TW202229309A - Compositions and methods for treatment of coronavirus infection - Google Patents

Abstract

Compounds and methods for the treatment of coronavirus infections are disclosed. Compounds according to Formula I as described herein, containing reactive functional groups for covalent binding to active site residues in target proteases, can be used for treating infection by SARS-CoV-2 and other coronaviruses.

Description

Compositions and methods for treatment of coronavirus infection

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). Initial symptoms of COVID-19 can include fever, fatigue, dry cough, pain, nasal congestion, runny nose, sore throat and diarrhea. Although approximately 80% of infected patients are expected to recover from the disease without special treatment, as of September 2021, approximately 1 in 6 patients are severely ill and have difficulty breathing, and approximately 4.5 million people have died worldwide. Other long-term sequelae have been reported, including neurological and cardiovascular conditions. Older adults and people with medical conditions such as high blood pressure, heart problems, and diabetes are at higher risk for serious illness. Similar to other coronaviruses, SARS-CoV-2 expresses a viral protease called 3-chymotrypsin-like cysteine protease (3CLpro), also known as major protease (Mpro), which is Lifecycle and replication are critical.

Cross-references to related applications

This application claims priority to US Provisional Patent Application No. 63/082,305, filed September 23, 2020, which is incorporated herein by reference in its entirety.

Provided herein are compounds useful as inhibitors of the major protease of coronaviruses, which exhibit higher potency in enzyme inhibition and increased efficacy in viral replication assays than previously known compounds. These compounds are useful in the treatment of infections with SARS-Co-V2 and previously known coronaviruses such as MERS and SARS, or viruses that may emerge in the future. Covalent binding of these compounds to catalytic residues in target proteases provides longer occupancy times at the active site and sustained protease inhibition. Because high levels of protease inhibition are generally required to effectively block viral replication, sustained protease inhibition is superior to the activity of protease inhibitors characterized by rapid reversible binding. I. Definition _

As used herein, the term "alkyl" alone or as part of another substituent refers to a straight or branched chain, saturated aliphatic radical having the specified number of carbon atoms. _Alkyl groups can _include any _{number of carbons , such as C1-2 , C1-3 , C1-4 , C1-5 , C1-6 , C1-7 , C1-8 , C1-9 , C1-10 , C2-3 , C2-4 , C2-5 , C2-6 , C3-4 , C3-5 , C3-6 , C4-5 , C4-6 and _ _ _ _ _ _ _ _ _ _} C _{5 - 6} . By way _of example _{, C1-6} alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, tertiary butyl, pentyl, isobutyl Amyl, hexyl, etc. Alkyl can also refer to alkyl groups having up to 20 carbon atoms, such as, but not limited to, heptyl, octyl, nonyl, decyl, and the like. Alkyl groups can be substituted or unsubstituted. Unless otherwise specified, "substituted alkyl" may be substituted with one or more groups selected from the group consisting of halo, hydroxy, amino, alkylamino, amido, amido, nitro, cyano and alkoxy.

As used herein, the term "alkoxy," alone or as part of another substituent, refers to a group of formula -OR, wherein R is an alkyl group.

As used herein, the term "cycloalkyl," alone or as part of another substituent, refers to a saturated or partially unsaturated, monocyclic, fused ring containing from 3 to 12 ring atoms, or the indicated number of atoms. Combined double-ring or bridged multi-ring ring assembly. _{Cycloalkyl groups} can _include any _{number of carbons , such as C3-6 , C4-6 , C5-6 , C3-8 , C4-8 , C5-8 , C6-8 , C3-9} , C _{3 - 10} , C _{3 - 11} and C _{3 - 12} . Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic cycloalkyl rings include, for example, norbornane, [2.2.2]bicyclooctane, decalin, and adamantane. Cycloalkyl groups can also be partially unsaturated, having one or more double or triple bonds in the ring. Representative partially unsaturated cycloalkyl groups include, but are not limited to: cyclobutane, cyclopentane, cyclohexane, cyclohexanediones (1,3- and 1,4-isomers), cyclohexane Heptane, cycloheptane, cyclooctane, cyclooctane (1,3-isomer, 1,4-isomer and 1,5-isomer), norbornane and norbornadiane . When the cycloalkyl group is a saturated monocyclic _{C3-8 cycloalkyl} group _, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. When the cycloalkyl group is a saturated monocyclic _{C3-6 cycloalkyl} group _, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups can be substituted or unsubstituted. Unless otherwise specified, "substituted cycloalkyl" may be substituted with one or more groups selected from the group consisting of halo, hydroxy, amino, alkylamino, amido, amido, nitro, cyano group and alkoxy group. The term "lower cycloalkyl" refers to cycloalkyl groups having three to seven carbons, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

As used herein, the terms "halo" and "halogen" alone or as part of another substituent refer to a fluorine, chlorine, bromine or iodine atom.

As used herein, the term "haloalkyl" alone or as part of another substituent refers to an alkyl group in which some or all of the hydrogen atoms are replaced by halogen atoms. Like alkyl, haloalkyl can have any suitable number _of carbon atoms _, such as _C1-6 . By way of example, haloalkyl includes trifluoromethyl, fluoromethyl, and the like. In some cases, the term "perfluoro" may be used to define a compound or group in which all hydrogens are replaced by fluorine. For example, perfluoromethyl refers to 1,1,1-trifluoromethyl.

As used herein, the term "aryl" alone or as part of another substituent refers to an aromatic ring system having any suitable number of carbon ring atoms and any suitable number of rings. Aryl groups can include any suitable number _of carbon ring atoms, such as _C6 , _C7 , C8, _C9 , _C10 , _C11 , _C12 , _C13 , _C14 , _C15 , or _{C16 , and C6-10} , C _{6 - 12} or C _{6 - 14} . Aryl groups can be monocyclic, fused to form bicyclic groups (eg, benzocyclohexyl) or tricyclic groups, or linked by bonds to form biaryl groups. Representative aryl groups include phenyl, naphthyl, and biphenyl. Other aryl groups include benzyl, which has a methylene linking group. Some aryl groups have 6 to 12 ring members, such as phenyl, naphthyl, or biphenyl. Other aryl groups have 6 to 10 ring members, such as phenyl or naphthyl. Some other aryl groups have 6 ring members, such as phenyl. Aryl groups can be substituted or unsubstituted. Unless otherwise specified, "substituted aryl" may be substituted with one or more groups selected from the group consisting of halo, hydroxy, amino, alkylamino, amido, amido, nitro, cyano and alkoxy.

As used herein, the term "heteroaryl" alone or as part of another substituent refers to a monocyclic or fused bicyclic or tricyclic aromatic ring assembly containing 5 to 16 ring atoms, wherein 1 to 5 One ring atom is a heteroatom such as N, O or S. Additional heteroatoms may also be suitable, including, but not limited to, B, Al, Si, and P. Heteroatoms can be oxidized to form certain moieties such as (but not limited to) -S(O)- and -S(O) _2- . _Heteroaryl groups can include any _{number of ring atoms , such as C5-6 , C3-8 , C4-8 , C5-8 , C6-8 , C3-9 , C3-10 , C3- 11} or _{C3-12 , wherein} at least one carbon atom is replaced by a heteroatom. Heteroaryl groups can include any suitable number of heteroatoms, such as 1, 2, 3, 4, or 5, or 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, 2 to 5, 3 to 4, or 3 to 5. For example, a heteroaryl group can be a _C5-8 heteroaryl _in which 1 to 4 carbon ring atoms are replaced by heteroatoms; or a _C5-8 heteroaryl _in which ₁ to ₃ carbon ring atoms are replaced by heteroatoms substitution; or C5-6 heteroaryl in which _{1 to 4} carbon ring atoms are replaced by heteroatoms; or C5-6 heteroaryl in which _{1 to 3} carbon ring atoms are replaced by heteroatoms. Heteroaryl groups can include groups such as: pyrrole, pyridine, imidazole, pyrazole, triazole, tetrazole, pyridine, pyrimidine, pyridox, tris(1,2,3-isomer, 1,2 , 4-isomer and 1,3,5-isomer), thiophene, furan, thiazole, isothiazole, oxazole and isoxazole. Heteroaryl groups can also be fused with aromatic ring systems such as benzene rings to form members including, but not limited to, benzopyrroles such as indole and isoindole, benzopyridines such as quinoline and isoindole quinoline), benzopyridine (quinoline), benzopyrimidine (quinazoline), benzopyridine (such as quinoline and quinoline), benzothiophene and benzofuran. Other heteroaryl groups include heteroaryl rings linked by bonds, such as bipyridine. Heteroaryl groups can be substituted or unsubstituted. Unless otherwise specified, "substituted heteroaryl" may be substituted with one or more groups selected from the group consisting of halo, hydroxy, amino, alkylamino, amido, amido, nitro, cyano group and alkoxy group.

Heteroaryl groups can be attached via any position on the ring. For example, pyrrole includes 1-pyrrole, 2-pyrrole and 3-pyrrole, pyridine includes 2-pyrrole, 3-pyridine and 4-pyridine, imidazole includes 1-imidazole, 2-imidazole, 4-imidazole and 5-imidazole, Pyrazole includes 1-pyrazole, 3-pyrazole, 4-pyrazole and 5-pyrazole, triazole includes 1-triazole, 4-triazole and 5-triazole, tetrazole includes 1-tetrazole and 5- -Tetrazoles, pyrimidines include 2-pyrimidine, 4-pyrimidine, 5-pyrimidine and 6-pyrimidine, pyrimidines include 3-pyrimidines and 4-pyrimidines, 1,2,3-tripyrimidines include 4-tripyrimidines and 5 -Three𠯤, 1,2,4-Three𠯤 including 3-Three Thiophene, furan including 2-furan and 3-furan, thiazole including 2-thiazole, 4-thiazole and 5-thiazole, isothiazole including 3-isothiazole, 4-isothiazole and 5-isothiazole, oxazole including 2-thiazole oxazole, 4-oxazole and 5-oxazole, isoxazole includes 3-isoxazole, 4-isoxazole and 5-isoxazole, indole includes 1-indole, 2-indole and 3-indole Indole, isoindole includes 1-isoindole and 2-isoindole, quinoline includes 2-quinoline, 3-quinoline and 4-quinoline, isoquinoline includes 1-isoquinoline, 3-isoquinoline Line and 4-isoquinoline, quinazoline includes 2-quinazoline and 4-quinazoline, ethylene includes 3-ethylene and 4-ethylene, benzothiophene includes 2-benzothiophene and 3-benzene thiophene, and benzofuran includes 2-benzofuran and 3-benzofuran.

Some heteroaryl groups include heteroaryl groups having 5 to 10 ring members and 1 to 3 ring atoms (including N, O, or S), such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyridine, pyrimidine, Pap, tris (1,2,3-isomer, 1,2,4-isomer and 1,3,5-isomer), thiophene, furan, thiazole, isothiazole, oxazole, isothiophene oxazole, indole, isoindole, quinoline, isoquinoline, quinoline, quinazoline, quinoline, quinoline, benzothiophene and benzofuran. Other heteroaryl groups include heteroaryl groups having 5 to 8 ring members and 1 to 3 heteroatoms, such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyridine, pyrimidine, pyridine, tris(1, 2,3-isomer, 1,2,4-isomer and 1,3,5-isomer), thiophene, furan, thiazole, isothiazole, oxazole and isoxazole. Some other heteroaryl groups include heteroaryl groups having 9 to 12 ring members and 1 to 3 heteroatoms, such as indole, isoindole, quinoline, isoquinoline, quinoline, quinazoline, quinoline , quinoline, benzothiophene, benzofuran and bipyridine. Other heteroaryl groups include heteroaryl groups having 5 to 6 ring members and 1 to 2 ring atoms including N, O or S, such as pyrrole, pyridine, imidazole, pyrazole, pyridine, pyrimidine, pyridoxine, Thiophene, furan, thiazole, isothiazole, oxazole and isoxazole.

Some heteroaryl groups include from 5 to 10 ring members and heteroatoms that are nitrogen only isomer, 1,2,4-isomer and 1,3,5-isomer), indole, isoindole, quinoline, isoquinoline, quinoline, quinazoline, quinoline and quinoline morpholino. Other heteroaryl groups include 5 to 10 ring members and heteroatoms that are only oxygen, such as furan and benzofuran. Some other heteroaryl groups include 5 to 10 ring members and heteroatoms that are only sulfur, such as thiophene and benzothiophene. Other heteroaryl groups include 5 to 10 ring members and at least two heteroatoms, such as imidazole, pyrazole, triazole, pyridine, pyrimidine, pyridine, tris(1,2,3-isomer, 1,2,3-isomer) 2,4-isomer and 1,3,5-isomer), thiazoles, isothiazoles, oxazoles, isoxazoles, quinoxalines, quinazolines, oxazolines and oxalines.

啶、吡唑啶、咪唑啶、哌𠯤(1,2-異構體、1,3-異構體及1,4-異構體)、環氧乙烷、氧雜環丁烷、四氫呋喃、氧雜環己烷(四氫哌喃)、氧雜環庚烷、環硫乙烷、硫雜環丁烷、硫雜環戊烷(四氫噻吩)、噻𠮿(四氫噻喃)、㗁唑啶、異㗁唑啶、噻唑啶、異噻唑啶、二氧雜環戊烷、二硫雜環戊烷、𠰌啉、硫代𠰌啉、二氧雜環己烷或二噻𠮿。雜環基亦可與芳環或非芳環系統稠合以形成包括(但不限於)吲哚啉之成員。雜環基可未經取代或經取代。除非另外說明，否則「經取代之雜環基」可經選自以下之一或多個基團取代：鹵基、羥基、胺基、側氧基(=O)、烷胺基、醯胺基、醯基、硝基、氰基及烷氧基。 As used herein, the term "heterocyclyl," alone or as part of another substituent, refers to a saturated ring system having 3 to 12 ring members and 1 to 4 heteroatoms of N, O, and S. Additional heteroatoms may also be suitable, including, but not limited to, B, Al, Si, and P. Heteroatoms can be oxidized to form certain moieties such as (but not limited to) -S(O)- and -S(O) _2- . _Heterocyclyl can include any _{number of ring atoms , such as C3-6 , C4-6 , C5-6 , C3-8 , C4-8 , C5-8 , C6-8 , C3- 9} , C _{3 - 10} , C _{3 - 11} or C _{3 - 12} , wherein at least one carbon atom is replaced by a heteroatom. Any suitable number of carbon ring atoms in a heterocyclyl group may be replaced by a heteroatom, such as 1, 2, 3, or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4, or 3 to 4. Heterocyclyl groups can include groups such as aziridine, azetidine, pyrrolidine, piperidine, azepane, azetidine,

pyridine, pyrazolidine, imidazolidine, piperidine (1,2-isomer, 1,3-isomer and 1,4-isomer), ethylene oxide, oxetane, tetrahydrofuran, Oxane (tetrahydropyran), oxetane, thiane, thietane, thiolane (tetrahydrothiophene), thiophene (tetrahydrothiopyran), 㗁oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, dioxolane, dithiacyclopentane, pyridine, thiothiazolidine, dioxane or dithiazolidine. Heterocyclyl groups can also be fused with aromatic or non-aromatic ring systems to form members including, but not limited to, indolines. Heterocyclyl groups can be unsubstituted or substituted. Unless otherwise specified, "substituted heterocyclyl" may be substituted with one or more groups selected from the group consisting of halo, hydroxy, amino, pendant oxy (=O), alkylamino, amido , acyl, nitro, cyano and alkoxy groups.

A heterocyclyl group can be attached via any position on the ring. For example, aziridine can be 1-aziridine or 2-aziridine, azetidine can be 1-azetidine or 2-azetidine, pyrrolidine can be 1- Pyrrolidine, 2-pyrrolidine or 3-pyrrolidine, piperidine can be 1-piperidine, 2-piperidine, 3-piperidine or 4-piperidine, pyrazolidine can be 1-pyrazolidine, 2-piperidine Pyrazolidine, 3-pyrazolidine or 4-pyrazolidine, imidazolidine can be 1-imidazolidinium, 2-imidazolidinium, 3-imidazolidinium or 4-imidazolidinium, piperidine can be 1-piperidine, 2 -piperazolidine, 3-piperazolidine or 4-piperazolidine, tetrahydrofuran can be 1-tetrahydrofuran or 2-tetrahydrofuran, oxazolidine can be 2-oxazolidine, 3-oxazolidine, 4-oxazolidine or 5-oxazolidine oxazolidine, isoxazolidine can be 2-isoxazolidine, 3-isoxazolidine, 4-isoxazolidine or 5-isoxazolidine, thiazolidine can be 2-thiazolidine, 3-thiazole pyridine, 4-thiazolidine, or 5-thiazolidine, the isothiazolidine may be 2-isothiazolidine, 3-isothiazidine, 4-isothiazolidine, or 5-isothiazidine, and the oxaline may be 2-oxathiazidine , 3-𠰌line or 4-𠰌line.

When the heterocyclyl group includes 3 to 8 ring members and 1 to 3 heteroatoms, representative members include, but are not limited to, pyrrolidine, piperidine, tetrahydrofuran, oxane, tetrahydrothiophene, thiazole, Pyrazolidine, imidazolidine, piperidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiazolidine, thiazolidine, dioxane and dithiazolidine. Heterocyclyl groups can also form rings with 5 to 6 ring members and 1 to 2 heteroatoms, representative members of which include, but are not limited to, pyrrolidine, piperidine, tetrahydrofuran, tetrahydrothiophene, pyrazolidine, imidazole pyridine, piperidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine and oxazolidine.

As used herein, the term "carbonyl" alone or as part of another substituent refers to -C(O)-, that is, double bonded to an oxygen and to two other groups in a moiety having a carbonyl group bonded carbon atoms.

As used herein, the term "amino" refers to the _-NR2 moiety, where each R group is H or alkyl. The amine moiety can be ionized to form the corresponding ammonium cation.

As used herein, the term "sulfonyl" refers to a _-SO2R moiety, where the R group is an alkyl, haloalkyl, or aryl group. The amine moiety can be ionized to form the corresponding ammonium cation. "Alkylsulfonyl" refers to an amine moiety in which the R group is an alkyl group.

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

As used herein, the term "cyano" refers to a carbon atom triple bonded to a nitrogen atom (ie, -C≡N).

As used herein, the term "carboxy" refers to the -C(O)OH moiety. Carboxyl moieties can be ionized to form the corresponding carboxylate anions.

As used herein, the term "amido" refers to a -NRC(O)R or _-C (O)NR2 moiety, where each R group is H or alkyl.

As used herein, the term "nitro" refers to the _-NO2 moiety.

As used herein, the term "pendant oxy" refers to an oxygen atom that is double bonded to a compound (ie, O=).

As used herein, the term "pharmaceutically acceptable excipient" refers to a substance that facilitates administration of an active agent to an individual. "Pharmaceutically acceptable" means that the excipient is compatible with the other ingredients in the formulation and is not harmful to its recipient. Pharmaceutical excipients suitable for use in the present invention include, but are not limited to, binders, fillers, disintegrants, lubricants, slip agents, coatings, sweeteners, flavoring agents and pigments.

As used herein, the term "salt" refers to an acid or base salt of an active agent such as a protease inhibitor. Acid salts of alkaline active agents include inorganic acid salts (eg, using hydrochloric acid, hydrobromic acid, phosphoric acid, and the like) and organic acid salts (eg, using acetic acid, propionic acid, glutamic acid, citric acid) and its analogs). Quaternary ammonium salts can be formed using reagents such as methyl iodide, ethyl iodide, and the like. It is understood that pharmaceutically acceptable salts are non-toxic.

The acidic active agent can be contacted with a base to obtain basic salts such as alkali metal salts and alkaline earth metal salts such as sodium, lithium, potassium, calcium, magnesium and ammonium salts such as ammonium, trimethylammonium, Diethylammonium salt and para-(hydroxymethyl)-methyl-ammonium salt.

The neutral form of the active agent can be regenerated by contacting the salt with a base or acid and isolating the parent compound as desired in a conventional manner. In some embodiments, the parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents, but in other respects the salt form may be equivalent to the parent form of the compound.

As used herein, the term "treat/treatment/treating" refers to successful treatment or amelioration of any marker of an injury, lesion, condition or symptom (eg, viral infection), including any objective or subjective parameter, such as remission Alleviate symptoms or make patients more tolerant of symptoms, injuries, lesions, or conditions; reduce the rate of symptom development; reduce the frequency or duration of symptoms or conditions; or, in some cases, prevent the onset of symptoms. Treatment or amelioration of symptoms can be based on any objective or subjective parameter, including, for example, the results of a physical examination.

As used herein, the terms "effective amount" and "therapeutically effective amount" refer to a dose of a compound (such as a 3CL ^pro inhibitor) that produces a therapeutic effect in a subject to which it is administered. The precise dose will depend on the purpose of treatment, and can be determined by those skilled in the art using known techniques (see, eg, Lieberman, Pharmaceutical Dosage Forms (Vol. 1-3, 1992); Lloyd, The Art , Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); Goodman and Gilman , The Pharmacological Basis of Therapeutics , 11th edition, 2006, edited by Brunton, McGraw-Hill; and Remington : The Science and Practice of Pharmacy , 21st edition, 2005 , edited by Hendrickson, Lippincott, Williams & Wilkins).

