＝4.4Hz,1H),4.08-3.88(m,1H),3.73(br d,J＝7.1Hz,1H),3.10(br s,2H),2.41(br s,1H),2.23(br s,3H),1.84(br s,2H),1.71(br s,2H),1.59-1.33(m,9H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.66-11.52(m,1H),9.12-8.85(m,1H),7.64(d,J＝7.8Hz,1H),7.56-7.49(m,1H),7.32-7.25(m,1H),7.16(s,1H),7.11-7.01(m,1H),5.11-4.94(m,1H),4.42-4.19(m,1H),3.98(dd,J＝6.7,10.0Hz,1H),3.80-3.69(m,1H),3.12-2.97(m,2H),2.38(br d,J＝4.5Hz,1H),2.27-2.13(m,3H),1.88-1.61(m,4H),1.54(br d,J＝4.8Hz,5H),1.44-1.23(m,4H)

Isomer 2 (rt=1.583 min) (1S, 3ar,7 as) -2- (7-chloro-1H-indole-2-carbonyl) -N- [ (1S) -1-cyano-2- [ (3S) -2-oxo-3-piperidinyl as a white solid]Ethyl group]-1, 3a,4,5,6,7 a-octahydroisoindole-1-carboxamide (99.03 mg,199.66umol,23.87% yield, 100% purity). MS (ESI) m/z 496.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )(273+80K)δ＝11.11(br s,1H),8.77(br s,1H),7.60(br s,1H),7.26(br d,J＝7.3Hz,2H),7.17-6.98(m,2H),5.00(br s,1H),4.32(br s,1H),3.93(s,1H),3.69(br s,1H),3.10(br s,2H),2.44-2.18(m,4H),1.80(br s,2H),1.68(br s,2H),1.54(br s,5H),1.36(br s,4H)

¹ H NMR(400MHz,DMSO-d ₆ )δ＝11.59(br s,1H),9.22-8.93(m,1H),7.64(d,J＝7.8Hz,1H),7.58-7.46(m,1H),7.34-7.24(m,1H),7.16(s,1H),7.11-6.99(m,1H),5.13-4.90(m,1H),4.46-4.24(m,1H),3.98(dd,J＝6.7,10.0Hz,1H),3.78-3.53(m,1H),3.15-2.91(m,2H),2.43-2.15(m,4H),1.96-1.75(m,2H),1.70-1.48(m,7H),1.46-1.24(m,4H)

EXAMPLE 310 Synthesis of viral protease inhibitor Compound 1286

Step 1: (S) -2-amino-3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propionic acid methyl ester hydrochloride

A solution of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propanoate (250 mg,800.36umol,1 eq.) in HCl/MeOH (5 mL, 4M) was stirred at 20℃for 1h. After the reaction was completed, the mixture was concentrated in vacuo to give methyl (S) -2-amino-3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propionate hydrochloride (200 mg, crude material, HCl) as a yellow solid.

Step 2: (S) -2- ((S) -2- ((tert-Butoxycarbonyl) amino) -4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

To (S) -2-amino-3- ((R) -5-oxo-4-azaspiro [2.4] at 20 ℃C]To a solution of methyl hept-6-yl) propionate hydrochloride (200 mg,804.16 mol,1 eq, HCl) and (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoic acid (217.00 mg,884.58 mol,1.1 eq) in DCM (10 mL) was added DMAP (196.49 mg,1.61mmol,2 eq) and EDCI (308.32 mg,1.61mmol,2 eq) and the mixture was stirred at 20 ℃ for 1h. After completion of the reaction, the mixture was adjusted to pH-1 with aqueous HCl (15 mL,1 m), followed by addition of water (10 mL) and extraction with DCM (9 mL x 3), followed by saturated NaHCO ₃ (15 mL) the organic phase was adjusted to pH 7, then concentrated in vacuo and passed through a column (SiO ₂ Ninhydrin, PE: ea=1:0 to 0:1) to give (2S) -2- [ [ (2S) -2- (tert-butoxycarbonylamino) -4, 4-dimethyl-pentanoyl as a white solid]Amino group]-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]Propionate (350 mg,732.58umol,91.10% yield, 92% purity). MS (ESI) m/z 440.2[ M+H ]] ⁺

Step 3: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

(2S) -2- [ [ (2S) -2- (tert-Butoxycarbonylamino) -4, 4-dimethyl-pentanoyl]Amino group]-3- [ (6R) -5-oxo-4-azaspiro [2.4]]Hept-6-yl]A solution of propionate (160 mg, 364.01. Mu. Mol,1 eq.) in HCl/MeOH (3 mL, 4M) was stirred at 20℃for 1h. After completion of the reaction, the mixture was concentrated in vacuo to give (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4] as a white solid]Methyl hept-6-yl) propionate (100 mg, crude, HCl). MS (ESI) m/z 340.2[ M+H ]] ⁺

Step 4: (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propanoic acid methyl ester

To (S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4]]To a solution of methyl hept-6-yl) propionate (90 mg,239.43umol,1 eq, HCl) and 7-chloro-5-methoxy-1H-indole-2-carboxylic acid (75.63 mg,335.20umol,1.4 eq) in DCM (5 mL) was added DMAP (58.50 mg,478.86umol,2 eq) and EDCI (91.80 mg,478.86umol,2 eq), and the mixture was stirred at 20 ℃ for 1H. After completion of the reaction, water (15 mL) was added to the mixture and extracted with DCM (5 mL x 3), followed by Na ₂ SO ₄ Dried, filtered and concentrated in vacuo and purified by preparative TLC (SiO ₂ Ea=1) purification to give (S) -2- ((S) -2-amino-4, 4-dimethylpentanamido) -3- ((R) -5-oxo-4-azaspiro [ 2.4) as a white solid]Methyl hept-6-yl) propionate (80 mg,131.62umol,54.97% yield, 90% purity). MS (ESI) m/z 547.2[ M+H ]] ⁺

Step 5: n- ((S) -1- (((S) -1-amino-1-oxo-3- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide

