Functionalized peptides as antiviral agents

Abstract

The present invention discloses a compound of formula (I) and a pharmaceutically acceptable salt thereof, which inhibits coronavirus replication activity. The present invention further relates to a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof, and a method for treating or preventing coronavirus infection in a subject in need thereof, comprising administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof to the subject. JPEG2023535701000100.jpg34166

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 054,048, filed July 20, 2020, the entire teachings of which are incorporated herein by reference.

[0002] The present invention relates to compounds and methods for inhibiting coronavirus replication activity by contacting 3C-like protease (sometimes referred to as "3CLpro," "main protease," or "Mpro") with a therapeutically effective amount of a 3C-like protease inhibitor. The present invention further relates to pharmaceutical compositions containing coronavirus 3C-like protease inhibitors in mammals by administering an effective amount of such coronavirus 3C-like protease inhibitors. [Background technology]

[0003] Coronaviruses are a family of enveloped, single-stranded, positive-stranded RNA viruses classified in the order Nidovirales. The Coronaviridae family has been known for over 60 years and includes pathogens of many animal species, including humans, horses, cattle, pigs, birds, cats, and monkeys. For example, the isolation of the prototype murine coronavirus strain JHM was reported in 1949. Coronaviruses are common viruses that generally cause mild to moderate upper respiratory tract illness in humans and are named for the crown-like spikes on their envelope surface. There are four major subgroups known as alpha-, beta-, gamma-, and delta-coronaviruses, with the first coronaviruses identified in the mid-1960s. Coronaviruses known to infect humans include alphacoronavirus 229E, NL63; betacoronavirus OC43, HKU1; SARS-CoV (the coronavirus that causes severe acute respiratory syndrome or SARS); and MERS-CoV (the coronavirus that causes Middle East respiratory syndrome or MERS). Humans are commonly infected with human coronaviruses 229E, NL63, 0C43, and HKU1, and symptoms typically include a brief, mild-to-moderate upper respiratory illness, such as runny nose, cough, sore throat, and fever. Occasionally, human coronaviruses cause lower respiratory illness, such as pneumonia, which is more common in people with cardiopulmonary disease, compromised immune systems, or the elderly. Transmission of common human coronaviruses is not fully understood. However, human coronaviruses likely spread from infected individuals to others through the air via coughing or sneezing, and through close personal contact, such as touching or shaking hands. These viruses can also be spread by touching a contaminated object or surface and then touching the mouth, nose, or eyes.

[0004] Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. The genomic RNA of CoVs has a 5' cap structure and a 3' poly(A) tail and contains at least six open reading frames (ORFs). The first ORF (ORF 1a / b) directly translates two polyproteins: pp1a and pp1ab. These polyproteins are processed into 16 nonstructural proteins by a 3C-like protease (3CLpro), also known as the main protease (Mpro). These nonstructural proteins are responsible for the production of subgenomic RNAs that encode four structural proteins, namely, the envelope, membrane, spike, and nucleocapsid proteins, among other accessory proteins. Consequently, it is understood that 3C-like proteases play a critical role in the coronavirus life cycle.

[0005] 3CLpro is a cysteine ​​protease responsible for most cleavage events within the precursor polyprotein. Active 3CLpro is a homodimer containing two protomers and characterized by a Cys-His dimer located between domains I and II. 3CLpro is conserved among coronaviruses, and several common features are shared among 3CLpro substrates in different coronaviruses. The lack of a human homolog of 3CLpro makes it an ideal antiviral target. Although compounds have been reported to inhibit 3CLpro activity, they have not been approved for coronavirus therapy. (See International Publication Nos. WO2018042343, WO2018023054, WO2005113580, and WO2006061714.)

[0006] Due to this high unmet clinical need, more effective treatments for coronavirus infections are needed. The present invention describes methods for preparing and using compounds believed to inhibit the coronavirus life cycle. Compounds of this type can be used to treat coronavirus infections and reduce the incidence of disease complications, such as organ failure or death.

[0007] There is a need in the art for novel therapeutic agents that treat, alleviate, or prevent coronavirus infections, which, when administered to coronavirus-infected patients as monotherapy or in combination with other coronavirus or adjunctive therapies, result in significantly improved prognosis, reduced disease progression, and increased seroconversion rates. Summary of the Invention

[0008] The present invention relates to novel antiviral compounds, pharmaceutical compositions containing such compounds, and methods for treating or preventing viral (particularly coronavirus) infections in subjects in need of such treatment with these compounds. The compounds of the present invention inhibit proteins encoded by coronaviruses or interfere with the coronavirus life cycle and are also useful as antiviral agents. Additionally, the present invention provides methods for preparing these compounds.

[0009] The present invention provides compounds represented by formula (I), and pharmaceutically acceptable salts, N-oxides, esters and prodrugs thereof: [ka] During the ceremony, A is, 1) optionally substituted -C1-C8 alkyl; 2) optionally substituted -C3-C 12 cycloalkyl, 3) optionally substituted 3- to 12-membered heterocycloalkyl; 4) optionally substituted aryl, and 5) optionally substituted heteroaryl; L1 is -C(R 11 R 12 )- and L2 is -C(R 11 R 12 )- and n1 is 0, 1, 2, 3 or 4; X is optionally substituted -C1-C6 alkyl, -CN, -C(O)R 15 , C(O)NR 13 R 14 , or C(O)C(O)NR 13 R 14 and Each Q is -C(R 11 'R 12 ')- and n2 is 0, 1, 2, 3 or 4, preferably n2 is not 0; Each R 11 , R 11 ', R 12 and R 12 ' are independently selected from the following: 1) hydrogen, 2) halogens, 3)-OR 16 , 4)-SR 16 , 5)-NR 13 R 14 , 6)-OC(O)NR 13 R 14 , 7) optionally substituted -C1-C6 alkyl; 8) optionally substituted -C3-C8 cycloalkyl; 9) optionally substituted 3- to 8-membered heterocycloalkyl; 10) optionally substituted aryl, and 11) optionally substituted heteroaryl; Alternatively, R 11 and R 12 together with the carbon atoms to which they are attached form an optionally substituted 3- to 8-membered carbocyclic or heterocyclic ring; Alternatively, if n1 is not 0, two adjacent R 11 groups, taken together with the carbon atoms to which they are attached, form an optionally substituted 3- to 8-membered carbocyclic or heterocyclic ring; Alternatively, n1 is 2, 3, or 4, and R 11 The groups, together with the carbon atoms to which they are attached, form an optionally substituted bridging moiety, in this embodiment: [ka] is preferably an optionally substituted 6- to 12-membered bridged heterocyclic ring system, R 13 and R 14 are each independently selected from: 1) hydrogen, 2) optionally substituted -C1-C6 alkyl; 3) optionally substituted -C3-C8 cycloalkyl; 4) optionally substituted 3- to 8-membered heterocycloalkyl; 5) optionally substituted aryl; 6) optionally substituted arylalkyl; 7) optionally substituted heteroaryl; 8) optionally substituted heteroarylalkyl; 9)-C(O)R 15 , 10)-S(O)2R 16 , and 11)-NH2, Alternatively, R 13 and R 14 taken together with the nitrogen atom to which they are attached form an optionally substituted 3- to 8-membered heterocycle.

[0010] R 15 is selected from the following: 1) hydrogen, 2) halogens, 3)-OH, 4) optionally substituted -C1-C6 alkyl; 5) optionally substituted -C1-C6 alkoxy; 6) optionally substituted -C3-C8 cycloalkyl; 7) optionally substituted 3- to 8-membered heterocycloalkyl; 8) optionally substituted aryl; 9) optionally substituted arylalkyl; 10) optionally substituted heteroaryl, and 11) optionally substituted heteroarylalkyl; R 16 is selected from the following: 1) hydrogen, 2)-OH, 3) optionally substituted -C1-C6 alkyl; 4) optionally substituted -C3-C8 cycloalkyl; 5) optionally substituted 3- to 8-membered heterocycloalkyl; 6) optionally substituted aryl; 7) optionally substituted arylalkyl; 8) optionally substituted heteroaryl, and 9) optionally substituted heteroarylalkyl. DETAILED DESCRIPTION OF THE INVENTION

[0011] One embodiment of the present invention is a compound of formula (I) above or a pharmaceutically acceptable salt thereof.

[0012] In certain embodiments of compounds of Formula (I), R 13 and R 14 are each independently hydrogen, optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C(O)R 15 , -S(O)2R 16 and NH2, or R 13 and R 14together with the nitrogen atom to which they are attached form an optionally substituted 3- to 8-membered heterocycle, and R 15 is selected from hydrogen, halogen, —OH, optionally substituted —C1-C6 alkyl, optionally substituted —C1-C6 alkoxy, optionally substituted —C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R 16 is selected from hydrogen, —OH, optionally substituted —C1-C6 alkyl, optionally substituted —C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0013] In certain embodiments of compounds of Formula (I), X is —CN.

[0014] In certain embodiments of compounds of Formula (I), X is —C(O)CHOC(O)R 21 , -C(O)CH2C(O)2R 21 , -C(O)CH2OR 21 or C(O)CH2R 22 and R 21 is hydrogen, optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 22 is halogen or NR 13 R 14 is.

[0015] In certain embodiments of compounds of Formula (I), X is —C(O)C(O)NHR 21 and R 21 is as defined above. Preferably, R 21 is optionally substituted benzyl, optionally substituted methyl, optionally substituted isopropyl, optionally substituted t-butyl, or optionally substituted cyclohexyl.

[0016] In certain embodiments of compounds of Formula (I), X is —C(O)R 21 and R 21 is defined earlier.

[0017] In certain embodiments of compounds of Formula (I), X is —CHR 21 OC(O)R 21 , -CHR 21 C(O)2R 21 , -CHR 21 (OR 21 ), or CH(OR 21 )2 and R 21 is defined earlier.

[0018] In certain embodiments, X is —CN, —C(O)H, [ka] In a preferred embodiment, X is —CN.