As used herein, the term "individual" refers to animals, such as mammals, including but not limited to primates (eg, humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, Mice and similar animals. II . Protease inhibitor compounds

， 及其醫藥學上可接受之鹽， 其中： R ¹係選自由以下組成之群：C _{6 - 10}芳基、5員至12員雜芳基及C _{1 - 6}鹵烷基，其中C _{6 - 10}芳基經一或多個R ^1a取代，且5員至12員雜芳基視情況經一或多個R ^1b取代； 各R ^1a獨立地為鹵素； 各R ^1b係獨立選自由鹵素、C _{1 - 3}烷基及C _{1 - 3}鹵烷基組成之群； R ²係選自由以下組成之群：吡咯啶基、哌啶基、氮雜環庚烷基及-CH ₂C(O)NH ₂，其中吡咯啶基、哌啶基及氮雜環庚烷基視情況經一或多個側氧基部分取代； R ³係選自由H及C _{1 - 6}烷基組成之群； R ⁴係選自由C _{1 - 6}烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^4a取代； 各R ^4a係獨立選自由C _{1 - 6}烷基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^4b取代； 各R ^4b為獨立選擇之鹵素； R ⁵係選自由H及C _{1 - 6}烷基組成之群； 或R ⁴及R ⁵共同形成視情況經一或多個R ^4a取代之單環或雙環5員至12員雜環基； R ⁶係選自由以下組成之群：C _{6 - 10}芳基、5員至12員雜芳基、-OR ⁷、-NHR ⁷、-NHC(O)OR ⁷及-CHR ⁷NHC(O)R ⁷，其中C _{6 - 10}芳基、5員至12員雜芳基視情況經一或多個R ^6a取代； 各R ^6a係獨立選自由C _{1 - 6}烷基、C _{1 - 6}烷氧基、羥基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群； 或R ⁵及R ⁶與其所附接之原子共同形成視情況經-NHR ⁷或-NHC(O)OR ⁷取代之5員至12員雜環基； R ⁷係選自由C _{1 - 6}烷基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^7a取代； 各R ^7a係獨立選自由以下組成之群：鹵素、C _{1 - 6}烷基、C _{6 - 10}芳基及C _{3 - 8}環烷基，其中C _{1 - 6}烷基、C _{6 - 10}芳基及C _{3 - 8}環烷基視情況經一或多個R ^7b取代；及 各R ^7b為獨立選擇之鹵素。 Provided herein are compounds of formula I:

, and pharmaceutically acceptable salts thereof, wherein: R ¹ is selected from the group consisting of C _{6 - 10} aryl, 5- to 12-membered heteroaryl and C _{1 - 6} haloalkyl, wherein C _{6 - 10} -aryl is substituted with one or more R ^1a , and 5- to 12-membered heteroaryl is optionally substituted with one or more R ^1b ; each R ^1a is independently halogen; each R ^1b is independently selected from halogen, _The group consisting _{of C1-3} alkyl _{and C1-3} haloalkyl _; R2 is selected from the group consisting of pyrrolidinyl, piperidinyl, azepanyl and ^-CH2C ₍ O) NH ₂ , wherein pyrrolidinyl, piperidinyl and azepanyl are optionally substituted with one or more pendant oxy moieties; R ³ is selected from the group consisting _of H and C _1-6 alkyl ^; R ₄ is selected from the group consisting _of C _1-6 alkyl and C _6-10 aryl, each _of which is optionally substituted with _one or more R ^4a _; each _R ^4a is independently selected from C _1-6 alkyl _, C _{3 -} The group consisting of ₈ cycloalkyl and C _6-10 aryl, each _of which is optionally substituted by _one or more R ^4b ; each R ^4b is independently selected halogen _; R ⁵ is selected from H _and C _1-6 alkyl or R ⁴ and R ⁵ together form a monocyclic or bicyclic 5- to 12-membered heterocyclic group substituted by one or more R ^4a as appropriate; R ⁶ is selected from the group consisting of: C _{6 - 10} Aryl, 5- to 12-membered heteroaryl, -OR ⁷ , -NHR ⁷ , -NHC(O)OR ⁷ and -CHR ⁷ NHC(O)R ⁷ , wherein C _{6 -10} aryl, 5- to ₁₂ -membered Member heteroaryl is optionally substituted with one or more R ^6a _{; each R 6a is independently selected from C 1-6 alkyl, C 1-6 alkoxy} ^, _{hydroxy , C 3-8 cycloalkyl and C 6 -} A group consisting of ₁₀ aryl groups; or R ⁵ and R ⁶ and the atoms to which they are attached together form a 5- to 12-membered heterocyclic group substituted by -NHR ⁷ or -NHC(O)OR ⁷ as appropriate; R ⁷ is selected _The group consisting _{of C1-6} alkyl _{, C3-8} cycloalkyl _{and C6-10} aryl, each of which is optionally substituted with _one or more R ^7a _; each R ^7a is independently selected from the group consisting of : halogen, C _1-6 alkyl _, C _6-10 aryl _and C _{3-8 cycloalkyl ,} wherein _{C 1-6 alkyl ,} C _{6-10 aryl and C 3-8 cycloalkyl} are _optionally One or more R ^7b is substituted; and each R ^7b is independently selected halogen.

Some embodiments of the present invention provide compounds of formula I and pharmaceutically acceptable salts thereof, wherein: R ¹ is selected from the group consisting of C _6-10 aryl, 5- to _{12 -} membered heteroaryl, and C _1-6 haloalkyl _, wherein C6-10 aryl is substituted with _one or more R ^1a , and 5- _{to 12 -} membered heteroaryl is optionally substituted with one or more R ^1b ; each R ^1a is independently halogen ; Each R ^1b is independently selected from the group consisting _of halogen, C _1-3 alkyl _and C _1-3 haloalkyl ^{; R 2} _{is selected} from the group consisting of: pyrrolidinyl, piperidinyl, azepan Alkyl and _-CH2C (O) _NH2 , wherein pyrrolidinyl, piperidinyl and azepanyl are optionally substituted with one or more pendant oxy moieties; ^R3 is selected from H and _{C1 -} the group consisting of ₆ alkyl groups; R ⁴ is selected from the group consisting of C _{1 -6} alkyl groups and C _{6 - 10} aryl groups _, each of which is optionally substituted with one or more R ^4a ; each R ^4a is independently selected from _The group consisting _of C _1-6 alkyl, C _3-8 cycloalkyl _and C _6-10 aryl, each _of which is optionally substituted by _one or more R ^4b ; each R ^4b is an independently selected halogen ^{; R 5} _is is selected from the group consisting of H and C _{1 -6} alkyl; R ⁶ is selected from the group consisting of C _{6 - 10} aryl, 5- to 12 _- membered heteroaryl, -OR ⁷ , -NHR ⁷ and -NHC (O) ^OR7 ; or ^R5 and R6 together with the atoms to which they are attached form a ^5- to 12-membered heterocyclyl optionally substituted with ^-NHR7 or -NHC(O) ^OR7 ; ^R7 is selected from _The group consisting _of C _1-6 alkyl and C _6-10 aryl, each _of which is optionally substituted with _one or more R ^7a _; each R ^7a is independently selected _from C _1-6 alkyl _, C _{6-10 aryl and C3-8cycloalkyl} , each of which is optionally substituted with one or more _R7b ; and ^{each R7b is} independently selected halogen.

， 其中下標m為1、2或3。在一些實施例中，R ²係選自由以下組成之群：2-側氧基吡咯啶-3-基、5-側氧基吡咯啶-3-基及2,5-二側氧基吡咯啶-3-基。 In some embodiments, R ² is -CH ₂ C(O)NH ₂ . In some embodiments, R ² is pyrrolidinyl, piperidinyl, or azepanyl. In some embodiments, R ² is substituted with one or more pendant oxy moieties. Examples of R groups include ⁽ but are not limited to):

, where the subscript m is 1, 2, or 3. In some embodiments, R ² is selected from the group consisting of 2-oxypyrrolidin-3-yl, 5-oxypyrrolidin-3-yl, and 2,5-dioxypyrrolidine -3-base.

_In some embodiments _, R ¹ is C _1-6 haloalkyl. In such embodiments, R ¹ can be, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trichloroethyl , 2,2,2-trifluoroethyl, pentachloroethyl, pentafluoroethyl, 1,1,1,3,3,3-hexachloropropyl, 1,1,1,3,3,3 - Hexafluoropropyl or an analogue thereof.

In some embodiments, R ¹ is a 5- to 12-membered heteroaryl group optionally substituted with one or more R ^1b . R ¹ can be, for example, isoxazolyl, oxazolyl, imidazolyl, pyrazolyl, pyridyl, pyridyl, pyridyl, pyridyl, or pyridyl. In some embodiments, R ¹ is pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl , pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl or pyrimidin-6-yl. In some embodiments, R ¹ is selected from isoxazol-3-yl, pyridin-3-yl, pyridin-4-yl, 2,6-lutidine-5-yl, and 2-methylpyrimidine- 5-base. In some embodiments, R ¹ is selected from isoxazol-3-yl, pyridin-3-yl, pyridin-4-yl, 2,6-lutidine-5-yl, and 2-methylpyrimidine- 5-base.

In some embodiments, R ¹ is phenyl substituted with 1-5 independently selected halo. The -OR ¹ moiety can be, for example, 2-halophenoxy; 3-halophenoxy; 4-halophenoxy; 2,3-dihalophenoxy; 2,4-dihalophenoxy; 5-dihalophenoxy; 2,6-dihalophenoxy; 3,4-dihalophenoxy; 3,5-dihalophenoxy; 2,3,6-trihalophenoxy; 2,3,5-trihalophenoxy; or 2,3,5,6-tetrahalophenoxy. In some embodiments, each halogen in the halophenoxy moiety can independently be fluorine, chlorine, or bromine.

In some embodiments, R ¹ is selected from the group consisting of phenyl, hexafluoroisopropyl, 2,6-dimethylpiperidin-4-yl, 2-methylpyrimidin-5-yl, and isopropyl Oxazol-3-yl wherein the phenyl group is substituted with 1-5 independently selected halogens.

， 及其醫藥學上可接受之鹽。 In some embodiments, compounds having the structure of Formula Ia are provided:

, and their pharmaceutically acceptable salts.

， 及其醫藥學上可接受之鹽，其中下標n為1至5範圍內之整數。 In some embodiments, compounds having the structure of Formula Ib are provided:

, and pharmaceutically acceptable salts thereof, wherein the subscript n is an integer in the range of 1 to 5.

In some embodiments, the subscript n is 3. In some embodiments, the subscript n is 4. In some embodiments, each R ^1a is fluoro.

In some embodiments, R ³ is H. In some embodiments, R ³ is methyl (-CH ₃ ), ethyl (-CH ₂ CH ₃ ), n-propyl, isopropyl, n-butyl, isobutyl, tertiary butyl, tertiary Butyl, n-pentyl, branched pentyl, n-hexyl, branched hexyl. ^In some embodiments, R5 is H. In some embodiments, R ⁵ is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tertiary butyl, tertiary butyl, n-pentyl, branched pentyl, n-hexyl, branched chain hexyl. ^In some embodiments, R5 is H or _-CH3 .

_In some embodiments, R ⁴ is C _1-6 alkyl _, optionally substituted with one or more R ^4a . _In some embodiments _, R ^4a is C _3-8 cycloalkyl or C _{6-10 aryl} substituted with halogen _. In some embodiments, R ⁴ and R ⁵ together form a monocyclic or bicyclic 5- to 12-membered heterocyclyl optionally substituted with one or more R ^4a . In some embodiments, R ⁴ is methyl substituted with cycloalkyl. ^R4 can be, for example, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl or cyclohexylmethyl. In some embodiments, R ⁴ is unsubstituted C _1-6 alkyl (eg, n _- propyl, isopropyl, n _- butyl, isobutyl, tertiary butyl, or tertiary butyl).

In some embodiments, R ⁶ is selected from the group consisting of indol-2-yl, benzofuran-2-yl, and benzyloxy, each of which is unsubstituted or substituted with one or more R ^6a . In some embodiments, R ⁶ is unsubstituted indol-2-yl or indol- _{2-yl substituted with C 1-6 alkoxy .} In some embodiments, R is ⁴ -alkoxyindol-2-yl, 5-alkoxyindol-2-yl, 6-alkoxyindol-2-yl, or 7-alkoxy Indol-2-yl (eg 4-methoxyindol-2-yl, 5-methoxyindol-2-yl, 6-methoxyindol-2-yl or 7-methoxyindol-2-yl dol-2-yl).

In some embodiments, R ⁶ is unsubstituted benzofuran-2 _- yl or C _1-6 alkoxy substituted benzofuran- ₂ -yl. In some embodiments, R ⁶ is 4-alkoxybenzofuran-2-yl, 5-alkoxybenzofuran-2-yl, 6-alkoxybenzofuran-2-yl, or 7- Alkoxybenzofuran-2-yl (eg 4-methoxybenzofuran-2-yl, 5-methoxybenzofuran-2-yl, 6-methoxybenzofuran-2-yl or 7-methoxybenzofuran-2-yl).

In some embodiments, R ⁵ and R ⁶ together with the atoms to which they are attached form a 5- to 12-membered heterocyclyl group optionally substituted with -NHR ⁷ or -NHC(O)OR ⁷ . For example, R ⁵ and R ⁶ can together form pendant oxydihydropyrrolyl, pendant oxydihydropyridyl, pendant oxydihydroimidazolyl, pendant oxydihydropyrazolyl, pendant oxydihydro Triazolyl, pendant oxydihydropyridine or pendant oxydihydrotrizyl. In some embodiments, R ⁵ and R ⁶ together with the atoms to which they are attached form 3-pendant oxy-3,4-dihydropyridine-2-yl or 2-pendant oxy-1,2-dihydro Pyridin-3-yl, each substituted with ^-NHR7 or -NHC(O) ^OR7 .

_In some embodiments, R ⁶ is -CHR ⁷ NHC(O)R ⁷ , wherein R ⁷ is C _1-6 alkyl, each _of which is optionally substituted with halogen. _In some embodiments, R ⁶ is -OR ⁷ _, wherein R ⁷ is C _3-8 cycloalkyl optionally substituted with halogen.

In some embodiments, compounds of Formula I, Formula Ia, and/or Formula Ib and pharmaceutically acceptable salts thereof are provided, wherein: R ³ is H _; R ⁴ is unsubstituted C _1-6 alkyl ₍ For example, n-propyl, isopropyl, n-butyl, isobutyl, tertiary or tertiary butyl) _{, C3-8cycloalkyl -} methyl (eg, cyclopropylmethyl, cyclobutylmethyl) (eg, halobenzyl); R is H; and R is indol ^{- 2} -yl or benzo Furan- _{2-yl, each of which is optionally substituted with one or more C 1-6 alkoxy} groups ₍ eg, methoxy).

In some embodiments, compounds of Formula I and/or Formula Ia and pharmaceutically acceptable salts thereof are provided, wherein: R ¹ is 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2 ,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl Phenyl, 2,3,6-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,5,6-tetrafluorophenyl, 1,1,1,3,3,3- hexafluoroisopropyl, 2,6-dimethylpiperidin-4-yl, or isoxazol- ₃ -yl; R ³ is H; R ⁴ is unsubstituted C _1-6 alkyl ₍ eg, n-propyl, isopropyl, n-butyl, isobutyl, tertiary or tertiary butyl), C _{3-8 cycloalkyl -} methyl (eg, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl or cyclohexylmethyl) or optionally substituted benzyl (eg, halobenzyl); R is H; and R is indol ^{- 2} -yl or benzofuran- _{2-yl groups, each of which is optionally substituted with one or more C 1-6 alkoxy} groups ₍ eg, methoxy).

， 下標m為1、2或3； R ¹為2-氟苯基、3-氟苯基、4-氟苯基、2,3-二氟苯基、2,4-二氟苯基、2,5-二氟苯基、2,6-二氟苯基、3,4-二氟苯基、3,5-二氟苯基、2,3,6-三氟苯基、2,3,5-三氟苯基、2,3,5,6-四氟苯基、1,1,1,3,3,3-六氟異丙基、2,6-二甲基哌啶-4-基或異㗁唑-3-基； R ³為H； R ⁴為未經取代之C _{1 - 6}烷基(例如，正丙基、異丙基、正丁基、異丁基、二級丁基或三級丁基)、C _{3 - 8}環烷基-甲基(例如，環丙基甲基、環丁基甲基、環戊基甲基或環己基甲基)或視情況經取代之苯甲基(例如，鹵基苯甲基)； R ⁵為H；及 R ⁶為吲哚-2-基或苯并呋喃-2-基，其各自視情況經一或多個C _{1 - 6}烷氧基(例如甲氧基)取代。 In some embodiments, there are provided ^compounds of Formula I, and pharmaceutically acceptable salts thereof, wherein: R is

, the subscript m is 1, 2 or 3; R ¹ is 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2,3,6-trifluorophenyl, 2,3 ,5-trifluorophenyl, 2,3,5,6-tetrafluorophenyl, 1,1,1,3,3,3-hexafluoroisopropyl, 2,6-dimethylpiperidine-4 -yl or isoxazol- ₃ -yl; R ³ is H; R ⁴ is unsubstituted C _1-6 alkyl (for example, n _- propyl, isopropyl, n-butyl, isobutyl, secondary butyl or tertiarybutyl) _{, C3-8cycloalkyl -} methyl (eg, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, or cyclohexylmethyl), or optionally substituted benzene Methyl (eg, halobenzyl); R ⁵ is H; and R ⁶ is indol-2-yl or benzofuran- _{2 -} yl, each optionally via one or more C _1-6 alkanes Oxy (eg methoxy) substituted.

In some embodiments, there are provided compounds of Formula I and pharmaceutically acceptable salts thereof, wherein: R ¹ is 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluoro Phenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2, 3,6-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,5,6-tetrafluorophenyl, 1,1,1,3,3,3-hexafluoroisopropyl , 2,6-dimethylpiperidin-4-yl or isoxazol-3-yl; R ² is 2-oxypyrrolidin-3-yl, R ³ is H; R ⁴ is isobutyl or cyclopropylmethyl; ^R5 is H; and R6 is indol- ² -yl or 4-methoxyindol-2-yl.

， 及其醫藥學上可接受之鹽。 In some embodiments, the compound is selected from the group consisting of:

, and their pharmaceutically acceptable salts.

， 或其醫藥學上可接受之鹽。 In some embodiments, the compound is:

, or a pharmaceutically acceptable salt thereof.

， 及其醫藥學上可接受之鹽。 In some embodiments, the compound is selected from the group consisting of:

, and their pharmaceutically acceptable salts.

， 或其醫藥學上可接受之鹽。 In some embodiments, the compound is:

, or a pharmaceutically acceptable salt thereof.

The compounds of the present invention may be further substituted. In general, the term "substituted", whether or not preceded by the term "optional", means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be selected from the group specified by more than one When a substituent of a group is substituted, the substituent at each position may be the same or different. Combinations of substituents are generally those that result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to a compound that does not occur when subjected to conditions that allow its production, detection, and, in certain embodiments, its recovery, purification, and use for one or more of the purposes disclosed herein substantially changed. In general, "substituted" as used herein does not encompass substitutions and/or alterations of key functional groups used to identify a molecule, such as the substitution of a "substituted" functional group into a different functional group. For example, "substituted phenyl" must still contain a phenyl moiety and, in this definition, cannot be modified by substitution to become, for example, cyclohexyl.

Examples of suitable monovalent substituents on substitutable carbon atoms for "optionally substituted" groups are independently: halogen ^; _- ( _CH2 ) _0-4Rα ^; _{- ( CH2 ) 0-4ORα} ; -O(CH ₂ ) _{0 - 4} R ^α , -O-(CH ₂ ) _{0 - 4} C(O)OR ^α ; -(CH ₂ ) _{0 - 4} CH(OR ^α ) ₂ ; -(CH ₂ ) _{0 - 4} SR ^α ; -(CH ₂ ) _{0 - 4} Ph, wherein Ph is a phenyl that can be substituted by R ^α ; -(CH ₂ ) _{0 - 4} O(CH ₂ ) _{0 - 1} phenyl, the phenyl Can be substituted by R ^α ; -CH=CHPh, wherein Ph is phenyl that can be substituted by R ^α ; -(CH ₂ ) _{0 - 4} O(CH ₂ ) _{0 - 1} -Py, wherein Py is substituted by R ^α -NO ₂ ; -CN; -N ₃ ; -(CH ₂ ) _{0 - 4} N(R ^α ) ₂ ; -(CH ₂ ) _{0 - 4} N(R ^α )C(O)R ^α ; -N(R ^α )C(S)R ^α ;-(CH ₂ ) _{0 - 4} N(R ^α )C(O)NR ^α ₂ ;-N(R ^α )C(S)NR ^α ₂ ;-( CH ₂ ) _{0 - 4} N(R ^α )C(O)OR ^α ;-N(R ^α )N(R ^α )C(O)R ^α ;-N(R ^α )N(R ^α )C(O ) ^NRα2 ;-N( ^Rα )N( ^Rα )C(O) ^ORα _{;-( CH2 ) 0-4C} (O) _Rα ^; -C(S) ^Rα ;-( _CH2 ) _{0 - 4} C(O)OR ^α ; -(CH ₂ ) _{0 - 4} C(O)SR ^α ; -(CH ₂ ) _{0 - 4} C(O)OSiR ^α ₃ ; -(CH ₂ ) _{0 - 4} OC(O)R ^α ; -OC(O)(CH ₂ ) _{0 - 4} SR-SC(S)SR ^α ; -(CH ₂ ) _{0 - 4} SC(O)R ^α ; -(CH ₂ ) _{0 - 4} C(O)NR ^α ₂ ; -C(S)NR ^α ₂ , -C(S)SR ^α ; -SC(S)SR ^α , -(CH ₂ ) _{0 - 4} OC(O)NR ^α ₂ ; -C(O)N( ^ORα ) ^Rα ;-C(O)C(O) ^Rα ;-C(O) _CH2C (O) ^Rα ;-C( ^NORα ) ^Rα ;-( CH ₂ ) _{0 - 4} SSR ^α ; -(CH ₂ ) _{0 - 4} S(O) ₂ R ^α ; -(CH ₂ ) _{0 - 4} S(O) ₂ OR ^α ; -(CH ₂ ) _{0 - 4} OS(O) ₂ R ^α ; -S(O) ₂ NR ^α ₂ ; -(CH ₂ ) _{0 - 4} S(O)R ^α ;- N( ^Rα )S(O) ^{2NRα2 ;} -N(Rα)S(O) ^{2Rα ;} -N( ^ORα ₎ ^Rα ; _-C (NH) _{NRα2 ;-P} (O ) ₂ Rα ^;-P (O) ^Rα2 ;-OP(O) ^Rα2 _; -OP(O) _{( ORα ) 2 ;} ^{-SiRα3 ;-(} C _1-4 linear or branched chain ) alkylene)-ON(R ^α ) ₂ ; or _- (C _1-4 straight or branched chain)alkylene)-C(O)ON(R ^α _{) 2 .} Each R ^α is independently: hydrogen; C _{1 -8} alkyl; -CH ₂ Ph, -O( _{CH 2 ) 0 - 1} Ph; -CH ₂ -(5- to 6-membered heteroaryl); C _{3 - 8 -} cycloalkyl; C6-10 aryl; 4- to ₁₀ -membered heterocyclyl; or _6- to 10-membered heteroaryl; and each ^Rα may be further substituted as described below.

