CN117136178A

CN117136178A - SARS-COV-2 Mpro inhibitor compounds

Info

Publication number: CN117136178A
Application number: CN202180084687.XA
Authority: CN
Inventors: 迈尔斯·斯图尔特·康格里夫; 约翰·安德鲁·克里斯托夫; 马克·皮克沃思; 克里斯·德格拉夫; 艾丽西亚·佩雷斯·伊格鲁埃洛; 乔纳森·斯蒂芬·梅森; 桑托什·S·库尔卡尼
Original assignee: Hepares Treatment Co ltd
Current assignee: Hepares Treatment Co ltd
Priority date: 2020-12-18
Filing date: 2021-12-20
Publication date: 2023-11-28
Also published as: GB202107385D0

Abstract

The disclosure herein relates to compounds of formula (1'), or salts thereof, and their use in the treatment of SARS-CoV-2 and related viruses and SARS-CoV-2:Mpro-related diseases:therein A, Q, X, Z, L, R ² 、R ³ And R is ⁹ As defined herein.

Description

SARS-COV-2 Mpro inhibitor compounds

Technical Field

The present application relates to novel compounds and their use as inhibitors of SARS-CoV-2 main protease (Mpro). The compounds described herein are useful for treating SARS-CoV-2 and related viruses and conditions associated with SARS-CoV-2:Mpro. The application also relates to pharmaceutical compositions comprising these compounds, the preparation of these compounds and compositions, and the use of these compounds and compositions in the treatment of SARS-CoV-2 and related viruses and disorders associated with SARS-CoV-2:Mpro. These compounds and compositions may be useful in preventing death or complications from chronic underlying diseases or complications in patients infected with SARS-CoV-2 and related viruses.

Background

Coronaviruses exist in nature for a long time and are transmitted to humans by co-occurrence of humans and animals, and often cause mild respiratory diseases such as cold at the time of infection. However, in the last two decades, the outbreak of new human coronavirus infections leading to severe respiratory diseases has become a major problem for global health problems. This includes severe acute respiratory syndrome coronavirus (SARS-CoV) outbreaks in 2002 to 2004, middle east respiratory syndrome coronavirus (MERS-CoV) outbreaks in 2012 to 2015, and coronavirus strain severe acute respiratory syndrome type 2 coronavirus (SARS-CoV-2) that has recently emerged in 2019, and is the causative agent of the outbreak of coronavirus disease (covd-19) viral pneumonia in 2019 to 2020. Although the tragedy and widespread impact of these incidents and the periodic appearance of new human coronaviruses have increased the likelihood of future outbreaks, we have not yet performed effective antiviral treatments against coronavirus infections.

SARS-CoV-2 has packaged a large RNA genome of 30kb, two thirds of which encode two multimeric proteins pp1a and pp1b (Hegyi et al Journal of General Virology (3): 595-99). These polyproteins are processed into 16 nonstructural proteins (nsps) that are released from the long polypeptide chain by two viral cysteine proteases, papain-like protease (nsp 3) and 3C-like protease (np 5). The 3C-like protease, also known as the main protease (Mpro), cleaves the viral polyprotein at 11 sites to produce 12 nonstructural proteins (nsp 5-16). Included among these nsps are those involved in replication and transcription mechanisms, such as RNA-dependent RNA polymerase (nsp 12) and helicase (nsp 13). The important role of Mpro in viral replication has been demonstrated in mutagenesis experiments (Kim et al virology 208 (1): 1-8;Stobart et al.Journal of Virology 86 (9): 4801-10), which makes it an attractive target for designing inhibitors for the treatment of coronavirus infections. Furthermore, there is no human protease with similar cleavage specificity, so selective inhibitors of Mpro are likely to be non-toxic (Anand et al 2003.Science 300 (5626): 1763-67).

The use of protease inhibitors to treat viral diseases is a precedent (Bacon et al, the New England Journal of Medicine 364 (13): 1207-17) and the similarity of the SARS-CoV-2 Mpro active site to other viral proteases has driven the success of determining clinically approved drugs that can be reused to treat COVID-19 (Riva et al, nature, 586:113-119). 18 viral protease inhibitors designed for the treatment of Human Immunodeficiency Virus (HIV) and Hepatitis C Virus (HCV) were screened and the anti-HCV drug boceprevir and preclinical inhibitor GC376 against Feline Infectious Peritonitis Virus (FIPV) were identified as inhibitors of SARS-CoV-2 Mpro (Fu et al Nature Communications 11 (1): 4417). Although GC376 inhibited the recombinant protease activity (IC ₅₀ =0.15 μm) to boceprevir (IC ₅₀ =8 μm) is stronger, GC376 shows side effects in tests performed on cats, causing potential safety problems (peerrsen et al journal of Feline Medicine and Surgery (4): 378-92). In another study, boceprevir was also identified as an inhibitor of SARS-CoV-2Mpro along with telaprevir, although both agents inhibited SARS-CoV-2Mpro, IC ₅₀ The value is>1. Mu.M (Anson et al 2020.Doi:10.21203/rs.3.Rs-26344/v 1). In addition to SARS-CoV-2Mpro, the inhibitory efficacy of boceprevir and telaprevir from the Mpro proteases of other eight coronaviruses (including SARS, MERS, HKU, HKU4, HKU5, NL63, FIPV and IBV) was also evaluated. In this selection, boceprevir was able to inhibit all the tested coronavirus proteases except NL63, and telaprevir showed similar broad spectrum activity, inhibitory activity in SARS, HKU4, HKU5, NL63 and IBV. Although these drugs have insufficient antiviral activity on SARS-CoV-2Mpro for clinical development, their ability to inhibit broad-spectrum proteases highlights the design potential of broad-spectrum antiviral drugs that can treat not only SARS-CoV-2 infection but also other human coronaviruses and potentially new coronaviruses that may occur in the future.

Similar sequences between the active sites of SARS-CoV and SARS-CoV-2 Mpro have also been developed for the identification of the SARS-CoV-2 Mpro inhibitor PF-07304114, a phosphate prodrug of PF-00835231 was originally designed for the treatment of SARS-CoV (Boras et al BioRxiv, 2020.09.12.293498). PF-00835231 inhibits SARS-CoV-2 Mpro, its K _i 0.27nM and shows broad inhibitory activity against another 10 coronavirus strains, K _i The values were between 0.03nM and 4nM. This translates into-1. Mu.M activity in a cell-based live virus assay. The activity of PF-00835231 in combination with remdesired (a nucleoside RNA dependent RNA polymerase inhibitor) was also evaluated as an antiviral agent targeting different aspects of the viral replication process, which may produce synergistic effects in combination. Indeed, PF-00835231 and remdesivir show a synergistic or additive effect in cell-based antiviral assays, indicating that the combination of an Mpro inhibitor with an antiviral drug and other modes of action may show clinical benefit.

In 2020, the crystal structure of the complex of SARS-CoV-2 Mpro and N3 (a Michael acceptor inhibitor) was published (Jin et al Nature 582 (7811): 289)-93) to enable virtual screening and structure-based drug design (SBDD) of SARS-CoV-2 Mpro inhibitors. This SBDD achievement includes the design of peptidomimetic alpha-keto amides as broad spectrum inhibitors of coronaviruses and enteroviruses, the two most promising inhibitors exhibiting 0.71. Mu.M to 12.27. Mu.M IC in a recombinant inhibition assay of enterovirus EV-A71 and CVB3 and coronavirus SARS-CoV and NL63 proteases ₅₀ Values (Zhang et al 2020.Journal of Medicinal Chemistry 63 (9): 4562-4578). The observed activity in the recombinant protease assay approximately matches the antiviral activity observed in the cell-based live virus assay, IC in both systems ₅₀ Values are within 10-fold, indicating that good activity in protease inhibition assays is a good indicator of antiviral activity.

Currently, there are no targeted therapeutic agents available for the treatment of covd-19, and effective therapeutic options remain very limited. Despite the extensive research activities being conducted and the extensive clinical trials being conducted, only remdesivir and favipiravir are approved in selected countries for limited use in the treatment of SARS-CoV-2 infection, but have limited efficacy (Zhou et al ACS Pharmacology & Translational Science 3 (5): 813-834). There is a need for targeted therapeutic agents for the treatment of SARS-CoV-2 infection, for which reason SARS-CoV-2 Mpro represents an attractive drug target for SARS-CoV-2. The compounds disclosed herein have been shown to be inhibitors of SARS-CoV-2 Mpro and thus represent potential candidates for the treatment of coronavirus infections and related disorders, including but not limited to COVID-19.

Disclosure of Invention

The present invention provides compounds having activity as SARS-CoV-2:Mpro inhibitors.

Provided are compounds of formula (1') or salts thereof:

wherein;

a is selected from:

q is CN or a group of the formula:

x is C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated or unsaturated hydrocarbon radicals, or X and R ⁹ To form optionally 1 to 3 fluorine atoms or 1 to 3C _1-3 Alkyl substituted C _3-6 A cycloalkyl ring;

y is O or NOR ¹⁶ ；

T ¹ Is CR (CR) ⁸ Or N;

T ² is CR (CR) ⁷ Or N;

T ³ is CR (CR) ⁶ Or N;

T ⁴ is CR (CR) ⁵ Or N;

T ⁵ is CR (CR) ⁴ Or N;

z is a 5-or 6-membered heterocyclic ring optionally substituted with oxo or with 1 to 6 fluorine atoms, or Z is C optionally substituted with 1 to 6 fluorine atoms _3-6 Cycloalkyl, or Z is- (CH) ₂ ) _p CONHR ¹³ Or Z is- (CH) ₂ ) _p CO ₂ R ¹³ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

R ¹ And R is ^1a Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Are linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms;

R ² and R is ³ Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group;

R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ and R is ⁸ Independently H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group;

R ⁹ is H or is linked to X to form a group optionally containing 1 to 3 fluorine atoms or 1 to 3C _1-3 Alkyl substituted C _3-6 A cycloalkyl ring;

R ¹¹ and R is ¹² Independently H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group; or R is ¹¹ And R is ¹² To form a cyclopropyl ring;

R ¹³ 、R ¹⁴ and R is ¹⁵ Independently H, or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group;

R ¹⁶ is H or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group;

p and m are independently 0 to 3;

wherein when R is ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

-CR ¹¹ ＝CR ¹² -, wherein R is ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The non-uniformity is H;

or-O-CHR ¹¹ -, wherein R is ³ Other than H, or where R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

The compounds of the invention are useful as SARS-CoV-2:Mpro inhibitors. The compounds of the invention are useful for treating SARS-CoV-2 and related viruses or diseases or conditions associated with SARS-CoV-2:Mpro. The compounds of the present invention are useful for preventing death or complications from chronic underlying diseases or complications in patients infected with SARS-CoV-2 and related viruses. Such chronic underlying diseases or complications may include, for example, hypertension, obesity, chronic pulmonary diseases (TB, asthma and cystic fibrosis), diabetes and cardiovascular diseases (coronary heart disease, congenital heart disease and heart failure). The compounds of the invention are useful for the preparation of medicaments. These compounds or medicaments are useful for treating, preventing, ameliorating, controlling or reducing the risk of SARS-CoV-2 and related viruses and diseases or conditions involving SARS-CoV-2:Mpro. These compounds or medicaments are useful for treating, preventing, ameliorating, controlling or reducing the risk of chronic underlying diseases or complications in patients infected with SARS-CoV-2 and related viruses.

The compounds of the present invention may be used as a single agent or in combination with one or more additional agents. The compounds of the invention are useful for treating SARS-CoV-2 and related viruses or conditions or symptoms associated therewith.

Detailed description of the invention

The present invention relates to novel compounds. The invention also relates to the use of the novel compounds as inhibitors of SARS-CoV-2:Mpro. The invention also relates to the use of the novel compounds in the preparation of a medicament for the inhibition of SARS-CoV-2:Mpro. The invention also relates to compounds, compositions and medicaments useful for treating SARS-CoV-2 and related viruses or conditions or symptoms associated therewith.

Provided are compounds of formula (1') or salts thereof:

wherein;

a is selected from:

q is CN or a group of the formula:

x is C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated or unsaturated hydrocarbon radicals, or X and R ⁹ To form optionally 1 to 3 fluorine groupsAtoms or 1 to 3C _1-3 Alkyl substituted C _3-6 A cycloalkyl ring;

y is O or NOR ¹⁶ ；

T ¹ Is CR (CR) ⁸ Or N;

T ² is CR (CR) ⁷ Or N;

T ³ is CR (CR) ⁶ Or N;

T ⁴ is CR (CR) ⁵ Or N;

T ⁵ is CR (CR) ⁴ Or N;

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

p and m are independently 0 to 3;

wherein when R is ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

Also provided are compounds of formula (1 b) or salts thereof:

wherein;

q is CN or a group of the formula:

y is O or NOR ¹⁶ ；

Z is a 5-or 6-membered heterocyclic ring optionally substituted by oxo or by 1 to 6 fluorine atoms, or Z is optionally substituted by 1 to 6Fluorine atom substituted C _3-6 Cycloalkyl, or Z is- (CH) ₂ ) _p CONHR ¹³ Or Z is- (CH) ₂ ) _p CO ₂ R ¹³ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ and R is ⁸ Independently H, halogen, CO ₂ R ¹⁴ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group;

R ¹¹ and R is ¹² Independently H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group;

R ¹³ 、R ¹⁴ and R is ¹⁵ Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group;

p and m are independently 0 to 3;

wherein when R is ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

-CR ¹¹ ＝CR ¹² -wherein R is ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The non-uniformity is H;

or-O-CHR ¹¹ -wherein R is ³ Other than H, or where R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

Also provided are compounds of formula (1 c) or salts thereof:

therein Z, Q, X, L, T ¹ 、R ² 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁹ As defined herein.

Also provided are compounds of formula (1) or salts thereof:

wherein;

y is O or NOR ¹⁶ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ ；

R ¹ Is H or C optionally substituted by 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon groups;

p and m are independently 0 to 3;

Wherein when R is ¹ Is C optionally substituted by 1 to 6 fluorine atoms _1-6 When saturated hydrocarbon groups, L is:

-CHR ¹¹ -CHR ¹² -；

Provided are compounds of formula (1 i):

wherein:

x is C optionally substituted with 1 to 6 fluorine atoms _1-8 Saturated or unsaturated hydrocarbon radicals, or X and R ⁹ To form optionally 1 to 3 fluorine atoms or 1 to 3C _1-3 Alkyl substituted C _3-6 A cycloalkyl ring;

y is O or NOR ¹⁶ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

p and m are independently 0 to 3.

Also provided are compounds of formulae (1 ' a), (1 ' ai) and (1 ' aii) and salts thereof:

therein A, X, Z, Y, L, R ¹ 、R ^1a 、R ² 、R ³ And R is ⁹ As defined herein.

Also provided are compounds of formulae (1 ' aa), (1 ' aai) and (1 ' aaii) and salts thereof:

Also provided are compounds of formulae (1 ca), (1 cai) and (1 caii) and salts thereof:

therein X, Z, Y, L, T ¹ 、R ¹ 、R ^1a 、R ² 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁹ As defined herein.

Also provided are compounds of formulae (1 caa), (1 caai) and salts thereof:

Also provided are compounds of formulae (1 a), (1 ai) and (1 aii) and salts thereof:

therein X, Z, Y, L, R ¹ 、R ^1a 、R ² 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ And R is ⁹ As defined herein.

Also provided are compounds of formulae (2 a), (2 ai) and (2 aii) and salts thereof:

therein X, Z, L, R ¹ 、R ^1a 、R ² 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (3 a), (3 ai) and (3 aii) and salts thereof:

therein Z, L, R ¹ 、R ^1a 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (3 b), (3 bi) and (3 bii) and salts thereof:

wherein the method comprises the steps of

Y is O or NOH;

z is a 5-or 6-membered heterocyclic ring optionally substituted with oxo or with 1 to 6 fluorine atoms, or Z is C optionally substituted with 1 to 6 fluorine atoms _3-6 Cycloalkyl;

l is-CH=CH-, -CH ₂ -CH ₂ -or-O-CH ₂ -；

R ¹ And R is ^1a Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-4 Saturated hydrocarbon groups;

R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ and R is ⁸ Independently H or halogen.

Also provided are compounds of formulae (3 c), (3 ci) and (3 cii) and salts thereof:

/>

wherein the method comprises the steps of

Y is O or NOH;

l is-CH=CH-, -CH ₂ -CH ₂ -or-O-CH ₂ -；

R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ and R is ⁸ Independently H or halogen.