[0019] In certain embodiments of compounds of Formula (I), A is derived by removal of a hydrogen atom from one of the following, and is optionally substituted: [ka]

[0020] In certain embodiments of compounds of Formula (I), A is selected from the following groups, wherein A is optionally substituted: [ka]

[0021] Preferably, A has 0, 1 or 2 substituents. Preferably, the substituents are independently selected from fluoro, chloro, hydroxy, methoxy, fluoromethoxy, difluoromethoxy and trifluoromethoxy.

[0022] In certain embodiments of compounds of Formula (I), A is —CHR 23 and R 23 Ha-NR 13 R 14 , optionally substituted -C3-C 12 It is cycloalkyl, optionally substituted 3- to 12-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0023] In certain embodiments of compounds of Formula (I), A is -CR 23 R 24 R 25 where R 24 is hydrogen, halogen, optionally substituted -C1-C6 alkyl, optionally substituted -C1-C6 alkoxy, optionally substituted -C3-C 12 cycloalkyl, optionally substituted 3- to 12-membered heterocycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl; R 25 is hydrogen or halogen, and R 23 is as defined above.

[0024] In certain embodiments of compounds of Formula (I), A is —C(NR 13 R 14 )R 24 R 25 and R 13 , R 14 , R 24 and R 25 is as defined above.

[0025] In certain embodiments of compounds of Formula (I), A is [ka] where R 14 , R 15 , R 24 and R 25 is as defined above. In certain embodiments, R14 and R 25 is hydrogen and R 24 is C1-C6-alkyl, preferably t-butyl. 15 is preferably benzyl, C1-C6-alkyl or C3-C8-cycloalkyl.

[0026] In certain embodiments of compounds of Formula (I), at least one Q is -CH2-. In certain embodiments of compounds of Formula (I), all Q are -CH2-.

[0027] In certain embodiments, the compound of formula (I) is represented by one of formulas (II-1)-(II-2), or a pharmaceutically acceptable salt thereof: [ka] In the formula, A, L1, L2, n, R 11 ' and X are as defined above.

[0028] In certain embodiments, the compound of formula (I) is represented by one of formulas (II-1a) to (II-2a), or a pharmaceutically acceptable salt thereof: [ka] wherein A, L1, L2, n, and X are as defined above.

[0029] In certain embodiments, the compound of formula (I) is represented by one of formulas (III-1)-(III-2), or a pharmaceutically acceptable salt thereof: [ka] where A, Q, n2 and X are as defined above, and R 17 is halogen, -OR 16 , -SR 16 , -NR 13 R 14 , -OC(O)NR13 R 14 , optionally substituted —C1-C6 alkyl, optionally substituted —C3-C8 cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, wherein m1 is 0, 1, 2, or 3, and m2 is 0, 1, 2, 3, or 4. Preferably, m1 is 0 and m2 is 0.

[0030] In certain embodiments, the compound of formula (I) is represented by one of formulas (IV-1) to (IV-4), or a pharmaceutically acceptable salt thereof: [ka] In the formula, A, R 17 , R 11 ', m1, m2 and X are as defined above.

[0031] In certain embodiments, the compound of formula (I) is represented by one of formulas (IV-1a) to (IV-4a), or a pharmaceutically acceptable salt thereof: [ka] wherein A and X are as defined above.

[0032] In certain embodiments, the compound of formula (I) is represented by one of formulas (V-1) to (V-4), or a pharmaceutically acceptable salt thereof: [ka] where each T is CR 11 and each U is -C(R 11 R 12 )-, and each V is -O-, -S-, -C(R 11 R 12 )- or N(R 13 R 14)-, m is 0, 1 or 2, m' is 0, 1, 2 or 3, and A, Q, n2, R 11 , R 12 , R 13 , R 14 and X is as defined above.

[0033] In certain embodiments, the compound of formula (I) is represented by one of formulas (VI-1) to (VI-6), or a pharmaceutically acceptable salt thereof: [ka] In the formula, A, R 11 ', V, m and X are as defined above.

[0034] In certain embodiments, the compound of formula (I) is represented by one of formulas (VI-1a) to (VI-8a), or a pharmaceutically acceptable salt thereof: [ka] wherein A and X are as defined above.

[0035] In certain embodiments, the compound of formula (I) is represented by one of formulas (VII-1) to (VII-6), or a pharmaceutically acceptable salt thereof: [ka] In the formula, A, R 11 ', V, m and X are as defined above.

[0036] In certain embodiments, the compound of formula (I) is represented by one of formulas (VII-1a) to (VII-6a), or a pharmaceutically acceptable salt thereof: [ka] wherein A and X are as defined above.

[0037] In certain embodiments, the present invention relates to compounds of formula (VI-1) to (VI-4) or formula (VII-1) to (VII-4), and pharmaceutically acceptable salts thereof: [ka] is selected from the following groups: [ka] Each of these groups is optionally substituted.

[0038] In certain embodiments, the present invention relates to compounds of formula (VI-5) to (VI-6) or formula (VII-5) to (VII-6), and pharmaceutically acceptable salts thereof: [ka] is selected from the following groups: [ka] Each of these groups is optionally substituted.

[0039] In certain embodiments, the present invention relates to compounds of Formula (VI-1) to (VI-6) or Formula (VII-1) to (VII-6), and pharmaceutically acceptable salts thereof, wherein A is selected from the following groups: [ka] Each of these groups is optionally substituted. Preferably, A has 0, 1, or 2 substituents. Preferably, the substituents are independently selected from fluoro, chloro, hydroxy, methoxy, fluoromethoxy, difluoromethoxy, and trifluoromethoxy.

[0040] In a particular embodiment, the present invention relates to a compound of one of formulas (VI-1) to (VI-6) and formula (VII-1) to (VII-6), or a pharmaceutically acceptable salt thereof, wherein A is —CHR 23 and R 23 is defined earlier.

[0041] In a particular embodiment, the present invention relates to a compound of one of formulas (VI-1) to (VI-6) and formulas (VII-1) to (VII-6), or a pharmaceutically acceptable salt thereof, wherein X is —CN, —C(O)CHOC(O)R 21 , -C(O)CH2C(O)2R 21 , -C(O)CH2OR 21 , -C(O)CH2R 22 , -C(O)C(O)NHR 21 , -C(O)R 21 , -CHR 21 OC(O)R 21 , -CHR 21 C(O)2R 21 , -CHR 21 (OR 21 ), or CH(OR 21 )2 and R 21 and R 22 is as defined above,

[0042] In certain embodiments, the compound of formula (I) is represented by one of formulas (VIII-1) to (VIII-12), or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 31 is hydrogen, -F, -Cl, -OCH3, or OCHF2, and X is as defined above. Preferably, X is -CN, -C(O)CH2OC(O)R 21 , -C(O)CH2C(O)2R 21 , -C(O)CH2OR 21 , -C(O)CH2R 22 , -C(O)C(O)NHR 21 , -C(O)R 21 , -CHR 21 OC(O)R21 , -CHR 21 C(O)2R 21 , -CHR 21 (OR 21 ), or CH(OR 21 )2 and R 21 and R 22 is as defined above.

[0043] In certain embodiments, the compound of formula (I) is represented by one of formulas (IX-1) to (IX-8), or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 32 is hydrogen, -F, Cl, -CH3, -CF3 or OR, R 33 -Cl, -Br, -OR 21 , -NHR 21 or OC(O)R 21 and R 34 is R 21 and preferably R 34 is optionally substituted -C1-C6 alkyl, optionally substituted -C3-C8 cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl, more preferably R 34 is benzyl, cyclohexyl, isopropyl, t-butyl or optionally substituted methyl, R 21 and A is as defined above.

[0044] In certain embodiments, the compound of formula (I) is represented by one of formulas (IX-1) to (IX-8), or a pharmaceutically acceptable salt thereof, wherein R 32 , R 33 and R 34 is as defined above, and A is selected from the group [ka] Each of these is optionally substituted. Preferably, A has 0, 1 or 2 substituents. Preferably, the substituents are independently selected from fluoro, chloro, hydroxy, methoxy, fluoromethoxy, difluoromethoxy and trifluoromethoxy.

[0045] In certain embodiments, the compound of formula (I) is represented by one of formulas (IX-1) to (IX-8), wherein A is —CHR 23 and R 23 , R 32 , R 33 and R 34 is as defined above.

[0046] In certain embodiments, the compound of formula (I) is represented by formula (Xa), or a pharmaceutically acceptable salt thereof, wherein A and X are as defined above. [ka]

[0047] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xa) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 1. [Table 1-1] [Table 1-2]

[0048] In certain embodiments, the compound of formula (I) is represented by formula (Xb), or a pharmaceutically acceptable salt thereof, wherein A and X are as defined above. [ka]

[0049] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xb) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 2. [Table 2-1] [Table 2-2]

[0050] In certain embodiments, the compound of formula (I) is represented by formula (Xc), or a pharmaceutically acceptable salt thereof, wherein A and X are defined above. [ka]

[0051] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xc) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 3. [Table 3-1] [Table 3-2]

[0052] In certain embodiments, the compound of formula (I) is represented by formula (Xd), or a pharmaceutically acceptable salt thereof, wherein A and X are defined above. [ka]

[0053] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xd) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 4. [Table 4-1] [Table 4-2] [Table 4-3]

[0054] In certain embodiments, the compound of formula (I) is represented by formula (Xe), or a pharmaceutically acceptable salt thereof, wherein A and X are defined above. [ka]

[0055] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xe) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 5. [Table 5-1] [Table 5-2]

[0056] In certain embodiments, the compound of formula (I) is represented by formula (Xf), or a pharmaceutically acceptable salt thereof, wherein A and X are defined above. [ka]

[0057] Representative compounds of the present invention include, but are not limited to, compounds of formula (Xf) and pharmaceutically acceptable salts thereof, where A and X are depicted for each compound in Table 6. [Table 6-1] [Table 6-2]

[0058] In certain embodiments, the compound of formula (I) is represented by one of formulas (XI-1) to (XI-8), or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 13 , R 14 , R 24 , R 25 and X is as defined above.