(S) -2- ((S) -2-amino-4, 4-dimethylpentanoylamino) -3- ((R) -5-oxo-4-azaspiro [2.4]]Methyl hept-6-yl) propionate (70 mg,127.96umol,1 eq.) in NH ₃ Solution in MeOH (4 mL, 7M)Stirred at 30℃for 16h. After completion of the reaction, the mixture was concentrated in vacuo to give N- ((S) -1- (((S) -1-amino-1-oxo-3- ((R) -5-oxo-4-azaspiro [ 2.4) as a white solid]Hept-6-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (70 mg, crude material). MS (ESI) m/z 532.2[ M+H ]] ⁺

Step 6: 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5-oxo-4-azaspiro [2.4] hept-6-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -5-methoxy-1H-indole-2-carboxamide

To N- ((S) -1- (((S) -1-amino-1-oxo-3- ((R) -5-oxo-4-azaspiro [ 2.4)]To a solution of hept-6-yl) propan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -7-chloro-5-methoxy-1H-indole-2-carboxamide (60 mg,112.78umol,1 eq.) in DCM (2 mL) was added a berges reagent (80.62 mg,338.33umol,3 eq.) and the mixture stirred at 30 ℃ for 2H. After the reaction was completed, the mixture was quenched with water (0.5 mL) and was purified by blowing N ₂ Dried and purified by preparative HPLC (column Waters Xbridge BEH C, 100 x 25mM x 5um; mobile phase: [ water (10 mM NH ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:30% -60%,10 min) to give 7-chloro-N- ((S) -1- (((S) -1-cyano-2- ((R) -5-oxo-4-azaspiro [ 2.4) as a white solid ]Hept-6-yl) ethyl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -5-methoxy-1H-indole-2-carboxamide (15 mg,28.89umol,25.62% yield, 99% purity). MS (ESI) m/z 514.1[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δppm 11.55(s,1H),9.02(d,J＝8.0Hz,1H),8.66(d,J＝8.0Hz,1H),7.82-7.73(m,1H),7.18-7.11(m,2H),7.00(d,J＝2.2Hz,1H),4.95(q,J＝7.8Hz,1H),4.61-4.53(m,1H),3.78(s,3H),2.64-2.58(m,1H),2.24-2.15(m,1H),1.99-1.91(m,2H),1.90-1.81(m,1H),1.77-1.65(m,2H),0.94(s,9H),0.75-0.67(m,1H),0.57-0.43(m,3H)。

EXAMPLE 311 Synthesis of viral protease inhibitor Compound 3075

Step 1: ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) carbamic acid tert-butyl ester

(2S) -2- (tert-Butoxycarbonylamino) -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (10 g,31.81mmol,1 eq.) in NH ₃ A solution in MeOH (80 mL) was stirred at 80℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in DCM (10 mL) and concentrated twice under reduced pressure to give N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a pale yellow gum]Methyl group]-2-oxo-ethyl]Tert-butyl carbamate (8.9 g, crude) and used directly in the next step.

Step 2: (S) -2-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) propanamide

A solution of tert-butyl N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] carbamate (8.9 g,29.73mmol,1 eq.) in 4MHCl/EtOAc (40 mL) was stirred at 25℃for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in toluene (10 mL) and concentrated twice under reduced pressure to give (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionamide (5.13 g, crude material, HCl) as a white solid and used directly in the next step.

Step 3: ((S) -1- ((1R, 2S, 5S) -2- (((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) carbamoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hex-3-yl) -3, 3-dimethyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester

To (1R, 2S, 5S) -3- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butanoyl]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxylic acid (0.5 g,1.36mmol,1 eq.) and (2S) -2-amino-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a solution of propionamide (405.57 mg,2.04mmol,1.5 eq.) in DMF (5 mL) was added PyBOP (776.78 mg,1.49mmol,1.1 eq.) and cooled to-30at-30deg.C, et is added to the mixture ₃ N (274.62 mg,2.71mmol,377.75uL,2 eq.). The mixture was stirred at-30℃for 2h. After completion, the reaction mixture was quenched with water (10 mL) and then extracted with ethyl acetate (6 mL x 2). The combined organic layers were washed with brine (10 ml x 3), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ The residue was purified with petroleum ether/ethyl acetate=20/1 to 0/1) to give N- [ (1S) -1- [ (1R, 2S, 5S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid ]Methyl group]-2-oxo-ethyl]Carbamoyl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-3-carbonyl]-2, 2-dimethyl-propyl]Tert-butyl carbamate (0.5 g,909.59umol,67.03% yield). MS (ESI) m/z 436.2[ M+H ]] ⁺

Step 4: (1R, 2S, 5S) -3- ((S) -2-amino-3, 3-dimethylbutyryl) -N- ((S) -1-amino-3- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) -1-oxopropan-2-yl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

N- [ (1S) -1- [ (1R, 2S, 5S) -2- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]Carbamoyl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-3-carbonyl]-2, 2-dimethyl-propyl]A mixture of tert-butyl carbamate (0.5 g,909.59umol,1 eq.) in HCl/EtOAc (4M, 200mL,879.52 eq.) was stirred at 20deg.C for 2h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. Next, the mixture was dissolved in toluene (10 mL) and concentrated twice under reduced pressure to give (1R, 2S, 5S) -3- [ (2S) -2-amino-3, 3-dimethyl-butyryl as a white solid]-N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ]Methyl group]-2-oxo-ethyl]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide (540 mg, crude material, HCl) and used directly in the next step. MS (ESI) m/z 450.3[ M+H ]] ⁺

To (1R, 2S, 5S) -3- [ (2S) -2-amino-3, 3-dimethyl-butyryl]-N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-6, 6-dimethyl-3-azabicyclo [3.1.0]To a solution of hexane-2-carboxamide (0.44 g,905.26umol,1 eq, HCl) in DCM (10 mL) was added DIEA (351.00 mg,2.72mmol,473.04ul,3 eq) and cooled to 0 ℃, followed by TFAA (152.11 mg,724.21umol,100.73ul,0.8 eq). The mixture was stirred at 0℃for 1h. After completion, the combined reaction mixture was poured into NaHCO ₃ Aqueous solution (20 mL) and extracted with DCM (10 mL x 2). The combined organic layers were purified by Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give (1R, 2S, 5S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid ]Methyl group]-2-oxo-ethyl]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide (350 mg, crude, 70.86% yield) and used directly in the next step. MS (ESI) m/z 546.1[ M+H ]] ⁺