Also provided are compounds of formulae (4 a), (4 ai) and (4 aii) and salts thereof:

/>

therein X, L, R ¹ 、R ^1a 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (5 a) and (5 ai) and salts thereof:

therein X, Z, L, R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (6 a) and (6 ai) and salts thereof:

therein X, Z, R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (7 a) and (7 ai) and salts thereof:

therein Z, R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (8 a) and (8 ai) and salts thereof:

therein X, R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (9 a), (9 ai) and (9 b) and salts thereof:

/>

Wherein R is ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

Also provided are compounds of formulae (2 av) to (9 bv) and salts thereof:

/>

therein X, Z, Y, L, R ¹ 、R ² 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

In the compounds herein, Q may be CN. Q may be a group of the formula:

q may be selected from the group consisting of:

q may beQ may be->

In the compounds herein, a may be selected from:

wherein T is ¹ 、T ² 、T ³ 、T ⁴ 、T ⁵ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ As defined herein.

A may be selected from:

in the compounds herein, T ¹ Can be CR ⁸ Or N. T (T) ¹ Can be CR ⁸ 。T ¹ May be N.

In the compounds herein, T ² Can be CR ⁷ Or N. T (T) ² Can be CR ⁷ 。T ² May be N.

In the compounds herein, T ³ Can be CR ⁶ Or N. T (T) ³ Can be CR ⁶ 。T ³ May be N.

In the compounds herein, T ⁴ Can be CR ⁵ Or N. T (T) ⁴ May beCR ⁵ 。T ⁴ May be N.

In the compounds herein, T ⁵ Can be CR ⁴ Or N. T (T) ⁵ Can be CR ⁴ 。T ⁵ May be N.

In the compounds herein, X is C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated or unsaturated hydrocarbon radicals, or X and R ⁹ To form optionally 1 to 3 fluorine atoms or 1 to 3C _1-3 Alkyl substituted C _3-6 Cycloalkyl rings. X may be C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated or unsaturated hydrocarbon groups. X may be C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, branched C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, C optionally substituted by 1 to 6 fluorine atoms _3-6 Cycloalkyl, C optionally substituted with 1 to 6 fluorine atoms _3-6 Cycloalkenyl, -CH optionally substituted with 1 to 6 fluorine atoms ₂ -C _3-5 Cycloalkyl or-CH optionally substituted with 1 to 6 fluorine atoms ₂ -C _3-5 A cycloalkenyl group. X may be C _1-6 Alkyl or branched C _1-6 An alkyl group. X may be selected from-CH ₂ CHF ₂ Isobutyl, neopentyl, -CH ₂ -cyclobutyl, cyclobutylmethyl, -CH ₂ -cyclopropyl, cyclopropylmethyl, -CH ₂ -difluorocyclobutyl, -CH ₂ -bicyclo [1.1.1]Amyl, -CH ₂ Cyclopentenyl groups. X may be-CH ₂ -cyclopropyl. X may be cyclopropylmethyl.

X may be the same as R ⁹ To form a group optionally containing 1 to 3 fluorine atoms or 1 to 3C atoms _1-3 Alkyl substituted C _3-6 Cycloalkyl rings. X may be the same as R ⁹ To form a group optionally containing 1 to 3 fluorine atoms or 1 to 3C atoms _1-3 An alkyl-substituted cyclobutyl ring. X may be the same as R ⁹ To form a cyclobutyl ring optionally substituted by 1 to 3 methyl groups. X may be the same as R ⁹ Are connected to form

X may be selected from the group consisting of:

x may beX may be +.>

In the compounds herein, R ⁹ Is H or is linked to X to form a group optionally containing 1 to 3 fluorine atoms or 1 to 3C _1-3 Alkyl substituted C _3-6 Cycloalkyl rings. R is R ⁹ May be H. R is R ⁹ Can be linked to X to form a cyclobutyl ring, optionally containing 1 to 3 fluorine atoms or 1 to 3C atoms _1-3 Alkyl substitution. R is R ⁹ May be linked to X to form a cyclobutyl ring optionally substituted by 1 to 3 methyl groups. R is R ⁹ Can be connected with X to form

In the compounds herein, Z may be a 5-or 6-membered heterocyclic ring optionally substituted with oxo or with 1 to 6 fluorine atoms, and Z may be C optionally substituted with 1 to 6 fluorine atoms _3-6 Cycloalkyl, Z can be- (CH) ₂ ) _p CONHR ¹³ Z may be- (CH) ₂ ) _p CO ₂ R ¹³ . Z may be a pyrrolidine ring optionally substituted with oxo or with 1 to 6 fluorine atoms. Z may be a pyrrolidone ring optionally substituted with 1 to 6 fluorine atoms. Z may be a 2-pyrrolidone ring optionally substituted with 1 to 6 fluorine atoms. Z may be a 2-pyrrolidone ring. Z may be selected from pyrrolidone, cyclopentane, cyclopropane, pyridine, -CH ₂ CONH ₂ And pyrazoles. Z may be pyrrolidone. Z may be 2-pyrrolidone.

Z may be selected from the group consisting of:

z may beZ may be->

In the compounds herein, p may be 0-3.p may be 0, 1, 2 or 3.p may be 0.p may be 1.p may be 2.p may be 3.

In the compounds herein, R ¹³ 、R ¹⁴ And R is ¹⁵ May independently be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group.

In the compounds herein, R ¹³ May be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹³ May be H or methyl. R is R ¹³ May be H.

In the compounds herein, R ¹⁴ May be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹⁴ May be H or methyl. R is R ¹⁴ May be H.

In the compounds herein, R ¹⁵ May be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹⁵ May be H or methyl. R is R ¹⁵ May be H.

In the compounds herein, Y may be O or NOR ¹⁶ . Y may be O. Y may be NOR ¹⁶ . Y may be NOH.

In the compounds herein, R ¹⁶ Is H or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹⁶ May be H.

In the compounds herein, L is selected from-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -and-O-CHR ¹¹ -a linking group. L may be-CH=CH-, -CH ₂ CH ₂ -、-CH ₂ -CH(CH ₂ CH ₃ )-、-CH ₂ -CH(CH ₂ CO ₂ H)-、-CH ₂ -CH(CH ₃ )-、-CH ₂ -CH(CF ₃ )-、-OCH ₂ -、-OCH(CH ₃ ) -or

L may be-CH=CH-, -CH ₂ CH ₂ -、-CH ₂ -CH(CH ₂ CH ₃ )-、-CH ₂ -CH(CH ₂ CO ₂ H) -or-OCH ₂ -. L may be-CHR ¹¹ -CHR ¹² -. When R is ¹ And R is ^1a When both are H, L may be-CR ¹¹ ＝CR ¹² -. When R is ¹ And R is ^1a When the unevenness is H, L may be-CR ¹¹ ＝CR ¹² -, wherein R is ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The unevenness is H. When R is ¹ And R is ^1a When the non-uniformity is H, L may be-O-CHR ¹¹ -. When R is ¹ And R is ^1a When the non-uniformity is H, L may be-O-CHR ¹¹ -wherein R is ³ Other than H, or wherein R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

L may be selected from the group consisting of:

Unless otherwise indicated, the L groups defined herein may take any possible orientation with respect to their point of attachment to the remainder of the molecule. For example, wherein embodiments 82 and 91 include L defined as-OCH ₂ -, where examples 86 and 88 include L being defined as-CH ₂ -CH(CF ₃ ) (Table 1).

In the compounds herein, R ¹¹ And R is ¹² Can be independently H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 Alkyl groupThe method comprises the steps of carrying out a first treatment on the surface of the Or R is ¹¹ And R is ¹² May be linked to form a cyclopropyl ring. R is R ¹¹ And R is ¹² Can be independently H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group; or R is ¹¹ And R is ¹² May be linked to form a cyclopropyl ring. R is R ¹¹ And R is ¹² Can be independently H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹¹ And R is ¹² Can be independently H, methyl, ethyl or-CH ₂ CO ₂ H。R ¹¹ Can be H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹¹ Can be H, methyl, ethyl or-CH ₂ CO ₂ H。R ¹² Can be H, - (CH) ₂ ) _m CO ₂ R ¹⁵ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ¹² Can be H, methyl, ethyl or-CH ₂ CO ₂ H。R ¹¹ Can be combined with R ¹² To form a cyclopropyl ring. R is R ¹² Can be combined with R ¹¹ To form a cyclopropyl ring. R is R ¹¹ Can be combined with R ¹² To form a cyclopropyl ring. R is R ¹² Can be combined with R ¹¹ To form a cyclopropyl ring.

In the compounds herein, m may be 0 to 3.m may be 0, 1, 2 or 3.m may be 0.m may be 1.m may be 2.m may be 3.p and m may independently be 0-3.

In the compounds herein, R ¹ And R is ^1a May independently be H or C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a May be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ¹ And R is ^1a C which may independently be H, optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, branched C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, C optionally substituted by 1 to 6 fluorine atoms _3-6 Cycloalkyl or optionally is covered by1 to 6 fluorine atom substituted-CH ₂ -C _3-6 Cycloalkyl groups. R is R ¹ And R is ^1a Can be independently C _1-6 Alkyl, branched C _1-6 Alkyl, C _3-6 Cycloalkyl or-CH ₂ -C _3-6 Cycloalkyl groups. R is R ¹ And R is ^1a Can be independently selected from H, methyl, ethyl, isopropyl, cyclopropyl, tert-butyl, isobutyl, cyclopropylmethyl and-CH ₂ And (3) cyclopropyl. R is R ¹ And R is ^1a May be H. R is R ¹ Can be combined with R ^1a To form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ¹ Can be combined with R ^1a To form a 3-to 6-membered saturated ring. R is R ¹ Can be combined with R ^1a To form an aziridine ring. R is R ¹ Can be combined with R ^1a To form an azetidine ring. R is R ¹ Can be combined with R ^1a To form a pyrrolidine ring. R is R ¹ Can be combined with R ^1a To form a piperidine ring.

In the compounds herein, R ¹ May be H or C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a May be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ¹ May be C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, branched C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, C optionally substituted by 1 to 6 fluorine atoms _3-6 Cycloalkyl, or-CH optionally substituted with 1 to 6 fluorine atoms ₂ -C _3-6 Cycloalkyl groups. R is R ¹ May be C _1-6 Alkyl, branched C _1-6 Alkyl, C _3-6 Cycloalkyl or-CH ₂ -C _3-6 Cycloalkyl groups. R is R ¹ Can be selected from H, methyl, ethyl, isopropyl, cyclopropyl, tert-butyl, isobutyl, cyclopropylmethyl and-CH ₂ And (3) cyclopropyl. R is R ¹ May be H. R is R ¹ Can be cyclopropyl. R is R ¹ Can be combined with R ^1a To form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ¹ Can be combined with R ^1a To form a 3-to 6-membered saturated ring. R is R ¹ Can be combined with R ^1a Linking to form an aziridine ring。R ¹ Can be combined with R ^1a To form an azetidine ring. R is R ¹ Can be combined with R ^1a To form a pyrrolidine ring. R is R ¹ Can be combined with R ^1a To form a piperidine ring.

In the compounds herein, R ^1a May be H or C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a May be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ^1a May be C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, branched C optionally substituted with 1 to 6 fluorine atoms _1-6 Alkyl, C optionally substituted by 1 to 6 fluorine atoms _3-6 Cycloalkyl, or-CH optionally substituted with 1 to 6 fluorine atoms ₂ -C _3-6 Cycloalkyl groups. R is R ^1a May be C _1-6 Alkyl, branched C _1-6 Alkyl, C _3-6 Cycloalkyl or-CH ₂ -C _3-6 Cycloalkyl groups. R is R ^1a Can be selected from H, methyl, ethyl, isopropyl, cyclopropyl, tert-butyl, isobutyl, cyclopropylmethyl and-CH ₂ -cyclopropyl. R is R ^1a May be H. R is R ^1a Can be combined with R ¹ To form a 3-to 6-membered saturated ring optionally containing additional heteroatoms. R is R ^1a Can be combined with R ¹ To form a 3-to 6-membered saturated ring. R is R ^1a Can be combined with R ¹ To form an aziridine ring. R is R ^1a Can be combined with R ¹ To form an azetidine ring. R is R ^1a Can be combined with R ¹ To form a pyrrolidine ring. R is R ^1a Can be combined with R ¹ To form a piperidine ring.

When R is ¹ And R is ^1a When joined together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms, the optional additional heteroatoms may be selected from N, O, S and oxidized forms thereof.

R ¹ And R is ^1a May independently be H or selected from the group consisting of:

or R is ¹ And R is ^1a May be linked to form a ring such that the group NR ¹ R ^1a The method comprises the following steps:

R ¹ may be H or may be selected from the group consisting of:

R ^1a may be H or may be selected from the group consisting of:

in the compounds herein, R ² And R is ³ Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ² And R is ³ Can be independently H, methyl or CH ₂ CF ₃ 。R ² And R is ³ May be H. R is R ¹ 、R ² And R is ³ May be H.

In the compounds herein, R ² May be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ² May be H. R is R ¹ And R is ² May be H.

In the compounds herein, R ³ May be H or C optionally substituted with 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ³ Can be H, methyl or CH ₂ CF ₃ 。R ³ May be H.

In the compounds herein, R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Independently H, halogenPlain, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Can be independently selected from H, halogen, CO ₂ R ¹⁴ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Can be independently selected from H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Can be independently selected from H, cl, F, CF ₂ H and CO ₂ H。R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Can be independently selected from H, cl, F and CO ₂ H。R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ May be H. R is R ⁴ May be F, R ⁵ May be H, R ⁶ Can be Cl, and R ⁷ And R is ⁷ May be H.

R ⁴ Can be selected from H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁴ Can be selected from H, cl, F, CF ₃ 、CF ₂ H and CO ₂ H。R ⁴ May be H.

R ⁵ Can be selected from H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁵ Can be selected fromSelf H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。R ⁵ Can be selected from H, cl, F, CF ₃ 、CF ₂ H and CO ₂ H。R ⁵ May be H.

R ⁶ Can be selected from H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁶ Can be selected from H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。R ⁶ Can be selected from H, cl, F, CF ₃ 、CF ₂ H and CO ₂ H。R ⁶ May be H.

R ⁷ Can be selected from H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁷ Can be selected from H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。R ⁷ Can be selected from H, cl, F, CF ₃ 、CF ₂ H and CO ₂ H。R ⁷ May be H.

R ⁸ Can be selected from H, halogen, CN, CO ₂ R ¹⁴ 、OR ¹⁴ 、SO ₂ R ¹⁴ 、SONHR ¹⁴ 、OSO ₂ R ¹⁴ 、PO(R ¹⁴ ) ₂ 、SF ₅ Or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group. R is R ⁸ Can be selected from H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。R ⁸ Can be selected from H, cl, F, CF ₃ 、CF ₂ H and CO ₂ H。R ⁸ May be H.

In the compounds herein, the a moiety may be selected from the group consisting of:

in the compounds herein, the following moieties:

may be selected from the group consisting of:

in the compounds herein, when R ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

For those wherein L is-CHR ¹¹ -CHR ¹² -a compound, R ¹ And R is ^1a C which may independently be H, optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a May be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms.

For those wherein L is-CHR ¹¹ -CHR ¹² -a compound, R ¹ And R is ^1a May be H.

For those wherein L is-CHR ¹¹ -CHR ¹² -a compound, R ¹ And R is ^1a May be C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a May be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms.

For those wherein L is-CR ¹¹ ＝CR ¹² -a compound, R ¹ And R is ^1a C which may independently be H, optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Can be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms, provided that R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The unevenness is H.

For those wherein L is-CR ¹¹ ＝CR ¹² -a compound, R ¹ And R is ^1a May be H.

For those wherein L is-CR ¹¹ ＝CR ¹² -a compound, R ¹ And R is ^1a May independently be C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Can be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms, provided that R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The unevenness is H.

For those wherein L is-O-CHR ¹¹ -a compound, R ¹ And R is ^1a C which may independently be H, optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Can be linked together to form a composition optionally comprising an additional3 to 6 membered saturated rings of heteroatoms of (2), provided that R ³ Is other than H, or provided that R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

For those wherein L is-O-CHR ¹¹ -a compound, R ¹ And R is ^1a May be H.