Examples of suitable monovalent substituents on _Rα are independently: halogen, - ⁽ _{CH2 )} ^0-2Rβ _{;-( CH2 ) 0-2OH ;-( CH2 )} ^0-2ORβ _{; -} (CH ₂ ) _{0 - 2} CH(OR ^β ) ₂ ; -CN; -N ₃ ; -(CH ₂ ) _{0 - 2} C(O)R ^β ; -(CH ₂ ) _{0 - 2} C(O)OH; -(CH ₂ ) _{0 - 2} C(O)OR ^β ; -(CH ₂ ) _{0 - 2} SR ^β ; -(CH ₂ ) _{0 - 2} SH; -(CH ₂ ) _{0 - 2} NH ₂ ; -(CH ₂ ) _{0 - 2} NHR ^β ; -(CH ₂ ) _{0 - 2} NR ^β ₂ ; -NO ₂ ; SiR ^β ₃ ; -OSiR ^β ₃ ; -C(O)SR ^β ; -(C _{1 - 4} linear or or ^{-SSR β ; wherein each R β is independently} selected from: C _{1 -4} alkyl; _{-CH 2} Ph; -O(CH ₂ ) _{0 - 1} Ph; _C3-8 cycloalkyl; C6-10 aryl; _4- to _{10 -} membered heterocyclyl; or _6- to ₁₀ -membered heteroaryl. Suitable divalent substituents on the saturated carbon atoms of Rα include ⁼ O and =S.

Examples of suitable divalent substituents on saturated carbon atoms of "optionally substituted" groups include the following: =O;=S;= ^NNRγ2 ;=NNHC(O) _Rγ ;= ^NNHC (O) OR ^γ ;=NNHS(O) ₂ R ^γ ;=NR ^γ ;=NOR ^γ ;-O(C(R ^γ ₂ )) _{2 - 3} O-; or -S(C(R ^γ ₂ )) _{2 - 3} S-; wherein each individual occurrence of ^Rγ is selected from: hydrogen _{; C1-8} alkyl, which may be substituted as defined below _{; C3-8} cycloalkyl _{; C6-10} aryl _{; 4 -} membered to 10-membered heterocyclyl; or 6- to 10-membered heteroaryl. Suitable divalent substituents bonded to the ortho-substitutable carbon of an "optionally substituted" group include: -O(CR ^β ₂ ) _{2 -3} O-; wherein each individual occurrence of _R ^β is selected from : hydrogen; _C1-8 alkyl which may be substituted as hereinafter defined; _C3-8 cycloalkyl _; C6-10 aryl; _4- to _{10 -} membered heterocyclyl; or _{6- to 10} -membered heteroaryl base.

Examples of suitable substituents on the alkyl group of ^Rγ include: halogen; ^-Rδ ; -OH; ^-ORδ ; -CN; -C(O)OH; -C(O) ^ORδ ; _-NH2 ; -NHR ^δ ; -NR ^δ ₂ ; or -NO ₂ ; wherein each R ^δ is independently: C _{1 -4} alkyl; _{-CH 2} Ph; -O(CH ₂ ) _{0 - 1} Ph; 4 to 10 members Heterocyclyl; or 6- to 10-membered heteroaryl.

Examples of suitable substituents on substitutable nitrogens of "optionally substituted" groups include: ^-Rε ; ^-NRε2 ; _-C (O) ^Rε ;-C(O) ^ORε ;-C (O)C(O) ^Rε ;-C(O) _CH2C (O) ^Rε ;-S(O) _2Rε ; ^-S (O) ^2NRε2 _{; -C} (S) ^NRε ₂ ; -C(NH)NR ^ε ₂ ; or -N(R ^ε )S(O) ₂ R ^ε ; wherein each R ^{ε is} independently: hydrogen _{; C1-8} alkyl substituted as defined _below ; _C3-8 cycloalkyl; C6-10 aryl; _4- to _{10 -} membered heterocyclyl; or _6- to ₁₀ -membered heteroaryl.

Examples of suitable substituents on the alkyl group of R ^ε are independently: halogen; ^-Rδ ;-OH; ^-ORδ ;-CN;-C(O)OH;-C(O) ^ORδ ;-NH ₂ ; ^-NHRδ ; ^-NRδ2 ; or _{-NO2 ;} wherein each ^Rδ is independently _{: C1-4} alkyl _{; -CH2Ph ;} -O( _{CH2 ) 0-1Ph ; C6- 10} -membered aryl; 4- to 10-membered heterocyclyl; or 6- to 10-membered heteroaryl.

Compounds of the invention can be prepared as outlined in Scheme 1 and as described in the Examples below. Protected aspartic acid (i) can be reacted with bromoacetonitrile and cyclized to form the corresponding pendant oxypyrrolidone (iii), followed by installation of an α-halomethylene group, which is displaced by alcohol to obtain Oxymethyl ketone (v). Removal of the protecting group yields amine (vi), which can be further processed in a subsequent acylation step. Process 1

Starting materials and reagents for the preparation of the compounds of the present invention can be obtained from commercial suppliers, or by methods known to those skilled in the art according to references (such as Fieser and Fieser , Reagents for Organic Synthesis , Vols . 1-28 ( Wiley, 2016); March, Advanced Organic Chemistry , 7th Ed. (Wiley, 2013) ; and Larock, Comprehensive Organic Transformations , 2nd Ed. ( Wiley, 1999)). If desired, the starting materials and intermediates of the reaction may be isolated and purified using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such substances can be characterized using conventional means including measuring physical constants and obtaining spectral data.

鹽(例如嘧啶鎓

鹽(諸如HATU)、四甲基銨鹽、雙吡咯啶銨鹽、雙哌啶銨鹽、咪唑鎓

鹽、衍生自 N , N , N'-三甲基- N'-苯脲之

鹽、基於N-𠰌啉基之銨/

偶合試劑、銻酸

鹽等)、有機磷試劑(例如次膦酸及磷酸衍生物)、有機硫試劑(例如磺酸衍生物)、三𠯤偶合試劑(例如2-氯-4,6-二甲氧基-1,3,5-三𠯤、氯化4-(4,6-二甲氧基-1,3,5-三𠯤-2-基)-4甲基𠰌啉鎓、四氟硼酸4-(4,6-二甲氧基-1,3,5-三𠯤-2-基)-4甲基𠰌啉鎓等)、吡啶鎓偶合試劑(例如向山試劑(Mukaiyama's reagent)、吡啶鎓四氟硼酸偶合試劑等)、聚合物載體試劑(例如與聚合物結合之碳化二亞胺、與聚合物結合之TBTU、與聚合物結合之2,4,6-三氯-1,3,5-三𠯤、與聚合物結合之HOBt、與聚合物結合之HOSu、與聚合物結合之IIDQ、與聚合物結合之EEDQ等)及其類似物。或者，醯化可使用活化羧酸衍生物，諸如酸酐、混合酸酐、酸氯化物或活化酯(例如五氟苯基酯或 N-羥基丁二醯亞胺基酯)進行。 The acylation step can be carried out with one or more coupling agents including, for example, carbodiimides (eg N , N' -dicyclohexylcarbodiimide (DCC), N , N' -dicyclopentylcarbodiimide Amines, N , N' -diisopropylcarbodiimide (DIC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), etc.), phosphonium salts (HOBt , PyBOP, HOAt, etc.), ammonium/

salts (e.g. pyrimidinium

Salts (such as HATU), tetramethylammonium salt, bispyrrolidine ammonium salt, bispiperidine ammonium salt, imidazolium

salt, derived from N , N , N' -trimethyl- N' -phenylurea

Salt, ammonium based on N-𠰌olinyl group/

Coupling reagent, antimonic acid

salts, etc.), organophosphorus reagents (such as phosphinic acid and phosphoric acid derivatives), organosulfur reagents (such as sulfonic acid derivatives), tris coupling reagents (such as 2-chloro-4,6-dimethoxy-1, 3,5-Tris(2-yl)-4-(4,6-dimethoxy-1,3,5-tris(2-yl)-4-methyl(4-(4,4-)-(4-)-(4-(4,4-)) 6-Dimethoxy-1,3,5-tri((2-yl)-4-methyl(()-pyridinium, etc.), pyridinium coupling reagent (such as Mukaiyama's reagent), pyridinium tetrafluoroboronic acid coupling reagent etc.), polymeric carrier reagents (e.g., polymer-bound carbodiimide, polymer-bound TBTU, polymer-bound 2,4,6-trichloro-1,3,5-tris-tris, and polymer-bound HOBt, polymer-bound HOSu, polymer-bound IIDQ, polymer-bound EEDQ, etc.) and the like. Alternatively, acylation can be carried out using activated carboxylic acid derivatives, such as anhydrides, mixed anhydrides, acid chlorides, or activated esters (eg, pentafluorophenyl esters or N -hydroxysuccinimidyl esters).

基-2-磺醯基(Mts)；4-甲氧基-2,3,6-三甲基苯磺醯基(Mtr)；乙醯胺基；鄰苯二醯亞胺基；及其類似基團。羥基保護基團之實例包括(但不限於)：苯甲基；三級丁基；三苯甲基；三級丁基二甲基矽烷基(TBDMS；TBS)；4,5-二甲氧基-2-硝基苯甲氧基羰基(Dmnb)；炔丙氧基羰基(Poc)；及其類似基團。其他醇保護基團及胺保護基團為熟習此項技術者已知的，包括例如由Green及Wuts ( Protective Groups in Organic Synthesis , 第 4 版2007, Wiley-Interscience, New York)描述之保護基團。可使用標準條件移除保護基團以恢復原始官能基，以用於其他合成處理。 Amines and other functional groups can be protected from undesirable side reactions during various synthetic steps. Examples of amine protecting groups include, but are not limited to: benzyloxycarbonyl; 9-intenylmethoxycarbonyl (Fmoc); tertiary butoxycarbonyl (Boc); allyloxycarbonyl (Alloc); p- Tosylsulfonyl (Tos); 2,2,5,7,8-pentamethyl-6-sulfonyl (Pmc); 2,2,4,6,7-pentamethyl-2,3- Dihydrobenzofuran-5-sulfonyl (Pbf);

yl-2-sulfonyl (Mts); 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr); acetamido; phthalimido; and the like group. Examples of hydroxyl protecting groups include, but are not limited to: benzyl; tert-butyl; trityl; tert-butyldimethylsilyl (TBDMS; TBS); 4,5-dimethoxy -2-Nitrobenzyloxycarbonyl (Dmnb); propargyloxycarbonyl (Poc); and the like. Other alcohol protecting groups and amine protecting groups are known to those skilled in the art and include, for example, protecting groups described by Green and Wuts ( Protective Groups in Organic Synthesis , 4th edition 2007, Wiley - Interscience, New York) . The protecting group can be removed using standard conditions to restore the original functional group for other synthetic treatments.

Unless stated to the contrary, the reactions described herein are carried out at atmospheric pressure at temperatures ranging from about -78°C to about 250°C. For example, the reaction can be carried out at about 0°C to about 125°C, or at about room temperature (or ambient temperature), eg, about 20°C. In some embodiments, the reaction is carried out at about 0°C, 20°C, 25°C, 90°C, 100°C, 110°C, 125°C, 150°C, 175°C, or 200°C. In some embodiments, the reaction is initiated at a first temperature (eg, about -78°C or about 0°C) and allowed to warm to a second, higher temperature (eg, about 20°C or about 25°C). Those skilled in the art will appreciate that various modifications can be made to the procedures described herein. III . Pharmaceutical compositions

In some embodiments, the 3CL ^pro inhibitor is administered as a pharmaceutical composition comprising at least one pharmaceutically acceptable excipient and the 3CL ^pro inhibitor or a pharmaceutically acceptable salt thereof. The 3CL ^pro inhibitor can be administered to the individual prior to administration of the one or more additional active agents, after administration of the one or more additional active agents, or concurrently with the administration of the one or more additional active agents. The 3CL ^pro inhibitor can be administered in a composition separate from one or more additional active agents, or in a composition containing one or more additional active agents. Also provided herein are compositions comprising: (i) one or more 3CL ^pro inhibitors; (ii) one or more pharmaceutically acceptable excipients; and optionally (iii) one or more additional active agents , which are each independently an anti-inflammatory, analgesic, antiviral, antitussive, or CYP3A4 inhibitor. Compositions can be formulated, for example, for oral, intravenous, intramuscular, intraperitoneal, subcutaneous, intrathecal, intraarterial, intranasal, or rectal administration vote.

Pharmaceutical compositions can be prepared by any method well known in the art of pharmacy and drug delivery. In general, the preparation of compositions includes the step of bringing into association the active ingredient with a carrier that contains one or more accessory ingredients. Pharmaceutical compositions are generally prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers, or both, and then, if necessary, shaping the product into the desired formulation. The compositions are preferably prepared and/or packaged in unit dosage form.

Pharmaceutical compositions can be in a form suitable for oral use. Compositions suitable for oral administration include, but are not limited to, lozenges, dragees, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups, elixirs, solutions , Buccal patch, oral mucilage, chewable tablet, chewable tablet, foamable powder and foamable tablet. Such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents, antioxidants, and preservatives to provide pharmaceutically aesthetically pleasing and palatable formulations.

Tablets usually contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients including: inert diluents such as cellulose, silica, alumina, calcium carbonate, sodium carbonate , glucose, mannitol, sorbitol, lactose, calcium phosphate and sodium phosphate; granulating and disintegrating agents such as corn starch and alginic acid; binding agents such as polyvinylpyrrolidone (PVP), cellulose, polyethylene glycol (PEG), starch, gelatin and acacia; and lubricants such as magnesium stearate, stearic acid and talc. Tablets may be uncoated, or coated (enteric or otherwise) by known techniques to delay disintegration and absorption in the gastrointestinal tract, and thereby provide a sustained action over a longer period of time. For example, time delay materials such as glyceryl monostearate or glyceryl distearate may be employed. Tablets may also be coated with a semipermeable membrane and, optionally, a polymeric osmotic agent to form an osmotic pump composition for controlled release, according to known techniques. The compositions for oral administration may be formulated as hard gelatin capsules, wherein the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin; or as soft gelatin capsules, wherein the active ingredient is mixed with Water or an oily medium such as peanut oil, liquid paraffin or olive oil.

Pharmaceutical compositions may also be in the form of injectable aqueous or oily solutions or suspensions. Sterile can be formulated using non-toxic parenterally acceptable vehicles including water, Ringer's solution and isotonic sodium chloride solution and acceptable solvents such as 1,3-butanediol Injectable preparations. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Aqueous suspensions contain the active agent in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include, but are not limited to: suspending agents such as sodium carboxymethylcellulose, methylcellulose, oily propylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, tragacanth ( gum tragacanth) and gum acacia; dispersing or wetting agents such as lecithin, polyoxyethylene stearate and polyethylene sorbitan monooleate; and preservatives such as ethyl, n- Propyl and parabens. Oily suspensions can be formulated by suspending the active ingredient in vegetable oils such as peanut oil, olive oil, sesame oil, or coconut oil or mineral oils such as liquid paraffin. Oily suspensions may contain a thickening agent, such as beeswax, hard paraffin, or cetyl alcohol. These compositions can be preserved by adding antioxidants such as ascorbic acid. Dispersible powders and granules, suitable for preparation of an aqueous suspension by the addition of water, can contain the active ingredient in admixture with a dispersing agent, wetting agent, suspending agent, or combinations thereof.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil, such as olive or flower oil; or a mineral oil, such as liquid paraffin; or a mixture of these. Suitable emulsifiers can be naturally occurring gums such as acacia or tragacanth; naturally occurring phospholipids such as soy lecithin; esters or partial esters derived from fatty acids and hexitols such as sorbitan mono Oleates; and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate.

Transdermal delivery can be achieved by means of iontophoretic patches and the like. The active ingredient may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the active agent with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols. IV . METHODS FOR THE TREATMENT OF CORONAVIRUS INFECTIONS

Also provided herein are methods for treating coronavirus infections. The infection can be, but is not limited to, SARS-CoV-2 infection, SARS-CoV infection, MERS-CoV infection, HCoV-229E infection, HCoV-OC43 infection, or HCoV-NL63 infection. Such methods include administering to an individual in need thereof a therapeutically effective amount of a compound as described above. In some embodiments, the individual has coronavirus disease 2019 (COVID-19). Compounds can act as 3CL ^pro inhibitors to slow or prevent replication of SARS-CoV-2 in an individual.

In some embodiments, the individual is a human, an agricultural animal (eg, livestock, such as cattle, sheep, pigs, or the like), or a companion animal (eg, a pet, such as a cat or dog). In some embodiments, the individual is a human over 50 years of age.

The 3CL ^pro inhibitor compounds of the present invention, as well as other active agents used in combination therapy as described herein, may be administered orally, intravenously, intramuscularly, intraperitoneally, subcutaneously, intrathecally, intraarterially, intranasally, rectally, or Administer to the subject via other means, if indicated. In some embodiments, the 3CL ^pro inhibitor is administered orally or via injection. In the methods provided herein, the active agent can be administered in any suitable dose. Generally, 3CL ^pro inhibitors or other active agents (eg, CYP3A4 inhibitors) are administered at doses in the range of about 0.1 mg to about 1000 mg per kilogram of body weight of the subject (ie, about 0.1-1000 mg/kg). The dosage of the 3CL ^pro inhibitor compound can be, for example, about 0.1-1000 mg/kg, or about 1-500 mg/kg, or about 25-250 mg/kg, or about 50-100 mg/kg. The dose of 3CL ^pro inhibitor can be about 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 , 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg/kg. In some embodiments, the 3CL ^pro inhibitor is administered in an amount ranging from about 0.1 mg/kg/day to about 100 mg/kg/day. In some embodiments, the 3CL ^pro inhibitor is administered in an amount ranging from about 0.1 mg/kg/day to about 1.0 mg/kg/day. In some embodiments, the individual is administered one or more (eg, two or three) doses of 10-100 mg (eg, 15-40 mg or 45-85 mg) of a 3CL ^pro inhibitor per day. In some embodiments, the total daily dose of the 3CL ^pro inhibitor ranges from about 10 to about 100 mg (eg, 15-40 mg, or 45-85 mg) per day.

The dosage may vary depending on the needs of the patient, the severity of the infection, the route of administration and the particular formulation to be administered. The dose administered to the patient should be sufficient to produce a beneficial therapeutic response in the patient. The size of the dose will also be determined by the presence, nature and extent of any adverse side effects that accompany the administration of the drug in a particular patient. Determination of the appropriate dose for a particular situation is within the skill of the standard practitioner. The total dose can be divided into aliquots and administered in aliquots over a period suitable for the treatment of COVID-19.

The 3CL ^pro inhibitor or other active agent may be administered over a period of time that will also vary depending on the severity of COVID-19 and the overall condition of the individual to whom the active agent is administered. Administration may be, for example, hourly, every 2 hours, every three hours, every four hours, every six hours, every eight hours, or twice a day (including every 12 hours) or any intermediate time interval thereof. Administration can be performed once a day, or every 36 hours or 48 hours, once a week, twice a week, or three times a week. Following treatment, individuals can be monitored for changes in symptoms and for relief of symptoms of COVID-19. The dose of active agent may be increased in cases where the individual does not respond significantly to a particular dose level, or may be decreased if symptom relief is observed, or if COVID-19 has abated, or if unacceptable side effects are found at a particular dose dose. Treating COVID-19 according to the methods of the present invention may include alleviating one or more symptoms including, but not limited to, fever, cough, and shortness of breath. In some embodiments, treatment of COVID-19 prevents serious, life-threatening illnesses, such as pneumonia.

The methods and compositions described herein can also be administered prophylactically to individuals at risk of contracting SARS-CoV-2 to reduce the risk of developing COVID-19.

In some embodiments, the level of SARS-CoV-2 3CL ^pro activity in the individual can be reduced by about 25% to about 95% when treating an individual according to the methods of the present invention. For example, 3CL ^pro activity in an individual can be reduced by about 35% to about 95% compared to the corresponding level of 3CL ^pro activity prior to the first administration of the active agent (eg, 24 hours prior to the first administration of the active agent). , or about 40% to about 85%, or about 40% to about 80%.