Step 6: (1R, 2S, 5S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

To (1R, 2S, 5S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]To a solution of hexane-2-carboxamide (0.3 g,549.86umol,1 eq.) in DCM (5 mL) was added the Bunges reagent (393.10 mg,1.65mmol,3 eq.). The mixture was stirred at 20℃for 2h. After completion, the reaction mixture was quenched with water (0.5 mL) at 20 ℃ and with N ₂ The system was blow-dried to give a residue. By preparative HPLC (column: waters Xbridge BEH C, 100X 30mM X10 um; mobile phase: [ water (10 mM NH) ₄ HCO ₃ )-ACN]The method comprises the steps of carrying out a first treatment on the surface of the B%:35% -60%,8 min) to obtain white solid (1R, 2S, 5S) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Ethyl group]-3- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide (91.51 mg,173.45umol,31.54% yield, 100% purity). MS (ESI) m/z 528.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.41(br d,J＝7.7Hz,1H),8.99(d,J＝8.4Hz,1H),7.83(s,1H),4.97-4.88(m,1H),4.40(br d,J＝7.1Hz,1H),4.20-4.14(m,1H),3.90(br dd,J＝5.5,10.4Hz,1H),3.68(br d,J＝10.6Hz,1H),2.21-2.08(m,2H),1.99(br dd,J＝8.8,12.3Hz,1H),1.75(ddd,J＝5.7,10.3,13.5Hz,1H),1.59-1.51(m,2H),1.29(d,J＝7.7Hz,1H),1.20-1.16(m,3H),1.10(s,3H),1.05-1.01(m,3H),1.01-0.95(m,9H),0.86-0.83(m,3H)

Step 7: (1R, 2S, 5S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-carboxamide

By SFC (column: DAICEL CHIRALPAK AD-H (250 mm. Times.30 mm,5 um); mobile phase: [ Neu-IPA)]The method comprises the steps of carrying out a first treatment on the surface of the B%:5% -15%,15 min) further isolating (1R, 2S, 5S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0]Hexane-2-carboxamide to give (1R, 2S, 5S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxopyrrolidin-3-yl) ethyl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [3.1.0] as a white solid]Hexane-2-carboxamide isomer 1 (5.1 mg,9.67umol,5.74% yield). MS (ESI) m/z 528.2[ M+H ] ] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.50-9.36(m,1H),9.09-8.96(m,1H),7.98-7.82(m,1H),4.98-4.87(m,1H),4.40(br d,J＝5.5Hz,1H),4.21-4.15(m,1H),3.97-3.85(m,1H),3.73-3.60(m,1H),2.53-2.52(m,1H),2.15-2.08(m,1H),2.07-1.93(m,1H),1.84-1.70(m,1H),1.58-1.47(m,2H),1.35-1.27(m,1H),1.22-1.17(m,3H),1.16-1.08(m,3H),1.05-1.00(m,3H),1.00-0.94(m,9H),0.86-0.81(m,3H)。

Obtaining (1R, 2S, 5S) -N- ((S) -1-cyano-2- ((R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) ethyl) -3- ((S) -3, 3-dimethyl-2- (2, 2-trifluoroacetamido) butanoyl) -6, 6-dimethyl-3-azabicyclo [ 3.1.0) as a white solid]Hexane-2-carboxamide isomer 2 (61.8 mg,117.14umol,69.59% yield). MS (ESI) m/z 528.2[ M+H ]] ⁺

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.41(br d,J＝7.7Hz,1H),8.99(br d,J＝8.2Hz,1H),7.84(s,1H),4.96-4.89(m,1H),4.40(br d,J＝7.1Hz,1H),4.17(s,1H),3.90(br dd,J＝5.4,10.5Hz,1H),3.68(br d,J＝10.6Hz,1H),2.52(d,J＝2.0Hz,1H),2.19-2.10(m,1H),1.99(br dd,J＝8.7,12.5Hz,1H),1.75(ddd,J＝5.6,10.1,13.3Hz,1H),1.59-1.51(m,2H),1.29(d,J＝7.7Hz,1H),1.19-1.16(m,3H),1.10(s,3H),1.02(s,3H),0.98(s,9H),0.86-0.84(m,3H)

EXAMPLE 312 Synthesis of viral protease inhibitor Compound 3073

Step 1: (2S) -2- [ [ (3S) -2- [ (2S) -2- (tert-Butoxycarbonylamino) -3, 3-dimethyl-butyryl ] -2-azaspiro [4.5] decane-3-carbonyl ] amino ] -3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] propionic acid methyl ester

At 25 ℃, to (2S) -2- [ [ (3S) -2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]To a mixture of methyl propionate (200 mg,480.82umol,1 eq, HCl) in DCM (10 mL) was added DMAP (117.48 mg,961.65umol,2 eq), (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butanoic acid (111.21 mg,480.82umol,1 eq) and EDCI (184.35 mg,961.65umol,2 eq). The mixture was stirred at 25℃for 60min. After completion, the reaction mixture was quenched with 20mL H ₂ O was diluted and extracted with 50mL EA (25 mL x 2). The combined organic layers were washed with 25mL brine (25 mL x 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue. By column chromatography (SiO ₂ Petroleum ether/ethyl acetate=10/1 to 1-1) The residue was purified to give 180mg of crude material. Preparative HPLC (column: waters Xbridge BEH C18 100X 30mm X10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN) by neutral conditions]The method comprises the steps of carrying out a first treatment on the surface of the B%:50% -80%,10 min) to purify the crude material. Obtaining the compound (2S) -2- [ [ (3S) -2- [ (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butyryl) as a white solid]-2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl propionate (70 mg,118.09umol,24.56% yield, 100% purity). MS (ESI) m/z 593.4[ M+H ]] ⁺

Step 2: n- [ (1S) -1- [ (3S) -3- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carbonyl ] -2, 2-dimethyl-propyl ] carbamic acid tert-butyl ester

To (2S) -2- [ [ (3S) -2- [ (2S) -2- (tert-butoxycarbonylamino) -3, 3-dimethyl-butyryl]-2-azaspiro [4.5]]Decane-3-carbonyl]Amino group]-3- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]NH was added to methyl propionate (70 mg,118.09umol,1 eq) ₃ MeOH (7M, 14.00mL,829.87 eq.). The mixture was stirred at 25℃for 16h. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. Obtaining the compound N- [ (1S) -1- [ (3S) -3- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Methyl group]-2-oxo-ethyl]Carbamoyl group]-2-azaspiro [4.5]]Decane-2-carbonyl]-2, 2-dimethyl-propyl]Tert-butyl carbamate (68 mg,115.34umol,97.67% yield, 98% purity). MS (ESI) m/z 578.4[ M+H ]] ⁺