For those wherein L is-O-CHR ¹¹ -a compound, R ¹ And R is ^1a Can independently be C optionally substituted with 1 to 6 fluorine atoms _1-6 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Can be linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms, provided that R ³ Is other than H, or provided that R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

The compound may be selected from any one of the compounds of examples 1 to 93 shown in table 1 or a salt thereof.

The compound may be a compound selected from the group consisting of:

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

3- ((S) -2-cinnamamido-3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

benzyl (2S) -1- ((4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamate;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2- (hydroxyimino) -4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

n- (tert-butyl) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -4-cyclopropyl-2-oxobutanamide;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -4-cyclopentyl-2-oxobutanamide;

3- ((S) -3-cyclopropyl-2- (3-phenylpropionamido) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

(2S) -N- (1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylbutyramide) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylbutyramide) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylpentanamido) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylpentanamido) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -4, 4-trifluoro-3-phenylbutyramide) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -4, 4-trifluoro-3-phenylbutyramide) pentanamide;

(2S) -N- (4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -4-methylpentanamide;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -N-ethyl-2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

3- ((S) -3-cyclopropyl-2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -N-cyclopropyl-2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

(2S) -N- (4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -N- (4- (ethylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

3- ((S) -3-cyclopropyl-2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) propanamido) -N-ethyl-2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

n-cyclopropyl-3- ((S) -3-cyclopropyl-2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

(2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -N- (4- (ethylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide;

benzyl (((2S) -3-cyclopropyl-1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -1-oxopropan-2-yl) carbamate;

n-cyclopropyl-3- ((S) -3-cyclopropyl-2- (3- (2, 4-dichlorophenyl) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide;

3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclobutylpropionamido) -N-cyclopropyl-2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butyramide;

(3S) -N- ((2S) -3-cyclopropyl-1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -1-oxopropan-2-yl) -3-phenylpentanamide;

(3R) -N- ((2S) -3-cyclopropyl-1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -1-oxopropan-2-yl) -3-phenylpentanamide;

(2S) -N- (4- (azetidin-1-yl) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-difluorophenyl) acrylamide) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chlorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (2-chloro-4-fluorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (4-fluorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chloro-3-fluorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (5-chloropyridin-2-yl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (5-fluoropyridin-2-yl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (4- (difluoromethyl) phenyl) acrylamido) -4, 4-dimethylpentanamide;

(1 r,2 r) -N- ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -2-phenylcyclopropane-1-carboxamide;

(1S, 2S) -N- ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) -2-phenylcyclopropane-1-carboxamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (2, 4-difluorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (2, 4-difluorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (2, 4-dichlorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (2, 4-dichlorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -2- ((S) -3- (4-chloro-2-fluorophenyl) pentanoylamino) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) - (2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanoamide;

(2S) -2- ((R) -3- (4-chloro-2-fluorophenyl) pentanoylamino) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanoamide;

(2S) -2- ((S) -3- (4-chlorophenyl) valeramido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((R) -3- (4-chlorophenyl) valeramido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (4-fluorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (4-fluorophenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (azetidin-1-yl) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3- (2- (trifluoromethoxy) phenyl) pentanamido) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3- (2- (trifluoromethoxy) phenyl) pentanamido) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (2- (difluoromethoxy) phenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S, -3- (2- (difluoromethoxy) phenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chloro-2-cyanophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (4-chloro-3-cyanophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((E) -3- (2-chloro-4-cyanophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3- (3- (trifluoromethoxy) phenyl) butyrylamide) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3- (3- (trifluoromethoxy) phenyl) butyrylamide) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (3- (difluoromethoxy) phenyl) butyramide) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (3- (difluoromethoxy) phenyl) butyrylamino) -4, 4-dimethylpentanamide;

(2S) -2- ((R) -3- (2-chloro-4- (methylsulfonyl) phenyl) pentanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((S) -3- (2-chloro-4- (methylsulfonyl) phenyl) pentanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamido;

3-chloro-4- ((3R) -1- (((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -1-oxopent-3-yl) phenyl methanesulfonate;

3-chloro-4- ((3S) -1- (((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) amino) -1-oxopent-3-yl) phenyl methanesulfonate;

(2S) -2- ((R) -3- (2-chloro-4- (dimethylphosphoryl) phenyl) pentanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((S) -3- (2-chloro-4- (dimethylphosphoryl) phenyl) pentanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((E) -3- (4- (pentafluoro-l 6-sulfanyl) phenyl) acrylamido) pentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (4-methoxyphenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (4-methoxyphenyl) pentanamido) -4, 4-dimethylpentanamide;

(2S) -2- (3- (4-chloro-2- (trifluoromethoxy) phenyl) propanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- (3- (4-chloro-2- (difluoromethoxy) phenyl) propanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- (3- (4-chloro-2-cyanophenyl) propanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- (3- (2-chloro-4-cyanophenyl) propanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- (3- (4-chloro-3- (trifluoromethoxy) phenyl) propanamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -N- (1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- (3- (2, 4-dichlorophenyl) propanamido) -4, 4-dimethylpentanamide;

2, 4-dichlorobenzyl ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate;

4-chloro-2-fluorobenzyl ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate;

(S) -1- (4-chloro-2-fluorophenyl) ethyl ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate;

(R) -1- (4-chloro-2-fluorophenyl) ethyl ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) carbamate;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -3- (2, 4-difluorophenyl) -4, 4-trifluorobutyramide) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (2, 4-difluorophenyl) -4, 4-4-trifluorobutyramide) -4, 4-dimethylpentanamide;

(2S) -2- ((R) -2-benzyl-3, 3-trifluoropropionamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(2S) -2- ((S) -2-benzyl-3, 3-trifluoropropionamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide;

(1S, 2S) -2- (4-chlorophenyl) -N- ((2S) -1- ((4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -4, 4-dimethyl-1-oxopent-2-yl) cyclopropane-1-carboxamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- (2, 4-dichlorophenoxy) acetamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -2- (2, 4-dichlorophenoxy) propanamido) -4, 4-dimethylpentanamide;

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((R) -2- (2, 4-dichlorophenoxy) propanamido) -4, 4-dimethylpentanamide.

The compounds of the invention may be present in the form of prodrugsAt the point. By "prodrug" is meant, for example, any compound that is converted in vivo to a biologically active compound of the invention. For example, some prodrugs are esters or phosphates (e.g., physiologically acceptable metabolically labile esters) of the active compound. During metabolism, an ester group (-C (=o) OR a phosphate group (P (=o (OH) ₂ -OR) is cleaved to yield the active agent. Such esters may be formed by esterification, for example, where appropriate, of the hydroxyl groups present in the parent compound with any other pre-protected reactive groups present in the parent compound, followed by deprotection as desired. Other functional groups present in the active compound, such as amide groups or amino groups, may be used to form the prodrug. In addition, some prodrugs are enzymatically activated to produce the active compound, or compounds that produce the active compound in a further chemical reaction (e.g., as in ADEPT, GDEPT, LIDEPT, etc.). For example, the prodrug may be a sugar derivative or other glycoside conjugate, or may also be an amino acid ester derivative.

Thus, there is provided a prodrug of a compound as defined herein, wherein the compound comprises a functional group which can be converted under physiological conditions to form a hydroxyl, amide or amino group. Suitable prodrugs may, for example, include compounds wherein the Z group is functionalized with a group that is cleaved in vivo to release the active compound.

For example, in prodrugs of compounds, Z may be the following group:

wherein R is ^P Is any group that can be cleaved in vivo to provide a compound in which Z is:

R ^P can be selected from:

for example, in prodrugs of compounds, Z may be selected from:

further embodiments include the use of a compound of the invention or a salt thereof or a pharmaceutical composition comprising a compound of the invention as a SARS-CoV-2:Mpro inhibitor. The compounds of the invention are useful as SARS-CoV-2:Mpro inhibitors. The compounds of the invention are useful for treating SARS-CoV-2 or a disease or condition associated with SARS-CoV-2:Mpro. The compounds of the invention are useful for preventing death or complications arising from chronic underlying diseases or complications in patients infected with SARS-CoV-2. Such chronic underlying diseases or complications may include, for example, hypertension, obesity, chronic pulmonary diseases (TB, asthma and cystic fibrosis), diabetes and cardiovascular diseases (coronary heart disease, congenital heart disease and heart failure). The compounds of the invention are useful for the preparation of medicaments. The compounds or medicaments are useful for treating, preventing, ameliorating, controlling or reducing the risk of diseases or conditions in which SARS-CoV-2 and SARS-CoV-2:Mpro are involved. The compounds or medicaments are useful for treating, preventing, ameliorating, controlling or reducing the risk of chronic underlying diseases or complications in patients suffering from SARS-CoV-2 infection.

The compounds of the present application may be used as a single agent or in combination with one or more additional agents. The compounds of the application are useful for treating SARS-CoV-2 or a disorder or condition associated therewith.

As provided herein, the compounds described herein or salts thereof, and the compositions described herein may be administered with agents that treat any of the diseases and conditions disclosed herein.

Definition of the definition

In the present application, the following definitions apply unless otherwise indicated.

The term "SARS-CoV-2:Mpro inhibitor" as used herein refers to any compound that binds to and modulates SARS-CoV-2:Mpro function.

The term "treatment" in connection with the use of any of the compounds described herein, including compounds of formula (1 b), is used to describe any form of intervention in which the compound is administered to a subject suffering from, or at risk of suffering from, or at potential risk of suffering from, the disease or disorder. Thus, the term "treatment" includes prophylactic (preventative) treatment and treatment of measurable or detectable symptoms of a disease or disorder.

The term "therapeutically effective amount" (e.g., in connection with a method of treating a disease or disorder) refers to an amount of a compound that is effective to produce a desired therapeutic effect. For example, if the condition is pain, then the therapeutically effective amount is an amount sufficient to provide the desired degree of pain relief. The desired degree of pain relief may be, for example, complete elimination of pain or a reduction in the severity of pain.

Unless otherwise indicated, terms such as "bicyclic", "hydrocarbon", "heterocyclic", "carbocycle", "alkyl", "cycloalkyl" and "halogen" are used in their conventional sense (as defined in IUPAC Gold Book). "optionally substituted with …" for any group means that the group may be substituted with one or more substituents, which may be the same or different, if desired.

To the extent that any of the compounds described have chiral centers, the scope of applicability of the invention extends to all optical isomers of those compounds, whether in racemic form or as resolved enantiomers. The invention described herein relates to all crystalline forms, solvates and hydrates of any of the disclosed compounds, regardless of how prepared. To the extent that any of the compounds disclosed herein have an acid or base center, such as a carboxylate or amino group, all salt forms of the compounds are included herein. In the case of pharmaceutical use, the salt should be considered as a pharmaceutically acceptable salt.

Salts or pharmaceutically acceptable salts that may be mentioned include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reacting the free acid or free base form of the compound with one or more equivalents of a suitable acid or base, optionally in a solvent, or in a salt-insoluble medium, and then removing the solvent or medium using standard techniques (e.g. in vacuo, by freeze drying or filtration). Salts may also be prepared by exchanging the counter ion of the compound in salt form with another counter ion, for example using a suitable ion exchange resin.

Examples of pharmaceutically acceptable salts include acid addition salts derived from inorganic and organic acids, and salts derived from metals such as sodium, magnesium, potassium and calcium.

Examples of acid addition salts include acid addition salts formed with the following acids: acetic acid, 2-dichloroacetic acid, adipic acid, alginic acid, arylsulfonic acid (e.g., benzenesulfonic acid, 2-naphthalenesulfonic acid, 1, 5-naphthalenedisulfonic acid, and p-toluenesulfonic acid), ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzoic acid, 4-acetamidobenzoic acid, butyric acid, (+) camphoric acid, camphorsulfonic acid, (+) - (1S) -camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexylsulfamic acid, dodecylsulfuric acid, 1, 2-ethanedisulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid (e.g., D-gluconic acid), glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), alpha-oxoglutarate, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isethionic acid, lactic acid (e.g., (+) -L-lactic acid and (±) -DL-lactic acid), lactobionic acid, maleic acid, malic acid (e.g., (-) -L-malic acid), malonic acid, (±) -DL-mandelic acid, metaphosphoric acid, methanesulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid (e.g., (+) -L-tartaric acid), thiocyanic acid, undecylenic acid, and valeric acid.

Any solvate of these compounds and salts thereof is also included. Preferred solvates are those formed by incorporating into the solid state structure (e.g., crystalline structure) of the compounds of the present invention a non-toxic pharmaceutically acceptable solvent molecule (hereinafter referred to as solvating solvent). Examples of such solvents include water, alcohols (e.g., ethanol, isopropanol, and butanol), and dimethylsulfoxide. Solvates may be prepared by recrystallising the compounds of the invention from a solvent or solvent mixture containing a solvating solvent. Whether a solvate has formed in any given case can be determined by analysis of the crystals of the compound using well known standard techniques such as thermogravimetric analysis (TGA), differential Scanning Calorimetry (DSC) and X-ray crystallography.

The solvate may be a stoichiometric or non-stoichiometric solvate. The specific solvate may be a hydrate, and examples of the hydrate include a hemihydrate, a monohydrate, and a dihydrate. For a more detailed discussion of solvates and methods for preparing and characterizing them, see Bryn et al, solid-State Chemistry of Drugs, second Edition, published by SSCI, inc of West Lafayette, IN, USA,1999,ISBN0-967-06710-3.

The term "pharmaceutical composition" in the context of the present invention refers to a composition comprising an active agent and further comprising one or more pharmaceutically acceptable carriers. Depending on the mode of administration and the nature of the dosage form, the composition may further comprise an ingredient selected from the group consisting of: such as diluents, adjuvants, excipients, carriers, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, antibacterial agents, antifungal agents, lubricants, and dispersing agents. The composition may take the form: such as tablets, dragees, powders, elixirs, syrups, liquid preparations including suspensions, sprays, inhalants, tablets, troches, emulsions, solutions, cachets, granules, capsules and suppositories, and liquid preparations for injection including liposomal preparations.

The compounds of the invention may contain one or more isotopic substitutions and reference to a particular element includes within its scope all isotopes of that element. For example, references to hydrogen include within its scope ¹ H、 ² H (D) and ³ h (T). Similarly, references to carbon and oxygen include within their scope, respectively ¹² C、 ¹³ C and C ¹⁴ C and C ¹⁶ O and ¹⁸ o. In a similar manner, whenever a particular functional group is referred to, isotopic variations are also included within its scope unless the context indicates otherwise. For example, references to alkyl groups such as ethyl or alkoxy groups such as methoxy also include variants in which one or more hydrogen atoms in the group are in the deuterium or tritium isotope form, e.g., all five hydrogen atoms in the ethyl group are in the deuterium isotope form (perdeuteroethyl) or all three hydrogen atoms in the methoxy group are in the deuterium isotope form (tridecylmethoxy). Isotopes may be radioactive or non-radioactive.

The therapeutic dosage may vary depending on the requirements of the patient, the severity of the condition being treated and the compound being used. Determination of the appropriate dosage for a particular situation is within the skill of the art. Typically, treatment is initiated with a smaller dose than the optimal dose of the compound. Thereafter, the dosage is increased in small increments until the optimal effect is reached. For convenience, the total daily dose may be administered in several portions of the day, if desired.

Of course, the size of the effective dose of the compound will vary with the severity of the condition being treated and the particular compound and its route of administration. The selection of an appropriate dosage is well within the ability of one of ordinary skill in the art without undue burden. Generally, the daily dose may range from about 10 μg to about 30mg per kilogram body weight of the human and non-human animals, preferably from about 50 μg to about 30mg per kilogram body weight of the human and non-human animals, such as from about 50 μg to about 10mg per kilogram body weight of the human and non-human animals, such as from about 100 μg to about 30mg per kilogram body weight of the human and non-human animals, such as from about 100 μg to about 10mg per kilogram body weight of the human and non-human animals, and most preferably from about 100 μg to about 1mg per kilogram body weight of the human and non-human animals.

Pharmaceutical preparation

Although the active compounds may be administered alone, they are preferably present in the form of a pharmaceutical composition (e.g., formulation).

Thus, in some embodiments of the present invention, a pharmaceutical composition is provided comprising at least one compound of the present invention and at least one pharmaceutically acceptable excipient.