In some embodiments, the methods further comprise administering to the subject analgesics (including anti-inflammatory analgesics), antiviral agents, and antitussives, or a combination thereof. Examples of non-steroidal anti-inflammatory agents (NSAIDs) include, but are not limited to, aceclofenac, 5-aminosalicylic acid, aspirin, celecoxib, dexibuprofen, Diclofenac, diflunisal, etodolac, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen ), indomethacin, ketoprofen, ketorolac, loxoprofen, mefenamic acid, nabumetone, naphthalene Naproxen, nimesulide, sulindac and their pharmaceutically acceptable salts. NSAIDs are effective in relieving symptoms such as fever and pain. Other analgesics such as paracetamol (acetaminophen) can also be administered with the 3CL ^pro inhibitor. Examples of other antiviral agents include, but are not limited to, protease inhibitors (eg, ritonavir, lopinavir, saquinavir, indinavir, or the like) analogs), nucleic acid polymerase inhibitors (eg, acyclovir, foscarnet, ganciclovir, ribavirin, or analogs thereof), interferon, target Antibodies or other biological agents that bind or invade coronaviruses and other small molecules suitable for the treatment of coronaviruses. Examples of antitussives include, but are not limited to, codeine, hydrocodone, benzonatate, dextromethorphan, and chlophedianol. In some embodiments, the methods include administering to the individual a CYP3A4 inhibitor. Administration of a CYP3A4 inhibitor prevents premature metabolism of the 3CL ^pro inhibitor and increases plasma concentration levels following oral or another route of administration. Examples of CYP3A4 inhibitors include, but are not limited to, aprepitant, azamulin, boceprevir, chlorzoxazone, cilostazol , cimetidine, ciprofloxacin, clotrimazole, cobicistat, conivaptan, crizotinib, cyclosporine cyclosporine, diltiazem, dronedarone, erythromycin, fluconazole, fluvoxamine, fosaprepitant, grapefruit juice, imatinib, istradefylline, itraconazole, ivacaftor, ketoconazole, lomitapide, posa Posaconazole, ranitidine, ranolazine, ritonavir (as appropriate with danoprevir, dasabuvir, elvitegravir) ), indinavir, lopinavir, ombitasvir, paritaprevir, saquinavir, tipranavir, and combinations thereof administered together), teravir telaprevir, telithromycin, ticagrelor, tofisopam, troleandomycin, verapamil, and voriconazole. It will be appreciated that active agents can be classified into more than one class; for example, ritonavir can be classified as a protease inhibitor and a CYP3A4 inhibitor.

Also provided herein are methods for inhibiting SARS-CoV-2 3CL ^pro . Such methods include contacting 3CL ^pro with an effective amount of a compound as described herein. Inhibiting 3CL ^pro typically involves contacting 3CL ^pro with an amount of a protease inhibitor sufficient to reduce 3CL ^pro activity compared to 3CL ^pro activity in the absence of the protease inhibitor. For example, contacting 3CL ^pro with a protease inhibitor can result in about 1% to about 99% inhibition of 3CL ^pro (that is, the activity of the 3CL ^pro that is inhibited is 99% of the activity of the 3CL ^pro in the absence of the compound. to within 1%). 3CL ^pro inhibition level can be from about 1% to about 10%, or about 10% to about 20%, or about 20% to about 30%, or about 30% to about 40%, or about 40% to about 50%, or about 50% to about 60%, or about 60% to about 70%, or about 70% to about 80%, or about 80% to about 90%, or about 90% to about 99%. 3CL ^pro inhibition levels can range from about 5% to about 95%, or about 10% to about 90%, or about 20% to about 80%, or about 30% to about 70%, or about 40% to about 60% Inside. In some embodiments, contacting 3CL ^pro with a protease inhibitor as described herein will result in complete (ie, 100%) inhibition of 3CL ^pro . Inhibition of 3CL ^pro according to the methods of the present invention can be performed in vitro or in vivo (eg, following administration of a protease inhibitor to an individual during the treatment of COVID-19). V. EXAMPLES Example 1. Preparation of N -(( S ) -3 - cyclopropyl - 1 - oxy - 1 - ((( S ) -3 - oxy - 1 - (( S ) -2- pendant oxypyrrolidin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) ) amino ) prop - 2 - yl ) benzofuran - 2 - methyl _ _ _ _ _ Amide ( 101 )

Compound 2 : To a mixture of compound 1 (10 g, 36.32 mmol, 1 equiv) in THF (100 mL) was added LiHMDS (1 M, 78.45 mL, 2.16 equiv) dropwise at -78 °C under _N2 . The mixture was stirred at -78°C for 1 hour. Compound 1A (6.53 g, 54.48 mmol, 3.63 mL, 1.5 equiv) was then added dropwise to the mixture. The mixture was stirred at -78°C for 2 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.43) showed that the reaction was complete. To the mixture was added MeOH (30 mL), and then the mixture was poured into a mixed solution (5 mL of HOAc/40 mL of THF), followed by stirring the resulting mixture at -78°C for 30 minutes. The mixture was poured into water (100 mL) and ethyl acetate (100 mL). The combined organic phases were washed with brine (100 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 0/1). Compound 2 (9 g, 28.63 mmol, 78.83% yield) was obtained as a yellow oil.

Compound 3 : To a mixture of compound 2 (15.37 g, 48.90 mmol, 1 equiv) in MeOH (160 mL) was added CoCl2.6H2O (6.98 _g , 29.34 mmol, 0.6 equiv) at ₀ °C. Next, NaBH4 ₍ 11.15 g, 294.85 mmol, 6.03 equiv) was added to the mixture at 0 °C. The mixture was stirred at 25°C for 12 hours. TLC (petroleum ether/ethyl acetate=0:1, Rf ₌ 0.43) indicated the formation of a new spot. The reaction mixture was quenched by the addition of _NH4Cl (200 mL), and then the mixture was filtered and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layers were washed with saturated aqueous NaHCO ₃ (200 mL), saturated aqueous Na ₂ SO ₃ (300 mL) and saturated brine (100 mL), dried over Na ₂ SO ₄ , filtered and the filtrate concentrated under reduced pressure, A residue is obtained. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 0/1). Compound 3 (8 g, 27.94 mmol, 57.14% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, chloroform- d ) δ 4.30-4.48 (m, 1 H) 3.60-3.84 (m, 3 H) 3.24-3.42 (m, 2 H) 2.36-2.54 (m, 2 H) 2.06- 2.24 (m, 1 H) 1.69-1.94 (m, 3 H) 1.44 (s, 9 H).

Compound 4 : To a solution of DIPA (14.00 g, 138.31 mmol, 19.55 mL, 5.5 equiv) in THF (50 mL) at 0 °C was added n-BuLi (2.5 M, 55.32 mL, 5.5 equiv). The mixture was stirred at 0°C for 1 hour. And then the solution was added to compound 3 (7.2 g, 25.15 mmol, 1 equiv) and chloro(iodo)methane (24.39 g, 138.31 mmol, 10.04 mL, 5.5 equiv) in THF (10 mL) at -78°C solution, and the mixture was stirred at -78°C for 3 hours. TLC (petroleum ether:ethyl acetate=0:1, Rf ₌ 0.45) showed that the reaction was complete. The reaction mixture was quenched by the addition of saturated aqueous _NH4Cl (30 mL), and then extracted with 300 mL (100 mL x 3) of ethyl acetate. The combined organic layers were washed with saturated aqueous Na ₂ SO ₃ (30 mL), saturated aqueous NaHCO ₃ (30 mL) and saturated brine (30 mL), dried over Na ₂ SO ₄ , filtered and the filtrate concentrated under reduced pressure, A residue is obtained. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 0/1). Compound 4 (1.3 g, 4.27 mmol, 16.96% yield) was obtained as a yellow oil.

Compound 5 : To a solution of compound 4 (1 g, 3.28 mmol, 1 equiv) in DMF (10 mL) at 0 °C was added K2CO3 ( _906.96 mg, 6.56 mmol, ₂ equiv), KI (1.09 g, 6.56 mmol, 2 equiv) and compound 5A (817.37 mg, 4.92 mmol, 1.5 equiv). The mixture was stirred at 25°C for 12 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.43) showed that the reaction was complete. The residue was diluted with _H2O (20 mL) and extracted with 90 mL (30 mL x 3) EtOAc. The combined organic layers were washed with 60 mL (20 mL x 3) of brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=3/1 to 1/1). Compound 5 (1.3 g) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol-d4) δ 7.03-7.20 (m, 1 H) 4.97-5.50 (m, 2 H) 4.23-4.39 (m, 1 H) 3.33-3.37 (m, 2 H) 2.29- 2.52 (m, 2 H) 1.95-2.01 (m, 1 H) 1.72-1.92 (m, 2 H) 1.45 (s, 10 H).

Compound 6 : A mixture of compound 5 (800.00 mg, 1.84 mmol, 1 equiv) in TFA (1 mL) and DCM (5 mL) was stirred at 25 °C. The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether:ethyl acetate=3:1, Rf ₌ 0.43) showed the formation of new spots and the completion of the reaction. The reaction mixture was concentrated under reduced pressure to give a residue. Compound 6 (( S )-3-(( S )-2-amino-3-oxy-4-(2,3,5,6-tetrafluorophenoxy)butyl) was obtained as a yellow oil ) pyrrolidin-2-one, 1.2 g, crude material, TFA).

Compound 8 : To a stirred solution of compound 7 (1.5 g, 11.61 mmol, 1 equiv) in MeOH (15 mL) was added _SOCl2 (4.15 g, 34.84 mmol, 2.53 mL) dropwise at 0 °C under _N2 , 3 equivalents). The resulting mixture was stirred at 70°C for 3 hours. The reaction mixture was concentrated under reduced pressure to give a yellow solid. And the solid was triturated with petroleum ether (10 mL×2) to obtain an off-white solid. And this solid was used in the next step without further purification. Compound 8 (1.95 g, 10.85 mmol, 93.46% yield, HCl) was obtained as an off-white solid. 1H NMR (400 MHz, methanol-d4) δ =4.16-4.07 (m, 1H), 3.85 (s, 3H), 1.97-1.85 (m, 1H), 1.83-1.70 (m, 1H), 0.88-0.72 ( m, 1H), 0.63-0.54 (m, 2H), 0.22-0.12 (m, 2H).

Compound 9 : To a solution of Compound 8 (0.2 g, 1.40 mmol, 1 equiv) in DMF (2 mL) was added HATU (798.48 mg, 2.10 mmol, 1.5 equiv), Compound 8A (227.00 mg, 1.40 mmol, 1 equiv) ) and DIPEA (361.87 mg, 2.80 mmol, 487.70 μL, 2 equiv), and the mixture was stirred at 25 °C for 12 h. TLC (petroleum ether:ethyl acetate=3:1, Rf ₌ 0.84) showed the formation of new spots and the completion of the reaction. The residue was poured into water (3 mL) and ethyl acetate (3 mL). The aqueous phase was extracted with ethyl acetate (1 mL×3). The combined organic phases were washed with brine (1 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 3/1). Compound 9 (0.3 g, 1.04 mmol, 74.58% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.73 (d, J =7.9 Hz, 1 H) 7.61 (d, J =8.4 Hz, 1 H) 7.53 (s, 1 H) 7.43-7.49 (m, 1 H) 7.24-7.40 (m, 1 H) 4.73 (dd, J = 8.4, 5.6 Hz, 1 H) 3.76 (s, 3 H) 1.69-2.00 (m, 2 H) 0.80-0.95 (m, 1 H) ) 0.41-0.55 (m, 2H)-0.01-0.29 (m, 2H).

Compound 10 : To a solution of compound 9 (0.28 g, 974.56 μmol, 1 equiv) in MeOH (2.5 mL) and H ₂ O (0.5 mL) at 25 °C was added LiOH.H ₂ O (81.78 mg, 1.95 mmol, 2 equiv). The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.26) showed the formation of new spots and the completion of the reaction. The mixture was adjusted to pH=8 with 1N HCl. The reaction mixture was then concentrated under reduced pressure to remove solvent. The residue was diluted with 6 mL (2 mL x 3) _H2O and extracted with 12 mL (4 mL x 3) EtOAc. The combined organic layers _were dried over _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 10 (0.2 g, 731.84 mol, 75.09% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.74 (d, J =7.9 Hz, 1 H) 7.62 (d, J =8.3 Hz, 1 H) 7.53 (s, 1 H) 7.44-7.49 (m, 1 H) 7.22-7.40 (m, 1 H) 4.72 (dd, J = 8.3, 5.5 Hz, 1 H) 1.75-1.97 (m, 2 H) 0.79-0.91 (m, 1 H) 0.46-0.59 (m, 2H) 0.04-0.25 (m, 2H).

N -(( S ) -3 - cyclopropyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 - (( S ) -2 - oxypyrrolidine - 3- yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl ) benzofuran - 2 - carboxamide ( 101 ) : at 0 _ _ _ _ To a solution of compound 10 (81.39 mg, 181.57 μmol, TFA) in DMF (2 mL) was added HATU (125.22 mg, 329.33 μmol, 1.5 equiv) at °C. To the mixture was added compound 6 (0.06 g, 219.55 μmol, 1 equiv) and DIPEA (85.12 mg, 658.66 μmol, 114.72 μL, 3 equiv) at 25°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was partitioned between _H2O (12 mL) and EtOAc (9 mL). The organic phase was separated, washed with 9 mL (3 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. By preparative HPLC (column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 μm; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 40%- 60%, 8 min) to purify the residue. Compound 101 (29 mg, 49.19 μmol, 22.41% yield, 100% purity) was obtained as a yellow oil. LCMS (ESI): m/z: _{[ M +} H] calcd for _C29H28F4N3O6 : 590.2; found 590.2; RT= _3.006 min. The isomer ratio of isomers observed was 2.45:10.58:1 as determined by supercritical fluid chromatography (SFC). ¹ H NMR (400 MHz, chloroform- d ) δ 8.59-9.26 (m, 1 H) 7.62-7.70 (m, 1 H) 7.52 (d, J =8.1 Hz, 1 H) 7.46 (s, 1 H) 7.35 -7.45 (m, 2 H) 7.30 (d, J = 7.9 Hz, 1 H) 6.71-6.88 (m, 1 H) 5.79-6.18 (m, 1 H) 5.00-5.26 (m, 2 H) 4.66-4.93 (m, 2 H) 3.30-3.43 (m, 2 H) 2.38-2.56 (m, 2 H) 1.99-2.17 (m, 2 H) 1.82-1.95 (m, 3 H) 0.71-0.92 (m, 1 H) ) 0.55 (br d, J = 7.9 Hz, 2 H) 0.09-0.28 (m, 2 H). Example 2. Preparation of (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidine - 3 -yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pent - 2 - yl ) carbamate benzyl ( 102 ) _ _ _ _ _ _

Compound 12 : To a solution of compound 5 (1.88 g, 7.09 mmol, 1 equiv) in DMF (20 mL) at 25 °C was added EDCI (1.70 g, 8.87 mmol, 1.25 equiv) and HOBt (1.20 g, 8.87 mmol, 1.25 equiv), the solution was stirred at 25 °C for 0.5 h. This solution is named A. To a solution of compound 11 (2.13 g, 7.09 mmol, 1 equiv, TFA) in DMF (5 mL) was added DIEA (3.67 g, 28.38 mmol, 4.94 mL, 4 equiv) at 0 °C in a separatory flask ), and the solution was stirred at 0 °C for 0.5 h. This solution is named B. Solution B was then added to solution A. The mixture was stirred at 25°C for 16 hours. The solution was poured into water (100 mL) and ethyl acetate (100 mL). The resulting mixture was extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with brine (100 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether:ethyl acetate=1/0 to 1/1) to give compound 12 (1.7 g, 3.92 mmol, 55.28% yield) as a white solid . ¹ H NMR (400 MHz, chloroform- d ) δ 7.88 (br d, J=6.4 Hz, 1 H) 7.28-7.41 (m, 5 H) 6.06 (s, 1 H) 5.39 (br d, J=8.4 Hz , 1 H) 5.10 (s, 2 H) 4.43-4.53 (m, 1 H) 4.26-4.37 (m, 1 H) 3.64-3.81 (m, 3 H) 3.29-3.39 (m, 2 H) 2.34-2.50 (m, 1 H) 2.10-2.25 (m, 1 H) 1.81-1.97 (m, 2 H) 1.62-1.76 (m, 3 H) 1.44-1.59 (m, 1 H) 0.83-1.04 (m, 6 H) ).

Compound 13 : To a mixture of DIPA (1.28 g, 12.69 mmol, 1.79 mL, 5.5 equiv) in THF (3 mL) at 0 °C was added n-BuLi (2.5 M, 5.08 mL, 5.5 equiv). The mixture was stirred at 0°C for 60 minutes. To the mixture was added chloro(iodo)methane (2.24 g, 12.69 mmol, 920.93 μL, 5.5 equiv) and a solution of compound 12 (1 g, 2.31 mmol, 1 equiv) in THF (2 mL) at -78 °C . The resulting mixture was then stirred at -78°C for 3 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.51) showed that the reaction was complete. The reaction mixture was quenched by the addition of _NH4Cl (100 mL), and then extracted with 300 mL (100 mL x 3) of ethyl acetate. The combined organic layers were washed with NaHCO ₃ (100 mL), Na ₂ SO ₃ (100 mL) and saturated brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue, It was purified by column chromatography (SiO ₂ , petroleum ether:ethyl acetate=1/0 to 1/1) to obtain compound 13 (0.89 g, crude material) as a yellow oil.

(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidin - 3 - yl )- Benzyl 4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) carbamate : to compound 13 ( 400 mg , 885.07 μmol , 1 equiv) in DMF (8 mL) was added 2,3,5,6-tetrafluorophenol (146.99 mg, 885.07 μmol, 1 equiv), KI (146.92 mg, 885.07 μmol, 1 equiv) and K ₂ CO ₃ (244.64 mg, 1.77 mmol, 2 equiv). The mixture was stirred at 25°C for 12 hours. The mixture was poured into water (100 mL) and ethyl acetate (100 mL). The resulting mixture was extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with brine (100 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80 x 40 mm x 3 μm; mobile phase: [water (10 mM _{NH4HCO3 )} -ACN]; B%: 35%-65%, 8 minutes) was obtained as a yellow solid to obtain compound 102 (30 mg, 51.59 μmol, yield 5.83%, purity 100%). ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.22-7.47 (m, 5 H) 7.02-7.19 (m, 1 H) 4.95-5.25 (m, 3 H) 4.47-4.74 (m, 2 H) 4.07 -4.21 (m, 1 H) 3.12-3.26 (m, 2 H) 2.20-2.58 (m, 2 H) 2.01-2.15 (m, 1 H) 1.65-1.92 (m, 3 H) 1.42-1.64 (m, 2H) 0.76-1.04 (m, 6H). LCMS (ESI): m/z: _{[ M +} H] _calcd for _C28H32F4N3O6 : 581; found 581; RT=3.082 min. The observed isomer ratio was 2.36:1:1.89:2.02:12.07 as determined by SFC. Example 3. Preparation of ( 1 -(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrole pyridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) -2 - oxy - 1,2- _ _ _ _ _ _ _ _ _ tert- butyl dihydropyridin - 3 - yl ) carbamate ( 103 )

Compound 15 : To a solution of compound 14 (10 g, 90.82 mmol, 1 equiv) in THF (150 mL) was added Boc2O (19.82 _g , 90.81 mmol, 20.86 mL, 1 equiv) at 25°C. The mixture was stirred at 70°C for 4 hours. Boc2O (15.86 _g , 72.65 mmol, 16.69 mL, 0.8 equiv) was then added, and the mixture was refluxed at 70 °C for 12 h. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.39) showed the formation of new spots and the completion of the reaction. The reaction mixture was partitioned between _H2O (300 mL) and EtOAc (180 mL). The organic phase was separated, washed with 120 mL (40 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 0/1). Compound 15 was obtained as a yellow solid (19 g, 90.38 mmol, 99.52% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.93 (br s, 1 H) 7.79 (br d, J =6.4 Hz, 1 H) 7.69 (s, 1 H) 7.04 (dd, J =6.5, 1.5 Hz, 1 H) 6.21 (t, J = 6.9 Hz, 1 H) 1.45 (s, 9 H).

Compound 17 : Compound 16 (12.5 g, 95.29 mmol, ₁ equiv) was dissolved in _H2SO4 (500 mL) at 0 °C. To the solution was then added NaNO2 (65.75 _g , 952.94 mmol, 10 equiv) in _H2O (100 mL) dropwise. The resulting mixture was stirred at 0°C for 3 hours, and then warmed to 20°C and stirred at 20°C for 16 hours. TLC (petroleum ether:ethyl acetate=0:1, Rf ₌ 0.42) showed the formation of new spots and the completion of the reaction. The reaction mixture was partitioned between _H2O (600 mL) and EtOAc (600 mL). The organic phase was separated, washed with 300 mL (100 mL x 3) of brine, dried _{over Na2SO4} and concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 17 was obtained as a yellow oil (17 g, 128.63 mmol, 67.49% yield). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 3.94 (dd, J =8.7, 4.9 Hz, 1 H) 1.68-1.80 (m, 1 H) 1.37-1.47 (m, 2 H) 0.83-0.91 (m , 6H).

Compound 18 : To a solution of compound 17 (16 g, 121.07 mmol, 1 equiv) in MeOH (40 mL) and 2,2-dimethoxypropane (160 mL) was added TMSCl (1.32 g) at 25 °C , 12.11 mmol, 1.54 mL, 0.1 equiv). The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with _H2O (300 mL) and extracted with 150 mL (50 mL x 3) EtOAc. The combined organic layers were washed with 90 mL (30 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 18 (14 g, 95.77 mmol, 79.10% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, chloroform- d ) δ 4.22 (dd, J =7.7, 5.7 Hz, 1 H) 3.71-3.83 (m, 2 H) 1.80-1.94 (m, 1 H) 1.52-1.61 (m, 2H) 0.90-1.01 (m, 6H).

Compound 19: To a solution of compound 18 (20 g, 136.81 mmol, 1 equiv) in DCM (200 mL) was added 2,6-lutidine (15.39 g, 143.65 mmol) at 0 °C under _N2 , 16.73 mL, 1.05 equiv) and _Tf2O (57.90 g, 205.22 mmol, 33.86 mL, 1.5 equiv). The mixture was stirred at 0 °C under _N2 for 30 min. TLC (petroleum ether:ethyl acetate=5:1, Rf ₌ 0.81) showed the formation of new spots and the completion of the reaction. The reaction mixture was concentrated under reduced pressure to remove solvent. After washing with a mixture of brine and 1N HCl (3:1 v/v), the residue was extracted with EtOAc (300 mL), then dried _{over Na2SO4} , filtered and the filtrate concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 19 was obtained as a brown oil (40 g, crude).