[0059] In certain embodiments, the compound of formula (I) is represented by one of formulas (XII-1) to (XII-4), or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 14 , R 15 , R 24 , R 25 and X is as defined above.

[0060] In certain embodiments, the compound of formula (I) is represented by one of formulas (XIII-1) to (XIII-4), or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 15 , R 24 and X is as defined above. Preferably, X is —CN and R 24 is optionally substituted -C1-C6 alkyl, for example t-butyl.

[0061] It will be understood that the description of the invention herein is to be construed in accordance with the rules and principles of chemical bonding. In some cases, it may be necessary to remove a hydrogen atom to accommodate a substituent at any given position.

[0062] It will be further understood that compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic, diastereomeric, and optically active forms. It will also be understood that certain compounds of the present invention may exist in different tautomeric forms. All tautomers are considered to be within the scope of the present invention.

[0063] In certain embodiments, the present invention provides a method for treating or preventing a coronavirus infection in a subject, e.g., a human, in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. The coronavirus can be an alpha, beta, gamma, or delta coronavirus. In certain embodiments, the coronavirus is one that infects humans, such as coronavirus 229E, coronavirus NL63, coronavirus OC43, coronavirus HKU1, SARS-CoV-1, SARS-CoV-2, and MERS-CoV. In certain embodiments, the coronavirus is SARS-CoV-1, SARS-CoV-2, or MERS-CoV. Preferably, the coronavirus is SARS-CoV-2.

[0064] Embodiments of the invention provide for administering the compounds to a healthy patient or a patient infected with the virus as a single agent or in combination with another agent that (1) is effective in treating or preventing coronavirus infection, (2) improves immune response and robustness, or (3) reduces inflammation and / or pain.

[0065] The compounds described herein, or salts, solvates or hydrates thereof, are believed to have activity in preventing, arresting or reducing the effects of coronaviruses by inhibiting the viral 3C or 3C-like protease, thereby interfering with or preventing polyprotein processing of the translated viral genome in the host cell, rendering the virus replication-incompetent.

[0066] In a further aspect, the present invention provides a method for treating a respiratory disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. Such respiratory disorders include, but are not limited to, acute airway disease or chronic airway disease. Examples of such respiratory disorders include acute asthma, pulmonary disease secondary to environmental exposure, acute pulmonary infection, and chronic pulmonary infection.

[0067] The compounds of the present invention and any other pharmaceutically active agents may be administered together or separately, and if administered separately, administration may be simultaneous or sequential in any order. The amounts of the compounds of the present invention and other pharmaceutically active agents and the relative timing of administration are selected to achieve the desired combined therapeutic effect. The administration of the compounds of the present invention and their salts, solvates, or other pharmaceutically acceptable derivatives in combination with other therapeutic agents may be combined by simultaneous administration in (1) a single pharmaceutical composition containing both compounds, or (2) separate pharmaceutical compositions, each containing one of the compounds.

[0068] In another embodiment of the combination therapy, the administration of a compound of the invention allows for the administration of at least one additional therapeutic agent at a lower dose or frequency than would be required to achieve a similar result in the prophylactic treatment of a coronavirus infection in an individual in need thereof, than would be required if the additional therapeutic agent were administered alone.

[0069] It is to be understood that the compounds encompassed by the present invention are compounds that are suitably stable for pharmaceutical use.

[0070] definition Listed below are definitions of various terms used to describe this invention. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0071] As used herein, the term "aryl" refers to a monocyclic or polycyclic carbocyclic ring system containing at least one aromatic ring, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system containing at least one aromatic ring. A polycyclic aryl can contain fused rings, covalently bonded rings, or a combination thereof.

[0072] The term "heteroaryl," as used herein, refers to a monocyclic or polycyclic aromatic radical having one or more ring atoms selected from S, O, and N, with the remaining ring atoms being carbon, and any N or S contained within the ring being optionally oxidized. Heteroaryls include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimizolyl, benzoxazolyl, and quinoxalinyl. Polycyclic heteroaryls can contain fused rings, covalently bonded rings, or combinations thereof.

[0073] According to the present invention, aromatic groups can be substituted or unsubstituted.

[0074] The term "alkyl" as used herein refers to a saturated straight or branched chain hydrocarbon radical. 12 "Alkyl," "C2-C4 alkyl," or "C3-C6 alkyl" refer to alkyl groups containing 1 to 4, 1 to 6, 1 to 8, 1 to 12, 2 to 4, and 3 to 6 carbon atoms, respectively. Examples of C1-C8 alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, and octyl radicals.

[0075] The term "alkenyl" as used herein refers to a straight or branched chain hydrocarbon radical having at least one carbon-carbon double bond formed by the removal of a single hydrogen atom. 12 "Alkenyl," "C2-C4 alkenyl," "C3-C4 alkenyl," or "C3-C6 alkenyl" refers to an alkenyl group containing 2 to 8, 2 to 12, 2 to 4, 3 to 4, or 3 to 6 carbon atoms, respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 2-methyl-2-buten-2-yl, heptenyl, octenyl, and the like.

[0076] The term "alkynyl" as used herein refers to a straight or branched chain hydrocarbon radical having at least one carbon-carbon double bond formed by the removal of a single hydrogen atom. 12 "Alkynyl," "C2-C4 alkynyl," "C3-C4 alkynyl," or "C3-C6 alkynyl" refers to an alkynyl group containing 2 to 8, 2 to 12, 2 to 4, 3 to 4, or 3 to 6 carbon atoms, respectively. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 2-propynyl, 2-butynyl, heptynyl, octynyl, and the like.

[0077] The term "cycloalkyl" as used herein refers to a monocyclic or polycyclic saturated carbocyclic ring, or a fused, bridged, or spiro-system bicyclic or tricyclic group, in which the carbon atoms are optionally oxo-substituted or optionally substituted with an exocyclic olefinic double bond. Preferred cycloalkyl groups include C3-C 12 Cycloalkyl, C3-C6 cycloalkyl, C3-C8 cycloalkyl and C4-C7 cycloalkyl. 12Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, and the like.

[0078] The term "cycloalkenyl," as used herein, refers to a monocyclic or polycyclic carbocyclic ring, or a fused, bridged, or spiro-based bicyclic or tricyclic group having at least one carbon-carbon double bond, wherein the carbon atoms are optionally oxo-substituted or optionally substituted with an exocyclic olefinic double bond. Preferred cycloalkenyl groups include C3-C 12 Cycloalkenyl, C3-C8 cycloalkenyl or C5-C7 cycloalkenyl groups are included. 12 Examples of cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-2-enyl, bicyclo[4.2.1]non-3-en-12-yl, and the like.

[0079] As used herein, the term "arylalkyl" refers to a functional group in which an alkylene chain is attached to an aryl group, e.g., -CHCH-phenyl. The term "substituted arylalkyl" refers to an arylalkyl functional group in which the aryl group is substituted. Similarly, the term "heteroarylalkyl" refers to a functional group in which an alkylene chain is attached to a heteroaryl group. The term "substituted heteroarylalkyl" refers to a heteroarylalkyl functional group in which the heteroaryl group is substituted.

[0080] As used herein, unless otherwise specified, the term "alkoxy," used alone or in combination with other terms, refers to an alkyl group having the specified number of carbon atoms attached to the remainder of the molecule through an oxygen atom, such as, for example, methoxy, ethoxy, 2-propoxy, 2-propoxy (isopropoxy), and higher homologs and isomers. Preferred alkoxy is (C-C)alkoxy.

[0081] It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, and cycloalkenyl moiety described herein can also be an aliphatic or alicyclic group.

[0082] An "aliphatic" group is a non-aromatic moiety containing any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen, or other atoms, and optionally containing one or more units of unsaturation, e.g., double and / or triple bonds. Examples of aliphatic groups include alkyl, alkenyl, alkynyl, O, OH, NH, NH, C(O), S(O), C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH, S(O)NH, S(O)NH, NHC(O)NH, NHC(O)C(O)NH, NHS(O)NH, NHS(O)NH, C(O)NHS(O), C(O)NHS(O)NH, or C(O)NHS(O)NH, groups containing one or more functional groups, non-aromatic hydrocarbons (optionally substituted), and groups in which one or more carbon atoms of a non-aromatic hydrocarbon (optionally substituted) have been replaced by a functional group. The carbon atoms of an aliphatic group are optionally substituted with oxo. The aliphatic group may be linear, branched, or cyclic, or a combination thereof, and preferably contains from about 1 to about 24 carbon atoms, more typically from about 1 to about 12 carbon atoms. In addition to aliphatic hydrocarbon groups as used herein, aliphatic groups expressly include, for example, alkoxyalkyl, polyalkoxyalkyl, e.g., polyalkylene glycols, polyamines, and polyimines. The aliphatic group is optionally substituted.

[0083] The terms "heterocycle" or "heterocycloalkyl" can be used interchangeably and refer to non-aromatic rings or bicyclic or tricyclic groups of fused, bridged, or spiro systems, where (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur, and nitrogen, (ii) each ring system can be saturated or unsaturated, (iii) the nitrogen and sulfur heteroatoms can be optionally oxidized, (iv) the nitrogen heteroatom can be optionally quaternized, (v) any of the above rings can be fused to an aromatic ring, and (vi) the remaining ring atoms are carbon atoms that are optionally oxo-substituted or optionally substituted with an exocyclic olefinic double bond. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 2-oxa-7-azaspiro[4.4]nonanyl, 7-oxoxoxepan-4-yl, and tetrahydrofuryl. Such heterocyclic groups may be further substituted. Heteroaryl or heterocyclic groups may be C-linked or N-linked (where possible).

[0084] It is understood that any alkyl, alkenyl, alkynyl, alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphatic moiety, etc. described herein, when used as a link to connect two or more groups or substituents, may be a divalent or polyvalent group, which may be on the same atom or different atoms. One skilled in the art can readily determine the valency of any such group from the context in which it occurs.