Pharmaceutically acceptable excipients may be selected from, for example, carriers (e.g., solid, liquid, or semi-solid carriers), adjuvants, diluents (e.g., solid diluents such as fillers or fillers; and liquid diluents such as solvents and co-solvents), granulating agents, binders, glidants, coating agents, controlled release agents (e.g., release-delaying or release-delaying polymers or waxes), binders, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, tonicity adjusting agents, thickening agents, flavoring agents, sweeteners, pigments, plasticizers, taste masking agents, stabilizers, or any other excipient conventionally used in pharmaceutical compositions.

The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g., human subject) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each excipient must also be "acceptable", i.e., compatible with the other ingredients of the formulation.

Pharmaceutical compositions containing the compounds of the invention may be formulated according to known techniques, see for example Remington's Pharmaceutical Sciences, mack Publishing Company, easton, PA, USA. The pharmaceutical composition may be in any form suitable for oral, parenteral, intravenous, intramuscular, intrathecal, subcutaneous, topical, intranasal, intrabronchial, sublingual, buccal, ocular, aural, rectal, intravaginal or transdermal administration.

Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, troches, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, wafers or patches such as oral patches.

The composition may be a tablet composition or a capsule composition. Tablet compositions may contain unit doses of the active compound in association with inert diluents or carriers, such as sugars or sugar alcohols, e.g.; lactose, sucrose, sorbitol or mannitol; and/or non-sugar derived diluents such as sodium carbonate, calcium phosphate, calcium carbonate, or cellulose or derivatives thereof such as microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, and starches such as corn starch. Tablets may also contain such standard ingredients as binders and granulating agents (e.g., polyvinylpyrrolidone), disintegrating agents (e.g., swellable crosslinked polymers such as crosslinked carboxymethylcellulose), lubricating agents (e.g., stearates), preserving agents (e.g., parabens), antioxidants (e.g., BHT), buffering agents (e.g., phosphate or citrate buffers), and effervescent agents (e.g., citrate/bicarbonate mixtures). Such excipients are well known and need not be discussed in detail herein.

Tablets may be designed to release the drug upon contact with gastric fluid (immediate release tablets), or in a controlled manner over a prolonged period of time or in a specific region of the gastrointestinal tract (controlled release tablets).

The pharmaceutical compositions generally comprise about 1% (w/w) to preferably about 95% (w/w) of the active ingredient and 99% (w/w) to 5% (w/w) of a pharmaceutically acceptable excipient (e.g., as described above) or a combination of these excipients. Preferably, the composition comprises from about 20% (w/w) to about 90% (w/w) active ingredient and from 80% (w/w) to 10% (w/w) of a pharmaceutically acceptable excipient or combination of excipients. The pharmaceutical composition comprises from about 1% to about 95%, preferably from about 20% to about 90%, of the active ingredient. The pharmaceutical compositions according to the invention may be in the form of, for example, unit dosage forms, such as ampoules, vials, suppositories, pre-filled syringes, coatings (dragees), powders, tablets or capsules.

Tablets and capsules may contain, for example, 0-20% disintegrant, 0-5% lubricant, 0-5% glidant and/or 0-99% (w/w) filler/filler (depending on the dosage of the drug). They may also contain 0-10% (w/w) of a polymeric binder, 0-5% (w/w) of an antioxidant, 0-5% (w/w) of a pigment. In addition, sustained release tablets will typically contain from 0 to 99% (w/w) of a controlled release (e.g., delayed) polymer (depending on the dosage). Film coatings for tablets or capsules typically contain 0-10% (w/w) polymer, 0-3% (w/w) pigment and/or 0-2% (w/w) plasticizer.

The composition may be a parenteral composition. Parenteral formulations typically contain 0-20% (w/w) buffer, 0-50% (w/w) co-solvent and/or 0-99% (w/w) water for injection (WFI)

(depending on the dose and whether or not it is freeze-dried). Formulations of intramuscular depots may also contain 0-99%

(w/w) oil.

The pharmaceutical formulation may be presented to the patient in a "patient pack" which is a single package (typically a blister pack) comprising the entire course of treatment.

The compounds of the invention will typically be presented in unit dosage form and, as such, will typically contain sufficient compound to provide the desired level of biological activity. For example, the formulation may contain from 1 nanogram to 2 grams of active ingredient, for example from 1 nanogram to 2 milligrams of active ingredient. Within these ranges, a particular subrange of the compound is 0.1 mg to 2 g of active ingredient (typically 10 mg to 1 g, e.g., 50 mg to 500 mg), or 1 microgram to 20 mg (e.g., 1 microgram to 10 mg, e.g., 0.1 mg to 2 mg of active ingredient).

For oral compositions, unit dosage forms may contain from 1 mg to 2 g, more typically from 10 mg to 1 g, for example from 50 mg to 1 g, for example from 100 mg to 1 g of active compound.

The active compound is administered to a patient (e.g., a human or animal patient) in need thereof in an amount (effective amount) sufficient to achieve the desired therapeutic effect. The precise amount of compound administered can be determined by the attending physician according to standard procedures.

These compounds may be administered with other drugs, for example, other drugs used to treat SARS-CoV-2 subjects. These compounds may be co-administered with HIV drugs known to block cypP 450-mediated metabolism, e.g., ritonavir or lopinavir

Combination of ritonavir.

Detailed Description

Examples

The present invention will now be described by referring to examples shown in table 1 below, but the present invention is not limited thereto.

TABLE 1

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Preparation of the Compounds of the invention

Some of the compounds of the present invention and their derivatives or synthetic intermediates may be prepared according to synthetic methods known to those skilled in the art. In some embodiments, the present invention provides methods of preparing the compounds of the present invention. Certain compounds of the invention may be prepared according to the following methods.

Preparation of the Compounds of the invention

The compounds of the present invention may be prepared by those routes included in fig. 1. For details of many standard transformations (e.g., transformations in the following routes) and other transformations that can be used to make the same, see standard reference textbooks, e.g., "Organic Synthesis", M.B.Smith, mcGraw-Hill (1994) or "Advanced Organic Chemistry",4th edition,J.March,John Wiley&Sons (1992).

Ester derivatives of alpha-amino acids (e.g., methyl ester alpha-amino acid derivatives) are commercially available or may be prepared by standard transformations known to those skilled in the art, including transformations detailed in the synthesis of the intermediates and synthesis section of the examples below. The ester derivative of the α -amino acid can be coupled with a carboxylic acid to give the corresponding amide derivative (scheme 1, step i). The amide coupling reaction conditions are generally those employing one or more coupling agents, such as propylphosphonic anhydride (T3P) or HATU, with a suitable base such as DIPEA or ET ₃ N in a solvent such as DCM or DMF, usually at room temperature. Alternatively, the ester derivative of the α -amino acid may be coupled with a carbamoyl chloride (e.g., benzyl chloroformate) and a suitable base (e.g., DIPEA) in a solvent (e.g., DCM), typically at room temperature, to form the carbamate derivative. The ester functions present in the amide-or carbamate-forming product can then be hydrolyzed under acidic or basic conditions, for example in a solvent such as THF, meOH, 1, 4-dioxane or H ₂ The solvent for O or a mixture of these solvents is used as lithium hydroxide monohydrate, usually at room temperature (scheme 1, step ii). Hydrolysis to form carboxylic acids, which can then be reacted with ester derivatives of alpha-amino acids under the amide coupling conditions described above (scheme 1, step iii). The ester functionality can be reduced to a primary alcohol using standard reducing conditions, for example using a reducing agent (e.g. sodium borohydride) in a suitable solvent (e.g. THF or MeOH) and solvent combination, typically at room temperature (scheme 1, step iv). The oxidation of primary alcohols to aldehydes can be carried out using an oxidizing agent such as Dess-martin reagent (Dess-Martin Periodinane) (DMP) in a suitable solvent such as DCM, THF or DMSO or a mixture of these solvents, typically at room temperature (scheme 1, step v). The aldehyde product can be reacted with acetone cyanohydrin in a suitable base such as ET ₃ In the presence of N, in a suitable solvent such as DCM, usually at room temperature, to give a 1-cyano-1-hydroxy derivativeOrganisms (route 1, step vi). Cyano groups can be hydrolyzed under standard conditions, for example using aqueous hydrogen peroxide in the presence of a base such as potassium carbonate, in a suitable solvent such as DMSO, typically at room temperature. The compounds of the present invention can be synthesized using a final oxidation step using the conditions described above (scheme 1, step v) (scheme 1, step viii).

Further compounds of the invention may be synthesized from other compounds of the invention. For example, the ketone group in the ketoamide functionality of the compound may be reacted with a nucleophile (e.g., hydroxylamine hydrochloride) in the presence of a suitable base (e.g., potassium carbonate) in a suitable solvent (e.g., ethanol) at elevated temperatures (e.g., 70 ℃).

In a further route, compounds of the invention may be prepared from aldehyde intermediates obtained in route 1 (including steps i) to v). The aldehyde may be reacted with an isocyanate (e.g. an alkyl isocyanate) in the presence of acetic acid in a suitable solvent (e.g. DCM), typically at room temperature (scheme 2, step i). The acetic acid group of the 1-acetoxy-1-alkylamide product of this step can be hydrolyzed under the conditions described above (scheme 1, step ii) to produce a 1-hydroxy-1-alkylamide product (scheme 3, step ii). The compounds of the present invention can be synthesized using a final oxidation step using the conditions described above (scheme 1, step v) (scheme 1, step viii).

In a further route, compounds of the invention can be prepared from the aldehyde intermediates obtained in route 1 (including steps i) to v)). The aldehyde may be reacted with hydroxylamine hydrochloride in the presence of a base such as potassium carbonate in a suitable solvent such as ethanol at elevated temperature (e.g., 70 ℃). The resulting oxime is dehydrated, for example using a dehydrating agent such as methyl N- (triethylammonium sulfonyl) carbamate (Burgess reagent), and can be used to synthesize the compound of the present invention (scheme 4, step ii).

General procedure

If the preparation route is not included, the relevant intermediate is commercially available. Commercial reagents were used without further purification. Room temperature (rt) refers to about 20 ℃ to 27 ℃. Recording on a Bruker instrument at 300 or 400MHz ¹ H NMR spectrum. Chemical shift values are reported in parts per million (ppm)The (delta) -values are shown relative to tetramethylsilane. The following abbreviations are used for multiplicity of NMR signals: s=singlet, br=broad, d=doublet, t=triplet, q=quartet, quin=quintet, h=heptad, dd=doublet, dt=doublet, m=multiplet. Coupling constants are listed as J values in Hz. NMR and mass spectral results were corrected to account for background peaks. TLC for monitoring the reaction refers to TLC operated using silica gel as a stationary phase.

LCMS experiments were performed under the following conditions. Instrument: agilent Technologies 1290Infinity II Series LC/6125Quadrupole MSD SL (methods a to E, H and K), ELSD detector (Polymer Laboratories PL-ELS2100 ICE), UV inactive compound for use in method a; column: waters XBiridge C8.5 μm,4.6x50mm (method A), atlantis dC 18.5 μm,4.6x50mm (method B), zorbax XDB C18.5 μm,4.6x50mm (method C), zorbax extension C18.5 μm,4.6x50mm (method D), XBiridge C8.5 μm,4.6x50mm (method E), acquity BEH C18.7 μm,2.1x50mm (method H), xselect CSH C18.5 μm,4.6x50mm, CSH (surface charge hybridization) (method K); gradient [ time (min)/solvent B solution A (%)]:0.0/5, 2.5/95, 4.0/95, 4.5/5, 6.0/5 (solvent a=0.1% TFA in H) ₂ O: meCN (95:5); solvent b=0.1% TFA MeCN solution (method a)), 0.0/5, 2.5/95, 4.0/95, 4.5/5, 6.0/5 (solvent a=0.1% HCO) ₂ H of H ₂ O: meCN (95:5) solution; solvent b=mecn (method B)), 0.0/5, 2.5/95, 4.0/95, 4.5/5, 6.0/5 (solvent a=0.1% hco) ₂ H of H ₂ O: meCN (95:5) solution; solvent b=mecn (method C)), 0.0/10, 4.0/95, 5.0/10, 6.0/10 (solvent a= 770.1mg NH) ₄ A solution of OAC in 1L Milli-Q water; solvent b=mecn (method D)), 0.0/10, 4.0/95, 5.0/95, 5.5/10, 7.0/10 (solvent a= 790.1mg NH) ₄ HCO ₃ A solution in 1L Milli-Q water; solvent b=mecn (method E)), 0.0/5, 0.25/5, 2.5/100, 3.0/100, 3.1/5, 4.0/5 (solvent a= 770.1mg NH) ₄ A solution of OAC in 1L Milli-Q water; solvent b=mecn (method H)), 0.0/05, 2.5/95, 4.0/95, 4.5/05, 6.0/05 (solvent a=0.1% v/v tfa in Milli-Q aqueous solution; meCN solution of solvent b=0.1%tfa) (squareMethod K)); the sample injection amount is usually 1 mu L; UV detection: 210nm to 400nm (methods A-E, H and K); column temperature 25 ℃; flow rate: 1.5mL/min (method A, B, C, K), 1.2mL/min (method D, E), or 0.8mL/min (method H). LCMS data in the experimental section are given in the following format: mass ion, retention time.

Analytical SFC experiments were performed under the following conditions. Instrument: PIC-10 (manufacturer PIC Solution Inc); column: YMC Cellulose-SC 5 μm,4.5x250mm; mobile phase: 60% CO ₂ 40% cosolvent (0.5% isopropylamine in methanol); sample injection amount: 15. Mu.L; flow rate: 4mL/min; column temperature: 35 ℃.

Mass spectrum guided preparation HPLC (Mass directed preparative HPLC) was performed under the following conditions. Instrument: agilent Technologies 1260Infinity II Series LC. Method B: column: x Bridge C8 (19 mm. Times.150 mm), 5 μm; gradient [ time (min)/solvent B solution A (%) ]:0.0/10, 15/95, 18/95, 19/10, 21/10 (solvent a=0.1% HCO) ₂ H of H ₂ An O solution; solvent b=mecn). Method F: column: xselect CSH C18 (19 mm. Times.150 mm), 5 μm; gradient [ time (min)/solvent B solution A (%)]:0.0/10, 10/50, 12/100, 16/100, 18/10, 20/10 (solvent a=0.1% hco) ₂ H of H ₂ An O solution; solvent b=mecn).

Preparative HPLC was performed under the following conditions. Method B: instrument: agilent Technologies 1260Infinity II Series LC; column: YMC Exrs C18,5 μm,30x150mm; gradient [ time (min)/solvent B solution A (%)]:0.0/10, 20/95, 23/95, 24/10, 26/10 (solvent A:0.1% HCO) ₂ H of H ₂ O solution, solvent b=mecn).

Abbreviations (abbreviations)

Dcm=dichloromethane

Dmf=n, N-dimethylformamide

DMP = dess-martin reagent

Dipea=n, N-diisopropylethylamine

DMSO = dimethyl sulfoxide

EthOAc=ethyl acetate

h=h

HATU = 1- [ bis (dimethylamino) methylene ] -1H-1,2, 3-triazolo [4,5-b ] pyridinium 3-oxide hexafluorophosphate

HPLC = high performance liquid chromatography

L=l

Lc=liquid chromatography

Lihmds=lithium bis (trimethylsilyl) amide

Mecn=acetonitrile

min = min

Ms=mass spectrometry

Nmr=nuclear magnetic resonance

Rt or rt=room temperature

SFC = supercritical fluid chromatography

T3p=propylphosphonic anhydride

THF = tetrahydrofuran

TLC = thin layer chromatography

Uplc=ultra high performance liquid chromatography

The prefixes n-, s-, i-, t-and t-have their usual meanings: normal, secondary, abnormal and tertiary states.

Synthesis of intermediates

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

Step 1:to a suspension of (t-butoxycarbonyl) -L-glutamic acid dimethyl ester (CAS No.59279-60-6, 250g, 0.328 mol) in THF (2500 mL) at-78℃LiHMDS (1M solution in THF, 1997mL,1.997 mol) was added dropwise, and the resulting reaction mixture was stirred at-78℃for 1 hour, and then bromoacetonitrile (76.5 mL,1.086 mol) was added at the same temperature. The resulting reaction mixture was stirred at-78 ℃ for 3 hours; after complete disappearance of the starting material monitored by TLC, the reaction mixture was quenched by addition of saturated aqueous ammonium chloride (2L) at-78 ℃ and then warmed to room temperature. The resulting mixture was extracted with EtOAc (2X 2000 mL) and dried over anhydrous Na ₂ SO ₄ The combined organic layers were dried and concentrated in vacuo. Purification by gradient flash column chromatography using silica gel (100-200 mesh) eluting with 0-20% EtOAc in petroleum ether afforded (2S, 4R) -2- ((tert-butoxycarbonyl) amino) -4- (cyanomethyl) glutarate (207 g,0.65 mol) as a pale yellow gum. The reaction was carried out in 5 batches on the scale of 50 g.