Compound 20 : To a solution of compound 15 (2.20 g, 10.46 mmol, 0.97 equiv) in THF (40 mL) was added NaH (646.85 mg, 16.17 mmol, 60% pure, 1.5 equiv) at 0 °C for 30 minute. Compound 19 (3 g, 10.78 mmol, 1 equiv) in THF (50 mL) was then added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched by addition of _H2O (100 mL) at 0 °C and extracted with 90 mL (30 mL x 3) of EtOAc. The combined organic layers were washed with 90 mL (30 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 20 was obtained as a brown oil (1 g, 2.96 mmol, 27.41% yield). ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.98 (br d, J =7.7 Hz, 1 H) 7.26 (dd, J =7.0, 1.8 Hz, 1 H) 7.18-7.29 (m, 1 H) 6.39 (t, J = 7.2 Hz, 1 H) 5.48 (dd, J = 10.5, 5.0 Hz, 1 H) 3.72 (s, 3 H) 1.97-2.13 (m, 2 H) 1.50-1.58 (m, 9 H) 1.33-1.44 (m, 1 H) 0.90-0.96 (m, 1 H) 0.93 (dd, J = 11.1, 6.7 Hz, 6 H).

Compound 21 : To a solution of compound 20 (0.8 g, 2.36 mmol, 1 equiv) in MeOH (10 mL) and _H2O ( ₂ mL) at 20 °C was added LiOH.H2O (198.41 mg, 4.73 mmol, 2 equiv). The mixture was stirred at 20°C for 1 hour. TLC (petroleum ether:ethyl acetate=0:1, Rf ₌ 0.26) showed the formation of new spots and the completion of the reaction. The mixture was adjusted to pH=8 with 1N HCl. The reaction mixture was then concentrated under reduced pressure to remove solvent. The residue was diluted with _H2O (15 mL) and extracted with 12 mL (4 mL x 3) EtOAc. The combined organic layers _were dried over _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 21 (0.8 g, crude material) was obtained as a yellow oil. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.79 (s, 1 H) 7.35 (dd, J =7.0, 1.4 Hz, 1 H) 6.30 (t, J =7.2 Hz, 1 H) 5.35 (dd, J = 11.2, 4.4 Hz, 1 H) 2.04-2.14 (m, 1 H) 1.79-1.96 (m, 1 H) 1.41-1.50 (m, 9 H) 1.21-1.30 (m, 1 H) 0.81-0.87 ( m, 6H).

( 1 -(( S ) -4 - methyl - 1 - oxo - 1 -(((( S ) -3 - oxo - 1 - (( S ) -2 - oxopyrrolidine - 3- yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) -2 - oxy - 1,2 - dihydropyridine- _ _ _ _ _ _ _ _ _ 3 - yl ) carbamate tert- butyl ester ( 103 ) : To a mixture of compound 21 (0.1 g, 308.29 μmol, 1 equiv) in DMF (3 mL) at 0 °C was added HATU (175.83 mg, 462.43 μmol, 1.5 equiv) and the mixture was stirred at 0°C for 1 hour. Subsequently, compound 6 (138.20 mg, 308.29 μmol, 1 equiv, TFA) and DIPEA (79.69 mg, 616.58 μmol, 107.40 μL, 2 equiv) were added to the mixture at 25°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was partitioned between _H2O (12 mL) and EtOAc (9 mL). The organic phase was separated, washed with 9 mL (3 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. By preparative HPLC (basic conditions; column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 μm; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 50%-70%, 8 min) to purify the residue. Compound 103 (17 mg, 24.69 μmol, 8.01% yield, 93.03% purity) was obtained as a white solid. The observed isomer ratio was 3.49:1:8.71:5.25. LCMS (ESI): m/z: [M ₊ H] _calcd for _C30H37F4N4O7 : ₆₄₁ ; found 641; RT= _3.321 min. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.95 (br s, 1 H) 7.33 (br d, J =7.5 Hz, 1 H) 7.04-7.18 (m, 1 H) 6.32-6.42 (m, 1 H) H) 5.57-5.69 (m, 1 H) 4.98-5.22 (m, 1 H) 4.65 (br d, J = 11.2 Hz, 1 H) 3.35 (br s, 1 H) 3.19-3.30 (m, 2 H) 2.14-2.47 (m, 2 H) 1.94-2.12 (m, 2 H) 1.63-1.93 (m, 3 H) 1.47-1.56 (m, 9 H) 1.42 (br dd, J = 13.3, 6.7 Hz, 1 H ) 0.84-0.99 (m, 6 H). Example 4. Preparation of ( 4 -(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrole pyridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) -3 - oxy - 3,4- _ _ _ _ _ _ _ _ _ Dihydropyridine - 2 - yl ) tertiary butyl carbamate ( 104 )

Compound 23 : To a solution of compound 18 (1 g, 9.00 mmol, 1 equiv) in DMF (2 mL) was added NaH (359.97 mg, 9.00 mmol, 60% pure, 1 equiv) at 0 °C for 30 minute. Compound 22 (2.50 g, 9.00 mmol, 1 equiv) was then added and the mixture was stirred at 25°C for 12 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.43) showed that the reaction was complete. The reaction mixture was quenched by addition of _H2O (10 mL) at 0 °C and extracted with 30 mL (10 mL x 3) of EtOAc. The combined organic layers were washed with 9 mL of brine (3 mL x ₃ ), dried over _Na2SO4 , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 1/1). Compound 23 (1 g, 4.18 mmol, 46.44% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 6.64-6.85 (m, 2 H) 5.40 (dd, J =10.8, 4.9 Hz, 1 H) 3.64-3.79 (m, 3 H) 2.03-2.10 (m , 1 H) 1.92-1.98 (m, 1 H) 1.34-1.46 (m, 1 H) 0.81-1.00 (m, 6 H).

Compound 24 : To a solution of compound 23 (0.5 g, 2.09 mmol, 1 equiv) in THF (2 mL) at 0 °C was added TEA (422.91 mg, 4.18 mmol, 581.71 μL, 2 equiv), DMAP (25.53 mg, 208.97 μmol, 0.1 equiv) and Boc2O (912.13 mg, 4.18 mmol, 960.14 μL, ₂ equiv). The mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched by addition of _H2O (10 mL) at 25 °C, and then extracted with 15 mL (5 mL x 3) of EtOAc. The combined organic layers were washed with brine (10 mL), dried _{over Na2SO4} , filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=5/1 to 3/1). Compound 24 (0.4 g, 1.18 mmol, 56.40% yield) was obtained as a yellow oil.

Compound 25 : To a solution of compound 24 (250 mg, 736.62 μmol, 1 equiv) in MeOH (5 mL) and _H2O (1 mL) at 25°C was added _LiOH.H2O (61.82 mg, 1.47 mmol, 2 equiv). The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether/ethyl acetate=0:1, Rf ₌ 0.43) indicated the formation of a new spot. The mixture was adjusted to pH=8 with 1N HCl. The reaction mixture was then concentrated under reduced pressure to remove solvent. The residue was diluted with 3 mL (1 mL x 3) _H2O and extracted with 9 mL (3 mL x 3) EtOAc. The combined organic layers _were dried over _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 25 (0.2 g, 614.71 μmol, 83.45% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.67 (d, J =4.5 Hz, 1 H) 7.31 (d, J =4.5 Hz, 1 H) 5.36-5.63 (m, 1 H) 2.02-2.20 ( m, 2 H) 1.99 (s, 1 H) 1.39 (s, 9 H) 0.85-0.99 (m, 6 H).

( 4 -(( S ) -4 - methyl - 1 - oxo - 1 -((( S ) -3 - oxo - 1 - (( S ) -2 - oxopyrrolidine - 3- group ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) -3 - oxy - 3,4 - dihydropyridine _ _ _ _ _ _ _ _ _ tert- butyl - 2 - yl ) carbamate ( 104 ) : To a solution of compound 25 (97.34 mg, 299.16 μmol, 1 equiv) in DMF (1 mL) at 0 °C was added HATU (170.63 mg, 448.74 μmol, 1.5 equiv). The mixture was stirred at 0°C for 1 hour. To the mixture was added compound 6 (0.1 g, 223.07 μmol, 1 equiv, TFA) and DIPEA (77.33 mg, 598.33 μmol, 104.22 μL, 2 equiv) at 25°C. The mixture was stirred at 25°C for 12 hours. The residue was poured into water (3 mL) and ethyl acetate (3 mL). The aqueous phase was extracted with ethyl acetate (1 mL×3). The combined organic phases were washed with brine (1 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure. By preparative HPLC (column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 μm; mobile phase: [water (0.05% NH ₃ H ₂ O + 10 mM NH ₄ HCO ₃ )-ACN]; B%: 40%- 65%, 8 min) to purify the residue. Compound 104 was obtained as a yellow oil (5 mg, 7.48 μmol, 2.50% yield, 95.74% purity). LCMS (ESI): m/z: [M+H] _calcd for _C29H36F4N5O7 : _642.25 ; found _642.25 ; RT= _1.618 min. The purity in QC_LCMS was 95.74%. The ee value in SFC=55.62%. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.20 (d, J =4.9 Hz, 1 H) 6.96-7.11 (m, 2 H) 5.44-5.57 (m, 1 H) 5.04-5.18 (m, 2 H) 4.68 (dd, J = 10.9, 4.1 Hz, 1 H) 3.20-3.29 (m, 2 H) 2.38-2.48 (m, 1 H) 2.22 (br s, 1 H) 2.00-2.09 (m, 1 H) ) 1.85-1.98 (m, 3 H) 1.69-1.81 (m, 1 H) 1.54 (s, 9 H) 1.42-1.51 (m, 1 H) 1.42-1.51 (m, 1 H) 0.96 (br t, J =5.8 Hz, 5 H) 0.92-0.99 (m, 1 H). Example 5. Preparation of N -(( S ) -3- ( 3 - fluorophenyl ) -1 - oxy - 1 - ((( S ) -3 - oxy - 1 -( ( S ) -2- Pendant oxypyrrolidin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) prop - 2 - yl - 1H - indole - 2 _ _ _ _ _ -formamide ( 105 ) _

Compound 27 : To a solution of compound 26 (1.5 g, 8.19 mmol, 1 equiv) in MeOH (16 mL) was added _SOCl2 (1.95 g, 16.38 mmol, 1.19 mL, 2 equiv) dropwise at 0 °C, And the resulting mixture was stirred at 70°C for 3 hours. The reaction mixture was concentrated under reduced pressure to give a pale yellow solid. And the solid was washed with petroleum ether (20 mL×3) and dried under reduced pressure to obtain a white solid. And this solid was used in the next step without further purification. Compound 27 (1.7 g, 7.28 mmol, 88.85% yield, HCl) was obtained as a white solid. 1H NMR (400 MHz, methanol-d4) δ 7.44-7.36 (m, 1H), 7.12-7.03 (m, 3H), 4.36 (t, J=6.8 Hz, 1H), 3.82 (s, 3H), 3.27 ( d, J=6.2 Hz, 1H), 3.23-3.16 (m, 1H).

Compound 29 : To a solution of compound 28 (344.84 mg, 2.14 mmol, 1 equiv) in DMF (1 mL) was added HATU (1.22 g, 3.21 mmol, 1.5 equiv) at 0 °C. To the mixture was added compound 27 (0.5 g, 2.14 mmol, 1 equiv, HCl) and DIPEA (553.10 mg, 4.28 mmol, 745.41 μL, 2 equiv) at 25°C. The mixture was stirred at 25°C for 12 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.43) showed that the reaction was complete. The reaction mixture was quenched by adding 9 mL (3 mL x 3) _H2O at 0 °C and extracted with 9 mL (3 mL x 3) EtOAc. The combined organic layers were washed with 9 mL (3 mL x 3) of brine, dried over Na ₂ SO ₄ , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=5/1 to 1/1). Compound 29 (0.7 g, 2.06 mmol, 96.12% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, chloroform- d ) δ 7.46-7.55 (m, 1 H) 7.27-7.35 (m, 1 H), 7.22-7.26 (m, 1 H) 7.11-7.18 (m, 1 H) 6.90 -6.99 (m, 2 H) 6.83-6.90 (m, 2 H), 6.73 (br d, J = 7.6 Hz, 1 H) 5.00-5.26 (m, 1 H) 3.79 (s, 3 H) 3.16-3.42 (m, 2H).

Compound 30 : To a solution of compound 29 (0.4 g, 1.18 mmol, 1 equiv) in MeOH (2.5 mL) and _H2O (0.5 mL) was added _LiOH.H2O (98.64 mg, 2.35 mL) at 25 °C mmol, 2 equiv). The mixture was stirred at 25°C for 1 hour. The mixture was adjusted to pH=8 with 1N HCl. The reaction mixture was then concentrated under reduced pressure to remove solvent. The residue was diluted with 15 mL (5 mL x 3) _H2O and extracted with 12 mL (4 mL x 3) EtOAc. The combined organic layers _were dried over _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 30 (0.3 g, 919.34 μmol, 78.22% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.60 (d, J =8.1 Hz, 1 H) 7.41 (dd, J =8.4, 0.7 Hz, 1 H) 7.17-7.31 (m, 2 H) 7.01- 7.11 (m, 4 H) 6.92 (td, J = 8.6, 2.0 Hz, 1 H) 4.90 (br s, 1 H) 3.36 (dd, J = 13.9, 5.1 Hz, 1 H) 3.15 (dd, J = 13.9 , 9.3 Hz, 1 H).

N -(( S ) -3- ( 3 - fluorophenyl ) -1 - oxo - 1 - ((( S ) -3 - oxo - 1 -((( S ) -2 - oxopyrrole Iridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl - 1H - indole - 2 - carboxamide _ _ _ _ _ _ ( 105 ) : To a solution of compound 6 (122.92 mg, 274.20 μmol, TFA) in DMF (2 mL) was added HATU (174.78 mg, 459.67 μmol, 1.5 equiv) at 0 °C. At 25 °C, to Compound 30 (0.1 g, 306.45 μmol, 1 equiv) and DIPEA (118.82 mg, 919.34 μmol, 160.13 μL, 3 equiv) were added to the mixture. The mixture was stirred at 25°C for 12 hours. The reaction mixture was partitioned into H ₂ O (12 mL) and EtOAc (9 mL). Separate the organic phase, wash with 9 mL (3 mL x 3) of brine, dry over Na ₂ SO ₄ , filter and concentrate the filtrate under reduced pressure to give a residue. Prepare by Type HPLC (column: Phenomenex Gemini-NX 150 x 30 mm x 5 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 45%-75%, 9 minutes) to purify the residue. Obtained in yellow Compound 105 as an oil (9.6 mg, 14.94 μmol, 4.88% yield, 100% purity). LCMS (ESI): m/z: _calcd for [ _{M + H} ] for _C32H28F5N4O5 : 643.2; found 643.2; RT=4.438 min. Isomer ratio observed was 2.32:1.73:1:8.86:10.40. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.52-7.63 (m, 1 H) 7.35-7.44 (m, 1 H) 6.84-7.33 (m, 8 H) 4.90-5.27 (m, 2 H) 4.50 (dd, J =11.5, 3.1 Hz, 1 H) 4.46-4.81 (m, 1 H) 3.00-3.29 (m, 4 H) 2.19-2.54 (m, 1 H) 2.02-2.17 (m, 1 H) 1.82-2.01 (m, 1 H) 1.61-1.82 (m, 2 H). Example 6 . Preparation of N -(( S ) -3 - cyclopropyl - 1 - oxo - 1 -((( S ) -3 - oxo - 1 -((( S ) -2 - oxopyridine pyridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl ) -1H - indole - 2 - methyl _ _ _ _ _ _ Amide ( 106 )

Compound 8 : To a stirred solution of compound 7 (1.5 g, 11.61 mmol, 1 equiv) in MeOH (15 mL) was added _SOCl2 (4.15 g, 34.84 mmol, 2.53 mL) dropwise at 0 °C under _N2 , 3 equivalents). The resulting mixture was stirred at 70°C for 3 hours. The reaction mixture was concentrated under reduced pressure to give a yellow solid. And the solid was triturated with petroleum ether (10 mL×2) to obtain an off-white solid. And this solid was used in the next step without further purification. Compound 8 (1.95 g, 10.85 mmol, 93.46% yield, HCl) was obtained as an off-white solid. ¹ H NMR (400 MHz, methanol-d4) δ = 4.16-4.07 (m, 1H), 3.85 (s, 3H), 1.97-1.85 (m, 1H), 1.83-1.70 (m, 1H), 0.88-0.72 (m, 1H), 0.63-0.54 (m, 2H), 0.22-0.12 (m, 2H).

Compound 31 : To a solution of compound 8 (0.5 g, 2.78 mmol, 1 equiv, HCl) in DMF (5 mL) was added HATU (1.99 g, 5.24 mmol, 1.5 equiv) at 0 °C. To the mixture was added compound 28 (562.77 mg, 3.49 mmol, 1 equiv) and DIPEA (902.62 mg, 6.98 mmol, 1.22 mL, 2 equiv) at 25°C. The mixture was stirred at 25°C for 12 hours. TLC (petroleum ether:ethyl acetate=1:1, Rf ₌ 0.43) showed that the reaction was complete. The reaction mixture was quenched by addition of _H2O (5 mL) at 0 °C and extracted with 9 mL (3 mL x 3) of EtOAc. The combined organic layers were washed with 9 mL (3 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=5/1 to 1/1). Compound 31 (0.7 g, 2.44 mmol, 70.01% yield) was obtained as a yellow oil.

Compound 32 : To a solution of compound 31 (0.4 g, 1.40 mmol, 1 equiv) in MeOH (2.5 mL) and _H2O (0.5 mL) at 25 °C was added _LiOH.H2O (117.24 mg, 2.79 mmol, 2 equiv). The mixture was stirred at 25°C for 1 hour. The mixture was adjusted to pH=8 with 1N HCl. The reaction mixture was then concentrated under reduced pressure to remove solvent. The residue was diluted with _H2O (6 mL) and extracted with 12 mL (4 mL x 3) EtOAc. The combined organic layers _were dried over _Na2SO4 , filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was used in the next step without purification. Compound 32 (0.2 g, 734.49 μmol, 52.58% yield) was obtained as a yellow oil. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.62 (d, J =8.1 Hz, 1 H) 7.44 (d, J =8.3 Hz, 1 H) 7.01-7.26 (m, 3 H) 4.71 (br dd , J = 8.3, 5.4 Hz, 1 H) 1.70-1.91 (m, 2 H) 0.81-0.98 (m, 1 H) 0.39-0.54 (m, 2 H) 0.02-0.27 (m, 2 H).

N -(( S ) -3 - cyclopropyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 - (( S ) -2 - oxypyrrolidine - 3- yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl ) -1H - indole - 2 - carboxamide ( 106 ) _ _ _ _ _ _ : To a solution of compound 6 (147.31 mg, 440.69 μmol, 1.2 equiv) in DMF (2 mL) at 0 °C was added HATU (209.46 mg, 550.87 μmol, 1.5 equiv). To the mixture was added compound 32 (147.31 mg, 328.60 μmol, TFA) and DIPEA (142.39 mg, 1.10 mmol, 191.90 μL, 3 equiv) at 25°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was partitioned between _H2O (12 mL) and EtOAc (9 mL). The organic phase was separated, washed with 9 mL (3 mL x 3) of brine, dried _{over Na2SO4} , filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 150×30 mm×5 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 40%-70%, 9 minutes). Compound 106 was obtained as a yellow oil (15 mg, 23.78 μmol, yield 6.48%, purity 93.32%). LCMS (ESI): m/z: [M ₊ H] _calcd for _C29H29F4N4O5 : ₅₈₉ ; found 589; RT= _4.244 min. The observed isomer ratio was 1.18:1:2.50:3.29. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.50-7.71 (m, 1 H) 7.36-7.48 (m, 1 H) 7.12-7.26 (m, 2 H) 6.91-7.10 (m, 2 H) 5.07 -5.35 (m, 1 H) 4.54-4.77 (m, 2 H) 3.18-3.31 (m, 2 H) 2.03-2.68 (m, 3 H) 1.61-1.93 (m, 4 H) 0.71-0.95 (m, 1 H) 0.37-0.59 (m, 2 H) 0.02-0.28 (m, 2 H). Example 7. Preparation of N -(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidine -3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole - 2 - carboxamide _ _ _ _ _ _ _ ( 107 )

Compound 34 : To a solution of compound 28 (887.14 mg, 5.50 mmol, 1 equiv) in DCM (5 mL) was slowly added EDCI (1.16 g, 6.06 mmol, 1.1 equiv), HOBt (818.21 mg, 6.06 mmol, 1.1 equiv) ), and then DIPEA (2.49 g, 19.27 mmol, 3.36 mL, 3.5 equiv) was added dropwise at 0 °C, and the resulting mixture was stirred at 0 °C for 20 min. Subsequently, Compound 33 was added to the mixture at 0°C, and the resulting mixture was stirred at 25°C for 1 hour. TLC (petroleum ether/ethyl acetate=3/1, product Rf ₌ 0.3) showed starting material consumed and one major spot observed. To the mixture was added water (10 mL), and the resulting mixture was separated into layers. The inorganic phase was extracted with DCM (10 mL x 3), and the combined organic phases were concentrated under reduced pressure to give a yellow solid. And the solid was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=100/0-100/13) to obtain compound 34 (1 g, 3.47 mmol, yield 63.00%) as a yellow solid . ¹ H NMR (400 MHz, chloroform-d) δ 9.78 (s, 1H), 7.97 (d, J =8.0 Hz, 1H), 7.75 (d, J =8.3 Hz, 1H), 7.67-7.57 (m, 1H) ), 7.51-7.42 (m, 1H), 7.04 (d, J = 8.4 Hz, 1H), 5.28-5.18 (m, 1H), 4.12 (s, 3H), 2.13-2.10 (m, 1H), 2.05 ( s, 2H), 1.32 (t, J = 5.8 Hz, 6H).