Step 3: (3S) -2- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2-azaspiro [4.5] decane-3-carboxamide

To tert-butyl N- [ (1S) -1- [ (3S) -3- [ [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] carbamoyl ] -2-azaspiro [4.5] decane-2-carbonyl ] -2, 2-dimethyl-propyl ] carbamate (68 mg,117.70umol,1 eq) was added HCl/EtOAc (4M, 3.78mL,128.39 eq). The mixture was stirred at 25℃for 60min. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was used directly in the next step. The compound (3S) -2- [ (2S) -2-amino-3, 3-dimethyl-butyryl ] -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2-azaspiro [4.5] decane-3-carboxamide (60 mg,114.37umol,97.18% yield, 98% purity, HCl) was obtained as a white solid.

Step 4: (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] methyl ] -2-oxo-ethyl ] -2- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -2-azaspiro [4.5] decane-3-carboxamide

To (3S) -2- [ (2S) -2-amino-3, 3-dimethyl-butyryl at 0deg.C]-N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2-azaspiro [4.5]]To DCM (4 mL) of decane-3-carboxamide (40 mg, 77.81. Mu.mol, 1 eq., HCl) were added DIEA (30.17 mg, 233.42. Mu.mol, 40.66. Mu.L, 3 eq.) and TFAA (19.61 mg, 93.37. Mu.L, 12.99. Mu.L, 1.2 eq.) and the mixture was stirred at 0deg.C for 30min. After completion, by adding 2mL of H at 0deg.C ₂ O quench the reaction mixture and extract with 10mL EA (5 mL x 2). The combined organic layers were washed with 5mL brine (5 mL 1), dried over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to give a residue which is used directly in the next step. Obtaining the compound (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl) as a white solid]Methyl group]-2-oxo-ethyl]-2- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] ]Butyryl group]-2-azaspiro [4.5]]Decane-3-carboxamide (60 mg,73.22umol,94.10% yield, 70% purity). MS (ESI) m/z 574.2[ M+H ]] ⁺

Step 5: (3S) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl ] ethyl ] -2- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ] butanoyl ] -2-azaspiro [4.5] decane-3-carboxamide

To (3S) -N- [ (1S) -2-amino-1- [ [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl]Methyl group]-2-oxo-ethyl]-2- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-2-azaspiro [4.5]]To a mixture of decane-3-carboxamide (30 mg,52.30umol,1 eq.) in DCM (3 mL) was added the Bungeus reagent (24.93 mg,104.59umol,2 eq.). The mixture was stirred at 25℃for 3h. After completion, the reaction mixture was quenched with 5mL H ₂ O was diluted and extracted with 10mL DCM (5 mL x 2). The combined organic layers were concentrated by blow-drying to give a residue. Preparative HPLC (column: waters Xbridge BEH C18 100X 30mm X10 um; mobile phase: [ water (10 mM NH4HCO 3) -ACN) by neutral conditions]The method comprises the steps of carrying out a first treatment on the surface of the B%:40% -70%,10 min) of the purified residue. Obtaining the compound (3S) -N- [ (1S) -1-cyano-2- [ (3R) -5, 5-dimethyl-2-oxo-pyrrolidin-3-yl as a white solid ]Ethyl group]-2- [ (2S) -3, 3-dimethyl-2- [ (2, 2-trifluoroacetyl) amino ]]Butyryl group]-2-azaspiro [4.5 ]]Decane-3-carboxamide (10 mg,17.40umol,33.28% yield, 96.7% purity). MS (ESI) m/z 556.2[ M+H ]] ⁺

1H NMR(400MHz,DMSO-d6)δ＝8.98-8.29(m,2H),7.50(br s,1H),5.01-4.78(m,1H),4.56(s,1H),4.31(br t,J＝8.6Hz,1H),3.79(br d,J＝10.1Hz,1H),3.29(br d,J＝10.0Hz,1H),2.60(br dd,J＝5.4,8.9Hz,1H),2.26-2.03(m,3H),1.87-1.71(m,1H),1.64-1.26(m,12H),1.25-1.13(m,6H),1.03(s,9H)

EXAMPLE 313 evaluation of antiviral Activity of Compounds against COVID-19 (nCoV-2019, SARS-CoV 2) Mpro in an enzymatic assay

Compounds were assayed using standard methods to assess compound activity and IC50. As an example of an evaluation of SARS-CoV2 Mpro, C-His 6-tagged Mpro (NC_ 045512) was cloned, expressed and purified in E.coli (E.coli). The assay buffer contained 20mM Tris-HCl (pH 7.3), 100mM NaCl, 1mM EDTA, 5mM TCEP and 0.1% BSA. In the Mpro enzymatic assay, the final concentration of Mpro protein and substrate is 25nM and 25. Mu.M, respectively. The Km of the Mpro substrate of the protease is 13.5. Mu.M.

The compounds were added to the assay plate. For the 100% inhibition control (HPE, one hundred percent action), 1 μm GC376 was added. For no inhibition control (ZPE, zero percent action), no compound was added. Each activity test point has an associated background control to normalize the fluorescence interference of the compound.

IC50 values for compounds were calculated by GraphPad Prism software using a nonlinear regression model of log (inhibitor) versus response-variable slope (four parameters). Inhibition activity was calculated using the following formula and IC50 values were calculated using% inhibition data.

Inhibition% = [ (sample-average ZPE)/(average HPE-average ZPE)]*100％ ^#

^# HEP: one hundred percent of control was acted. Contains substrate+enzyme+1. Mu.M GC376.

ZPE: zero percent effectiveness control. Contains enzyme + substrate and no compound.

Sample: compound activity assay wells. Comprising a compound + enzyme + substrate.

BG: compound background control wells. Contains compound + substrate and no enzyme.

Example 314 evaluation of antiviral Activity of Compounds against human coronaviruses (HCov) 229E and OC43 in cytopathic effect (CPE) assay

Compounds were assayed against a variety of coronavirus strains (including HCoV 229E and OC43 strains) using standard methods. The antiviral activity of the compounds was calculated based on protection of virus-induced CPE at each concentration normalized by the virus control.