LCMS (method C) M/z 337.1 (M+Na) at 1.93 min.

¹ H NMR:(300MHz,DMSO-d ₆ )δ7.36(d,J＝8.1Hz,,1H),4.10-4.05(m,1H),3.66(s,3H),3.63(s,3H),2.87-2.73(m,3H),2.05-1.99(m,2H),1.39(s,9H)。

Step 2: to a stirred suspension of (2S, 4R) -2- ((tert-butoxycarbonyl) amino) -4- (cyanomethyl) glutaric acid (207 g,0.659 mol) in MeOH (2L) at 0deg.C was added CoCl ₂ .6H ₂ O (78.1 g, 0.399 mol) and then NaBH was added in portions over 30 minutes ₄ (149.6 g,3.955 mol) and the resulting reaction mixture was stirred at room temperature for 15 hours. After completion of the reaction monitored by TLC, the reaction mixture was concentrated in vacuo and 20% MeOH in DCM (3L) and H were added ₂ O (2L) and the mixture was stirred for 10 minutes. The resulting suspension was filtered through celite and rinsed with 20% MeOH in DCM (2L). The filtrate was transferred to a separatory funnel, the organic layer was separated, washed with brine solution (2L), and dried over anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Flash column chromatography purification by gradient using silica gel (100-200 mesh) eluting with 0-100% etoac in petroleum ether afforded methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) 2-oxopyrrolidin-3-yl) propionate (92 g,0.32 mol) as a pale yellow solid.

LCMS (method C) M/z 187.2 (M+H-100) at 1.34 minutes.

¹ H NMR:(300MHz,DMSO-d ₆ )δ7.64(s,1H),7.42(d,J＝7.8Hz,1H),4.06-4.02(m,1H),3.69(s,3H),3.16-3.08(m,2H),2.32-2.21(m,1H),2.16-2.11(m,1H),2.03-1.93(m,1H),1.69-1.66(m,2H),1.33(s,9H)。

Step 3: to a stirred solution of methyl (S) -2- ((tert-butoxycarbonyl) amino) -3- ((S) 2-oxopyrrolidin-3-yl) propionate (10 g,0.034 mol) in 1, 4-dioxane (50 mL) was added a solution of 4N HCl in dioxane (100 mL) at 0 ℃ and the resulting reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction monitored by TLC, the supernatant layer was decanted from the reaction mixture and the remaining viscous solid material was dried in vacuo to give methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate hydrochloride (intermediate 1,8.5g crude, 0.038 mol) as a yellow gum, which was used without further purification.

LCMS (method E) M/z 187.3 (M+H) at 0.82-1.19 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.67(s,3H),7.98(s,1H),4.19(t,J＝5.2Hz,1H),3.76(s,3H),3.21-3.18(m,2H),2.53-2.51(m,1H),2.28-2.27(m,1H),2.05-2.03(m,1H),1.90-1.89(m,1H),1.70-1.65(m,1H)。

Synthesis of examples

Example 1:3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide

Step 1: to a stirred solution of (S) -methyl 2-amino-3-cyclopropylpropionate hydrochloride (CAS No.206438-31-5, 17g,0.096 mol) and (E) -3- (4-chloro-2-fluorophenyl) acrylic acid (CAS No.312693-55-3, 16g,0.08 mol) in DCM (10 mL) was added DIPEA (59 mL,0.32 mol), followed by a 50% EtOAc (101.7 mL,0.16 mol) solution of T3P. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction monitored by TLC, the reaction mass was taken up in DCM (1L) and 10% nahco ₃ The aqueous solution (5L) was partitioned between. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP column (230-400 mesh) eluting with 0-40% EtOAc in petroleum ether afforded methyl (S, E) -2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionate (18 g,0.055 mol) as a white solid.

LCMS (method D) M/z 326.1 (M+H) at 3.07 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.67(d,J＝7.2Hz,1H),7.70(t,J＝8.4Hz,1H),7.56-7.53(m,1H),7.47(d,J＝16.0Hz,1H),7.39-7.36(m,1H),6.86(d,J＝15.6Hz,1H),4.45-4.44(m,1H),3.65(s,,3H),1.69-1.67(m,1H),1.59-1.57(m,1H),0.82-0.78(m,1H),0.44-0.40(m,2H),0.15-0.04(m,2H)。

Step 2: to (S, E) -methyl 2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionate (18 g,0.055 mol) in THF (200 mL), meOH (20 mL) and H at room temperature ₂ LiOH.H was added to the stirred solution in O (50 mL) ₂ O (6.92 g,0.165 mol). The resulting reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction by TLC, the reaction mixture was concentrated in vacuo, and the resulting residue was dissolved in H ₂ O (300 mL) was acidified to about pH 5 using 1.5N HCl, then extracted with 10% MeOH in DCM (2X 500 mL). The organic layers were separated, combined and taken up with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo to give (S, E) -2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionic acid (13 g,0.044 mol) as an off-white solid.

LCMS (method C) M/z 312.0 (M+H) at 2.06 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ7.83(d,J＝6.0Hz,1H),7.77-7.73(m,1H),7.51(d,J＝10.8Hz,1H),7.41-7.34(m,2H),7.03(d,J＝15.6Hz,1H),4.04-4.00(m,1H),1.65-1.52(m,2H),0.82-0.78(m,1H),0.30-0.29(m,2H),0.04-0.01(m,2H)。

Step 3: to a stirred solution of (S, E) -2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionic acid (10 g,0.0365 mol) and methyl (S) -2-amino-3- ((S) 2-oxopyrrolidin-3-yl) propionate hydrochloride (intermediate 1,8.5g,0.0365 mmol) in DCM (150 mL) was added DIPEA (31.7 mL,0.182 mol) followed by T3P (50% EtOAc solution, 34.8mL,0.054 mol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mixture was taken up in DCM (500 mL) and 10% nahco ₃ Aqueous solution (500 ml) was partitioned between. The organic layer was separated with 10% NaHCO ₃ Aqueous (500 mL) wash with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP chromatography column (230-400 mesh) eluting with 0-90% EtOAc in petroleum ether afforded methyl (S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (9.2 g,0.019 mmol) as an off-white solid.

LCMS (method C) M/z 480.2 (M+H) at 1.81 minutes.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.58(d,J＝7.8Hz,1H),8.42(d,J＝7.8Hz,1H),7.71-7.65(m,2H),7.56-7.52(m,1H),7.46-7.41(m,1H),7.39-7.36(m,1H),6.89(d,J＝15.9Hz,1H),4.50-4.35(m,2H),3.62(s,3H),3.15-3.09(m,2H),2.28-2.28(m,1H),2.09-2.03(m,2H),1.65-1.49(m,4H),0.80-0.79(m,1H),0.39-0.38(m,2H),0.10-0.09(m,2H)。

Step 4: to a stirred solution of methyl (S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -3- ((S) 2-oxopyrrolidin-3-yl) propanoate (9.2 g,0.019 mol) in THF (100 mL) was added MeOH (30 mL), followed by the addition of sodium borohydride (1.45 g,0.038 mol) in portions at 0 ℃. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction monitored by TLC, the reaction mixture was taken up in 20% meoh in DCM (500 mL) and H ₂ O (500 ml) was partitioned between. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP chromatography column (230-400 mesh) eluting with 0-6% MeOH in DCM afforded (E) -3- (4-chloro-2-fluorophenyl) -N- ((S) -3-cyclopropyl-1- (((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1-oxopropan-2-yl) acrylamide (6.4 g,0.014 mol) as an off-white solid.

LCMS (method C) M/z 452.2 (M+H) at 1.58 min.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.39(d,J＝8.1Hz,1H),7.83(d,J＝8.7Hz,1H),7.68-7.65(m,1H),7.55-7.46(m,2H),7.41-7.36(m,2H),6.90(d,J＝15.9Hz,1H),4.65(t,J＝5.7Hz,1H),4.44-4.42(m,1H),3.82-3.72(m,1H),3.26-3.06(m,4H),2.27-2.18(m,2H),1.78(t,J＝11.7Hz,1H),1.60-1.57(m,2H),1.47-1.39(m,2H),0.80-0.79(m,1H),0.38-0.37(m,2H),0.11-0.08(m,2H)。

Step 5: to a stirred solution of (E) -3- (4-chloro-2-fluorophenyl) -N- ((S) -3-cyclopropyl-1- (((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -1-oxopropan-2-yl) acrylamide (4.5 g,9.95 mmol) in DCM (100 mL) was added dess-martin reagent (6.34 g,14.9 mmol) in portions at room temperature. The resulting reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction by TLC, the reaction was monitored by 10% NaHCO ₃ The reaction mixture was quenched with aqueous solution (200 mL) and extracted with DCM (2×200 mL). The combined organic layers were treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP chromatography column (230-400 mesh) eluting with 0-6% meoh in DCM afforded (E) -3- (4-chloro-2-fluorophenyl) -N- ((2S) -3-cyclopropyl-1-oxo-1- ((1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) propan-2-yl) acrylamide (4.4 g,9.77 mmol) as an off-white solid.

LCMS (method E) M/z 450.0 (M+H) at 1.78-1.96 min.

Chiral SFC analysis (method 2): 1.73&2.01min (48.8% & 45.0%)

¹ H NMR:(300MHz,DMSO-d ₆ )δ9.43(s,0.5H),8.63(d,J＝7.6Hz,0.5H),8.50(d,J＝7.6Hz,0.5H),8.38(d,J＝7.6Hz,0.5H),7.69-7.65(m,2H),7.56-7.53(m,2H),7.47-7.37(m,2H),6.93-6.88(m,1H),4.51-4.41(m,1H),4.38-4.31(m,0.5H),4.20-4.17(m,0.5H),3.21-3.12(m,3H),2.25-2.15(m,1.5H),1.91-1.85(m,1.5H),1.69-1.51(m,3H),0.81-0.70(m,1H),0.42-0.39(m,2H),0.10-0.09(m,2H)。

Step 6: step 6: to a stirred solution of (E) -3- (4-chloro-2-fluorophenyl) -N- ((2S) -3-cyclopropyl-1-oxo-1- ((1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) propan-2-yl) acrylamide (4.4 g,9.7 mmol) in DCM (40 mL) at room temperature was added triethylamine (2.75 mL,19.5 mmol) followed by the addition ofAcetone cyanohydrin (1.66 mL,19.5 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction monitored by TLC, the reaction mixture was concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 50g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% meoh in DCM afforded (E) -3- (4-chloro-2-fluorophenyl) -N- ((2S) -1- (1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) acrylamide (2.7 g,5.66 mmol) as an off-white solid.

LCMS (method E) M/z 450.1 (M-CN) at 1.79-1.96 minutes.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.37-8.07(m,2H),7.60-7.24(m,5H),6.82-6.74(m,1H),6.64-6.55(m,1H),4.46-4.22(m,2H),3.91-3.85(m,1H),3.15-2.93(m,2H),2.33-2.23(m,1H),2.02-1.71(m,2H),1.52-1.31(m,4H),0.71-0.54(m,1H),0.29-0.27(m,2H),0.15-0.09(m,2H)。

Step 7: to a stirred solution of (E) -3- (4-chloro-2-fluorophenyl) -N- ((2S) -1- (1-cyano-1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) acrylamide (2.7 g,5.6 mmol) in DMSO (30 mL) was added K ₂ CO ₃ (2.34 g,16.9 mmol) and then 30% H was added ₂ O ₂ Aqueous (6.4 mL,56.6 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mixture was partitioned between 10% meoh in DCM (100 mL) and brine (100 mL). The aqueous layer was further extracted with 10% MeOH in DCM (3X 200 mL) and the combined organic layers were taken up with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP chromatography column (230-400 mesh) eluting with 0-10% meoh in DCM afforded 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamide) -2-hydroxy-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (960 mg,1.93 mmol) as an off-white solid.

LCMS (method E) M/z 495.0 (M+H) at 1.78 minutes.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.43(d,J＝7.6Hz,1H),7.71-7.23(m,8H),6.90(d,J＝21.2Hz,1H),5.66(d,J＝7.6Hz,1H),4.49-4.41(m,1H),4.11-4.09(m,1H),3.85-3.83(m,1H),3.12-3.04(m,2H),2.27-2.12(m,4H),1.60-1.37(m,3H),0.80-0.79(m,1H),0.38-0.37(m,2H),0.11-0.08(m,2H)。

Step 8: to a stirred solution of 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-hydroxy-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (960 mg,1.93 mmol) in DCM (40 mL) and DMSO (10 mL) at room temperature was added dess-martin reagent (1.22 g,2.895 mmol). The resulting reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction by TLC, 10% NaHCO was used ₃ The reaction mixture was quenched with aqueous (200 mL) and extracted with EtOAc (200 mL). The organic layer was separated with 10% NaHCO ₃ Aqueous (150 mL) wash with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 50g silica SNAP chromatography column (230-400 mesh) eluting with 0-10% meoh in DCM afforded 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (example 1, 400mg,0.811 mmol) as an off-white solid.

LCMS (method C) M/z 493.1 (M+H) at 1.21-1.44 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.61(d,J＝7.6Hz,0.5H),8.43-8.41(m,1H),8.04-8.09(m,0.6H),7.79-7.71(m,0.6H),7.69-7.66(m,1.7H),7.56-7.42(m,3.9H),7.39-7.36(m,0.9H),6.89(d,J＝16.0Hz,1H),6.12(d,J＝40.8Hz,0.7H),5.15-5.05(m,0.5H),4.61-4.45(m,1H),4.05-4.01(m,0.5H),3.15-3.12(m,2H),2.26-2.16(m,2H),1.82-1.53(m,5H),0.79-0.73(m,1H),0.40-0.36(m,2H),0.13-0.09(m,2H)。

Example 2:3- ((S) -2-cinnamamido-3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide

Step 1: to a stirred solution of (S) -methyl 2-amino-3-cyclopropylpropionate hydrochloride (CAS No.206438-31-5,3.34g,18.57 mmol) and cinnamic acid (CAS No.140-10-3,2.75g,18.57 mmol) in DCM (40 mL) was added DIPEA (14 mL,77.35 mmol) followed by T3P in 50% EtOAc (14.7 mL,23.20 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mixture was taken up in DCM (300 mL) and 10% NaHCO ₃ Aqueous solution (300 mL) was partitioned between. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP column (230-400 mesh) eluting with 0-40% EtOAc in petroleum ether afforded methyl (S) -2-cinnamamido-3-cyclopropylpropionate (4.40 g,16.1 mmol) as an off-white solid.

LCMS (method C) M/z 274.1 (M+H) at 2.10 min.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.52(d,J＝7.2Hz,1H),7.58(d,J＝6.3Hz,2H),7.48-7.39(m,4H),6.75(d,J＝15.9Hz,1H),4.42(q,J＝5.7Hz,1H),3.64(s,3H),1.71-1.54(m,2H),0.82-0.77(m,1H),0.43-0.41(m,2H),0.16-0.03(m,2H)。

Step 2: to methyl (S) -2-cinnamamido-3-cyclopropylpropionate (4.0 g,14.63 mmol) in 1, 4-dioxane (50 mL), meOH (10 mL), and H at room temperature ₂ LiOH.H was added to the stirred solution in O (10 mL) ₂ O (1.23 g,29.26 mmol). The resulting reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction monitored by TLC, the reaction mixture was concentrated in vacuo. Dissolving the obtained residue in H ₂ O (100 mL) was acidified to about pH 5 using 1.5N HCl, then extracted with 20% MeOH in DCM (150 mL). The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo to give (S) -2-cinnamamido-3-cyclopropylpropionic acid (3.30 g,12.7 mmol) as an off-white solid.