Compound 35 : To a solution of compound 34 (500 mg, 1.73 mmol, 1 equiv) in MeOH (2.5 mL) and _H2O (2.5 mL) at 25°C was added _LiOH.H2O (109.15 mg, 2.60 mmol, 1.5 equiv), and the resulting mixture was stirred at 25 °C for 12 h. The pH of the reaction mixture was adjusted to 2-3 with HCl (2 M) and then concentrated under reduced pressure to give a white solid. And the solid was dissolved in ethyl acetate (5 mL) and water (3 mL), and the resulting mixture was separated. The aqueous phase was extracted with ethyl acetate (3 mL×2), and the combined organic phases were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to give compound 35 (440 mg, 1.60 mmol, yield 92.50%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.64 (s, 1H), 11.56 (s, 1H), 8.57 (d, J =8.1 Hz, 1H), 7.73-7.55 (m, 1H), 7.51- 7.36 (m, 1H), 7.31-7.15 (m, 2H), 7.12-6.96 (m, 1H), 4.55-4.42 (m, 1H), 1.85-1.66 (m, 2H), 1.65-1.54 (m, 1H) ), 1.05-0.76 (m, 6H).

N -(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole - 2 - carboxamide ( 107 ) : _ _ _ _ _ _ To a solution of compound 35 (50 mg, 182.27 μmol, 1 equiv) in DMF (1 mL) at 0°C was added HATU (103.96 mg, 273.41 μmol, 1.5 equiv), DIPEA (82.45 mg, 637.95 μmol, 111.12 μL, 3.5 equiv) and compound 6 (85 mg, 229.28 μmol, 1.26 equiv, HCl, crude, calculated as 100% pure), and the resulting mixture was stirred at 25°C for 2 hours. TLC (petroleum ether/ethyl acetate = 1/2, product R _f = 0.5) showed that a new major spot was observed. The reaction mixture was diluted with water (2 mL), and the resulting mixture was extracted with ethyl acetate (2 mL x 3), and the combined organic phases were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to give a yellow oil . And the oil was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=100/10-0/100) to obtain a yellow oil. Purification by preparative HPLC (column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 μm; mobile phase: [water (10 mM _{NH4HCO3 )} -ACN]; B%: 30%-60%, 12 minutes) As an oil, compound 107 was obtained as a white solid (12 mg, 20.32 μmol, 11.15% yield, 100% purity). LCMS (ESI): m/z: [M ₊ H] _calcd for _C29H31F4N4O5 : ₅₉₁ ; found 591; RT= _2.570 min. ¹ H NMR (400 MHz, methanol-d ₄ ) δ 7.71-7.49 (m, 1H), 7.48-7.34 (m, 1H), 7.29-7.12 (m, 2H), 7.11-6.85 (m, 2H), 5.29 -5.12 (m, 1H), 4.75-4.61 (m, 2H), 4.45-4.07 (m, 1H), 3.30-3.19 (m, 2H), 2.70-2.37 (m, 1H), 2.52-2.36 (m, 1H), 2.34-2.22 (m, 1H), 2.18-2.02 (m, 1H), 1.96-1.64 (m, 5H), 1.14-0.85 (m, 6H). ee value = 30.16%. Example 8. Preparation of N - ( ( S ) -1 - ( ( ( S ) -4 - ( ( 1,1,1,3,3,3 - hexafluoropropan - 2 - yl ) oxy ) -3 - side _ _ _ Oxy - 1 -(( S ) -2 - oxypyrrolidin - 3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -1H -Indole - 2 - carboxamide ( 108 ) _

The title compound was prepared as described in Example 7 using ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoro in the final step Prop-2-yl)oxy)-3-side oxybutyl)pyrrolidin-2-one in place of compound 6 (( S )-3-(( S )-2-amino-3-sideoxy -4-(2,3,5,6-tetrafluorophenoxy)butyl)pyrrolidin-2-one). The product was a white solid (40 mg, 63.26 μmol, 28.48% yield, 93.703% purity). The observed isomer ratio was 2.25:1:14.05:15.68:28.93:10.56 as determined by SFC. LCMS (ESI): m/z: _[ M ₊ H] _calcd for _C26H31F6N4O5 : ₅₉₃ ; found 593. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.61 (d, J =8.0 Hz, 1 H) 7.44 (d, J =8.3 Hz, 1 H) 7.16-7.25 (m, 2 H) 7.03-7.11 ( m, 1 H) 4.91-5.11 (m, 2 H) 4.53-4.82 (m, 3 H) 3.16-3.29 (m, 2 H) 2.22-2.62 (m, 2 H) 1.97-2.13 (m, 1 H) 1.63-1.91 (m, 5 H) 0.89-1.11 (m, 6 H). Example 9. Preparation of N - ( ( S ) -3 - cyclopropyl - 1 - ( ( ( S ) -4 - ( ( 1,1,1,3,3,3 - hexafluoropropan - 2 - yl ) oxy _ _ _ yl ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidin - 3 - yl ) but - 2 - yl ) amino ) -1 - oxyprop - 2 - yl )- 1H - indole - 2 - carboxamide ( 109 )

The title compound was prepared as described in Example 6 using ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoro in the final step prop-2-yl)oxy)-3-pendoxobutyl)pyrrolidin-2-one in place of compound 6 . The product was obtained as a white solid (40 mg, 64.35 μmol, 19.47% yield, 95% purity). LCMS (ESI): m/z: _[ M ₊ H] _calcd for _C26H29F6N4O5 : 591; found _591.3 . The observed isomer ratio was 1:5.31:2.14:6.05 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.62 (d, J =8.0 Hz, 1 H) 7.44 (d, J =8.3 Hz, 1 H) 7.15-7.26 (m, 2 H) 7.02-7.10 ( m, 1 H) 4.99 (dt, J = 12.0, 6.0 Hz, 1 H) 4.57-4.82 (m, 5 H) 3.21-3.29 (m, 2 H) 2.20-2.66 (m, 2 H) 1.96-2.09 ( m, 1 H) 1.59-1.88 (m, 4 H) 0.87 (br s, 1 H) 0.52 (br s, 2 H) 0.21 (br s, 2 H). Example 10. Preparation of N -(( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidine -3 - yl ) -4- ( 2,3,6 - trifluorophenoxy ) butan - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole - 2 - carboxamide ( 110 ) _ _ _ _ _

The title compound was prepared as described in Example 7 using ( S )-3-(( S )-2-amino-3-oxy-4-(2,3,6-trifluorobenzene in the final step) oxy)butyl)pyrrolidin-2-one in place of compound 6 . The product was obtained as a white solid (17 mg, 28.17 μmol, 9.66% yield, 94.88% purity). LCMS (ESI): m/z: [ _{M +} H] calcd for _C29H32F3N4O5 : _573.3 ; found _573.3 . The observed isomer ratio was 2.5:3:1:10.4:14.2 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.92-1.07 (m, 6 H) 1.19-2.69 (m, 9 H) 3.18-3.30 (m, 2 H) 4.02-4.80 (m, 2 H) 5.00 -5.27 (m, 1 H) 6.57-6.99 (m, 2 H) 7.02-7.11 (m, 1 H) 7.12-7.27 (m, 2 H) 7.37-7.46 (m, 1 H) 7.50-7.66 (m, 1H). Example 11. Preparation of N -(( S ) -3 - cyclohexyl- 1 - oxy - 1 - ((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidine -3 - yl ) -4- ( 2,3,6 - trifluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl ) -1H - indole - 2 - carboxamide ( 111 ) _ _ _ _ _

The title compound was prepared as described in Example 7, using benzyl( S )-2-amino-3-cyclohexylpropionate in the first step in place of methyl L -leucine (compound 33 ) and 1H -Indole-2-carboxylic acid (compound 28 ) was coupled with ( S )-3-(( S )-2-amino-3-oxo-4-(2,3,6-) in the final step Trifluorophenoxy)butyl)pyrrolidin-2-one replaces compound 6 . The product was obtained as a white solid (2 mg, 2.97 μmol, 1.87% yield, 90.88% purity). LCMS (ESI): m/z: [ _{M +} H] calcd for _C32H36F3N4O5 : _613.3 ; found _613.3 . ee value = 84.98%. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.88-1.13 (m, 2 H) 1.15-1.26 (m, 2 H) 1.27-1.35 (m, 1 H) 1.40-1.57 (m, 1 H) 1.64 -1.88 (m, 7 H) 1.88-1.95 (m, 1 H) 2.06-2.16 (m, 1 H) 2.22-2.34 (m, 1 H) 2.35-2.53 (m, 1 H) 2.54-2.68 (m, 1 H) 3.18-3.29 (m, 2 H) 4.10-4.43 (m, 1 H) 4.63-4.68 (m, 1 H) 4.96-5.14 (m, 2 H) 6.81-7.03 (m, 2 H) 7.03- 7.11 (m, 1 H) 7.13-7.28 (m, 2 H) 7.32-7.48 (m, 1 H) 7.53-7.72 (m, 1 H). Example 12. Preparation of N - ( ( S ) -1 - ( ( ( S ) -4 - ( ( 1,1,1,3,3,3 - hexafluoropropan - 2 - yl ) oxy ) -3 - side _ _ _ Oxy - 1 -(( S ) -2 - oxypyrrolidin - 3 - yl ) but - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 -Methoxy - 1H - indole - 2 - carboxamide ( 112 ) _

The title compound was prepared as described in Example 7, using 4-methoxy-1H-indole-2-carboxylic acid in the first step instead of 1H-indole-2-carboxylic acid (compound 28 ) and L -leucine The methyl ester (compound 33 ) was coupled with ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropane- 2-yl)oxy)-3- pendant oxybutyl)pyrrolidin-2-one in place of compound 6 . The product was obtained as a white solid (15 mg, 22.89 μmol, 10.31% yield, 95.016% purity). The observed isomer ratio was 1:5.4:3.7:8.6 as determined by SFC. LCMS (ESI): m/z: _[ M ₊ H] _calcd for _C27H33F6N4O6 : 623 _; found 623. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.28 (s, 1 H) 7.10-7.21 (m, 1 H) 7.03 (br d, J =8.1 Hz, 1 H) 6.51 (br d, J =7.5 Hz, 1 H) 4.93-5.09 (m, 1 H) 4.48-4.82 (m, 4 H) 3.93 (s, 3 H) 3.25 (br d, J = 6.3 Hz, 2 H) 2.20-2.64 (m, 2 H) 1.95-2.12 (m, 1 H) 1.60-1.92 (m, 5 H) 0.90-1.09 (m, 6 H). Example 13. Preparation of 4 - methoxy- N - (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 -Pendant oxypyrrolidin - 3 - yl ) -4 - phenoxybutan - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole - 2 - carboxamide ( 113 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-3-oxy-4-phenoxybutyl)pyrrolidine-2 in the final step - Ketone in place of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2-yl)oxy)-3-side oxybutyl)pyrrolidin-2-one. The product was obtained as a yellow oil (0.07 g, 116.75 μmol, 21.97% yield, 91.5% purity). LCMS (ESI): m/z: [M ₊ H] calcd for _C30H37N4O6 : _{549.3 ;} found 549.3. The observed isomer ratio was 2.19:1:3.88 as determined by SFC. ¹ H NMR (400 MHz, chloroform- d ) δ 10.78 (br s, 1 H) 9.58-9.81 (m, 1 H) 8.57 (br d, J =6.0 Hz, 1 H) 7.28-7.33 (m, 1 H) ) 7.07-7.23 (m, 2 H) 6.66-7.04 (m, 5 H) 6.49 (d, J = 7.7 Hz, 1 H) 6.05-6.15 (m, 1 H) 4.51-5.01 (m, 4 H) 3.90 -4.05 (m, 3 H) 2.92-3.36 (m, 2 H) 2.52-2.66 (m, 1 H) 2.28-2.49 (m, 1 H) 2.02-2.12 (m, 1 H) 1.94 (dt, J = 10.4, 5.2 Hz, 1 H) 1.73-1.84 (m, 3 H) 1.22-1.55 (m, 1 H) 0.92-1.00 (m, 6 H). Example 14. Preparation of N -(( S ) -1 -((( S ) -4- ( 2 - fluorophenoxy ) -3 - oxy - 1 - (( S ) -2 - oxypyrrolidine -3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 - methoxy - 1H - indole - 2 - carboxamide ( 114 _ )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-4-(2-fluorophenoxy)-3-pendoxobutyl in the final step )pyrrolidin-2-one in place of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2-yl)oxy )-3-oxobutyl)pyrrolidin-2-one. The product was obtained as a yellow oil (10 mg, 16.64 μmol, 4.66% yield, 94.3% purity). LCMS (ESI): m/z: [M+H] calcd for _C30H36FN4O6 : _{567.3 ;} found _567.3 . The observed isomer ratio was 2.82:1:4.13 as determined by SFC. ¹ H NMR (400 MHz, chloroform- d ) δ 10.73 (br s, 1 H) 9.33-9.51 (m, 1 H) 8.58 (br d, J =5.4 Hz, 1 H) 7.19 (q, J =7.8 Hz , 1 H) 7.05-7.14 (m, 2 H) 6.91-7.04 (m, 3 H) 6.66-6.77 (m, 1 H) 6.39-6.53 (m, 1 H) 6.39-6.53 (m, 1 H) 5.80 -5.98 (m, 1 H) 4.89-4.93 (m, 1 H) 4.86 (d, J = 3.6 Hz, 2 H) 4.74-4.82 (m, 1 H) 3.92-4.01 (m, 3 H) 2.90-3.39 (m, 2 H) 2.60 (br d, J = 11.7 Hz, 1 H) 2.32-2.50 (m, 1 H) 2.00-2.08 (m, 1 H) 1.68-1.93 (m, 4 H) 1.24-1.40 ( m, 1 H) 0.99 (br d, J = 5.5 Hz, 6 H). Example 15. Preparation of N -(( S ) -1 -((( S ) -4- ( 2,6 - difluorophenoxy ) -3 - oxy - 1 - ( ( S ) -2 - oxy pyrrolidin - 3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 - methoxy - 1H - indole - 2 - carbamide Amine ( 115 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-4-(2,6-difluorophenoxy)-3-oxygen in the final step ylbutyl)pyrrolidin-2-one instead of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2-yl ) oxy)-3-pendant oxybutyl)pyrrolidin-2-one. The product was obtained as a yellow solid (55 mg, 85.33 mmol, 25.45% yield, 90.7% purity). LCMS (ESI): m/z: _[ M ₊ H] _calcd for _C30H35F2N4O6 : _585.3 ; found 585.3. The observed isomer ratio was 1:4.7:1:6.8 as determined by SFC. ¹ H NMR (400 MHz, chloroform- d ) δ 10.91 (br s, 1 H) 9.32-9.55 (m, 1 H) 8.44 (d, J =6.1 Hz, 1 H) 7.15-7.25 (m, 1 H) 7.09-7.14 (m, 1 H) 7.00-7.04 (m, 1 H) 6.87-6.95 (m, 2 H) 6.79 (br d, J = 8.1 Hz, 1 H) 6.44-6.55 (m, 1 H) 5.94 (s, 1 H) 4.90-5.01 (m, 2 H) 4.63-4.89 (m, 1 H) 3.93-3.99 (m, 3 H) 2.81-3.36 (m, 2 H) 2.22-2.61 (m, 2 H) ) 2.04-2.11 (m, 1 H) 1.69-1.92 (m, 4 H) 1.25-1.50 (m, 1 H) 0.91-1.03 (m, 6 H). Example 16. Preparation of 4 - methoxy- N - (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 -Pendant oxypyrrolidin - 3 - yl ) -4- ( 2,3,6 - trifluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole - 2- _ _ _ _ _ Formamide ( 116 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-3-oxy-4-(2,3,6-trifluorobenzene in the final step) Oxy)butyl)pyrrolidin-2-one instead of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2 -yl)oxy)-3-side oxybutyl)pyrrolidin-2-one. The product was obtained as a white solid (55 mg, 87.06 μmol, 18.73% yield, 95.39% purity). LCMS (ESI): m/z: _{[ M +} H] calcd for _C30H34F3N4O6 : _603.3 ; found 603.3. The observed isomer ratio was 6.9:1:31.9:5.6:45.3 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.91-1.10 (m, 6 H) 1.61-1.92 (m, 5 H) 2.04-2.65 (m, 3 H) 3.19-3.30 (m, 2 H) 3.89 -3.95 (m, 3 H) 3.99-4.44 (m, 1 H) 4.52-4.78 (m, 2 H) 4.98-5.25 (m, 1 H) 6.45-6.53 (m, 1 H) 6.55-7.07 (m, 3H) 7.09-7.31 (m, 2H). Example 17. Preparation of 4 - methoxy- N - (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 -Pendant oxypyrrolidin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) -1H - indole- _ _ _ _ _ _ _ 2 - Carboxamide ( 117 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-3-oxy-4-(2,3,5,6-tetrakis) in the final step Fluorophenoxy)butyl)pyrrolidin-2-one instead of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropane) -2-yl)oxy)-3-side oxybutyl)pyrrolidin-2-one. The product was obtained as a white solid (21 mg, 32.13 μmol, 11.91% yield, 94.94% purity). LCMS (ESI): m/z: _[ M ₊ H] calcd for _C30H33F4N4O6 : _621.3 ; found _621.3 . The observed isomer ratio was 2.6:9.9:1:7.9:2.6 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.94-1.06 (m, 6 H) 1.64-1.95 (m, 5 H) 2.00-2.17 (m, 1 H) 2.23-2.65 (m, 2 H) 3.21 -3.30 (m, 2 H) 3.86-3.95 (m, 3 H) 4.09-4.40 (m, 1 H) 4.54-4.79 (m, 2 H) 5.09-5.28 (m, 1 H) 6.45-6.57 (m, 1H) 6.89-7.32 (m, 4H). Example 18. Preparation of N -(( S ) -1 -(((( S ) -4- ( isoxazol - 3 - yloxy ) -3 - oxy - 1 - (( S ) -2 - oxy pyrrolidin - 3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 - methoxy - 1H - indole - 2 - carbamide Amines ( 118 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-4-(isoxazol-3-yloxy)-3-oxygen in the final step ylbutyl)pyrrolidin-2-one instead of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2-yl ) oxy)-3-pendant oxybutyl)pyrrolidin-2-one. The product was obtained as a yellow solid (50 mg, 87.41 μmol, 10.64% yield, 94.33% purity). LCMS (ESI): m/z: [M+H] _calcd for _C27H34N5O7 : _540.3 ; found _540.3 . The observed isomer ratio was 1.18:4.08:1:7.25:15.99 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.87-1.09 (m, 6 H) 1.57-1.94 (m, 5 H) 2.01-2.22 (m, 1 H) 2.25-2.69 (m, 2 H) 3.09 -3.26 (m, 2 H) 3.93 (s, 3 H) 4.52-4.73 (m, 2 H) 5.01-5.27 (m, 2 H) 5.98-6.23 (m, 1 H) 6.51 (d, J = 7.75 Hz , 1 H) 6.92-7.07 (m, 1 H) 7.12-7.19 (m, 1 H) 7.24-7.35 (m, 1 H) 8.35 (dd, J = 5.82, 1.81 Hz, 1 H). Example 19. Preparation of N -(( S ) -1 -((( S ) -4 - (( 2,6 - lutidine - 4 - yl ) oxy ) -3 - pendoxo - 1 - ( ( S ) -2 - oxypyrrolidin - 3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 - methoxy - 1H- _ Indole - 2 - carboxamide ( 119 )

The title compound was prepared as described in Example 12 using (S)-3-((S)-2-amino-4-((2,6-lutidine-4-yl)oxy in the final step ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3- Hexafluoroprop-2-yl)oxy)-3-side oxybutyl)pyrrolidin-2-one. The product was obtained as a yellow solid (23 mg, 36.50 μmol, 7.40% yield, 91.68% purity). ee value = 32.32%. LCMS (ESI): m/z: _[ M+H] calcd for _C31H40N5O6 : _578.3 ; found _578.3 . ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.88-1.07 (m, 6 H) 1.59-1.94 (m, 5 H) 1.96-2.25 (m, 2 H) 2.26-2.43 (m, 6 H) 2.43 -2.63 (m, 1 H) 3.17-3.30 (m, 2 H) 3.93 (s, 3 H) 4.27-4.70 (m, 2 H) 4.89-5.17 (m, 2 H) 6.51 (d, J = 7.75 Hz , 1 H) 6.54-6.72 (m, 2 H) 6.96-7.05 (m, 1 H) 7.09-7.19 (m, 1 H) 7.24-7.33 (m, 1 H). Example 20. Preparation of N -(( S ) -1 -(((( S ) -4- ( isoxazol - 4 - yloxy ) -3 - oxo - 1 - (( S ) -2 - oxo pyrrolidin - 3 - yl ) butan - 2 - yl ) amino ) -4 - methyl - 1 - oxypentan - 2 - yl ) -4 - methoxy - 1H - indole - 2 - carbamide Amine ( 120 )

The title compound was prepared as described in Example 12 using ( S )-3-(( S )-2-amino-4-(isoxazol-4-yloxy)-3- pendant oxygen in the final step ylbutyl)pyrrolidin-2-one instead of ( S )-3-(( S )-2-amino-4-((1,1,1,3,3,3-hexafluoropropan-2-yl ) oxy)-3-pendant oxybutyl)pyrrolidin-2-one. The product was a white solid (80 mg, 146.69 μmol, 21.55% yield, 98.94% purity). The observed isomer ratio was 6.13:23.98:33.41:1 as determined by SFC. LCMS (ESI): m/z: [M+H] _calcd for _C27H34N5O7 : _540.3 ; found _540.3 . ¹ H NMR (400 MHz, methanol- d ₄ ) δ 1.01 (dd, J =16.44, 6.14 Hz, 6 H) 1.60-1.94 (m, 5 H) 2.03-2.15 (m, 1 H) 2.25-2.36 (m , 1 H) 2.37-2.62 (m, 1 H) 3.20-3.30 (m, 2 H) 3.93 (s, 3 H) 4.46-4.69 (m, 2 H) 4.80-4.88 (m, 2 H) 6.51 (d , J = 7.67 Hz, 1 H) 7.03 (d, J = 8.33 Hz, 1 H) 7.12-7.18 (m, 1 H) 7.26-7.34 (m, 1 H) 8.31-8.40 (m, 1 H) 8.44- 8.56 (m, 1H). Example 21. Preparation of N -(( S ) -3 - cyclopropyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrole pyridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) prop - 2 - yl ) benzofuran - 2 - carboxamide ( 121 _ _ _ _ _ _ )