Reagents and instruments used in this assay include the luminescent cell viability assay kit CellTiter Glo (Promega) and the microplate reader Synergy2 (BioTek).

Virus-HCoV 229E

Cytopathic effect (CPE) was measured by CellTiter Glo according to the manufacturer's manual. The antiviral activity of the compounds was calculated based on protection of virus-induced CPE at each concentration normalized by the virus control.

Virus-Hcov OC43

The reference compound used was adefovir; detection reagent: cellTiter Glo. ) CPE was measured by CellTiter Glo according to the manufacturer's manual. The antiviral activity of the compounds was calculated based on protection of virus-induced CPE at each concentration normalized by the virus control.

The cytotoxicity of the compounds was assessed under the same conditions, but in parallel in the absence of viral infection. Cell viability was measured by CellTiter Glo. The antiviral activity and cytotoxicity of the compounds are expressed as% inhibition and% viability, respectively, and are calculated by the formulae.

The activity data are shown in tables 3, 4 and 5.

Table 3. Activity data for the compounds.

/>

A is greater than or equal to 30. Mu.M, B is greater than or equal to 10. Mu.M and less than 30. Mu.M, C is greater than or equal to 2. Mu.M and less than or equal to 10. Mu.M, and D is less than 2. Mu.M.

TABLE 4 Activity data for Compounds

/>

A is >30 mu M, B is >10 mu M and is less than or equal to 30 mu M, C is more than or equal to 2 mu M and is less than or equal to 10 mu M, and D is <2 mu M.

TABLE 5 Activity data for Compounds

Numbering of compounds	229E CC ₅₀
		130 (isomer 1)	A
135	A
		170	A

Incorporated by reference

All publications and patents mentioned herein, including those listed below, are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of conflict, the present disclosure, including any definitions herein, will control.

Equivalent(s)

Although specific embodiments of the disclosure have been discussed, the above specification is illustrative and not restrictive. Many variations of the disclosure will become apparent to those of ordinary skill in the art upon review of the specification. The full scope of the disclosure, and the full scope of equivalents thereof, and the specification, and such variations, should be determined with reference to the claims.

Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure.

Sequence listing

<110> America Shang Padi s bioscience Co

<120> cysteine protease inhibitors and methods of use thereof

<130> PARB-004WO

<140>

<141>

<150> 63/195,460

<151> 2021-06-01

<150> 63/179,128

<151> 2021-04-23

<150> 63/173,146

<151> 2021-04-09

<150> 63/172,478

<151> 2021-04-08

<150> 63/171,675

<151> 2021-04-07

<150> 63/129,018

<151> 2020-12-22

<150> 63/091,630

<151> 2020-10-14

<150> 63/067,669

<151> 2020-08-19

<150> 63/039,297

<151> 2020-06-15

<150> 63/036,866

<151> 2020-06-09

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 302

<212> PRT

<213> severe acute respiratory syndrome coronavirus 2 (Severe acute respiratory syndrome coronavirus 2)

<400> 1

Ala Gly Leu Arg Lys Met Ala Gln Pro Ser Gly Phe Val Glu Lys Cys

1 5 10 15

Val Val Arg Val Cys Tyr Gly Asn Thr Val Leu Asn Gly Leu Trp Leu

20 25 30

Gly Asp Ile Val Tyr Cys Pro Arg His Val Ile Ala Ser Asn Thr Thr

35 40 45

Ser Ala Ile Asp Tyr Asp His Glu Tyr Ser Ile Met Arg Leu His Asn

50 55 60

Phe Ser Ile Ile Ser Gly Thr Ala Phe Leu Gly Val Val Gly Ala Thr

65 70 75 80

Met His Gly Val Thr Leu Lys Ile Lys Val Ser Gln Thr Asn Met His

85 90 95

Thr Pro Arg His Ser Phe Arg Thr Leu Lys Ser Gly Glu Gly Phe Asn

100 105 110

Ile Leu Ala Cys Tyr Asp Gly Cys Ala Gln Gly Val Phe Gly Val Asn

115 120 125

Met Arg Thr Asn Trp Thr Ile Arg Gly Ser Phe Ile Asn Gly Ala Cys

130 135 140

Gly Ser Pro Gly Tyr Asn Leu Lys Asn Gly Glu Val Glu Phe Val Tyr

145 150 155 160

Met His Gln Ile Glu Leu Gly Ser Gly Ser His Val Gly Ser Ser Phe

165 170 175

Asp Gly Val Met Tyr Gly Gly Phe Glu Asp Gln Pro Asn Leu Gln Val

180 185 190

Glu Ser Ala Asn Gln Met Leu Thr Val Asn Val Val Ala Phe Leu Tyr

195 200 205

Ala Ala Ile Leu Asn Gly Cys Thr Trp Trp Leu Lys Gly Glu Lys Leu

210 215 220

Phe Val Glu His Tyr Asn Glu Trp Ala Gln Ala Asn Gly Phe Thr Ala

225 230 235 240

Met Asn Gly Glu Asp Ala Phe Ser Ile Leu Ala Ala Lys Thr Gly Val

245 250 255

Cys Val Glu Arg Leu Leu His Ala Ile Gln Val Leu Asn Asn Gly Phe

260 265 270

Gly Gly Lys Gln Ile Leu Gly Tyr Ser Ser Leu Asn Asp Glu Phe Ser

275 280 285

Ile Asn Glu Val Val Lys Gln Met Phe Gly Val Asn Leu Gln

290 295 300

Claims

1. A protease inhibitor compound represented by:

R ^3a Selected from the group consisting ofAnd 4-10 membered heterocycle, wherein the heterocycle may be optionally substituted with one, two or three substituents each independently selected from the group consisting of: hydroxy, C ₁ -C ₈ Alkoxy, oxo, and warhead a;

R ^3b selected from hydrogen and C ₁ -C ₈ An alkyl group; wherein R is ^3a And R is ^3b Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered heterocyclic ring, wherein the heterocyclic ring can optionally be one, two or three of them are each independently selected from C ₆ -C ₁₄ Aryl and warhead a substituents;