LCMS (method B) M/z 260.0 (M+H) at 2.10 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ12.70(s,1H),8.39(d,J＝8.0Hz,1H),7.59-7.57(m,2H),7.47-7.39(m,4H),6.78(d,J＝15.6Hz,1H),4.40(t,J＝2.8Hz,1H),1.66-1.59(m,2H),0.81-0.81(m,1H),0.44-0.41(m,2H),0.17-0.08(m,2H)。

Step 3-8: the title compound of example 2, 3- ((S) -2-cinnamamido-3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (25 mg,0.079 mmol) was prepared from step 2 product (3.30 g,12.7 mmol) and intermediate 1 (2.01 g,10.7 mmol) by steps 3-8 using the procedure detailed in example 1.

LCMS (method K) M/z 441.2 (M+H) at 1.60-1.71 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.60(d,J＝7.2Hz,0.5H),8.26-8.24(m,1H),8.03(s,0.5H),7.78(s,0.5H),7.68-7.51(m,3H),7.45-7.38(m,4H),7.30-7.21(m,1H),6.82-6.77(m,1H),6.16-6.07(m,0.5H),5.15-5.05(m,0.5H),4.61-4.45(m,1H),4.05-4.01(m,0.5H),3.15-3.12(m,2H),2.19-2.12(m,2H),1.95-1.53(m,5H),0.81-0.70(m,1H),0.42-0.37(m,2H),0.14-0.09(m,2H)。

Example 3: ((2S) -1- ((4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid benzyl ester

Step 1: to a stirred solution of methyl (S) -2-amino-3-cyclopropylpropionate (CAS No.732231-41-3,1g,6.98 mmol) in DCM (15 mL) was added DIPEA (3.6 mL,20.95 mmol) followed by benzyl chloroformate (1.42 g,8.38 mmol) at 0deg.C. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mass was taken up in DCM (20 mL) and H ₂ O (50 mL) between partitions. The organic layer was separated with 10% NaHCO ₃ Aqueous (30 mL) and brine (30 mL) were washed with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) with 0-20% EtOAc in petroleum ether(S) -2- (((benzyloxy) carbonyl) amino) -3-cyclopropylpropionic acid methyl ester (1.1 g,3.96 mmol) was obtained as an off-white solid.

LCMS (method A) M/z 278.2 (M+H) at 2.39 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ7.40-7.30(m,6H),5.17(s,2H),4.50-4.49(m,1H),3.63(s,3H),1.65-1.61(m,1H),1.48-1.46(m,1H),0.78(s,1H),0.42-0.37(m,2H),0.13-0.10(m,1H),0.02-0.01(m,1H)。

Step 2: to a stirred solution of methyl (S) -2- (((benzyloxy) carbonyl) amino) -3-cyclopropylpropionate (1.1 g,3.96 mmol) in THF (10 mL) was added MeOH (2 mL) and H ₂ O (5 mL) followed by addition of LiOH.H ₂ O (0.142 g,5.94 mmol). The resulting reaction mixture was stirred at room temperature for 30 minutes. After completion of the reaction by TLC, the reaction mixture was acidified to about pH 5 with 1.5N HCl (5 mL) and quenched with EtOAc (50 mL) and H ₂ O (50 mL) between partitions. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% meoh in DCM afforded methyl (S) -2- (((benzyloxy) carbonyl) amino) -3-cyclopropylpropionate (1.0 g,3.79 mmol) as an off-white solid.

LCMS (method C) M/z 264.1 (M+H) at 1.77 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ7.58(d,J＝8.0Hz,,1H),7.40-7.22(m,6H),5.13(s,2H),4.03-3.97(m,1H),1.63-1.59(m,1H),1.50-1.47(m,1H),0.79(t,J＝7.2Hz,1H),0.42-0.36(m,2H),0.16-0.01(m,2H)。

Step 3-8: the title compound of example 3, ((2S) -1- ((4-amino-3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) amino) -3-cyclopropyl-1-oxopropan-2-yl) carbamic acid benzyl ester (5 mg,0.015 mmol) was prepared from step 2 product (1.0 g,3.79 mmol) and intermediate 1 (0.84 g,4.55 mmol) by the procedure detailed in example 1 using step 3-8.

LCMS (method A) M/z 445.1 (M+H) at 1.95-2.09 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.43(d,J＝7.2Hz,0.5H),8.01-7.96(m,1H),7.77(s,0.5H),7.65(s,0.5H),7.59(s,0.5H),7.49-7.47(m,1H),7.37-7.31(m,5.5H),7.04(s,0.5H),5.08-5.03(m,2.5H),4.31-4.28(m,0.5H),4.15-4.05(m,1H),3.15-3.05(m,2H),2.29-1.79(m,3H),1.64-1.40(m,4H),0.75-0.71(m,1H),0.40-0.34(m,,2H),0.13-0.07(m,2H)。

Example 4:3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2- (oximido) -4- ((S) -2-oxopyrrolidin-3-yl) butanamide

To a stirred solution of 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (example 1,0.1g,0.20 mmol) in EtOH (10 mL) at room temperature was added K ₂ CO ₃ (55 mg,0.4 mmol) and hydroxylamine hydrochloride (28 mg,0.4 mol). The resulting reaction mixture was stirred at 70℃for 3 hours. After completion of the reaction monitored by UPLC-MS, the reaction mixture was filtered and rinsed with EtOH (5 mL). The filtrate was concentrated in vacuo and purified by mass directed preparative HPLC (method F) to give the title compound of example 4, 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2- (hydroxyimino) -4- ((S) -2-oxopyrrolidin-3-yl) butanamide as a mixture of isomers (off white solid, 10mg,0.019 mmol).

LCMS (method C) M/z 508.0 (M+H) at 2.04 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ11.98(s,1H),8.51-8.50(m,1H),8.02-8.00(m,1H),7.70-7.68(m,1H),7.59-7.56(m,2H),7.54-7.31(m,4H),6.88(d,J＝16.0Hz,1H),5.47-5.45(m,1H),4.42-4.39(m,1H),3.15-3.08(m,2H),2.26-2.23(m,3H),1.55-1.51(m,4H),0.79-0.74(m,1H),0.39-0.35(m,2H),0.12-0.10(m,2H)。

Example 5: n- (tert-butyl) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamidePhenyl) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide

Step 1: to a stirred solution of (E) -3- (4-chloro-2-fluorophenyl) -N- ((2S) -3-cyclopropyl-1-oxo-1- ((1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) amino) propan-2-yl) acrylamide (example 1, step 5 product, 430mg,0.955 mmol) in DCM (5 mL) was added AcOH (0.060 mL,1.051 mmol) followed by t-butyl isocyanide (0.12 mL,1.051 mmol) at 0deg.C. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction monitored by TLC, the reaction mass was concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% meoh in DCM afforded 1- (tert-butylamino) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -1-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl acetate (300 mg,0.50 mmol) as an off-white solid.

LCMS (method C) M/z 593.2 (M+H) at 2.34 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.43-8.37(m,1H),8.01-7.84(m,1H),7.69-7.66(m,1H),7.57-7.46(m,3H),7.42-7.36(m,2H),6.91-6.87(m,1H),4.92-4.76(m,1H),4.48-4.47(m,1H),4.35-4.21(m,1H),3.12-3.05(m,2H),2.21-2.19(m,2H),2.18-2.11(m,1H),2.10-2.06(m,3H),2.00-1.93(m,1H),1.56-1.45(m,3H),1.25-1.18(m,9H),0.85-0.65(m,1H),0.41-0.37(m,2H),0.12-0.08(m,2H)。

Step 2: to 1- (tert-butylamino) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -1-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl acetate (300 mg,0.629 mmol) in 1, 4-dioxane (3 mL), meOH (1 mL) and H ₂ LiOH.H was added to the stirred solution in O (2 mL) ₂ O (52.2 mg,1.25 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction was taken up in EtOAc (25 mL) and H ₂ O (25 ml) was partitioned between. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% meoh in DCM afforded N- (tert-butyl) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-hydroxy-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (200 mg,0.36 mmol) as an off-white solid.

LCMS (method A) M/z 551.2 (M+H) at 2.07 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.45-8.43(m,1H),7.73-7.67(m,1H),7.56-7.37(m,5H),7.02(s,1H),6.98-6.86(m,1H),5.77-5.74(m,1H),4.46-4.45(m,1H),4.19-4.01(m,1H),3.77-3.75(m,1H),3.29-3.05(m,2H),2.34-2.13(m,2H),2.05-1.95(m,1H),1.63-1.53(m,2H),1.49-1.39(m,1H),1.48-1.45(m,10H),0.79-0.69(m,1H),0.46-0.29(m,2H),0.15–0.10(m,2H)。

Step 3: to a stirred solution of N- (tert-butyl) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -2-hydroxy-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (200 mg,0.36 mmol) in DCM (10 mL) and DMSO (3 mL) at room temperature was added dess-martin reagent (230 mg,0.54 mmol) in portions. The resulting reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction by TLC, 10% NaHCO was added ₃ Aqueous (50 mL) and the mixture extracted with DCM (2X 50 mL). The combined organic layers were treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-6% meoh in DCM afforded N- (tert-butyl) -3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamide) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide (example 5, 80mg,0.15 mmol) as an off-white solid.

LCMS (method C) M/z 547.1 (M-H) at 2.19-2.62 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.58-8.56(m,1H),8.42-8.40(m,1H),7.92(s,1H),7.70-7.66(m,2H),7.56-7.53(m,1H),7.46-7.42(m,1H),7.39-7.36(m,1H),6.91-6.87(m,1H),5.01-5.10(m,1H),4.54-4.52(m,1H),3.18-3.12(m,2H),2.35-2.49(m,1H),2.37-2.34(m,,1H),2.08-1.92(m,1H),1.72-1.61(m,1H),1.61-1.50(m,3H),1.39-1.20(m,9H),0.85-0.70(m,1H),0.42-0.37(m,2H),0.14-0.09(m,2H)。

Example 6:3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -4-cyclopropyl-2-oxobutanamide

Step 1-6: the title compound of example 6, 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropyl-propionamido) -4-cyclopropyl-2-oxobutanamide (25 mg,0.05mmol, white solid) was prepared from (S, E) -2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionic acid (example 1, step 2 product, 2.0g,6.41 mmol) and (S) -methyl 2-amino-3-cyclopropylpropionate hydrochloride (CAS No.206438-31-5,1.38g,7.69 mmol) by steps 1-6 using the procedure detailed in example 1. Step 3 uses THF/DMSO (2:1, 15 mL) as solvent.

LCMS (method C) M/z 450.1 (M+H) at 1.79-2.06 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.50-8.39(m,2H),8.01(d,J＝4.8Hz,1H),7.74-7.66(m,2H),7.56-7.53(m,1H),7.48-7.43(m,1H),7.39-7.36(m,1H),6.94-6.88(m,1H),5.15-5.10(m,1H),4.66-4.58(m,1H),1.67-1.47(m,4H),0.84-0.74(m,2H),0.42-0.35(m,,4H),0.13-0.02(m,4H)。

Example 7:3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamido) -4-cyclopentyl-2-oxobutanamide

Step 1-6: the title compound of example 7, 3- ((S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionamide) -4-cyclopentyl-2-oxobutanamide (10 mg,0.02mmol, off-white solid) was prepared from (S, E) -2- (3- (4-chloro-2-fluorophenyl) acrylamido) -3-cyclopropylpropionic acid (example 1, step 2 product, 1.0g,3.21 mmol) and (S) -methyl 2-amino-3-cyclopentylpropionate hydrochloride (CAS No.1191996-99-2,0.79g,3.85 mmol) by steps 1-6 using the procedure detailed in example 1. Step 3 uses THF/DMSO (2:1, 15 mL) as solvent.

LCMS (method C) M/z 478.1 (M+H) at 2.00-2.28 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.48-8.34(m,2H),,8.02(s,1H),7.76(s,1H),7.70-7.65(m,1H),7.56-7.53(m,1H),7.45(d,,J＝16.0Hz,1H),7.39-7.36(m,1H),6.93-6.88(m,1H),5.05-5.05(m,1H),4.60-4.51(m,1H),1.90-1.44(m,11H),1.15-1.09(m,2H),0.87-0.76(m,1H),0.42-0.36(m,2H),0.12-0.07(m,2H)。

Example 8:3- ((S) -3-cyclopropyl-2- (3-phenylpropionamido) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide

Step 1: to a stirred solution of methyl (S) -2-amino-3-cyclopropylpropionate hydrochloride (CAS No.206438-31-5,2.38g,13.20 mmol) and 3-phenylpropionic acid (2.01 g,13.20 mmol) in DCM (30 mL) was added DIPEA (11.57 mL,66.00 mmol) followed by T3P (50% EtOAc solution, 12.68mL,19.90 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mixture was taken up in DCM (100 mL) and 10% nahco ₃ Aqueous solution (100 mL) was partitioned between. The organic layer was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 100g silica SNAP column (230-400 mesh) eluting with 0-10% EtOAc in petroleum ether afforded methyl (S) -3-cyclopropyl-2- (3-phenylpropionamido) propionate (2.12 g,7.69 mmol) as an off-white solid.

LCMS (method C) M/z 276.1 (M+H) at 2.04 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.27(s,1H),7.26-7.21(m,5H),4.30-4.28(m,1H),3.61(s,3H),2.81-2.80(m,2H),2.43-2.25(m,2H),1.59-1.44(m,2H),0.78-0.56(m,1H),0.37-0.35(m,2H),0.07-0.01(m,2H)。

Step 2: to methyl (S) -3-cyclopropyl-2- (3-phenylpropionamido) propionate (2.12 g,7.69 mmol) in 1, 4-dioxane (10 mL), meOH (5 mL), and H at room temperature ₂ LiOH.H was added to the stirred solution in O (5 mL) ₂ O (0.365 g,15.3 mmol). The resulting reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction monitored by TLC, the reaction mixture was concentrated in vacuo. Dissolving the obtained residue in H ₂ O (100 mL) was acidified to about pH 5 with 1.5N HCl and extracted with 20% MeOH in DCM (150 mL). The organic layer was separated with anhydrous Na ₂ SO ₄ Drying and concentration in vacuo afforded (S) -3-cyclopropyl-2- (3-phenylpropionamido) propionic acid (1.90 g,7.23 mmol) as an off-white solid, which was used without further purification.

LCMS (method H) M/z 262.3 (M+H) at 1.50 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ12.50(s,1H),8.13(d,J＝8.0Hz,1H),7.28-7.17(m,5H),4.25-4.24(m,1H),2.84-2.70(m,2H),2.56-2.31(m,2H),1.56-1.46(m,2H),0.81-0.65(m,1H),0.52-0.35(m,2H),0.15-0.09(m,2H)。

Step 3: to a stirred solution of (S) -3-cyclopropyl-2- (3-phenylpropionamido) propionic acid (1.90 g,7.20 mmol) and methyl (S) -2-amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate hydrochloride (intermediate 1,1.35g,7.20 mmol) in DCM (10 mL) was added Et ₃ N (4.05 mL,29.1 mmol) followed by HATU (4.14 g,10.9 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction by TLC, the reaction mixture was taken up in DCM (100 mL) and 10% nahco ₃ Aqueous solution (100 mL) was partitioned between. The organic layer was separated, washed with saturated brine (100 mL), and dried over anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Flash column chromatography (B) by gradient using 25g silica SNAP column (230-400 mesh)iotatage-Isolera) was eluted with 0-2% meoh in DCM to give methyl (S) -2- ((S) -3-cyclopropyl-2- (3-phenylpropionamido) propanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (1.24 g,2.88 mmol) as an off-white solid.

LCMS (method A) M/z 430.3 (M+H) at 1.83 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.47-8.45(m,1H),8.01-7.98(m,1H),7.65(s,1H),7.25-7.21(m,5H),4.50-4.23(m,2H),4.12-3.95(m,1H),3.45-3.75(m,3H),3.25-3.01(m,2H),2.80-2.78(m,2H),2.09-1.91(m,2H),1.60-1.41(m,4H),1.18(t,J＝9.2Hz,2H),0.75-0.58(m,1H),0.42-0.15(m,2H),0.13-0.09(m,2H)。

Step 4-8: the title compound 3- ((S) -3-cyclopropyl-2- (3-phenylpropionamido) propanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butanamide of example 8 (off-white solid, 20mg,0.045 mmol) was prepared from the product of step 3 (1.24 g,2.80 mmol) by step 4-8 using the procedure detailed in example 1. Example 8 was isolated after purification by preparative HPLC using method B.