The title compound was prepared as described in Example 6, using benzofuran-2-carboxylic acid in place of 1H-indole-2-carboxylic acid (compound 28 ) and ( S )-2-amino-3-cyclopropylpropionic acid methyl Ester (compound 8 ) coupling. The product was obtained as a yellow oil (29 mg, 49.19 μmol, 22.41% yield, 100% purity). LCMS (ESI): m/z: _{[ M +} H] calcd for _C29H28F4N3O6 : _590.2 ; found 590.2. The observed isomer ratio was 2.45:10.58:1 as determined by SFC. ¹ H NMR (400 MHz, chloroform- d ) δ 8.59-9.26 (m, 1 H) 7.62-7.70 (m, 1 H) 7.52 (d, J =8.1 Hz, 1 H) 7.46 (s, 1 H) 7.35 -7.45 (m, 2 H) 7.30 (d, J = 7.9 Hz, 1 H) 6.71-6.88 (m, 1 H) 5.79-6.18 (m, 1 H) 5.00-5.26 (m, 2 H) 4.66-4.93 (m, 2 H) 3.30-3.43 (m, 2 H) 2.38-2.56 (m, 2 H) 1.99-2.17 (m, 2 H) 1.82-1.95 (m, 3 H) 0.71-0.92 (m, 1 H) ) 0.55 (br d, J = 7.9 Hz, 2 H) 0.09-0.28 (m, 2 H). Example 22. Preparation of N -(( S ) -3 - cyclopropyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrole Iridin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) butan - 2 - yl ) amino ) propan - 2 - yl ) -N - methylbenzofuran - 2- _ _ _ _ _ _ _ Formamide ( 122 )

The title compound was prepared as described in Example 21 using (S)-methyl 3-cyclopropyl-2-(methylamino)propanoate in place of ( S )-2-amino-3-cyclopropylpropanoic acid The methyl ester is coupled with benzofuran-2-carboxylic acid. The product was obtained as a yellow oil (0.06 g, 94.44 μmol, 12.33% yield, 95% purity). LCMS (ESI): m/z: _{[ M +} H] calcd for _C30H30F4N3O6 : _604.3 ; found 604.3. ¹ H NMR (400 MHz, chloroform- d ) δ -0.20-0.23 (m, 2 H) 0.24-0.61 (m, 2 H) 0.63-0.91 (m, 1 H) 1.19-1.74 (m, 1 H) 1.79 -2.20 (m, 4 H) 2.23-2.65 (m, 2 H) 2.92-3.29 (m, 2 H) 3.33 (br s, 3 H) 4.64-4.95 (m, 1 H) 5.00-5.34 (m, 3 H) 6.78 (br d, J =8.50 Hz, 1 H) 7.28-7.34 (m, 1 H) 7.35-7.47 (m, 2 H) 7.54 (br d, J =7.00 Hz, 1 H) 7.67 (d, J = 7.75 Hz, 1 H) 8.37 (br s, 1 H). Example 23. Preparation of 4 - methoxy- N - (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 -Pendant oxypyrrolidin - 3 - yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) benzofuran - 2- _ _ _ _ _ _ _ Formamide ( 123 )

The title compound was prepared as described in Example 21 via coupling of 4-methoxybenzofuran-2-carboxylic acid with methyl L -leucine (Compound 1B ) followed by hydrolysis of the resulting methyl ester to give (4- Methoxybenzofuran-2-carbonyl) -L -leucine for use in the final step with compound 6 (( S )-3-(( S )-2-amino-3-pendoxyloxy- 4-(2,3,5,6-Tetrafluorophenoxy)butyl)-pyrrolidin-2-one) reaction. The product was obtained as a white solid (20 mg, 30.51 μmol, 5.65% yield, 94.81% purity). LCMS (ESI): m/z: [ _M +H] _calcd for _C30H32F4N3O7 : _622.2 ; found _622.2 . The observed isomer ratio was 5.9:23.7:1 as determined by SFC. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.92-1.09 (m, 6 H) 1.62-1.97 (m, 5 H) 2.04-2.18 (m, 1 H) 2.26-2.63 (m, 2 H) 3.96 (s, 3 H) 4.07-4.45 (m, 1 H) 4.56-4.76 (m, 2 H) 5.11-5.26 (m, 1 H) 6.64-6.88 (m, 1 H) 6.91-7.24 (m, 2 H) ) 7.33-7.44 (m, 1 H) 7.46-7.57 (m, 1 H). Example 24. Preparation of (( S ) -4 - methyl - 1 - oxy - 1 -((( S ) -3 - oxy - 1 -(( S ) -2 - oxypyrrolidine - 3 -yl ) -4- ( 2,3,5,6 - tetrafluorophenoxy ) but - 2 - yl ) amino ) pentan - 2 - yl ) carbamate 4,4 - difluorocyclohexyl ester ( 124 _ _ _ _ _ _ _ _ )

The title compound was prepared as described in Example 7 using 4,4-difluorocyclohexyl chloroformate in the first step in place of 1H-indole-2-carboxylic acid (compound 28 ). 4,4-Difluorocyclohexyl chloroformate was obtained in 68.55% yield via the reaction of 4,4-difluorocyclohexane-1-ol with triphosgene in dichloromethane containing pyridine (2.8 equiv.) . The title compound (10 mg, 16.41 μmol, 4.01% yield, 100% purity) was obtained as a white solid. LCMS (ESI): m/z: _{[ M} +H] calcd for _C27H34F6N3O6 : _{610.3 ;} found 610.3. The observed enantiomeric excess was 87.1% as determined by SF. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.83-1.10 (m, 6 H) 1.20-1.59 (m, 3 H) 1.65-2.19 (m, 12 H) 2.20-2.65 (m, 2 H) 2.97 -3.29 (m, 1 H) 3.34-3.49 (m, 1 H) 3.69-4.45 (m, 2 H) 4.63-4.81 (m, 1 H) 5.16 (d, J = 8.58 Hz, 1 H) 7.12 (ddd , J = 10.46, 7.24, 3.28 Hz, 1 H). Example 25. Preparation of ( 1R , 2S , 5S ) -3 - ( ( S ) -3,3 - dimethyl - 2- ( 2,2,2 - trifluoroacetamido ) butanoyl ) -6,6- _ _ _ _ _ _ _ Dimethyl - N - ( ( S ) -3 - oxy - 1 - ( ( S ) -2 - oxypyrrolidin - 3 - yl ) -4- ( 2,3,6 - trifluorophenoxy yl ) butan - 2 - yl ) -3 - azabicyclo [ 3.1.0 ] hexane - 2 - carboxamide ( 125 ) _ _ _ _

Compound 38 : To a mixture of compound 36 (2 g, 15.25 mmol, 1 equiv) in MeOH (9 mL) at 25 °C was added TEA (2.18 g, 21.55 mmol, 3.00 mL, 1.41 equiv) and compound 37 ( 2.03 g, 15.86 mmol, 1.60 mL, 1.04 equiv) and the mixture was stirred at 25 °C for 12 h. TLC (petroleum ether:ethyl acetate=0:1, Rf ₌ 0.43) showed that the reaction was complete. The residue was concentrated under reduced pressure to give compound 38 (3.2 g, 9.75 mmol, 63.91% yield, TEA) as a white solid. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 1.01 (s, 9 H) 4.17 (s, 1 H).

Compound 40 : To a mixture of compound 38 (1 g, 3.05 mmol, 1 equiv, TEA) in toluene (1 mL) and DMF (1 mL) at 25 °C was added compound 39 (187.91 mg, 913.60 μmol, 0.3 equiv, HCl), EDCI (758.94 mg, 3.96 mmol, 1.3 equiv) and NMM (770.07 mg, 7.61 mmol, 837.03 μL, 2.5 equiv). The mixture was stirred at 25°C for 5 hours. LCMS showed the reaction was complete. The mixture was filtered, and the filtrate was concentrated under reduced pressure. Purification by preparative HPLC (column: Phenomenex Gemini-NX C18 75 x 30 mm x 3 μm; mobile phase: [water (10 mM _{NH4HCO3 )} -ACN]; B%: 40%-60%, 12 minutes) The residue gave compound 40 (0.31 g, 819.27 μmol, 26.90% yield) as a white solid. ¹ H NMR (400 MHz, chloroform- d ) δ 0.98-1.04 (m, 7 H) 1.08 (s, 5 H) 1.49-1.52 (m, 1 H) 1.54-1.56 (m, 1 H) 3.72 (s, 1 H) 3.78 (s, 1 H) 3.94 (dd, J = 10.19, 4.06 Hz, 1 H) 4.09 (dd, J = 10.08, 5.48 Hz, 1 H) 4.30-4.49 (m, 1 H) 4.55-4.64 (m,1H) 6.79-6.92 (m,1H).

Compound 41 : To a mixture of compound 40 (0.3 g, 792.84 μmol, 1 equiv) in _H2O (1 mL) and MeOH (1 mL) at 25 °C was added _LiOH.H2O (49.91 mg, 1.19 mmol, 1.5 equiv), and the mixture was stirred at 25 °C for 12 h. LCMS showed the reaction was complete. The mixture was combined with 1M HCl (10 mL) and ethyl acetate (10 mL), and the aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with brine (10 mL x 3), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give compound 40 (0.21 g, 576.35 mmol, yield 72.69) as a white solid %).

( 1R , 2S , 5S ) -3 - ( ( S ) -3,3 - dimethyl - 2- ( 2,2,2 - trifluoroacetamido ) butyryl ) -6,6 - dimethyl- _ _ _ _ _ _ _ N - ( ( S ) -3 - oxy - 1 - ( ( S ) -2 - oxypyrrolidin - 3 - yl ) -4- ( 2,3,6 - trifluorophenoxy ) butan- _ 2 - yl ) -3 - azabicyclo [ 3.1.0 ] hexane - 2 - carboxamide ( 125 ) : Compound 42 was prepared as described for compound 6 in Example 1 using 2,3,6- Trifluorophenol replaces 2,3,4,6-tetrafluorophenol. To a solution of compound 41 (0.1 g, 274.45 μmol, 1 equiv) and compound 42 (118.10 mg, 274.45 μmol, 1 equiv, TFA) in DMF (1 mL) at 25°C was added HATU (208.71 mg, 548.91 μmol, 2 equiv) and DIPEA (141.89 mg, 1.10 mmol, 191.22 μL, 4 equiv). The mixture was stirred at 25°C for 1 hour. LCMS showed the reaction was complete. The mixture was filtered, and the filtrate was concentrated under reduced pressure. By preparative HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10 μm; mobile phase: [water( _{NH3H2O + NH4HCO3 )} -ACN]; B%: 25%-65%, 8 min), the residue was purified to give the product 125 (80 mg, 115.14 μmol, 41.95% yield, 95.37% purity) as a yellow solid. LCMS (ESI): m/z: _[ M ₊ H] calcd for _C30H37F6N4O6 : 663.3 _; found 663.3; RT= _3.152 min. The observed enantiomeric excess was 11.6% as determined by SF. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 0.92-1.09 (m,15H) 1.42-1.48 (m,1H) 1.57-1.65 (m,1H) 1.71-1.86 (m,1H) 1.86 -1.98 (m, 1 H) 2.01-2.16 (m, 1 H) 2.29-2.41 (m, 1 H) 2.60-2.70 (m, 1 H) 3.23-3.30 (m, 1 H) 3.32-3.37 (m, 1 H) 3.64-3.88 (m, 1 H) 3.93-4.13 (m, 1 H) 4.20-4.31 (m, 1 H) 4.32-4.43 (m, 1 H) 4.55-4.69 (m, 1 H) 4.69- 4.84 (m, 1 H) 5.07-5.12 (m, 1 H) 6.95-7.04 (m, 2 H). Example 26. Analysis of SARS - CoV2 3CL ^pro inhibition

30 mM stock solutions of compounds in DMSO were prepared. Compounds were tested for their efficacy in inhibiting the cysteine protease activity of SARS-CoV-2 Mpro by monitoring cleavage of the fluorescent substrate in a 10-dose _IC50 curve (see Reaction Biology SARS-CoV-2 MPro Protease Assay, Table of Contents No. Mpro) with 3-fold serial dilutions starting with a high test concentration of 10 μM. The final concentration of substrate was 5 μM. Protease activity was measured as a time-course increase in the fluorescent signal from the fluorescent peptide substrate, and the initial linear portion of the slope (signal/min) was analyzed. Data were evaluated in terms of Hill slope (% enzymatic activity of control without inhibitor as 100% activity) and _IC50 . Compound binding and inhibition were not affected by the presence or absence of a reducing agent (DTT), which was added to mimic the reducing environment of the cell and prevent oxidation of the protease active site. The activity was compared with the reference compound ( 2S)-2-((S ) -2-(((benzyloxy)carbonyl)amino)-4-methylpentamido)-1-hydroxy-3-( The activity of ( S )-2-oxypyrrolidin-3-yl)propane-1-sulfonate ( 001 ) was compared. As summarized in Table 1, these compounds are potent inhibitors of SARS-CoV-2 3CL ^pro , with a number of compounds exhibiting _IC50 values well below 15 nM. Table 1 compound 3CL ^pro inhibitory activity 102 ++ 103 + 104 ++ 105 +++ 106 +++ 107 +++ 108 + 109 + 110 +++ 112 + 113 ++ 114 ++ 115 ++ 116 +++ 117 +++ 121 ++ 122 +++ 123 ++ 124 ++ 125 ++ +++: 1 nM < _IC50 ≤ 15 nM ++: 15 nM < _IC50 ≤ 150 nM +: 150 nM < _IC50 < 300 nM Example 27. Major protease inhibition in various coronaviruses.

The _IC50 values of compound 107 against the major proteases of SARS-CoV2, 229E, OC43, MERS, SARS, HKU1 and NL63 coronaviruses were determined, using a FRET-based cytoplasmic cleavage assay to measure the resistance to each of these virus strains. Inhibition of one of the isolated Mpro/3CLpro proteases. Under the analytical conditions used, _IC50 values were measured in the range of 7.9 nM to 96 nM. The potency of compound 107 was approximately 2 to 3.5 times higher than that of comparator 001 against the major proteases of SARS-CoV2, 229E, MERS, HKU1 and NL63. The data indicate that these inhibitors have broad activity against the Mpro/3CLpro protease of most coronavirus strains. Example 28. Effect of Inhibitors on SARS - CoV - 2 Virus Replication

SARS-CoV-2 was prepared by passage of virus in Vero 76 cells using test medium in MEM supplemented with 2% FBS and 50 μg/mL gentamicin (USA-WA1/2020) raw material. Test compounds were dissolved in DMSO to make 50 mM stock solutions. Compounds were serially diluted from a (high) starting test concentration of 10 μM. Each dilution was added to 5 wells of a 96-well plate along with 80-100% confluent Vero 76 cells. Each dilution of three wells was infected with virus, and two wells were left uninfected to assess compound cytotoxicity. Six wells that were infected and untreated served as virus controls, and six wells that were uninfected and untreated served as cell controls. SARS-CoV-2 was prepared to achieve the lowest possible multiplication of infection (MOI) that would yield >80% cytopathic effect (CPE) within 5 days. This assay assesses multiple rounds of viral infection, replication, and viral production. Plates were incubated at 37±2°C, 5% CO ₂ . On day 5 post-infection, once the untreated virus control wells reached maximum CPE, the disks were stained with neutral red dye for approximately 2 hours (±15 minutes). The incorporated dye was extracted in 50:50 Sorensen citrate buffer/ethanol for >30 minutes and the optical density was read at 540 nm with a spectrophotometer. This assay evaluates virus-induced CPE and the ability of compounds to inhibit it. Optical densities were converted to percent cellular controls and normalized to virus controls, and then the concentration of test compound required to inhibit CPE by 50% ( _EC50 ) and 90% (EC90) was calculated by regression analysis.

For the virus yield reduction assay, on day 3 post-infection, supernatant fluid from wells treated with and without each compound at all concentrations tested was collected, followed by neutral red staining (3 wells pooled), and Virus titers were tested using a standard end-point dilution CCID50 assay and calculated using the Reed-Muench (1948) equation. The concentration of compound required to reduce virus yield by 1 log10 was calculated by regression analysis (EC90).

The compounds provided herein exhibited high potency in viral replication assays as assessed by _EC90 values. Unexpectedly, compound 102 exhibited a 3.7-fold improvement ( _EC90 ) in the viral replication assay compared to reference compound 001 , although the _IC50 for in vitro _3CL ^pro inhibition by compound 102 was higher than that of reference compound 001 about an order of magnitude. In the CPE-based viral replication assay described above, a similar increase in potency of compound 102 was observed compared to control compound 001.

The antiviral activities of the compounds described herein are summarized in Table 2. Virus-induced CPE assay _EC50 values and Virus Yield Reduction ( _VYR ) EC90 values (the concentration calculated to reduce viral yield by 1 log) were determined over three days. Also included are ₅₀ % cytotoxicity concentration values (CC50). The observed antiviral potency of the compounds is consistent with 3CLpro inhibitor activity. Antiviral activity was enhanced in the presence of CP-100356, a Pgp inhibitor. Compared to known comparators remdesivir and 001 , the compounds of the present invention exhibited significantly higher potency. The CC50 ( ₅₀ % cytotoxic concentration) value was determined to assess cytotoxicity. The _CC50 values determined for compounds 105 and 107 were in the range of 17-19 μM. The _CC50 values determined for other compounds in the table were greater than 30 μM. Table 2 compound +/- CP -100356 Pgp inhibitor EC ₅₀ EC ₉₀ 105 - † † 105 + ††† ††† 107 - † † 107 + ††† ††† 114 - † † 114 + †† † 116 - † † 116 + ††† ††† 121 - † † 121 + ††† †† remdesivir - † † †††: _IC50 / _EC90 < 500 nM ††: 500 nM ≤ IC ₅₀ /EC ₉₀ ≤ 1 µM †: 1 µM < _IC50 / _EC90 Example 29. Effects of inhibitors on SARS - CoV - 2 virus replication in human lung cells .

Compounds 107 , 110 , 116 and 117 were tested in a cell-based antiviral assay using the SARS CoV-2 reporter virus in which nanoluciferase is inserted into the viral genome at ORF7. The cell-based assay measures nanoluciferase activity as an exponential form of viral replication in the human lung cell line A549 cells expressing ACE2 at 72 hours post-infection.

Test article preparation: Working stock solutions of each compound were prepared by diluting stock solutions (10 mM in DMSO) to 33.3 μM (for compound 107 ) and 100 μM (for compounds 110 , 116 and 117 ). An 80 μL aliquot of each working stock sample was transferred to the wells of an empty ECHO plate (Labcyte Cat. No. P-05525). These samples were diluted 2-fold by transferring 40 μL of each sample to adjacent wells containing 40 μL of medium and mixing. This process was repeated to generate an additional 8 wells with serially diluted samples, each well containing a 2-fold diluted sample of the previous well. A 90-nL aliquot of each sample was then dispensed into the corresponding well of an analytical reserve tray (ARP) using an ECHO555 sonic liquid handling system. The final assay concentration range in the assay was 0.2-100 nM for compound 107 and 0.6-300 nM for compounds 110 , 116 and 117 .

Measurement of antiviral activity: A549 cells expressing ACE-2 were grown and split 1:6 twice a week in high glucose DMEM supplemented with 20% HI FBS, 1% NEAA, 100 μg/ ml of blasticidin. The blasticidin was removed from the medium at one passage prior to using the cells for the assay. On the day of the assay, cells were harvested in DMEM supplemented with 2% HI FBS, 1% HEPES, 1% Pen/Strep. In the BSL-2 laboratory, assay preparation discs (ARPs) pre-dosed with test compounds were prepared by adding 5 [mu]L assay medium to each well. The disks and cells were then transferred to the BSL-3 apparatus. A working stock of SARS CoV-2 Nanoluciferase Reporter Virus (NLRV) passaged five times in A549 cells expressing ACE2 was diluted in medium containing 160,000 cells per milliliter (MOI of about 0.002) 6000 times and agitate at 200 RPM for about 10 minutes. A 25 μL aliquot of virus-inoculated cells (4000 cells) was added to each well in rows 3 to 24 of the assay plate. Wells in rows 23-24 contained no test compound and only virus-infected cells as a 0% inhibition control. A 25 μL aliquot of cells was added to rows 1 to 2 of each plate (without test compound) prior to virus inoculation as a 100% inhibition control containing cells only. After incubating the plates for 72 hours at 37°C/5% CO2 and 90% humidity, 30 μL NanoGlo (Promega) was added to each well. After 10 min incubation at room temperature, luciferase activity was measured as an index of viral titer by reading luminescence using a BMG CLARIOstar plate reader (bottom reading). The disk was sealed with a clear cover and the surface was decontaminated, followed by fluorescence reading.

The _EC50 values for compounds 107 , 110 , 116 and 117 were determined in the range of 22 nM to 35 nM in A549 human lung cells. The _EC50 value of remdesivir determined using the same assay conditions was 130 nM. Example 30. Inhibitor pharmacokinetics and selectivity .