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^A independently at each occurrence selected from the group consisting of: halogen, cyano, hydroxy, oxo, SF ₅ 、-CH ₂ CF ₃ 、-CF ₃ 、-O-CF ₃ 、-O-CHF ₂ 、-S-CH ₃ 、-S(O) ₂ -CH ₃ 、-NH ₂ -O-phenyl, -O- (C) ₁ -C ₈ Alkyl) -phenyl, -NHC (O) R ^B 、-NHC(O)OR ^B 、-NHC(O)O-(C ₁ -C ₈ Alkyl) -R ^B 、-N(R ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) O-phenyl, -N (R) ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-C(O)-OC(CH ₃ ) ₃ 、C ₁ -C ₈ Alkyl, C ₂ -C ₁₀ Alkenyl, C ₂ -C ₁₀ Alkynyl, C ₁ -C ₈ Heteroalkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl) - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl) - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocyclyl, wherein R is ^B The alkyl, heterocyclyl, heteroaryl, or aryl may be optionally substituted with one, two, or three substituents each independently selected from the group consisting of: halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, SF ₅ 、-NH ₂ Hydroxyl and oxo;

or R is ^1a And R is ² Can be joined together to form, together with the carbon to which they are attached, a 4-10 membered monocyclic or bicyclic heterocycle having a ring nitrogen、NR ^G Or C ₃ -C ₁₀ Cycloalkyl, wherein the cycloalkyl or heterocycle may optionally be substituted on the free carbon by one, two or three groups each selected from R ^A Is substituted by a substituent of (a);

R ^x independently at each occurrence selected from the group consisting of: halogen, hydroxy, oxo, CF ₃ 、SF ₅ Cyano, -O- (R) ^xx )-OCH ₃ 、-OCHF ₂ 、-OCF ₃ 、-O-(C ₁ -C ₈ Alkyl), -C (O) O (CH) ₃ )、-N(R ^y ) ₂ 、-N(R ^y )C(O)R ^y 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) N (R) ^y ) ₂ 、-N(R ^y )(C ₁ -C ₈ Alkyl) C (O) OH, - (C ₁ -C ₈ Alkyl) - (C ₃ -C ₁₀ Cycloalkyl group, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₃ -C ₁₀ Cycloalkyl, C ₆ -C ₁₄ Aryl, -O-C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, and 4-10 membered heterocycle;

Wherein said alkyl group,Aryl, heterocycle or heteroaryl groups may optionally be substituted with one or more groups each independently selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl;

R ^G selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh substituted-C (=o) -C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl; -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ An aryl group); -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ An aryl group); -C (O) - (5-10 membered heteroaryl); -C (O) - (4-10 membered heterocyclyl); and-C (O) - (4-10 membered heterocyclyloxy); wherein the aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj Substitution;

R ^hh Independently at each occurrence selected from the group consisting of: halo, cyano, -NR ^m R ^m 、-NR ^m (C＝O)R ^m Phenyl, cycloalkyl, heterocyclyl and C ₁ -C ₆ An alkoxy group;

R ^jj independently at each occurrence selected from the group consisting of: halo, oxo, hydroxy, cyano, C ₁ -C ₆ Alkyl, C _1-6 Haloalkyl, C ₁ -C ₆ Alkoxy, C ₁ -C ₆ Haloalkoxy, C _3-6 Cycloalkyl, SF ₅ And NH ₂ ；

R ^y Independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CF ₃ 、-CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl), C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH;

a is a warhead; and is also provided with

X is selected from C (R) ^xy ) And N, wherein R is ^xy Selected from the group consisting of: H. d, -OH, -NH ₂ Halogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Haloalkyl and C ₁ -C ₈ An alkoxy group.

2. The compound of claim 1, wherein the compound is represented by:

3. the compound of claim 1, wherein the compound is represented by:

4. the compound of claim 1, wherein the compound is represented by:

5. the compound of claim 1, wherein the compound is represented by:

6. the compound of claim 1, wherein the compound is represented by:

7. the compound of claim 1, wherein the compound is represented by:

8. the compound of claim 1, wherein the compound is represented by:

9. the compound of claim 1, wherein the compound is represented by:

wherein pp is selected from 0, 1, 2 and 3.

10. The compound of claim 1, wherein the compound is represented by:

Wherein ss is selected from 0, 1, 2 and 3 and mm is selected from 1, 2 and 3.

11. The compound of any one of claims 1-6, 9, and 10, wherein a is selected from the group consisting of: cyano, -C (O) R ^D 、-C(O)CH ₂ N(R ^b R ^c )、-C(O)CH ₂ OC(O)R ^D 、-C(O)C(O)R ^D 、-(CH＝CH)C(O)OR ^D 、-(CH＝CCN)C(O)OR ^D 、-(CH＝CCN)C(O)(NH)R ^D 、-CH(CN)(OH)、-CH(CN)(NR ^b R ^c )、

Wherein the method comprises the steps of

R ^E independently at each occurrence selected from the group consisting of: c (C) ₁ -C ₈ Alkyl, C ₁ -C ₈ Alkoxy, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle and 5-10 membered heteroaryl, wherein R ^E Optionally substituted with one, two or three substituents each independently selected from the group consisting of: halogen, cyano, C ₁ -C ₈ Alkyl and C ₁ -C ₈ An alkoxy group;

R ^cc selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl, - (C) ₁ -C ₈ Alkyl) - (C ₆ -C ₁₄ Aryl group, C ₆ -C ₁₄ Aryl, 5-10 membered heteroaryl, - (C) ₁ -C ₈ Alkyl) - (5-10 membered heteroaryl), 5-10 membered heterocycle and-N (R) ^b R ^c ) Wherein R is ^b And R is ^c Each independently selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl and C ₃ -C ₆ Cycloalkyl, or R ^b And R is ^c Can be joined together to form, together with the nitrogen to which they are attached, a 5-10 membered heterocyclic ring;

R ^b And R is ^c Each selected from the group consisting of: hydrogen, -CH ₂ C(O)O(C ₁ -C ₈ Alkyl), -C (O) - (C) ₁ -C ₈ Alkyl), -S (O) ₂ -(C ₁ -C ₈ Alkyl group, C ₁ -C ₈ Alkyl, C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) -C ₆ -C ₁₄ Aryl, wherein said C ₁ -C ₈ The alkyl group may be optionally substituted with one or more substituents each independently selected from the group consisting of: halogen, C ₃ -C ₆ Cycloalkyl, C ₆ -C ₁₄ Aryl, 4-10 membered heterocycle, and 5-10 membered heteroaryl.