LCMS (method C) M/z 443.1 (M+H) at 1.69-1.84 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.58-8.46(m,1H),8.00-7.97(m,2H),7.76(s,1H),7.68(s,1H),7.27-7.14(m,5H),5.11-5.01(m,1H),4.44-4.25(m,1H),3.18-3.11(m,2H),2.81(t,J＝8.0Hz,2H),2.47-2.46(m,2H),2.18-2.17(m,2H),1.91-1.85(m,1H),1.65-1.41(m,,4H),0.65-0.58(m,1H),0.37-0.32(m,2H),0.10-0.02(m,2H)。

Example 9:(2S) -N- (1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide

Step 1: to a stirred solution of (E) -3- (2, 4-dichlorophenyl) acrylic acid (2.50 g,11.5 mmol) and (S) -2-amino-4, 4-dimethylvaleric acid methyl ester hydrochloride (2.70 g,13.8 mmol) in DCM (40 mL) was added DIPEA (6.00 m)L,34.6 mmol) and then T3P (50% EtOAc solution, 10.0mL,17.3 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours, then at EtOAc (300 mL) and H ₂ O (200 mL) was partitioned between. The phases were separated with 10% NaHCO ₃ The organic layer was washed with aqueous solution (200 mL) and brine (200 mL), and dried over anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 50g silica SNAP column (230-400 mesh) eluting with a solution of 0-8% etoac in petroleum ether afforded methyl (S, E) -2- (3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanoate (3.37 g,9.40 mmol) as a white solid.

LCMS (method C) M/z 357.8 (M+H) at 2.69 min.

¹ H NMR:(300MHz,DMSO-d ₆ )δ8.66(d,J＝8.0Hz,1H),7.75-7.66(m,3H),7.53-7.50(m,1H),6.76(d,J＝16.0Hz,1H),4.50-4.45(m,1H),3.65(s,3H),1.76-1.57(m,2H),0.92-0.90(m,9H)。

Step 2: liOH.H ₂ O (0.59 g,14.1 mmol) was added to (S, E) -2- (3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanoic acid methyl ester (3.37 g,9.40 mmol) in THF (40 mL), meOH (5 mL) and H ₂ O (10 mL) and the mixture was stirred at room temperature for 1 hour. After concentration in vacuo, the residue obtained is dissolved in H ₂ O (100 mL) was acidified to about pH 5 using 1.5N HCl and extracted with 20% MeOH in DCM (150 mL). The organic phase was separated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo to give (S, E) -2- (3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanoic acid (3.10 g,9.00 mmol) as a white solid.

LCMS (method C) M/z 344.0 (M+H) at 2.33 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ12.90(s,1H),8.48(d,J＝8.4Hz,1H),7.74-7.72(m,2H),7.66(d,J＝15.6Hz,1H),7.52-7.50(m,1H),6.79(d,J＝15.6Hz,1H),4.39-4.35(m,1H),1.74-1.70(m,1H),1.59-1.53(m,1H),0.91(s,9H)。

Step 3: to (S, E) -2- (3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanoic acid (3.10 g,9.00 mmol) and (S) -2-To a stirred solution of methyl amino-3- ((S) -2-oxopyrrolidin-3-yl) propionate hydrochloride (intermediate 1,3.00g,13.5 mmol) in DCM (30 mL) was added DIPEA (4.70 mL,27.2 mmol) followed by T3P (50% EtOAc solution, 8.00mL,13.5 mmol). After stirring at room temperature for 16 hours, the reaction mixture was taken up in EtOAc (150 mL) and H ₂ O (150 mL) between partitions. 10% NaHCO for organic layer ₃ Aqueous (100 mL) and saturated brine (50 mL) were washed with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% meoh in DCM afforded methyl (S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propionate (1.50 g,2.92 mmol) as a pale yellow solid.

LCMS (method C) M/z 511.7 (M+H) at 2.15 minutes.

Step 4: to a stirred solution of methyl (S) -2- ((S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamido) -3- ((S) -2-oxopyrrolidin-3-yl) propanoate (1.50 g,2.92 mmol) in THF (20 mL) at 0deg.C was added MeOH (2 mL) followed by NaBH in portions ₄ (166 mg,4.39 mmol). After stirring at room temperature for 2 hours, the reaction mixture was quenched with brine solution (50 mL) and extracted with EtOAc (100 mL). Anhydrous Na for organic phase ₂ SO ₄ Dried and concentrated under reduced pressure to give (S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4, 4-dimethylpentanamide (1.2 g,2.47 mmol) as an off-white solid.

LCMS (method C) M/z 484.1 (M+H) at 1.68 minutes.

Step 5: dess-martin reagent (1.57 g,3.71 mmol) was added to a solution of (S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -N- ((S) -1-hydroxy-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4, 4-dimethylvaleramide (1.20 g,2.47 mmol) in DCM (15 mL) at 0 ℃. After stirring for 1 hour at room temperature, 10% NaHCO was added ₃ Aqueous (50 mL) and EtOAc (200 mL) and the phases were separated. The organic phase was treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-3% meoh in DCM afforded (2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethyl-N- (1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) pentanamide (0.75 g,1.55 mmol) as an off-white solid.

LCMS (method E) M/z 482.2 (M+H) at 1.90-2.10 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ9.41-9.38(m,1H),8.65(d,J＝7.2Hz,1H),8.51(d,J＝8.4Hz,1H),7.79-7.63(m,3H),7.53-7.50(m,2H),6.83-6.78(m,1H),4.51-4.50(m,1H),4.48-4.30(m,1H),3.21-3.09(m,2H),2.50-2.41(m,1H),2.34-2.26(m,1H),2.13-2.12(m,3H),1.72-1.49(m,2H),0.93-0.91(m,9H)。

Step 6: NH is added to ₂ OH.HCl (57 mg,0.82 mmol) and K ₂ CO ₃ (171 mg,1.24 mmol) was added to a solution of (2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethyl-N- (1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) pentanamide (0.20 g,0.41 mmol) in EtOH (10 mL). After stirring at 70℃for 16 hours, the reaction mixture was cooled to room temperature and quenched with EtOAc (10 mL) and H ₂ O (10 mL) between partitions. The organic phase was treated with anhydrous Na ₂ SO ₄ Dried, concentrated in vacuo, and purified by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh), eluting with 0-5% meoh in DCM to give (2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -N- (1- (hydroxyimino) -3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4, 4-dimethylvaleramide (150 mg,0.30 mmol) as an off-white solid.

LCMS (method E) M/z 497.1 (M+H) at 2.09 minutes.

Step 7: methyl N- (triethylammonium sulfonyl) carbamate (0.071 g,0.30 mmol) was added to a stirred solution of (2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -N- (1- (hydroxyimino) -3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) -4, 4-dimethylvaleramide (0.15 g,0.30 mmol) in PhMe (50 mL) and the resulting reaction mixture stirred at 80℃for 16 hours. In EtOAc (50 mL) and H ₂ O (50 ml) betweenAfter compounding, the organic phase was taken up in anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. The resulting crude material was purified by gradient flash column chromatography (Biotage-Isolera) using a 10g silica SNAP column (230-400 mesh), eluting with 0-5% MeOH in DCM, followed by mass directed preparative HPLC (method B). The pure fractions were concentrated in vacuo, then taken up in EtOAc (10 mL) and H ₂ O (10 ml) was partitioned between. The organic phase was treated with anhydrous Na ₂ SO ₄ Dried, concentrated in vacuo and lyophilized to give the title compound of example 9, (2S) -N- (1-cyano-2- ((S) -2-oxopyrrolidin-3-yl) ethyl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide (10 mg,0.02 mmol) as an off-white solid.

LCMS (method C) M/z 479.1 (M+H) at 2.59 minutes.

Chiral SFC analysis (method 2): 1.82&3.69 min (47.6% & 49.1%).

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.99(d,J＝8.0Hz,1H),8.55(d,J＝8.4Hz,1H),7.73-7.65(m,4H),7.54-7.51(m,1H),6.78(d,J＝15.6Hz,1H),4.96-4.94(m,1H),4.46-4.45(m,1H),3.14-3.10(m,,2H),2.15-2.09(m,2H),1.73-1.50(m,5H),0.93(s,9H)。

Example 10:(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide

Step 1: acOH (0.1 g,1.65 mmol) followed by cyclopropylisonitrile (55 mg,0.82 mmol) was added to a stirred solution of (2S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethyl-N- (1-oxo-3- ((S) -2-oxopyrrolidin-3-yl) propan-2-yl) pentanamide (example 9, step 5 product, 0.40g,0.82 mol) in DCM (15 mL). After stirring at room temperature for 16 hours, the reaction mixture was concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP chromatography column (230-400 mesh) eluting with 0-5% MeOH in DCM afforded 1- (cyclopropylamino) -3- ((S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanoylamino) -1-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl acetate (380 mg,0.62 mmol) as an off-white solid.

LCMS (method C) M/z 609.1 (M+H) at 1.91 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.40(d,J＝8.4Hz,1H),8.08-8.03(m,2H),7.89(d,J＝9.2Hz,1H),7.72-7.63(m,3H),7.57-7.50(m,1H),6.81(d,J＝16.0Hz,1H),5.76-5.66(m,1H),4.76-4.74(m,1H),4.74-4.68(m,1H),3.27-3.12(m,2H),2.12-2.05(m,4H),2.00-1.92(m,3H),1.54-1.49(m,4H),0.92-0.89(m,9H),0.63-0.57(m,2H),0.44-0.42(m,2H)。

Step 2: meOH (3 mL) and H ₂ O (5 mL), then LiOH.H ₂ O (399mg, 0.93 mmol) was added to a stirred solution of 1- (cyclopropylamino) -3- ((S) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamido) -1-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl acetate (380 mg,0.62 mmol) in THF (5 mL) and the reaction mixture stirred at room temperature for 1 hour. EtOAc (50 mL) and H were added ₂ O (50 mL), separate phases with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo to give (2S) -N- (4- (cyclopropylamino) -3-hydroxy-4-oxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide (350 mg,0.61 mmol) as an off-white solid.

LCMS (method C) M/z 567.1 (M+H) at 1.75 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.47-8.44(m,1H),7.78-7.64(m,3H),7.58-7.50(m,2H),7.46-7.43(m,1H),7.34-7.33(m,1H),6.88-6.77(m,1H),5.82-5.63(m,2H),4.06-4.03(m,1H),3.83-3.81(m,1H),3.13-3.04(m,2H),2.68-2.67(m,1H),2.12-2.05(m,2H),2.00-1.90(m,1H),1.54-1.50(m,4H),0.92-0.83(m,9H),0.59-0.55(m,2H),0.48-0.46(m,2H)。

Step 3: dess-Martin reagent (399mg, 0.92 mmol) was added to (2S) -N- (4- (cyclopropylamino) -3-hydroxy-4-oxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2) at 0deg.C4-dichlorophenyl) acrylamido) -4, 4-dimethylvaleramide (350 mg,0.61 mmol) in DCM (10 mL). After stirring for 1 hour at room temperature, 10% NaHCO was added ₃ Aqueous (20 mL) and the reaction mixture was extracted with EtOAc (2X 50 mL). The combined organic layers were treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. After purification by mass-directed preparative HPLC (method B), the pure fractions were concentrated in vacuo and purified in 10% NaHCO ₃ Partitioned between aqueous (25 mL) and EtOAc (50 mL). Anhydrous Na for organic phase ₂ SO ₄ The title compound of example 10 was dried and concentrated in vacuo to give (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((E) -3- (2, 4-dichlorophenyl) acrylamido) -4, 4-dimethylpentanamide (145 mg,0.25 mmol) as a white solid.

LCMS (method E) M/z 565.2 (M+H) at 2.05-2.23 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.73-8.66(m,2H),8.41(d,J＝8.4Hz,1H),7.72-7.64(m,3H),7.56-7.50(m,1H),7.34-7.33(m,1H),6.81(d,J＝16.0Hz,1H),4.98-4.90(m,1H),4.56-4.55(m,1H),3.18-3.12(m,2H),2.91-2.86(m,1H),2.74-2.73(m,2H),2.53-2.50(m,2H),2.22-2.17(m,1H),1.70-1.63(m,2H),0.92-0.83(m,9H),0.67-0.63(m,2H),0.59-0.57(m,2H)。

Example 11:(2S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylvaleramide

Step 1-8: the title compound of example 11 was prepared in analogy to the procedure detailed above, as white solid (2S) -2- ((E) -3- (4-chloro-2-fluorophenyl) acrylamido) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethylpentanamide (17 mg,0.03 mmol).

LCMS (method A) M/z 549.0 (M+H) at 1.87-2.04 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.73-8.62(m,1H),8.46-8.40(m,1H),7.75-7.65(m,2H),7.55-7.52(m,1H),7.49-7.42(m,1H),7.38-7.33(m,1H),6.86-6.80(m,1H),6.25-6.14(m,1H),5..10-5.00(m,1H),4.61-4.59(m,1H),3.18-3.11(m,2H),2.74-2.67(m,1H),2.18-2.15(m,2H),1.67-1.50(m,3H),0.97-0.91(m,2H),0.86-0.82(m,9H),0.66-0.51(m,4H)。

Example 12:(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylbutyramide) pentanamide

Step 1-8: the title compound of example 12 was prepared in analogy to the procedure detailed above, as off-white solid (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylbutyramide) pentanamide (9.9 mg,0.02 mmol).

LCMS (method A) M/z 513.3 (M+H) at 1.74-1.92 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.73-8.71(m,1H),8.48-8.46(m,1H),8.03-8.01(m,1H),7.72-7.69(m,1H),7.29-7.15(m,,5H),4.95-4.90(m,1H),4.37-4.36(m,1H),3.18-3.15(m,4H),2.68-2.67(m,1H),2.40-2.36(m,2H),2.30-2.10(m,1H),1.90-1.88(m,1H),1.68-1.56(m,3H),1.46-1.40(m,1H),1.17-1.14(m,3H),0.90-0.85(m,,9H),0.66-0.57(m,4H)。

Example 13:(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylbutyramide) pentanamide

Step 1-8: example 1 was prepared using a procedure similar to that detailed above3, (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylbutyramide) pentanamide (10 mg,0.02 mmol) as an off-white solid.

LCMS (method A) M/z 513.3 (M+H) at 1.78-1.96 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.74-8.71(m,1H),8.58-8.48(m,1H),8.06-7.97(m,1H),7.72-7.70(m,1H),7.30-7.15(m,,5H),5.05-5.01(m,1H),4.31-4.29(m,1H),3.20-3.20(m,3H),2.74-2.73(m,1H),2.39-2.34(m,2H),2.19-2.14(m,1H),1.91-1.86(m,1H),1.76-1.50(m,3H),1.36-1.32(m,2H),1.28-1.25(m,3H),0.89-0.80(m,,9H),0.67-0.64(m,2H),0.59-0.58(m,2H)。

Examples 14 and 15:(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylpentanamido) pentanamide and (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylpentanamido) pentanamide

Step 1: to a stirred solution of 3-phenylpentanoic acid (2.00 g,11.2 mmol) and methyl (S) -2-amino-4, 4-dimethylvalerate hydrochloride (3.29 g,16.8 mmol) in DCM (30 mL) at 0deg.C was added DIPEA (5.80 mL,33.7 mmol) followed by dropwise addition of T3P (50% EtOAc solution, 10.6mL,16.8 mol). After stirring at room temperature for 16 hours, the reaction mixture was taken up in EtOAc (200 mL) and H ₂ O (200 mL) was partitioned between each phase and the phases separated. With 10% NaHCO ₃ The organic phase was washed with aqueous (200 mL) and brine (100 mL). The organic phase was treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP column (230-400 mesh) eluting with 0-14% EtOAc in petroleum ether afforded methyl (2S) -4, 4-dimethyl-2- (3-phenylpentanamido) pentanoate as an off-white solid as a mixture of diastereomers.

Diastereomers were separated by preparative chiral SFC purification using the following procedure. Instrument: PIC 100 (PIC Solution, inc.); column: chiralcel OX-H (250X 30) mm,5 μm; mobile phase: CO ₂ 0.5% isopropyl amine in methanol (80:20); flow rate: 70g/min; back pressure: 100bar; wavelength: 210nm; cycle time: 4.5 minutes.