Compounds 107 and 116 were administered intravenously (1 mg/kg), orally (5 mg/kg), subcutaneously (5 mg/kg) and intranasally (5 mg/kg) to laboratory mice. Compounds were formulated for IV and oral administration in DMSO/PEG300/solutol/water (v:v:v:v, 5:20:5:70) and in 0.5% HPMC ( 4000 cps), 0.5% polysorbate, 10 mM PBS (pH 7.4) for subcutaneous and intranasal administration. For compound 107 , _Cmax values of 51.4 ng/mL (po), 424 ng/mL (sc) and 737 ng/mL (in) were observed; for compound 116 , 538 ng/mL (sc) and 443 ng were observed _Cmax value per mL (in). The data indicate that this series of compounds provides in vivo effective plasma concentrations for the treatment of coronavirus infection.

Compounds 107 , 110 , 116 , and 117 were evaluated for selectivity and represent inhibitors in this series of compounds. Selectivity was analyzed using an industry standard poor enzyme panel screening of enzymes as well as a panel of cellular proteases. Screening was performed at a single concentration (10 μM) for each compound. High selectivity levels of SARS-CoV-2 3Clpro/Mpro proteases against off-target targets were identified. Off-target activity was identified at any level only in the case of a set of metabolic enzymes (Cytochrome P450 1A2/3A4/2C19 and UGT1A1) and a small subset of the cysteine cathepsin family of proteases and calpain-1. No other off-target activities were identified in the case of other cellular proteases or pharmacologically important enzymes. VI . Exemplary Embodiments

， 或其醫藥學上可接受之鹽， 其中： R ¹係選自由以下組成之群：C _{6 - 10}芳基、5員至12員雜芳基及C _{1 - 6}鹵烷基，其中C _{6 - 10}芳基經一或多個R ^1a取代，且5員至12員雜芳基視情況經一或多個R ^1b取代； 各R ^1a獨立地為鹵素； 各R ^1b係獨立選自由鹵素、C _{1 - 3}烷基及C _{1 - 3}鹵烷基組成之群； R ²係選自由以下組成之群：吡咯啶基、哌啶基、氮雜環庚烷基及-CH ₂C(O)NH ₂，其中吡咯啶基、哌啶基及氮雜環庚烷基視情況經一或多個側氧基部分取代； R ³係選自由H及C _{1 - 6}烷基組成之群； R ⁴係選自由C _{1 - 6}烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^4a取代； 各R ^4a係獨立選自由C _{1 - 6}烷基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^4b取代； 各R ^4b為獨立選擇之鹵素； R ⁵係選自由H及C _{1 - 6}烷基組成之群； 或R ⁴及R ⁵共同形成視情況經一或多個R ^4a取代之單環或雙環5員至12員雜環基； R ⁶係選自由以下組成之群：C _{6 - 10}芳基、5員至12員雜芳基、-OR ⁷、-NHR ⁷、-NHC(O)OR ⁷及-CHR ⁷NHC(O)R ⁷，其中C _{6 - 10}芳基、5員至12員雜芳基視情況經一或多個R ^6a取代； 各R ^6a係獨立選自由C _{1 - 6}烷基、C _{1 - 6}烷氧基、羥基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群； 或R ⁵及R ⁶與其所附接之原子共同形成視情況經-NHR ⁷或-NHC(O)OR ⁷取代之5員至12員雜環基； R ⁷係選自由C _{1 - 6}烷基、C _{3 - 8}環烷基及C _{6 - 10}芳基組成之群，其各自視情況經一或多個R ^7a取代； 各R ^7a係獨立選自由以下組成之群：鹵素、C _{1 - 6}烷基、C _{6 - 10}芳基及C _{3 - 8}環烷基，其中C _{1 - 6}烷基、C _{6 - 10}芳基及C _{3 - 8}環烷基視情況經一或多個R ^7b取代；及 各R ^7b為獨立選擇之鹵素。 2. 如實施例1之化合物或其醫藥學上可接受之鹽，其中R ²係選自由以下組成之群：2-側氧基吡咯啶-3-基、5-側氧基吡咯啶-3-基及2,5-二側氧基吡咯啶-3-基。 3. 如實施例1或實施例2之化合物，其中R ¹係選自由以下組成之群：苯基、六氟異丙基、2,6-二甲基哌啶-4-基、2-甲基嘧啶-5-基及異㗁唑-3-基，其中苯基經1-5個獨立選擇之鹵素取代。 4. 如實施例1至3中任一項之化合物，其具有式Ia之結構：

， 或其醫藥學上可接受之鹽。 5. 如實施例1至4中任一項之化合物，其具有式Ib之結構：

， 或其醫藥學上可接受之鹽，其中下標n為1至5範圍內之整數。 6. 如實施例1至5中任一項之化合物或其醫藥學上可接受之鹽，其中下標n為4或3。 7. 如實施例1至6中任一項之化合物或其醫藥學上可接受之鹽，其中各R ^1a為氟。 8. 如實施例1至7中任一項之化合物或其醫藥學上可接受之鹽，其中R ₃為H。 9. 如實施例1至8中任一項之化合物或其醫藥學上可接受之鹽，其中R ⁴為C _{1 - 6}烷基，其視情況經一或多個R ^4a取代。 10. 如實施例9之化合物或其醫藥學上可接受之鹽，其中R ^4a為C _{3 - 8}環烷基或經鹵素取代之C _{6 - 10}芳基。 11. 如實施例1至10中任一項之化合物或其醫藥學上可接受之鹽，其中R ⁵為H或-CH ₃。 12. 如實施例1至11中任一項之化合物或其醫藥學上可接受之鹽，其中R ⁶係選自由吲哚-2-基、苯并呋喃-2-基及苯甲氧基組成之群，其各自視情況經一或多個R ^6a取代。 13. 如實施例12之化合物或其醫藥學上可接受之鹽，其中R ⁶經C _{1 - 6}烷氧基取代。 14. 如實施例1至9中任一項之化合物或其醫藥學上可接受之鹽，其中R ⁵及R ⁶與其所附接之原子共同形成視情況經-NHR ⁷或-NHC(O)OR ⁷取代之5員至12員雜環基。 15. 如實施例14之化合物或其醫藥學上可接受之鹽，其中R ⁵及R ⁶與其所附接之原子共同形成3-側氧基-3,4-二氫吡𠯤-2-基或2-側氧基-1,2-二氫吡啶-3-基，其各自經-NHR ⁷或-NHC(O)OR ⁷取代。 16. 如實施例1之化合物，其係選自本文中所揭示之任何化合物或其醫藥學上可接受之鹽。 17. 如實施例1之化合物，其係選自本文中所揭示之化合物或其醫藥學上可接受之鹽之任何單一對映異構體或非對映異構體。 18. 一種醫藥組合物，其包含如實施例1至17中任一項之化合物及醫藥學上可接受之賦形劑。 19. 一種用於治療冠狀病毒感染之方法，該方法包含向有需要之個體投與治療有效量之如實施例1至17中任一項之化合物或其醫藥學上可接受之鹽，或治療有效量之如實施例18之醫藥組合物。 20. 如實施例19之方法，其中該冠狀病毒係選自由以下組成之群：SARS-CoV-2、SARS-CoV、MERS-CoV、HCoV-229E、HCoV OC43及HCoV NL63。 21. 如實施例20之方法，其中該冠狀病毒為SARS-CoV-2。 22. 如實施例19至21中任一項之方法，其中該個體患有冠狀病毒疾病2019 (COVID-19)。 23. 如實施例19至22中任一項之方法，其中該化合物係經口、經鼻內或經由注射投與。 24. 如實施例19至23中任一項之方法，其進一步包含向該個體投與選自由以下組成之群的一或多種藥劑：消炎劑、鎮痛劑、抗病毒劑及鎮咳劑。 25. 如實施例19至24中任一項之方法，其進一步包含向該個體投與CYP3A4抑制劑。 26. 如實施例19至25中任一項之方法，其中該個體為人類、農業動物或伴侶動物。 27. 一種抑制冠狀病毒主要蛋白酶之方法，該方法包含使該蛋白酶與有效量之如實施例1至17中任一項之化合物接觸。 28. 如實施例27之方法，其中該冠狀病毒主要蛋白酶為SARS-CoV-2 3CL ^pro。 Exemplary embodiments provided according to the subject matter disclosed in the present invention include (but are not limited to) various technical solutions and the following examples: 1. A compound of formula I:

, or a pharmaceutically acceptable salt thereof, wherein: R ¹ is selected from the group consisting of C _{6 - 10} aryl, 5- to 12-membered heteroaryl and C _{1 - 6} haloalkyl, wherein C _{6 - 10} -aryl is substituted with one or more R ^1a , and 5- to 12-membered heteroaryl is optionally substituted with one or more R ^1b ; each R ^1a is independently halogen; each R ^1b is independently selected from halogen, _The group consisting _{of C1-3} alkyl _{and C1-3} haloalkyl _; R2 is selected from the group consisting of pyrrolidinyl, piperidinyl, azepanyl and ^-CH2C ₍ O) NH ₂ , wherein pyrrolidinyl, piperidinyl and azepanyl are optionally substituted with one or more pendant oxy moieties; R ³ is selected from the group consisting _of H and C _1-6 alkyl ^; R ₄ is selected from the group consisting _of C _1-6 alkyl and C _6-10 aryl, each _of which is optionally substituted by _one or more R ^4a _; each _R ^4a is independently selected from C _1-6 alkyl _, C _{3 -} The group consisting of ₈ cycloalkyl and C _6-10 aryl, each _of which is optionally substituted with _one or more R ^4b ; each R ^4b is independently selected halogen _; R ⁵ is selected from H _and C _1-6 alkyl or R ⁴ and R ⁵ together form a monocyclic or bicyclic 5- to 12-membered heterocyclic group substituted by one or more R ^4a as appropriate; R ⁶ is selected from the group consisting of: C _{6 - 10} Aryl, 5- to 12-membered heteroaryl, -OR ⁷ , -NHR ⁷ , -NHC(O)OR ⁷ and -CHR ⁷ NHC(O)R ⁷ , wherein C _{6 - 10} aryl, 5- to 12-membered Member heteroaryl is optionally substituted with one or more R ^6a _{; each R 6a is independently selected from C 1-6 alkyl, C 1-6 alkoxy} ^, _{hydroxy , C 3-8 cycloalkyl and C 6 -} A group consisting of ₁₀ aryl groups; or R ⁵ and R ⁶ and the atoms to which they are attached together form a 5- to 12-membered heterocyclic group substituted by -NHR ⁷ or -NHC(O)OR ⁷ as appropriate; R ⁷ is selected _The group consisting _{of C1-6} alkyl _{, C3-8} cycloalkyl _{and C6-10} aryl, each of which is optionally substituted with _one or more R ^7a _; each R ^7a is independently selected from the group consisting of : halogen, C _1-6 alkyl _, C _6-10 aryl _and C _{3-8 cycloalkyl ,} wherein _{C 1-6 alkyl ,} C _{6-10 aryl and C 3-8 cycloalkyl} are _optionally One or more R ^7b is substituted; and each R ^7b is independently selected halogen. 2. The compound of embodiment 1 or a pharmaceutically acceptable salt thereof, wherein R ² is selected from the group consisting of: 2-oxypyrrolidin-3-yl, 5-oxypyrrolidin-3 -yl and 2,5-dioxypyrrolidin-3-yl. 3. The compound of embodiment 1 or embodiment 2, wherein R ¹ is selected from the group consisting of: phenyl, hexafluoroisopropyl, 2,6-dimethylpiperidin-4-yl, 2-methyl pyrimidin-5-yl and isoxazol-3-yl wherein the phenyl group is substituted with 1-5 independently selected halogens. 4. The compound of any one of embodiments 1 to 3, which has the structure of formula Ia:

, or a pharmaceutically acceptable salt thereof. 5. The compound of any one of embodiments 1 to 4, which has the structure of formula Ib:

, or a pharmaceutically acceptable salt thereof, wherein the subscript n is an integer in the range of 1 to 5. 6. The compound of any one of embodiments 1 to 5 or a pharmaceutically acceptable salt thereof, wherein subscript n is 4 or 3. 7. The compound of any one of embodiments 1 to 6, or a pharmaceutically acceptable salt thereof, wherein each R ^1a is fluorine. 8. The compound of any one of embodiments 1 to 7 or a pharmaceutically acceptable salt thereof, wherein R ₃ is H. 9. The compound of any one of embodiments 1 to ₈ , or a pharmaceutically acceptable salt thereof, wherein R ⁴ is C _1-6 alkyl _, optionally substituted with one or more R ^4a . _10. The compound of embodiment ₉ or a pharmaceutically acceptable salt thereof _, wherein R ^4a is C _3-8 cycloalkyl or C _6-10 aryl substituted by halogen _. 11. The compound of any one of embodiments 1 to 10 or a pharmaceutically acceptable salt thereof, wherein R ⁵ is H or -CH ₃ . 12. The compound of any one of embodiments 1 to 11 or a pharmaceutically acceptable salt thereof, wherein R ⁶ is selected from the group consisting of indol-2-yl, benzofuran-2-yl and benzyloxy groups, each of which is optionally substituted with one or more R ^6a . 13. The compound of embodiment ₁₂ or a pharmaceutically acceptable salt thereof _, wherein R ⁶ is substituted with C _1-6 alkoxy. 14. The compound of any one of embodiments ¹ to ⁹ , or a pharmaceutically acceptable salt thereof, wherein R and R and the atoms to which they are attached together form optionally ^-NHR or -NHC(O) 5- to 12-membered heterocyclyl substituted by OR ⁷ . 15. The compound of embodiment 14 or a pharmaceutically acceptable salt thereof, wherein R ⁵ and R ⁶ and the atoms to which they are attached together form 3-side oxy-3,4-dihydropyridine-2-yl or 2-pendant oxy-1,2-dihydropyridin-3-yl, each of which is substituted with -NHR ⁷ or -NHC(O)OR ⁷ . 16. The compound of embodiment 1, which is selected from any compound disclosed herein or a pharmaceutically acceptable salt thereof. 17. The compound of Embodiment 1 selected from any single enantiomer or diastereomer of a compound disclosed herein or a pharmaceutically acceptable salt thereof. 18. A pharmaceutical composition comprising the compound of any one of embodiments 1 to 17 and a pharmaceutically acceptable excipient. 19. A method for treating coronavirus infection, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof as any one of embodiments 1 to 17, or a treatment An effective amount of the pharmaceutical composition as in Example 18. 20. The method of embodiment 19, wherein the coronavirus is selected from the group consisting of SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-229E, HCoV OC43 and HCoV NL63. 21. The method of embodiment 20, wherein the coronavirus is SARS-CoV-2. 22. The method of any one of embodiments 19 to 21, wherein the individual suffers from coronavirus disease 2019 (COVID-19). 23. The method of any one of embodiments 19 to 22, wherein the compound is administered orally, intranasally, or via injection. 24. The method of any one of embodiments 19 to 23, further comprising administering to the individual one or more agents selected from the group consisting of anti-inflammatory agents, analgesics, antiviral agents, and antitussives. 25. The method of any one of embodiments 19 to 24, further comprising administering to the individual a CYP3A4 inhibitor. 26. The method of any one of embodiments 19 to 25, wherein the individual is a human, an agricultural animal, or a companion animal. 27. A method of inhibiting the major protease of a coronavirus, the method comprising contacting the protease with an effective amount of the compound of any one of embodiments 1 to 17. 28. The method of embodiment 27, wherein the major protease of the coronavirus is SARS-CoV-2 3CL ^pro .

Although the foregoing has been described in detail with the aid of illustrations and examples for purposes of clarity and understanding, those skilled in the art will appreciate that certain changes and modifications can be practiced within the scope of the appended claims. Additionally, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference was individually incorporated by reference.

Claims (28)

A compound of formula I:

, or a pharmaceutically acceptable salt thereof, wherein: R ¹ is selected from the group consisting of: substituted phenyl, unsubstituted phenyl, other C _{6 - 10} aryl, 5-membered to 12-membered heterocyclic Aryl _and C _1-6 haloalkyl groups, wherein: phenyl is substituted with 4, _{3 , 2} or 1 R ^1a , C _6-10 aryl is substituted with one or more R ^1a , and 5- to 12-membered heterocycles Aryl is optionally substituted with one or more R ^1b ; each R ^1a is independently halogen; each R ^1b is independently selected from the group consisting _of halogen, C _1-3 alkyl _and C _1-3 haloalkyl ^; _{R 2} is selected from the group consisting of pendant oxypyrrolidinyl, dioxypyrrolidinyl, pyrrolidinyl, piperidinyl, azepanyl, and _-CH2C (O) _NH2 , wherein piper Aridyl and azepanyl are optionally substituted with one or more pendant oxy moieties; R ³ is selected from the group consisting _of H and C _1-6 alkyl ^{; R 4} _is selected from the group consisting of: _One or more R ^4a substituted propyl, unsubstituted _propyl , C _1-2 alkyl _, C _4-6 alkyl _and C _{6-10 aryl ,} wherein C _1-2 alkyl _{, C 4 - 6} alkyl _and C _6-10 aryl are optionally substituted by _one or more R ^4a _; each _R ^4a is independently selected _from C _1-6 alkyl _, C _{3-8 cycloalkyl and} C _6-10 aryl group, each of which is optionally substituted with one or more R ^4b ; each R ^4b is independently selected halogen; R ⁵ is selected from the group consisting _of H and C _1-6 alkyl ^{; or R 4 and R 5} _form ^together A monocyclic or bicyclic 5- to 12-membered heterocyclic group optionally substituted with one or more R ^4a ; R ⁶ is selected from the group consisting of: indolyl substituted with R ^6a , other 5- to 12-membered heterocyclic groups Aryl, C _{6 - 10} aryl, -OR ⁷ , -NHR ⁷ , -NHC(O)OR ⁷ and -CHR ⁷ NHC(O)R ⁷ , wherein C _{6 - 10} aryl and 5- to 12-membered hetero group Aryl is optionally substituted with one or more R ^6a _; each _R ^6a is independently selected from C _1-6 alkoxy _, C _1-6 alkyl, hydroxy _, C _{3-8 cycloalkyl and C 6-10 aryl} or R ⁵ and R ⁶ and the atoms to which they are attached together form a 5- to 12-membered heterocyclic group substituted with -NHR ⁷ or -NHC(O)OR ⁷ as appropriate; R ⁷ is selected from C _The group consisting _{of 1-6} alkyl _{, C3-8} cycloalkyl _{and C6-10} aryl, each of which is optionally substituted with _one or more R ^7a _; each R ^7a is independently selected from the group consisting of: halogen _, C _1-6 alkyl group _, C _6-10 aryl group _and C _3-8 cycloalkyl group _, wherein C _{1-6 alkyl} group _{, C 6-10 aryl group and C 3-8 cycloalkyl group are} optionally _modified by _one or more multiple R ^7b substitutions; and Each R ^7b is an independently selected halogen. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ² is selected from the group consisting of: 2-oxypyrrolidin-3-yl, 5-oxypyrrolidin-3-yl and 2,5-dioxypyrrolidin-3-yl. The compound of claim 1, wherein R ¹ is selected from the group consisting of phenyl, hexafluoroisopropyl, 2,6-dimethylpiperidin-4-yl, 2-methylpyrimidin-5-yl and isoxazol-3-yl, wherein the phenyl group is substituted with 1-5 independently selected halogens. As the compound of claim 1, it has the structure of formula Ia:

, or a pharmaceutically acceptable salt thereof. Such as the compound of claim 1, it has the structure of formula Ib:

, or a pharmaceutically acceptable salt thereof, wherein the subscript n is an integer in the range of 1 to 5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein subscript n is 4 or 3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein each R ^1a is fluorine. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ³ is H. The compound of claim _{1 or a pharmaceutically acceptable salt thereof, wherein R 4 is C 1-6 alkyl} ^optionally _substituted with one or more R ^4a . The compound of claim ₉ or a pharmaceutically acceptable salt thereof, wherein R ^4a is C _{3 -8} cycloalkyl or C _{6 - 10} aryl substituted by halogen. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ⁵ is H or -CH ₃ . The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ⁶ is selected from the group consisting of indol-2-yl, benzofuran-2-yl and benzyloxy, each of which is optionally One or more R ^6a substitutions. The compound of claim ₁₂ or a pharmaceutically acceptable salt thereof _, wherein R ⁶ is substituted with C _1-6 alkoxy. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ⁵ and R ⁶ together with the atoms to which they are attached form 5 to 12 members substituted by -NHR ⁷ or -NHC(O)OR ⁷ as appropriate Member heterocyclyl. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein R ⁵ and R ⁶ together with the atoms to which they are attached form 3-side oxy-3,4-dihydropyridine-2-yl or 2 -Pendant oxy-1,2-dihydropyridin-3-yl, each of which is substituted with ^-NHR7 or -NHC(O) ^OR7 . The compound of claim 1, which is selected from the group consisting of:

, and their pharmaceutically acceptable salts. The compound of claim 1, which is selected from the group consisting of:

, and their pharmaceutically acceptable salts. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable excipient. A method for treating a coronavirus infection, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound of claim 1 or a pharmaceutically acceptable salt thereof. The method of claim 19, wherein the coronavirus is selected from the group consisting of SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-229E, HCoV OC43 and HCoV NL63. The method of claim 20, wherein the coronavirus is SARS-CoV-2. The method of claim 19, wherein the individual has coronavirus disease 2019 (COVID-19). The method of claim 19, wherein the compound is administered orally, intranasally or via injection. The method of claim 19, further comprising administering to the individual one or more agents selected from the group consisting of anti-inflammatory agents, analgesics, antiviral agents, and antitussives. The method of claim 19, further comprising administering to the individual a CYP3A4 inhibitor. The method of claim 19, wherein the individual is a human, agricultural animal or companion animal. A method of inhibiting the major protease of coronavirus, the method comprising contacting the protease with an effective amount of the compound of claim 1. The method of claim 27, wherein the major protease of the coronavirus is SARS-CoV-2 3CL ^pro .