12. The compound of claim 11, wherein a is selected from the group consisting of: -CN,

13. The compound of any one of claims 1, 4 and 9, wherein R ^1a Selected from the group consisting of:

14. the compound of any one of claims 1, 4 and 9, wherein R ^1a Is- (C) ₁ -C ₈ Alkyl) -R ¹ 。

15. The compound of any one of claims 1, 4 and 9, wherein R ^1b Is hydrogen.

16. The compound of claim 1, wherein R ^1a And R is ^1b Joined together to form

17. The compound of claim 1, wherein R ^3a Is a 4-10 membered heterocyclic ring substituted with A.

18. The compound of claim 1, wherein R ^3a Selected from the group consisting of:

19. the compound of any one of claims 1-8, wherein R ³ Is a 4-10 membered heterocycle.

20. The compound of any one of claims 1-8 and 10, wherein R ³ Selected from the group consisting of:

21. the compound of any one of claims 1-8, wherein R ² Selected from the group consisting of:

/>

22. the compound of claim 1, wherein R ^1a And R is ² Joined together to form a heterocycle selected from the group consisting of:

/>

wherein R is ^1b H.

23. The compound of any one of claims 1, 10 and 22, wherein R ^G Selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh substituted-C (=o) -C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl groups.

24. The compound of any one of claims 1, 10 and 22, wherein R ^G Selected from the group consisting of: -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ Aryl), -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ Aryl), -C (O) - (5-10 membered heteroaryl), -C (O) - (4-10 membered heterocyclyl) and-C (O) - (4-10 membered heterocyclyloxy); wherein the aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj And (3) substitution.

25. The compound of any one of claims 1, 10 and 22, wherein R ^G Selected from the group consisting of:

/>

26. a compound selected from the group consisting of:

/>

and pharmaceutically acceptable salts or stereoisomers thereof.

27. A pharmaceutical composition comprising a compound of any one of claims 1-26 and a pharmaceutically acceptable excipient.

28. A substantially reversible conjugate represented by:

wherein Cys is ₁₄₅ Is cysteine at position 145 or an equivalent active site on a CL or 3CL protease; IR is a viral protease inhibitor; and wherein the compound forming the conjugate comprises-CN warhead.

29. A method of ameliorating or treating a viral infection in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1-26.

30. The method of claim 29, wherein the viral infection is caused by a virus selected from the group consisting of: RNA viruses, DNA viruses, coronaviruses, papillomaviruses, pneumoviruses, picornaviruses, influenza viruses, adenoviruses, cytomegaloviruses, polyomaviruses, poxviruses, flaviviruses, alphaviruses, ebola viruses, measles viruses, enteroviruses, orthopneumoviruses, lentiviruses, arenaviruses, herpesviruses, and hepaciviruses.

31. The method of claim 29, wherein the viral infection is caused by a virus selected from the group consisting of: norwalk virus, feline calicivirus, MD145, murine norovirus, swine vesicular virus, rabbit hemorrhagic disease virus, enterovirus (EV) -68 virus, EV-71 virus, polio virus, coxsackievirus, foot and mouth disease virus, hepatitis A, porcine teschovirus, rhinovirus, human coronavirus, transmissible gastroenteritis virus, murine hepatitis virus, bovine coronavirus, feline transmissible peritonitis virus, and severe acute respiratory syndrome coronavirus.

32. The method of any one of claims 29-31, wherein the viral infection is a coronavirus infection.

33. The method of any one of claims 29-32, wherein the viral infection is a coronavirus selected from the group consisting of: 229Eα coronavirus, NL63 α coronavirus, OC43 β coronavirus, HKU1 β coronavirus, middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and SARS-CoV-2 (COVID-19).

34. The method of any one of claims 29-33, wherein the viral infection is SARS-CoV-2.

35. The method of claim 29 or 30, wherein the viral infection is an arenavirus infection.

36. The method of claim 35, wherein the arenavirus is selected from the group consisting of: the Huning virus, lassa virus, lu Yao virus, ma Qiubo virus and Sabina virus.

37. The method of claim 29 or 30, wherein the viral infection is an influenza infection.

38. The method of claim 37, wherein the influenza is influenza H1N1, H3N2, or H5N1.

39. A method of inhibiting viral transmission, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting viral release comprising administering to a patient carrying the virus a therapeutically effective amount of a compound of any one of claims 1-26, and/or contacting an effective amount of a compound of any one of claims 1-26 with a virally infected cell.

40. The method of any one of claims 29-39, further comprising administering another therapeutic agent.

41. The method of any one of claims 29-39, further comprising administering an additional antiviral therapeutic agent.

42. The method of claim 41, wherein the antiviral therapeutic agent is selected from the group consisting of: ribavirin, famprivir, ST-193, oseltamivir, zanamivir, pamil, danoprevir, ritonavir, radenpyr, bivalirudin, mitsubishi, etila, emtricitabine, tenofovir disoproxil, tenofovir alafenamide hemi-fumarate, abacavir, dolutegravir, efavirenz, elbavir, ledipavir, ganciclovir, sofosbuvir, bitaravir, dasabacavir, lamivudine, atazanavir, obetavir, lamivudine, stavudine, nevirapine, rilpivirine, ziprairivir, cimiravir, dacarbavir Grazopicir, pirenzvir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafavir, cabazitaxel, cytarabine, ecalciline, epigallocatechin gallate, itravir, fotif Sha Wei, gemcitabine, grifficine, isoprinosine, indinavir, malaevir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, nootkatole, pulefafur, PRO 140, raltegafur, prazidine, saquinavir, tebuvir, TNX-355, valacyclovir, VIR-576, and zalcitabine.

43. The method of claim 40, wherein the other therapeutic agent is selected from the group consisting of: protease inhibitors, fusion inhibitors, M2 proton channel blockers, polymerase inhibitors, 6-endonuclease inhibitors, neuraminidase inhibitors, reverse transcriptase inhibitors, acyclovir, protease inhibitors, arbidol, atazanavir, rituximab, boceprevir, cidofovir, bivalve, darunavir, docouabain Sha Nuo, alidomide, entry inhibitors, entecavir, famciclovir, fomivir, fosamprenavir, foscarnet phosphine ethanol, ganciclovir, ibacitabine, infliximab, iodate, imiquimod, inosine, integrase inhibitors, interferons, lopina Weiluo, moroxydine, ne Sha Wei, nucleoside analogues, penciclovir, praecox, podophyllotoxin, ribavirin, telanavir, trifluoretoside, tricxib, triamcinolone, telavancin, valacyclovir, valganciclovir, vec Wei Luoke, vidarabine, valamidine and zidovudine.