[0085] The term "substituted" refers to substitution where one, two, or more hydrogen atoms are independently replaced with a substituent, including, but not limited to: -F, -Cl, -Br, -I, -OH, -C 1- C12 -Alkyl, C2-C 12 -Alkenyl, C2-C 12 -Alkynyl, -C3-C 12 -Cycloalkyl, protected hydroxy, -NO2, -N3, -CN, -NH2, protected amino, oxo, thioxo, -NH-C 1- C 12 Alkyl, -NH-C2-C8-alkenyl, -NH-C2-C8-alkynyl, -NH-C3-C 12 -Cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, -OC 1- C 12 -Alkyl, -O-C2-C8-alkenyl, -O-C2-C8-alkynyl, -O-C3-C 12 -cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)-C 1- C 12 -Alkyl, -C(O)-C2-C8-alkenyl, -C(O)-C2-C8-alkynyl, -C(O)-C3-C 12 -Cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocycloalkyl, -CONH2, -CONH-C 1- C 12 -Alkyl, -CONH-C2-C8-alkenyl, -CONH-C2-C8-alkynyl, -CONH-C3-C 12 -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -OCO2-C 1- C 12 -Alkyl, -OCO2-C2-C8-alkenyl, -OCO2-C2-C8-alkynyl, -OCO2-C3-C 12 -cycloalkyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocycloalkyl, -CO2-C 1- C 12 Alkyl, -CO2-C2-C8 alkenyl, -CO2-C2-C8 alkynyl, CO2-C3-C 12-Cycloalkyl, -CO2-aryl, CO2-heteroaryl, CO2-heterocycloalkyl, -OCONH2, -OCONH-C 1- C 12 -Alkyl, -OCONH-C2-C8-alkenyl, -OCONH-C2-C8-alkynyl, -OCONH-C3-C 12 -cycloalkyl, -OCONH-aryl, -OCONH-heteroaryl, OCONH-heterocyclo-alkyl, -NHC(O)H, -NHC(O)-C 1- C 12 -Alkyl, -NHC(O)-C2-C8-alkenyl, -NHC(O)-C2-C8-alkynyl, -NHC(O)-C3-C 12 -cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclo-alkyl, -NHCO2-C 1- C 12 -Alkyl, -NHCO2-C2-C8-alkenyl, -NHCO2-C2-C8-alkynyl, -NHCO2-C3-C 12 -cycloalkyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocycloalkyl, -NHC(O)NH2, -NHC(O)NH-C 1- C 12 -Alkyl, -NHC(O)NH-C2-C8-alkenyl, -NHC(O)NH-C2-C8-alkynyl, -NHC(O)NH-C3-C 12 -cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH, -NHC(S)NH-C 1- C 12 -Alkyl, -NHC(S)NH-C2-C8-alkenyl, -NHC(S)NH-C2-C8-alkynyl, -NHC(S)NH-C3-C 12 -cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH2, -NHC(NH)NH-C 1- C 12-Alkyl, -NHC(NH)NH-C2-C8-alkenyl, -NHC(NH)NH-C2-C8-alkynyl, -NHC(NH)NH-C3-C 12 -cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C 1- C 12 -Alkyl, -NHC(NH)-C2-C8-alkenyl, -NHC(NH)-C2-C8-alkynyl, -NHC(NH)-C3-C 12 -cycloalkyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocycloalkyl, -C(NH)NH-C 1- C 12 -Alkyl, -C(NH)NH-C2-C8-alkenyl, -C(NH)NH-C2-C8-alkynyl, -C(NH)NH-C3-C 12 -cycloalkyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)-C 1- C 12 -Alkyl, -S(O)-C2-C8-alkenyl, -S(O)-C2-C8-alkynyl, -S(O)-C3-C 12 -Cycloalkyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocycloalkyl, -SO2NH2, -SO2NH-C 1- C 12 -Alkyl, -SO2NH-C2-C8-alkenyl, -SO2NH-C2-C8-alkynyl, -SO2NH-C3-C 12 -Cycloalkyl, -SO2NH-aryl, -SO2NH-heteroaryl, -SO2NH-heterocycloalkyl, -NHSO2-C 1- C 12 -Alkyl, -NHSO2-C2-C8-alkenyl, -NHSO2-C2-C8-alkynyl, -NHSO2-C3-C 12-cycloalkyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocycloalkyl, -CH2NH2, -CH2SO2CH3, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C3-C 12 -Cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -methoxyethoxy, -SH, -SC 1- C 12 -Alkyl, -S-C2-C8-alkenyl, -S-C2-C8-alkynyl, -S-C3-C 12 In certain embodiments, the substituents are halo, preferably Cl and F; 1- C4-Alkyl, preferably methyl and ethyl; halo-C 1- C4-Alkyl, such as fluoromethyl, difluoromethyl and trifluoromethyl; C2-C4-Alkenyl; Halo-C2-C4-Alkenyl; C3-C6-Cycloalkyl, such as cyclopropyl; C 1- C4-alkoxy, such as methoxy and ethoxy; halo-C 1- C4-alkoxy, for example, fluoromethoxy, difluoromethoxy and trifluoromethoxy, -CN; -OH; NH2; C 1- C4-Alkylamino; Di(C 1- and NO2. It is understood that aryl, heteroaryl, alkyl, etc. can be further substituted. In some cases, each substituent on the substituted moiety is optionally further substituted with one or more groups, each of which is selected from C1-C4-alkyl, CF3, -OCH3, -OCF3, -F, -Cl, -Br, -I, -OH, -NO2. 2、 Preferably, the substituted alkyl group is substituted with one or more halogen atoms, more preferably one or more fluorine or chlorine atoms.

[0086] The terms "halo" or "halogen," as used herein, alone or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom.

[0087] As used herein, the term "optionally substituted" means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional groups individually and independently selected from the groups described herein.

[0088] The term "hydrogen" includes hydrogen and deuterium. Furthermore, the recitation of an atom includes other isotopes of that atom so long as the resulting compound is pharmaceutically acceptable.

[0089] The term "hydroxy activating group," as used herein, refers to a labile chemical moiety known in the art to activate a hydroxyl group and release it during a synthetic procedure such as a substitution or elimination reaction. Examples of hydroxyl activating groups include, but are not limited to, mesylates, tosylates, triflates, p-nitrobenzoates, phosphonates, and the like.

[0090] As used herein, the term "activated hydroxyl" refers to a hydroxy group that is activated with a hydroxyl activating group as defined above, including, for example, a mesylate, tosylate, triflate, p-nitrobenzoate, or phosphonate group.

[0091] The term "hydroxy protecting group," as used herein, refers to a labile chemical moiety known in the art to protect a hydroxyl group from undesired reactions during synthetic procedures. After said synthetic procedures, the hydroxy protecting groups described herein can be selectively removed. Hydroxy protecting groups known in the art are described in T.H. Greene and P.G.M.Wuts, "Protecting Groups in Organic Synthesis ( Protective Groups in Organic Synthesis ), 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenyl-methyl(trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

[0092] The term "protected hydroxy" as used herein refers to a hydroxy group protected with a hydroxy protecting group as defined above, including, for example, a benzoyl, acetyl, trimethylsilyl, triethylsilyl, or methoxymethyl group.

[0093] The term "hydroxy prodrug group," as used herein, refers to a promoiety group known in the art to temporarily alter the physicochemical and therefore biological properties of the parent drug by covering or masking the hydroxy group. After synthetic procedures, the hydroxy prodrug groups described herein must be capable of reverting back to the hydroxy group in vivo. Hydroxy prodrug groups known in the art are described in Kenneth B. Sloan, "Prodrugs, Topical and Intraocular Drug Delivery ( Prodrugs,Topical and Ocular Drug Delivery )" (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992).

[0094] The term "amino-protecting group," as used herein, refers to a labile chemical moiety known in the art to protect amino groups against undesired reactions during synthetic procedures. After synthetic procedures, the amino-protecting groups described herein can be selectively removed. Hydroxy-protecting groups known in the art are described in T.H. Greene and P.G.M.Wuts, "Protecting Groups in Organic Synthesis ( Protective Groups in Organic Synthesis ), 3rd edition, John Wiley & Sons, New York (1999). Examples of amino-protecting groups include, but are not limited to, methoxycarbonyl, t-butoxycarbonyl, 12-fluorenyl-methoxycarbonyl, benzyloxycarbonyl, and the like.

[0095] The term "protected amino" as used herein refers to an amino group protected with an amino-protecting group as defined above.

[0096] The term "leaving group" means a functional group or atom that can be displaced by another functional group or atom in a substitution reaction, such as a nucleophilic substitution reaction. By way of example, representative leaving groups include chloro, bromo, and iodo groups, sulfonate ester groups (mesylate, tosylate, brosylate, nosylate, etc.), and acyloxy groups (acetoxy, trifluoroacetoxy, etc.).

[0097] As used herein, the term "aprotic solvent" refers to a solvent that is relatively inert to proton activity, i.e., does not act as a proton donor. Examples include, but are not limited to, hydrocarbons (such as hexane and toluene), halogenated hydrocarbons (such as methylene chloride, ethylene chloride, and chloroform), heterocyclic compounds (such as tetrahydrofuran and N-methylpyrrolidinone), and ethers (such as diethyl ether and bis-methoxymethyl ether). Such compounds are well known to those skilled in the art, and it will be apparent to those skilled in the art that individual solvents or mixtures thereof may be preferred for particular compounds and reaction conditions, depending on factors such as the solubility of the reagents, the reactivity of the reagents, and the preferred temperature range. Detailed descriptions of aprotic solvents can be found in organic chemistry textbooks or specialized treatises, such as "Organic Solvents Physical Properties and Purification Methods ( Organic Solvents Physical Properties and Methods of Purification )" 4th edition, edited by John A. Riddick et al., Vol. II, Techniques of Chemistry Series , John Wiley & Sons, NY, 1986.