The first eluting peak in the purification is referred to as elution 1 and the second eluting peak is referred to as elution 2. After separation, the eluate was concentrated in vacuo to give the step 1 eluted 1 product (1.0 g) and the step 1 eluted 2 product (0.8 g), which were used in the subsequent steps without determining absolute stereochemistry.

Step 1, elution 1:

LCMS (method A) M/z 320.4 (M+H) at 2.41 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.19(d,J＝7.6Hz,1H),7.30-7.26(m,2H),7.19-7.16(m,3H),4.28-4.23(m,1H),3.53(s,3H),2.93(d,J＝4.8Hz,1H),2.47-2.41(m,1H),2.36-2.31(m,1H),1.64-1.48(m,4H),0.87-0.84(m,9H),0.75-0.66(m,3H)。

Step 1, elution 2:

LCMS (method A) M/z 320.3 (M+H) at 2.36 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.16(d,J＝7.6Hz,1H),7.28-7.24(m,2H),7.18-7.14(m,3H),4.23-4.18(m,1H),3.60(s,3H),2.93-2.90(m,1H),2.42-2.34(m,2H),1.68-1.62(m,1H),1.55-1.49(m,2H),1.45-1.39(m,1H),0.75-0.73(m,9H),0.71-0.67(m,3H)。

Steps 2 to 8, using a procedure similar to that described above, the title compound of example 14 was prepared from step 1 eluting the 1 product (1.4 g) as an off-white solid, (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylpentanamido) pentanamide or (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylpentanamido) pentanamide (35 mg,0.07 mmol).

LCMS (method A) M/z 527.4 (M+H) at 1.83-2.00 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.71(d,J＝4.8Hz,1H),8.42(d,J＝6.8Hz,1H),7.98(d,J＝8.4Hz,1H),7.71-7.69(m,1H),7.28-7.24(m,2H),7.17-7.14(m,3H),4.94-4.89(m,1H),4.34-4.30(m,1H),3.25-3.09(m,2H),2.97-2.90(m,1H),2.67-2.67(m,1H),1.90-1.80(m,1H),1.66-1.53(m,5H),1.49-1.37(m,2H),0.87(s,9H),0.71-0.60(m,7H),0.59-0.56(m,3H)。

Steps 2 to 8, using a procedure similar to that described above, the title compound of example 15 was prepared from step 1 eluting the 2 product (1.0 g) as an off-white solid, (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -3-phenylpentanamido) pentanamide or (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -3-phenylpentanamido) penta-mide (30 mg,0.06 mmol).

LCMS (method A) M/z 527.4 (M+H) at 1.83-2.00 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.73-8.70(m,1H),8.50-8.45(m,1H),8.00-7.90(m,1H),7.70(d,J＝5.6Hz,1H),7.29-7.23(m,2H),7.17-7.13(m,3H),4.95-4.82(m,1H),4.25-4.13(m,1H),4.08-3.94(m,1H),3.22-3.04(m,2H),2.89-2.87(m,1H),2.45-2.29(m,2H),2.18-2.05(m,2H),1.89-1.77(m,1H),1.68-1.53(m,3H),1.50-1.39(m,2H),1.33-1.20(m,1H),0.89-0.84(m,3H),0.70-0.66(m,11H),0.53-0.53(m,2H)。

Examples 16 and 17:(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -4, 4-4-trifluoro-3-phenylbutyramide) pentanamide and (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -4, 4-trifluoro-3-phenylbutyramide) pentanamide

Step 1: to 4, 4-trifluoro-3-phenylbutyric acid (2.0 g,9.16 mmol) and (S) -2-amino-4, 4-dimethylvaleric acid methyl ester hydrochloride (2.18 g,11.0 mmol) at 0deg.CDIPEA (6.30 mL,33.7 mmol) was added to a stirred solution in DCM (30 mL), followed by T3P (50% EtOAc solution, 8.60mL,13.8 mmol). After stirring at room temperature for 16 hours, the reaction mixture was taken up in EtOAc (200 mL) and H ₂ O (200 mL) was partitioned between each phase and the phases separated. The organic phase was treated with 10% NaHCO ₃ Aqueous (200 mL) and brine (100 mL) were washed. The organic phase was treated with anhydrous Na ₂ SO ₄ Dried and concentrated in vacuo. Purification by gradient flash column chromatography (Biotage-Isolera) using 25g silica SNAP column (230-400 mesh) eluting with 0-14% EtOAc in petroleum ether afforded methyl (S) -4, 4-dimethyl-2- (4, 4-trifluoro-3-phenylbutyramide) valerate as an off-white solid as a mixture of diastereomers.

Diastereomers were separated by preparative chiral SFC purification using the following procedure. Instrument: PIC 100 (PIC Solution, inc.); column: (R, R) wheelk 250 x 30mm,5 μm; mobile phase: CO ₂ MeOH (90:10); flow rate: 70g/min; back pressure: 100bar; wavelength: 210nm; cycle time: 5.0 minutes.

The first eluting peak in the purification was designated elution 1 and the second eluting peak was designated elution 2. After separation, the eluate was concentrated in vacuo to give the step 1 eluted 1 product (1.0 g) and the step 1 eluted 2 product (0.8 g), which were used in the subsequent steps without determining absolute stereochemistry.

Steps 2 to 8, using a procedure similar to that described above, the title compound of example 16 was prepared from step 1 eluting the 1 product (1.0 g) as an off-white solid, (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -4, 4-trifluoro-3-phenylbutyramide) pentanamide or (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -4, 4-trifluoro-3-phenylbutyramide) pentanamide (5 mg,0.01 mmol).

LCMS (method A) M/z 567.3 (M+H) at 1.88-2.07 min.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.71-8.70(m,1H),8.55(d,J＝6.8Hz,1H),8.17(d,J＝8.4Hz,1H),7.72(d,J＝5.6Hz,1H),7.34-7.31(m,5H),4.98-4.97(m,1H),4.18-4.16(m,1H),4.02-4.01(m,2H),3.19-3.12(m,3H),2.95-2.89(m,2H),2.78-2.74(m,2H),2.18-2.17(m,2H),1.89-1.87(m,1H),1.63-1.57(m,3H),1.48-1.44(m,1H),1.19-1.17(m,1H),0.63-0.59(m,9H)。

Steps 2 to 8, using a procedure similar to that detailed above, the title compound of example 17 was prepared from step 1 eluting the 2 product (0.8 g) from step 1 as an off-white solid, (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((S) -4, 4-trifluoro-3-phenylbutyramide) pentanamide or (2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -4, 4-dimethyl-2- ((R) -4, 4-4-trifluoro-3-phenylbutyramide) pentanamide (40 mg,0.07 mmol).

LCMS (method A) M/z 567.3 (M+H) at 1.86-2.05 minutes.

¹ H NMR:(400MHz,DMSO-d ₆ )δ8.71(d,J＝4.4Hz,1H),8.47(d,J＝7.2Hz,1H),8.24-8.18(m,1H),7.74-7.69(m,1H),7.34-7.31(m,5H),4.89-4.85(m,1H),4.32-4.31(m,1H),4.04-4.02(m,1H),3.16-3.11(m,2H),2.85-2.67(m,3H),2.17-2.16(m,1H),2.05-2.02(m,1H),1.84-1.81(m,1H),1.65-1.54(m,2H),1.40-1.24(m,1H),1.22-1.19(m,1H),0.71(s,7H),0.59-0.56(m,6H)。

Examples 18 to 33 were synthesized using procedures similar to those detailed above.

Biological activity

Construct design of SARS-CoV-2 Mpro

SARS-CoV-2-Mpro (main protease/3C-like protease, uniProt ID: P0DTD 1) protein sequence up to and including its self-cleavage boundary, and the preceding N-terminal 5 amino acid residues, including P1 Glutamine residues, were codon optimized for E.coli expression and cloned into pET26b (Merck, # US 169862-3) or pGEX6P1 (Fisher Scientific, # 10350355) vectors using BamHI and XhoI sites. Thus, the expression construct has the native viral N-terminal sequence and a C-terminal modified 3C-protease cleavage site (LEVLFQGK), with an alternative lysine residue at the P2' position, followed by a polyhistidine (His-8) tag.

Protein expression and protein purification

Chemically active BL21 (DE 3) -RIL E.coli (Agilent, # 230240) cells were transformed with the relevant coronavirus Mpro construct and grown overnight at 37℃on LB agar plates supplemented with the appropriate antibiotics. All incubation steps were performed at 37 ℃ unless otherwise indicated. The colony scraping blade was grown in 15mL of antibiotic-supplemented LB medium for a period of about 2 hours, taking care not to exceed an Optical Density (OD) density of 2.0 measured at 600nm in a spectrophotometer. The preculture was used to inoculate 500mL of expression culture: LB medium for IPTG-induced expression or self-induced Superbroth medium (Formedium, # AIMSB 0210). Expression was induced in LB medium by addition of IPTG to a final concentration of 0.5mM at an OD of 0.7-1.0. Then incubated overnight at 18 ℃. In self-induced expression, once an OD of 0.7-1.0 was observed, the temperature was reduced to 18 ℃ and then grown overnight. Cells were harvested by centrifugation and frozen until use.

Resuspending the thawed cells in a resuspension buffer: 20mM Tris-HCl pH 8.0, 150mM NaCl and DNase I (Merck# 4716728001) and cleaved by sonication. Lysates were clarified by centrifugation at 23,000rcf for 15 min at 4 ℃. The supernatant was loaded onto 5mL of NiNTA resin (Cytiva, # 17-5248-02) at a flow rate of 0.5 mL/min. The resin was washed with the same buffer containing 20mM imidazole as described above. The Mpro protein was eluted using the same buffer containing 250mM imidazole. Target proteins were further purified in resuspension buffer using Superdex S75 16/60pg (GE, # GE 28-9893-33) column. Protein purity was assessed by SDS-PAGE and identity was confirmed by mass spectrometry. The purified protein was concentrated and frozen until later use.

SARS-CoV-2Mpro enzyme assay

In a Fluorescence Resonance Energy Transfer (FRET) based enzyme assay, SARS-Cov-2Mpro activity was measured using the FRET substrate Dabcyl-KTSAVLQSGFRKM-E (Edans) -amide. In brief, 5. Mu.L of test compound (concentration range 100. Mu.M to 0.0017. Mu.M) was pre-incubated with 5. Mu.L of 20nM (final concentration) Mpro enzyme for 30 min at 30℃in assay buffer containing 20mM HEPES, 120mM NaCl, 0.4mM EDTA, 4mM DTT and 20% glycerol. By adding 10. Mu.L of 20. Mu.LThe reaction is initiated by the M (final concentration) FRET substrate (Dabcyl-KTSAVLQSGFRKM-E (Edans) -amide). The reaction was incubated for 1 hour and the resulting fluorescence intensity was measured at 30 ℃ at ex=360 nm/em=490 nm using a SPARK 20M plate reader (Tecan). Boscalid was used as a reference standard compound. Determination of pIC using 4PL GraphPad Prism ₅₀ And pKi, data expressed as mean n=2±sd.

TABLE 2

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Claims

1. A compound of formula (1') or a salt thereof:

wherein;

a is selected from:

q is CN or a group of the formula:

y is O or NOR ¹⁶ ；

T ¹ Is CR (CR) ⁸ Or N;

T ² Is CR (CR) ⁷ Or N;

T ³ is CR (CR) ⁶ Or N;

T ⁴ is CR (CR) ⁵ Or N;

T ⁵ is CR (CR) ⁴ Or N;

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

R ¹³ 、R ¹⁴ and R is ¹⁵ Independent and independentIs H, or C optionally substituted by 1 to 6 fluorine atoms _1-3 An alkyl group;

p and m are independently 0 to 3;

wherein when R is ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

2. The compound of claim 1, which is a compound of formula (1 c) or a salt thereof:

3. the compound of claim 1, which is a compound of formula (1 b) or a salt thereof:

wherein;

q is CN or a group of the formula:

y is O or NOR ¹⁶ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ -；

p and m are independently 0 to 3;

wherein when R is ¹ And R is ^1a When the unevenness is H, L is:

-CHR ¹¹ -CHR ¹² -；

4. The compound of claim 1, which is a compound of formula (1) or a salt thereof:

wherein;

y is O or NOR ¹⁶ ；

L is-CR ¹¹ ＝CR ¹² -、-CHR ¹¹ -CHR ¹² -or-O-CHR ¹¹ ；

p and m are independently 0 to 3;

-CHR ¹¹ -CHR ¹² -；

5. The compound of claim 1, which is a compound of formula (1 ai) or a salt thereof:

6. the compound according to any one of claims 1 to 5, wherein Y is O.

7. A compound according to any one of claims 1 to 3, wherein R ¹ And R is ^1a Independently H or selected from the group consisting ofIn (a):

or wherein R is ¹ And R is ^1a To form a ring, so that the group NR ¹ R ^1a The method comprises the following steps:

8. the compound of claim 1, which is a compound of formula (1 aii) or a salt thereof:

9. the compound according to any one of claims 1 to 8, wherein R ² H.

10. The compound according to any one of claims 1 to 9, wherein X is selected from the group consisting of:

11. The compound of claim 1, which is a compound of formula (3 bi), (3 bii), (3 ci) or (3 cii) and salts thereof:

wherein the method comprises the steps of

Y is O or NOH;

z is optionally oxo or quiltA 5-or 6-membered heterocyclic ring substituted with 1 to 6 fluorine atoms, or Z is C optionally substituted with 1 to 6 fluorine atoms _3-6 Cycloalkyl;

l is-CH=CH-, -CH ₂ -CH ₂ -or-O-CH ₂ -；

R ¹ And R is ^1a Independently H or C optionally substituted with 1 to 6 fluorine atoms _1-4 Saturated hydrocarbon radicals, or R ¹ And R is ^1a Are linked together to form a 3-to 6-membered saturated ring optionally containing additional heteroatoms;

R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ and R is ⁸ Independently H or halogen.

12. The compound according to any one of claims 1 to 11, wherein Z is selected from the group consisting of:

13. the compound of claim 1, which is a compound of formula (4 ai) or (4 aii) or a salt thereof:

14. the compound according to any one of claims 1 to 13, wherein L is-CHR ¹¹ -CHR ¹² -。

15. The compound of claim 1, wherein R ¹ And R is ^1a Are all H and L is-CR ¹¹ ＝CR ¹² -, or wherein R ¹ And R is ^1a Non-uniform H and L is-CR ¹¹ ＝CR ¹² -wherein R is ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ The unevenness is H.

16. The compound of claim 1, wherein R ¹ And R is ^1a Are all H and L is-O-CHR ¹¹ -, or wherein R ¹ And R is ^1a Non-uniform H and L is-O-CHR ¹¹ -wherein R is ³ Other than H, or wherein R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Is not H.

17. The compound of claim 1, wherein L is-ch=ch-, -CH ₂ CH ₂ -、-CH ₂ -CH(CH ₂ CH ₃ )-、-CH ₂ -CH(CH ₂ CO ₂ H)-、-OCH ₂ -、-CH ₂ -CH(CH ₃ )-、-CH ₂ -CH(CF ₃ )-、-OCH(CH ₃ ) -or

18. The compound according to any one of claims 1 to 17, wherein R ³ Is H, methyl or CH ₂ CF ₃ 。

19. The compound of claim 1, which is a compound of formula (9 b) or a salt thereof:

20. the compound according to any one of claims 1 to 19, wherein R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ And R is ⁸ Independently selected from H, CN, cl, F, CF ₂ H、OCH ₃ 、OCF ₃ 、OCF ₂ H、SO ₂ CH ₃ 、OSO ₂ CH ₃ 、PO(CH ₃ ) ₂ 、SF ₅ And CO ₂ H。

21. The compound of claim 20, wherein R ⁴ Is F, R ⁵ Is H, R ⁶ Is Cl, R ⁷ And R is ⁸ H.

22. The compound of claim 1, selected from the group consisting of compounds of:

(2S) -N- (4- (cyclopropylamino) -3, 4-dioxo-1- ((S) -2-oxopyrrolidin-3-yl) butan-2-yl) -2- ((S) -3- (2- (difluoromethoxy) phenyl) pentanamido) -4, 4-dimethylpentanamide;

23. A compound according to any one of claims 1 to 22, which has SARS-CoV-2Mpro inhibitor activity.

24. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 23 and a pharmaceutically acceptable excipient.

25. Use of a compound or composition according to any one of claims 1-24 for the treatment of SARS-CoV-2 or for the treatment of a condition associated with SARS-CoV-2:mpro.