44. The method of claim 41, wherein the additional antiviral therapeutic agent is selected from the group consisting of: lamivudine, interferon alpha, VAP anti-idiotype antibody, enfu Wei De, amantadine, rimantadine, pranolide, acyclovir, zidovudine, fomivir, morpholino, protease inhibitors, double stranded RNA activated caspase oligomers (DRACO), rifampicin, zanamivir, oseltamivir Weidan nopuvir, ritonavir, radciclovir, cobalastat, eptifibatide, emtricitabine, tenofovir disoproxil, tenofovir, valavermectin, abacavir, dulutegravir, efavirenz, ganciclovir, sofosbuvir, bitorubivir, dasabavir, lamivudine, atazanavir, obuwei, obutyvir, emtricitabine, tenofovir, valacil, valvulafir, and valvulafir lamivudine, stavudine, nevirapine, rilpivirine, palivir, cimlopivir, dacarbazine, pezopivir, pirenz-tavir, adefovir, amprenavir, an Puli, alavir, anti-goat antibodies, balafrevir, cabavir, cytarabine, eleganciclovir, epigallocatechin gallate, itraprine, ferti Sha Wei, gemcitabine, grifeis, isoprinosine, indinavir, malavir, metaxazone, MK-2048, nelfinavir, nevirapine, nifedipine, norelvir, praxafovir, PRO 140, raltegafur, prazidine, quinavir, telbivudine, TNX-355, valacyclovir, pir-576 and zanciclovir.

45. A method of prophylactically treating a patient at risk of a viral infection comprising administering to the patient an effective amount of a compound of any one of claims 1-26.

46. The method of claim 45, wherein the compound is administered prior to viral exposure.

47. The method of claim 45, wherein the compound is administered after viral exposure.

48. An engineered CL or 3CL viral protease, wherein:

wherein the sulfur atom at the cysteine residue and the nitrile of the exogenous nitrile modifier undergo a reaction to form a thioiminoester adduct, and wherein the engineered SARS-protease does not retain the protease activity of the unmodified CL or 2CL protease.

49. The engineered viral protease of claim 48, wherein the engineered viral protease substantially prevents viral replication of SARS-CoV 2.

50. The engineered viral protease of claim 48, wherein the CL or 3CL protease is represented by SEQ ID NO: 1.

51. The engineered viral protease of claim 48, wherein the exogenous nitrile modifier of SEQ ID NO. 1 enzymatically inhibits IC ₅₀ Less than 20 micromoles.

52. The engineered viral protease of claim 48, wherein the thioiminoester adduct produced by the in vivo reaction between the exogenous nitrile modifier and the cysteine at position 145 of SEQ ID NO:1 is represented by:

wherein the method comprises the steps of

IR is the exogenous nitrile modifier after undergoing the reaction.

53. An engineered SARS-COV2-3CL viral protease represented by SEQ ID No. 1, wherein the cysteine at position 145 of SEQ ID No. 1 has a non-naturally occurring covalent modification resulting from a reaction between an exogenous nitrile modifier and the cysteine at position 145 of SEQ ID No. 1, wherein the exogenous nitrile modifier is represented by:

wherein the sulfur atom at the cysteine residue and-C≡N of the exogenous nitrile modifier undergo a reaction to form a thioiminoester adduct, and wherein

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

R ^B Is C ₁ -C ₆ Alkyl (optionally one, two or three independently selected from halogen, -NR ^m R ^m and-NR ^m (C＝O)R ^m Substituent of the group consisting of R ^m At each occurrence selected from H or C _1-3 Alkyl (optionally substituted with one, two or three halo)); or 8-10 membered bicyclic heteroaryl (optionally one, two or three are each independently selected from halogenSubstituents for groups or methoxy);

R ^1a is H; or (b)

54. A compound represented by:

/>

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein

R ¹ Is C ₁ -C ₆ Alkyl or-CH ₂ -C _3-10 Cycloalkyl;

R ^G is-C (O) R ^B ；

R ^1a is H; or (b)

R ¹ And R is ^1a Together with the nitrogen and carbon to which they are attached form a group which is optionally substituted on the free carbon by one or two groups each selected from methyl, halo or CF ₃ Is substituted by substituents of (2)4-10 membered monocyclic, bicyclic or spiro heterocycle.

55. Sup>A compound represented by formulSup>A IV-Sup>A or formulSup>A IV-B:

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

R ^1b selected from hydrogen and C ₁ -C ₈ An alkyl group;

R ^3b selected from hydrogen and C ₁ -C ₈ Alkyl group；

Wherein the alkyl, aryl, heterocycle or heteroaryl may optionally be substituted with one or more groups each independently selected from oxo, halogen and C ₁ -C ₈ Substituent substitution of alkyl;

R ^G selected from the group consisting of: hydrogen; optionally by one, two or three R ^gg Substituted C _1-6 An alkyl group; optionally by one, two or three R ^hh Substituted-C(＝O)-C _1-6 An alkyl group; -C (=o) -C _3-6 Cycloalkyl; -C (O) - (C) ₂ -C ₁₀ Alkenyl) - (C ₆ -C ₁₄ An aryl group); -C (O) - (5-10 membered heteroaryl); -C (O) - (C) ₁ -C ₆ Alkyl) -O- (C ₆ -C ₁₄ An aryl group); -C (O) - (4-10 membered heterocyclyl); and-C (O) - (4-10 membered heterocyclyloxy); wherein the aryl, heterocyclyl or heteroaryl may optionally be substituted with one, two or three R ^jj Substitution;

R ^y Independently at each occurrence selected from the group consisting of: hydrogen, C ₁ -C ₈ Alkyl, C ₁ -C ₈ Heteroalkyl, -CF ₃ 、-CH ₂ CF ₃ 、C ₁ -C ₈ Alkoxy, - (C) ₁ -C ₈ Alkoxy) - (5-10 membered aryl)、C ₃ -C ₆ Cycloalkyl and- (C) ₁ -C ₈ Alkyl) COOH.