[0098] The term "protic solvent" as used herein refers to a solvent that tends to provide a proton, such as an alcohol, e.g., methanol, ethanol, propanol, isopropanol, butanol, t-butanol, etc. Such solvents are well known to those skilled in the art, and it will be apparent to those skilled in the art that individual solvents or mixtures thereof may be preferred for particular compounds and reaction conditions, depending on factors such as, for example, the solubility of the reagents, the reactivity of the reagents, and the preferred temperature range. A detailed description of protic solvents can be found in organic chemistry textbooks or specialized treatises, e.g., "Organic Solvent Physical Properties and Purification Methods ( Organic Solvents Physical Properties and Methods of Purification )" 4th edition, edited by John A. Riddick et al., Vol. II, Techniques of Chemistry Series , John Wiley & Sons, NY, 1986.

[0099] Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. As used herein, the term "stable" refers to compounds that have sufficient stability to permit their manufacture and that will maintain their integrity for a period of time sufficient to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0100] The synthesized compounds can be separated from the reaction mixture and further purified by methods such as column chromatography, high pressure liquid chromatography, or recrystallization. As can be appreciated by those skilled in the art, additional methods for synthesizing compounds of the formulae herein will be apparent to those skilled in the art. In addition, the various synthetic steps can be performed in an alternative sequence or order to obtain the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful for synthesizing the compounds described herein are known in the art and are described, for example, in R. Larock, "Comprehensive Organic Transformations ( Comprehensive Organic Transformations )」2 nd Ed. Wiley-VCH (1999); TW Greene and PG M Butts "Protective Groups in Organic Synthesis ( Protective Groups in Organic Synthesis ) 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser "Fieser and Fieser's Reagents for Organic Synthesis ( Fieser and Fieser's Reagents for Organic Synthesis)" John Wiley and Sons (1994); and L. Paquette, ed. "Encyclopedia of Reagents for Organic Synthesis ( Encyclopedia of Reagents for Organic Synthesis ) John Wiley and Sons (1995) and its successors.

[0101] As used herein, the term "subject" refers to an animal. Preferably, the animal is a mammal. More preferably, the mammal is a human. A subject also refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, fish, birds, etc.

[0102] The compounds of the present invention can be modified by appending appropriate functional groups to enhance selective biological properties. Such modifications are known in the art and can include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism, or alter excretion rate.

[0103] The compounds described herein contain one or more asymmetric centers and therefore give rise to enantiomers, diastereomers, and other stereoisomers that can be defined with respect to absolute stereochemistry as (R)- or (S)-, or (D)- or (L)-amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors by the procedures described above or by resolving the racemic mixtures. Resolution can be carried out in the presence of a resolving agent, by chromatography, or by repeated crystallization, or by some combination of these techniques known to those skilled in the art. Further details regarding resolution can be found in Jacques et al., "Enantiomers, Racemates, and Resolution ( Enantiomers, Racemates, and Resolutions)" (John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds, other unsaturation, or other centers of geometric asymmetry, unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers or cis and trans isomers. Likewise, all tautomeric forms are intended to be included. Tautomers may be cyclic or acyclic. Configurations of carbon-carbon double bonds appearing herein have been selected for convenience only and are not intended to designate a particular configuration unless expressly stated in text. Thus, a carbon-carbon double bond or carbon-heteroatom double bond shown arbitrarily herein as trans can be cis, trans, or a mixture of the two in any proportion.

[0104] Certain compounds of the present invention may also exist in different stable conformational forms that may be separable. Torsional asymmetry due to restricted rotation about an asymmetric single bond, for example, due to steric hindrance or ring strain, may allow for the separation of different conformers. The present invention includes each conformational isomer of these compounds and mixtures thereof.

[0105] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and that is commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., J. Pharmaceutical Sciences, 66:2-19 (1977), describe pharmaceutically acceptable salts in detail. Salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reacting the free base function with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts of amino groups formed with inorganic acids (such as hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids) or organic acids (such as acetic, maleic, tartaric, citric, succinic, or malonic acids), or formed using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate Representative alkali or alkaline earth metal salts include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed, where appropriate, using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates and arylsulfonates.

[0106] As used herein, the term "pharmaceutically acceptable ester" refers to an ester that hydrolyzes in vivo, including those that readily decompose in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic, and alkanedioic acids, with each alkyl or alkenyl moiety advantageously having 6 or fewer carbon atoms. Examples of specific esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, and ethylsuccinates.

[0107] Pharmaceutical Composition Pharmaceutical compositions of the present invention comprise a therapeutically effective amount of a compound of the present invention formulated together with one or more pharmaceutically acceptable carriers or excipients.

[0108] As used herein, the term "pharmaceutically acceptable carrier or excipient" means a non-toxic, inert solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose, sucrose, and the like; starches such as corn starch and potato starch; cellulose and its derivatives, for example, sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions. Also, other non-toxic compatible lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservatives, and antioxidants can also be present in the composition, according to the discretion of the formulator.

[0109] The pharmaceutical compositions of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir, preferably orally or by injection. The pharmaceutical compositions of the present invention can contain any conventional non-toxic pharmaceutically acceptable carrier, adjuvant, or vehicle. Optionally, the pH of the formulation may be adjusted with a pharmaceutically acceptable acid, base, or buffer to enhance the stability of the formulated compound or its delivery form. As used herein, the term "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques.

[0110] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid ester sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and perfumes.

[0111] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, such as a solution of 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, United States Pharmacopoeia (USP), and isotonic sodium chloride solution. Additionally, sterile, fixed oils are conventionally used as solvents or suspending media. For this purpose, any bland fixed oil can be used, including synthetic mono- or diglycerides. Additionally, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0112] Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0113] To prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. In this case, the rate of absorption of the drug depends on its rate of dissolution, which, in turn, may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot formulations are made by forming microencapsulated matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the ratio of drug to polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations have also been prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0114] Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing a compound of the invention with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ambient temperature but liquid at body temperature and will melt in the rectum or vaginal cavity and release the active compound.

[0115] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate, and / or with a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) humectants such as glycerol; d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarders such as paraffin; f) absorption accelerators such as quaternary ammonium compounds; g) humectants such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite clay; and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0116] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0117] Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulating art. The dosage forms may optionally contain opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0118] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and, if necessary, any necessary preservatives or buffers. Ophthalmic formulations, ear drops, eye ointments, powders, and solutions are also contemplated within the scope of the present invention.

[0119] The ointments, pastes, creams, and gels may contain, in addition to the active compounds of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0120] Powders and sprays can contain, in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain conventional propellants, such as chlorofluorohydrocarbons.

[0121] Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0122] For pulmonary delivery, the therapeutic compositions of the present invention are formulated and administered to a patient in solid or liquid particulate form by direct administration, e.g., inhalation, into the respiratory system. The solid or liquid particulate forms of active compounds prepared for practicing the present invention include particles of respirable size, i.e., particles small enough to pass through the mouth and larynx upon inhalation and enter the bronchi and alveoli of the lungs. The delivery of aerosolized therapeutic agents, particularly aerosolized antibiotics, is known in the art (see, e.g., US Pat. No. 5,767,068 to Van Devanter et al., US Pat. No. 5,508,269 to Smith et al., and WO 98 / 43650 to Montgomery, all of which are incorporated herein by reference).

[0123] Combination and Alternation Therapies The compounds of the invention may be used in combination with one or more antiviral or anti-inflammatory therapeutic agents useful in the prevention or treatment of viral diseases or associated pathophysiology. Thus, the compounds of the invention and their salts, solvates, or other pharmaceutically acceptable derivatives may be used alone or in combination with other antiviral or anti-inflammatory therapeutic agents.The compounds herein and pharmaceutically acceptable salts thereof may be used in combination with one or more other agents that may be useful in the prevention or treatment of: respiratory diseases, inflammatory diseases, autoimmune diseases, such as antihistamines, corticosteroids (e.g., fluticasone propionate, fluticasone furoate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furoate, triamcinolone, flunisolide), NSAIDs, leukotriene modulators (e.g., montelukast, zafirlukast, pranlukast), triprophallic acid inhibitors (e.g., fluticasone propionate, fluticasone furoate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furoate, triamcinolone, flunisolide), steroid ... triamcinolone, flunisolide), triprophallic acid inhibitors (e.g., fluticasone propionate, fluticasone furoate, triamcinolone, flunisolide), triprophallic acid inhibitors (e.g., fluticasone propionate, fluticasone furoate, triamcinolone, flunisolide), triprophallic acid inhibitors (e.g., fluticasone propionate, tase inhibitors, IKK2 inhibitors, p38 inhibitors, Syk inhibitors, protease inhibitors such as elastase inhibitors, integrin antagonists (e.g. beta-2 integrin antagonists), adenosine A2a agonists, mediator release inhibitors such as sodium cromoglycate, 5-lipoxygenase inhibitors (zyflo), DP1 antagonists, DP2 antagonists, PI3K delta inhibitors, ITK inhibitors, LP (lysophosphatidic) inhibitors or FLAP (5-lipoxygenase activating protein) inhibitors (e.g. , sodium 3-(3-(tert-butylthio)-1-(4-(6-ethoxypyridin-3-yl)benzyl)-5-((5-ethylpyridin-2-yl)methoxy)-1H-indol-2-yl)-2,2-dimethylpropanoate), bronchodilators (e.g., muscarinic antagonists, beta-2 agonists), methotrexate and similar agents; monoclonal antibody therapies such as anti-IgE, anti-TNF, anti-IL-5, anti-IL-6, anti-IL-12, anti-IL-1 and similar agents; cytokine receptor therapies, e.g., etanercept and similar agents; anti Non-specific immunotherapies (e.g., interferons or other cytokines / chemokines, chemokine receptor modulators such as CCR3, CCR4 or CXCR2 antagonists, other cytokine / chemokine agonists or antagonists, TLR agonists and similar agents), appropriate anti-infectives including antibiotics, antifungals, antiparasitics, antimalarials, antiprotozoals, antituberculous agents, and antivirals including those listed at https: / / www.drugs.com / drug-class / anti-infectives.html.In general, combination therapy is preferred over alternation therapy because it typically induces multiple simultaneous stresses on the virus.

[0124] While the present invention has been described in terms of various preferred embodiments, it is not intended to be limited thereto, but rather, those skilled in the art will recognize that variations and modifications can be made which are within the spirit of the invention and the scope of the appended claims.

[0125] Antiviral activity The inhibitory amount or dose of the compounds of the present invention can range from about 0.01 mg / kg to about 500 mg / kg, or alternatively from about 1 to about 50 mg / kg. The inhibitory amount or dose will also vary depending on the route of administration, as well as the possibility of co-administration with other drugs.

[0126] According to the therapeutic methods of the present invention, viral infections are treated or prevented in a patient, such as a human or another animal, by administering to the patient a therapeutically effective amount of a compound of the present invention in an amount and for a time necessary to achieve the desired result.

[0127] A "therapeutically effective amount" of a compound of the present invention refers to that amount of compound that confers a therapeutic effect on the treated subject, at a reasonable benefit / risk ratio applicable to any medical treatment. The therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., the subject shows signs of or feels an effect). Effective amounts of the compounds described above may range from about 0.1 mg / kg to about 500 mg / kg, preferably from about 1 to about 50 mg / kg. The effective dose will also vary depending on the route of administration and the possibility of co-administration with other drugs. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the time, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical arts.

[0128] The total daily dose of the compounds of this invention administered to a human or other animal in single or divided doses can be, for example, 0.01 to 50 mg / kg body weight or more, usually 0.1 to 25 mg / kg body weight. Single dose compositions may contain such amounts or submultiples thereof to make up the daily dose. In general, treatment regimens according to the present invention comprise administering to a patient in need of such treatment from about 10 mg to about 1000 mg of a compound of this invention per day, in single or multiple doses.

[0129] The compounds of the present invention described herein can be administered, for example, by injection, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in ophthalmic preparations, or by inhalation, at doses ranging from about 0.1 to about 500 mg / kg body weight, or at doses ranging from 1 mg to 1000 mg / dose, every 4 to 120 hours, or according to the requirements of the particular drug. The methods herein contemplate administration of an effective amount of a compound or compound composition to achieve the desired or stated effect. Typically, pharmaceutical compositions of the present invention are administered about 1 to about 6 times per day, or alternatively as a continuous infusion. Such administration can be used for chronic or acute treatment. The amount of active ingredient that can be combined with pharmaceutical excipients or carriers to produce a single dosage form varies depending on the host treated and the particular mode of administration. Typical preparations contain from about 5% to about 95% active compound (w / w). Alternatively, such preparations may contain from about 20% to about 80% active compound.

[0130] Lower or higher doses than those recited above may be required. The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptom, the patient's disposition to the disease, condition or symptom, and the judgment of the treating physician.

[0131] Once the patient's condition has improved, a maintenance dose of the compound, composition or combination of the present invention may be administered, if necessary. Thereafter, the dosage or frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the improved condition is maintained when the symptoms have been alleviated to a desired level. However, patients may require intermittent treatment on a long-term basis upon recurrence of disease symptoms.

[0132] When the compositions of the invention comprise a combination of a compound of the formulae described herein with one or more additional therapeutic or prophylactic agents, both the compound and the additional agents should be present at dosage levels that are about 1-100%, more preferably about 5-95%, of the dosage normally administered in a monotherapy regimen. The additional agents can be administered separately from the compounds of the invention as part of a multiple-dose regimen. Alternatively, these agents can be part of a single dosage form, mixed together with the compounds of the invention in a single composition.

[0133] "Additional therapeutic or prophylactic agents" include, but are not limited to, immunotherapy (e.g., interferon), therapeutic vaccines, antifibrotic agents, anti-inflammatory agents such as corticosteroids or NSAIDs, bronchodilators such as beta-2 adrenergic agonists and xanthines (e.g., theophylline), mucolytic agents, antimuscarinics, anti-leukotrienes, cell adhesion inhibitors (e.g., ICAM antagonists), antioxidants (e.g., N-acetylcysteine), cytokine agonists, cytokine antagonists, pulmonary surfactants, and / or antibacterial and antiviral agents (e.g., ribavirin and amantidine). The compositions of the present invention may also be used in combination with gene replacement therapy.

[0134] Abbreviation Abbreviations that may be used in the description of the following schemes and examples are as follows: acetyl Ac; acetic acid AcOH; di-tert-butyl-dicarbonate BocO; t-butoxycarbonyl Boc; benzoyl Bz; benzyl Bn; potassium tert-butoxide t-BuOK; aqueous sodium chloride brine; carbonyldiimidazole CDI; dichloromethane DCM or CH2Cl2; methyl CH3; acetonitrile CH3CN; cesium carbonate Cs2CO3; copper chloride (I) CuCl; copper iodide (I) CuI; dibenzylideneacetone dba; 1,8-diazomethane Bicyclo[5.4.0]undec-7-ene (DBU); Diethyl azodicarboxylate (DEAD); Diisopropyl azodicarboxylate (DIAD); N,N-Diisopropylethylamine (DIPEA) or (i-Pr)2EtN; 1,1,2-tris(acetyloxy)-1,2-dihydro-1,2-benziodoxol-3-(1H)-one (DMP) or Dess-Martin periodinane; 4-Dimethylaminopyridine (DMAP); 1,2-Dimethoxyethane (DME); N,N-Dimethylformamide (DMF); Dimethyl sulfoxide (DMSO); Ethyl acetate Chilled EtOAc; Ethanol EtOH; Diethyl ether Et2O; O-(7-Azabenzotriazol-2-yl)-N,N,N',N',-tetramethyluronium hexafluorophosphate HATU; Hydrogen chloride HCl; Potassium carbonate K2CO3; n-Butyllithium n-BuLi; 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone DDQ; Lithium diisopropylamide LDA; Lithium 2,2,6,6-tetramethyl-piperidinate LiTMP; Methanol MeOH; Magnesium Mg; Methoxymethyl MOM; Mesyl or SO2-CH3 is Ms; sodium bis(trimethylsilyl)amide NaHMDS; sodium chloride NaCl; sodium hydride NaH; sodium bicarbonate or sodium hydrogen carbonate NaHCO3; sodium carbonate Na2CO3; sodium hydroxide NaOH; sodium sulfate Na2SO4; sodium bisulfite or sodium hydrogen sulfite NaHSO3; sodium thiosulfate Na2S2O3; hydrazine NH2NH2; ammonium chloride NH4Cl; nickel Ni; hydroxyl OH; osmium tetroxide OsO4; triflate OTf; polyphosphate PPA;p-Toluenesulfonic acid PTSA; p-Toluenesulfonic acid pyridinium PPTS; Tetrabutylammonium fluoride TBAF; Triethylamine TEA or Et3N; Triethylsilyl TES; Triethylsilyl chloride TESCl; Triethylsilyl trifluoromethanesulfonate TESOTf; Trifluoroacetic acid TFA; Tetrahydrofuran THF; N,N,N',N'-Tetramethylethylenediamine TMEDA; Triphenylphosphine TPP or PPh3; Tosyl or SO2-C6H4CH3 is Tos or Ts; Tolylsulf Tosyl anhydride (Ts2O); p-tolylsulfonic acid (TsOH); palladium (Pd); phenyl (Ph); tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3); tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4); trans-dichlorobis-(triphenylphosphine)palladium(II) (PdCl2(PPh3)2); platinum (Pt); rhodium (Rh); room temperature (rt); ruthenium (Ru); tert-butyldimethylsilyl (TBS); trimethylsilyl (TMS); trimethylsilyl chloride (TMSCl).

[0135] Synthesis method The compounds and processes of the present invention will be better understood in connection with the following synthetic schemes, which illustrate how the compounds of the present invention can be prepared. These schemes are for illustrative purposes and are not intended to limit the scope of the invention. Particular solvents, reagents, or reaction conditions described herein may be substituted with equivalent, similar, or suitable solvents, reagents, or reaction conditions without departing from the general scope of the synthetic methods.

[0136] Scheme 1: [ka] As shown in Scheme 1, compounds such as 5 (Q1 is defined as Q, Q2 is defined as Q, R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Intermediate 1 (R1 is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle, and J is defined as an amino-protecting group) can react with nitrile 2 (X is defined as halogen, OMs, OAc, OTf, OTs, or OTf) in a carbon-carbon bond-forming reaction typically mediated by a base (denoted as [base]), including, but not limited to, LDA, LiHMDS, or LiTMP. Intermediate 3 can be reduced (denoted as [reduction]), typically mediated by a reducing agent, including, but not limited to, LiBH4 or NaBH4. Lactam 4 can be reacted in a deprotection step (denoted as [deprotection]) typically mediated by an acidic reagent, including but not limited to, TFA or HCl, to produce compound 5. Alternatively, lactam 4 can be reacted in a deprotection step (denoted as [deprotection]) mediated by a reducing agent, including but not limited to, hydrogen on palladium on carbon, to produce compound 5.

[0137] Scheme 2: [ka] As shown in Scheme 2, compounds such as 3 (Q1 is defined as Q, Q2 is defined as Q, and A, L1, and L2 are as defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Acid 1 can be reacted in a coupling reaction with amine 2 (X as defined above), typically mediated by a base (denoted as [base]) including, but not limited to, DIPEA, EtN, or DBU, and an activating agent (denoted as [activator]) including, but not limited to, HATU or EDC.

[0138] Scheme 3: [ka] As shown in Scheme 3, compounds such as 3 (where Q is defined as Q, Q is defined as Q, A, L, and L are as defined above, and R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Acid 1 can be reacted with amine 2 in a coupling reaction typically mediated by a base (denoted as [base]) including, but not limited to, DIPEA, EtN, or DBU, and an activating agent (denoted as [activator]) including, but not limited to, HATU or EDC.

[0139] Scheme 4: [ka] As shown in Scheme 4, compounds such as 4 (A, L1, and L2 are defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Amine 1 (R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be coupled with acid 2 (A, as defined above), typically mediated by a base (denoted as [Base]) including, but not limited to, DIPEA, EtN, or DBU, and an activating agent (denoted as [Activator]) including, but not limited to, HATU or EDC. Ester 3 can be reacted in a hydrolysis reaction (denoted as [Hydrolysis]), typically mediated by an acidic reagent including, but not limited to, TFA or HCl, to generate acid 4. Alternatively, ester 3 can be reacted in a hydrolysis reaction (denoted as [Hydrolysis]), typically mediated by a basic reagent including, but not limited to, NaOH, LiOH, or MeSnOH, to generate acid 4. Alternatively, ester 3 can be reacted in a hydrolysis reaction (denoted as [hydrolysis]) typically mediated by a reducing agent including, but not limited to, hydrogen on palladium on carbon to produce acid 4.

[0140] Scheme 4b: [ka] As shown in Scheme 4b, compounds such as 3 (A, L1, and L2 are as defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Amine 1 (R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be reacted with an activated form of acid 2 (A, as defined above) in a coupling reaction, where the acid activation occurs by reaction with an activating agent (denoted as [activating agent]) to generate an intermediate activated ester intermediate. The aforementioned coupling reaction between amine 1 and the activated form of acid 2 is mediated by a base (denoted as [base]), including but not limited to NaOH, NaHCO3, or KOH, to generate amide 3.

[0141] Scheme 5: [ka] As shown in Scheme 5, compounds such as 3 (Q1 is defined as Q, Q2 is defined as Q, and A, L1, and L2 are as defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Intermediate 1 (R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be reacted in a reduction reaction using reagents including, but not limited to, LiBH4 or NaBH4 (denoted as [reducing agent]) to produce alcohol 2, which can undergo an oxidation reaction using a group of reagents including, but not limited to, SO3-pyridine, DMP, or Ac2O / DMSO (denoted as [oxidizing agent]) to produce aldehyde 3.

[0142] Scheme 6: [ka] As shown in Scheme 6, compounds such as 2 (Q is defined as Q, Q is defined as Q, and A, L, and L are as defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Ester 1 (R is defined as H, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be reacted with ICHCl and a basic reagent (denoted as [base]), such as, but not limited to, LDA or nBuLi, to generate 2.

[0143] Scheme 7: [ka] As shown in Scheme 7, compounds such as 3 (where Q1 is defined as Q, Q2 is defined as Q, A, L1, and L2 are as defined above, and R is defined as optionally substituted alkyl, optionally substituted aryl, or optionally substituted heterocycle) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Halide 1 can be reacted with acid 2 and a basic reagent (denoted as [base]), including but not limited to CsF or NaF, to produce ester 3.

[0144] Scheme 8: [ka] As shown in Scheme 8, compounds such as 3 (Q1 is defined as Q, Q2 is defined as Q, and A, L1, and L2 are as defined above) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Halide 1 can be reacted with acid 2 and a basic reagent (denoted as [Base]) including, but not limited to, CsF or NaF to generate an intermediate ester, which can then be reacted in a hydrolysis reaction (denoted as [Hydrolysis]) mediated by a reagent including, but not limited to, K2CO3 or Cs2CO3 to generate alcohol 3.

[0145] Scheme 9: [ka] As shown in Scheme 9, compounds such as 3 (Q1 is defined as Q and Q2 is defined as Q) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Ester 1 (R is defined as an optionally substituted aryl or alkyl, and PG is defined as a nitrogen-based protecting group) can be reacted with ammonia to generate an intermediate amide, which can then undergo a dehydration reaction, shown as [dehydration], mediated by reagents including, but not limited to, Pd(CO2CF3)2 or TFAA, to generate nitrile 2, which can undergo a deprotection reaction, shown as [deprotection], mediated by reagents including, but not limited to, TFA, HCl, palladium, or platinum, to generate primary amine 3.

[0146] Scheme 10: [ka] As shown in Scheme 10, compounds such as 4 (where Q1 is defined as Q, Q2 is defined as Q, A, L1, and L2 are as defined above, and R is defined as optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted heterocycle) can be prepared according to the synthetic methods set forth herein or by similar methods known to those skilled in the art. Aldehyde 1 can be reacted with isonitrile 2 to form intermediate hydroxyamide 3. This intermediate can undergo an oxidation reaction, shown as [oxidation], mediated by reagents including, but not limited to, sulfur trioxide pyridine complex (Py-SO3), DMSO, oxalyl chloride, and / or acetic anhydride, to generate ketoamide 4.

[0147] All references cited herein, including but not limited to abstracts, articles, magazines, publications, texts, papers, internet websites, databases, patents, and patent publications, whether in print, electronic, computer-readable storage media, or other form, are expressly incorporated by reference in their entirety.

[0148] Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications, including but not limited to those relating to the chemical structures, substituents, derivatives, formulations and / or methods of the invention, can be made without departing from the spirit of the invention and the scope of the appended claims.

[0149] While the present invention has been described in terms of various preferred embodiments, it is not intended to be limited thereto, but rather, those skilled in the art will recognize that variations and modifications can be made which are within the spirit of the invention and the scope of the appended claims.

[0150] example The compounds and processes of this invention will be better understood in connection with the following examples, which are intended to be illustrative only and not to limit the scope of the invention. Starting materials were available from commercial suppliers or prepared by methods well known to those skilled in the art.

[0151] General conditions: Mass spectra were performed on an LC-MS system using electrospray ionization. These were Agilent 1290 Infinity II systems equipped with an Agilent 6120 Quadrupole detector. Spectra were acquired using a ZORBAX Eclipse XDB-C18 column (4.6 x 30 mm, 1.8 microns). Spectra were acquired at 298 K using a mobile phase of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). Spectra were acquired using the following solvent gradient: 0-1.5 min 5% (B), 1.5-4.5 min 5-95% (B), and 4.5-6 min 95% (B). The solvent flow rate was 1.2 mL / min. Compounds were detected at wavelengths of 210 nm and 254 nm. [M+H] + refers to the monoisotopic molecular weight.

[0152] NMR spectra were performed on a Bruker 400 MHz spectrometer. Spectra were measured at 298 K and referenced using the solvent peak. 1 1 H NMR chemical shifts are reported in parts per million (ppm).

[0153] Compounds were purified by reverse-phase high-performance liquid chromatography (RPHPLC) using a Gilson GX-281 automated liquid handling system. Unless otherwise noted, compounds were purified on a Phenomenex Kinetex EVO C18 column (250 x 21.2 mm, 5 microns). Unless otherwise noted, compounds were purified at 298 K using a mobile phase of water (A) and acetonitrile (B) with a gradient elution of 0% to 100% (B). The solvent flow rate was 20 mL / min, and compounds were detected at a wavelength of 254 nm.

[0154] Alternatively, compounds were purified by normal phase liquid chromatography (NPLC) using a Teledyne ISCO Combiflash purification system. Compounds were purified on REDISEP silica gel cartridges. Compounds were purified at 298 K and detected at a wavelength of 254 nm.

[0155] Example 1: Synthesis of (1S,3aR,6aS)-N-((S)-4-chloro-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0156] Step 1: A suspension of 4-methoxy-1H-indole-2-carboxylic acid (3.6 g) in DCM (94 mL) was cooled to 0 °C. Next, tert-butyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate oxalate (6.81 g) was added, followed by DMAP (0.690 g). EDC (7.22 g) was then added, and the reaction was stirred at 0 °C for 30 minutes, then at room temperature overnight. The mixture was then washed with water (1 × 30 mL), and the organic layer was dried over NaSO, filtered, and concentrated. The crude product was used directly in the next step.

[0157] Step 2: Trifluoroacetic acid (32.6 mL) was added to a solution of methyl (1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylate (7.24 g) in DCM (106 mL) at room temperature. The resulting solution was stirred at room temperature for 2 h and then concentrated. The crude residue was purified on silica gel (ethyl acetate:cyclohexane 0-100%) to give (1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (4 g, 58% yield over two steps).

[0158] Step 3: A vial was charged with (S)-3-((S)-2-amino-4-chloro-3-oxobutyl)pyrrolidin-2-one hydrochloride (150 mg), DMF (3 mL), HATU (239 mg), and (1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (212 mg) and then cooled to 0° C. Next, Hunig's base (261 mg) was added. The reaction mixture was stirred for 75 minutes, then diluted with ethyl acetate and washed three times with saturated NaHCO, water, and brine. The organic layer was dried over sodium sulfate, filtered, and concentrated. The residue was purified on silica gel to give (1S,3aR,6aS)-N-((S)-4-chloro-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide as a light brown foam (195 mg, 61% yield). ESI MS m / z=516.1 [M+H] + .

[0159] Example 2: Synthesis of (S)-3-((1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butyl 2,6-dichlorobenzoate [ka]

[0160] Step 1: A vial was charged with 2,6-dichlorobenzoic acid (62 mg), cesium fluoride (91 mg), and (1S,3aR,6aS)-N-((S)-4-chloro-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (140 mg). The vial was purged with nitrogen gas, and then DMF (2 mL) was added. The reaction mixture was heated at 65° C. for 1 hour. After cooling to room temperature, the reaction mixture was partitioned between ethyl acetate and water. The organic phase was washed with saturated NaHCO, water, and brine. The organic layer was concentrated, and the residue was purified on silica gel to give (S)-3-((1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butyl 2,6-dichlorobenzoate (115 mg, 63% yield). ESI MS m / z=670.1 [M+H] + .

[0161] Example 3: Synthesis of (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide. [ka]

[0162] Step 1: A 350 mL sealed tube was charged with a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-[(3S)-2-oxopyrrolidin-3-yl]propanoate (25.00 g, 87.312 mmol, 1.00 equiv.) in NH3(g) in MeOH (250 mL, 7 mol / L). The resulting solution was stirred at 70 °C for 16 h. The reaction was concentrated in vacuo. The residue was purified by silica gel column (DCM / MeOH = 1:0 to 10:1). This afforded 15 g (63.32%) of tert-butyl N-[(1S)-1-carbamoyl-2-[(3S)-2-oxopyrrolidin-3-yl]ethyl]carbamate as a white solid.

[0163] Step 2: A 3 L four-neck round-bottom flask purged and maintained with an inert atmosphere of nitrogen was charged with tert-butyl N-[(1S)-1-carbamoyl-2-[(3S)-2-oxopyrrolidin-3-yl]ethyl]carbamate (150.00 g, 552.859 mmol, 1.00 equiv.), dichloroacetonitrile (607.81 g, 5528.590 mmol, 10.00 equiv.) in ACN (900 mL), and water (900 mL). This was followed by the addition of Pd(COCF) (11.03 g, 33.172 mmol, 0.06 equiv.) at room temperature. The resulting solution was stirred at room temperature for 16 hours. The resulting mixture was extracted with DCM (3 × 600 mL). The combined organic layers were washed with brine (1 × 1 L) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (2:1). After concentration of the combined product fractions, the residue was triturated under DCM, and the resulting solid was isolated and dried under vacuum. This gave (51 g, 36.42%) tert-butyl ((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)carbamate as a white solid. ESI MS m / z=254.1 [M+H] +.1HNMR(CDCl3)δ 6.30(s,1H),5.90(s,1H),4.77-4.59(m,1H),3.48-3.31(m,2H),2.49(dddd,J=23.7,11.8,7.3,2.7 Hz,2H),2.38-2.23(m,1H),2.02-1.79(m,2H),1.48(s,9H).

[0164] Step 3: Trifluoroacetic acid (790 μL) was added to a solution of tert-butyl ((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)carbamate (20 mg, 0.079 mmol) and DCM (0.790 mL) at 22° C. After 15 min, the resulting solution was directly concentrated in vacuo. The residue was redissolved in methanol (2 mL) and concentrated in vacuo, then redissolved in ethyl acetate (2 mL) and concentrated once more. The crude (S)-2-amino-3-((S)-2-oxopyrrolidin-3-yl)propanenitrile 2,2,2-trifluoroacetate was used without further purification. 1HNMR(DMSO-d6)δ 8.94(bs,2H),4.80(dd,J=8.7,6.6 Hz,1H),3.24-3.16(m,2H),2.50(m,1H),2.30(dddd,J=12.1,8.8,5.6,3.4 Hz,1H),2.15(ddd,J=14.5,8.1,6.6 Hz,1H),1.97-1.91(m,1H),1.74(ddt,J=12.5,10.5,9.0 Hz,1H).

[0165] Step 4: 4-(Difluoromethoxy)-1H-indole-2-carboxylic acid (3.5 g, 15.41 mmol) was suspended in 25 mL of DCM and 4.5 mL of THF. Oxalyl chloride (1.618 mL, 18.49 mmol) was then added dropwise at 22 °C, followed by DMF (3 drops). After 15 min, the mixture became homogeneous, and TLC analysis (aliquot quenched with MeOH) indicated complete conversion. The resulting solution was directly concentrated to give crude 4-(difluoromethoxy)-1H-indole-2-carbonyl chloride as a red oil, which was used immediately without further purification.

[0166] Step 5: Ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride (9.57 g, 43.6 mmol) was suspended in EtOH (87 mL), and then 5 N aqueous sodium hydroxide (37 mL, 185 mmol) was added with vigorous stirring at 22° C. After 1 h, the ethanol was distilled in vacuo to give approximately 40 mL of a viscous aqueous solution containing sodium (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate, which was used directly.

[0167] Step 6: Crude 4-(difluoromethoxy)-1H-indole-2-carbonyl chloride (2.161 g, 8.8 mmol) was dissolved in THF (5 mL) at 22 °C. Then, an aqueous solution of sodium (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate (16 mL) was poured into the solution with vigorous stirring at 22 °C. After stirring for 1 h, 1 N HCl was added until the pH of the solution was approximately 1. The resulting aqueous suspension was extracted twice with DCM, and then the pooled organic fractions were dried over MgSO, filtered, and concentrated. The resulting oil was subjected to silica gel chromatography to give (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (2.67 g, 7.33 mmol, 83% yield) as a pale yellow solid. 1HNMR(DMSO-d6)δ 12.64(bs,1H),11.88(s,1H),7.38-7.29(m,1H),7.20(m,1H),7.00-6.95(m,1H),6.82(m,1H),4.36(d,J=3.6 Hz,1H),4.13(dd,J=10.5,8.2 Hz,1H),3.81(dd,J=10.5,4.3 Hz,1H),3.70(m,1H),2.90-2.79(m,1H),2.64(m,1H),2.02-1.90(m,1H),1.88-1.66(m,3H),1.58(m,4H).

[0168] Step 7: HATU (69.6 mg, 0.183 mmol) was added to a solution of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (57.8 mg, 0.159 mmol), (S)-2-amino-3-((S)-2-oxopyrrolidin-3-yl)propanenitrile 2,2,2-trifluoroacetate (32.6 mg, 0.122 mmol), DMF (0.407 mL) and EtN (150 μL, 1.08 mmol) at 22 °C. The resulting solution was stirred at 22 °C for 24 h and then directly purified via RPHPLC to give (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide as a white solid (2 mg). ESI MS m / z = 500.1 [M+H] + .

[0169] Example 4: Synthesis of (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide. [ka]

[0170] The synthesis of Example 4 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 4-(difluoromethoxy)-1H-indole-2-carboxylic acid was replaced with 4-methoxy-1H-indole-2-carboxylic acid.

[0171] Characterization data for Example 4 was obtained: ESI MS m / z=464.1 [M+H] +. 1HNMR (chloroform-d)δ 9.77(s,1H),8.18(d,J=7.1 Hz,1H),7.22(t,J=8.1 Hz,1H),7.11-7.00(m,2H),6.76(s,1H),6.50(d,J=7.7 Hz,1H),4.92(t,J=8.3 Hz,1H),4.54(d,J=3.2 Hz,1H),4.24(t,J=9.4 Hz,1H),3.96(s,3H),3.84(dd,J=10.8,4.8 Hz,1H),3.32(m,2H),3.04(m,1H),2.87(m,1H),2.65(m,1H),2.33(m,2H),2.11-1.50(m,10H).

[0172] Example 5: Synthesis of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0173] Step 1: Methyl (S)-2-amino-3-((S)-2-oxopyrrolidin-3-yl)propanoate hydrochloride (0.880 g, 3.95 mmol) and (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (1.01 g, 2.77 mmol) were dissolved in a mixture of DMF and DCM (1:1 v / v, 26 mL), and the resulting solution was cooled to 0 °C under a nitrogen atmosphere. HATU (1.126 g, 2.96 mmol) was then added in one portion. After stirring for 3 min, Hunig's base (1.656 mL, 9.48 mmol) was added dropwise. After stirring at 0 °C for 30 min, pre-cooled 1 N HCl (15 mL) was added, followed by water (40 mL). The resulting mixture was extracted twice with DCM. The pooled organic fractions were washed twice with saturated aqueous NaHCO3 and then brine, then dried over MgSO4 and concentrated. The resulting brown oil was purified by silica gel chromatography to afford (S)-2-((1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-3-((S)-2-oxopyrrolidin-3-yl)propanoate (1.42 g, 2.67 mmol, 96% yield) as a white foam. 1H NMR (chloroform-d) δ 9.87 (s, 1H), 7.85 (d, J = 7.2 Hz, 1H), 7.29 (d, J = 8.2 Hz, 1H), 7.20 (d, J = 8.0 Hz, 1H), 6.97 (s, 1H), 6.90-6.45 (m, 3H), 6.01 (s, 1H), 4.63 (app s, 1H), 4.56 (m, 1H), 4.22 (t, J = 9.4 Hz, 1H), 3.80 (dd, J = 10.5 Hz, 4.6 Hz,1H),3.73(m,4H),3.26(m,2H),3.00(m,1H),2.86(m,1H),2.55(m,1H),2.41-2.30(m,1H),2 .23-2.11(m,1H),2.04(m,2H),1.98-1.85(m,3H),1.85-1.72(m,3H),1.48(m,2H),1.43(m,1H).

[0174] Step 2: Methyl (S)-2-((1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-3-((S)-2-oxopyrrolidin-3-yl)propanoate (1.42 g, 2.67 mmol) was dissolved in THF (26.7 mL) and cooled to 0 °C with stirring under a nitrogen atmosphere. Lithium borohydride (2 M in THF, 6.67 mL, 13.33 mmol) was then added dropwise over 4-5 min. After 40 min at 0 °C, 1 M HCl (1 equiv., 13.5 mL) was then added slowly over 5 min. The resulting cloudy solution was partitioned between EtOAc and water, and the phases were separated. The aqueous phase was extracted three times with EtOAc, and the pooled organic fractions were dried over MgSO and concentrated. The residue was subjected to silica gel chromatography to give (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-hydroxy-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide (965 mg, 1.913 mmol, 71.7% yield) as a white solid. 1HNMR (chloroform-d) δ 10.45(s,1H),7.95(d,J=8.0 Hz,1H),7.29(app d,J=8.3 Hz,1H),7.19(t,J=8.0 Hz,1H),6.99(s,1H),6.85-6.47(m,2H),6.12(s,1H),4.51(d,J=3.4 Hz,1H),4.27(t,J=9.4 Hz,1H),4.01(m,1H),3.74(td,J=11.1,4.3 Hz,2H),3.61(dd,J=11.6,4.2 Hz,1H),3.22(m,2H),3.06-2.93(m,1H),2.75(m,1H),2.60-2.50(m,1H),2.34 (m,1H),2.17(m,3H),1.95-1.83(m,3H),1.83-1.67(m,3H),1.67-1.45(m,5H).

[0175] Step 3: (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-hydroxy-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide (0.958 g, 1.899 mmol) was dissolved in DCM (12.66 mL) and then cooled to 0 °C under a nitrogen atmosphere. Dess-Martin periodinane (1.127 g, 2.66 mmol) was then added in one portion. After 1 h, the resulting brown suspension was poured into saturated aqueous NaSO. The phases were separated, and the organic phase was washed with saturated aqueous NaHCO and then brine before being dried over MgSO and concentrated. The residue was then subjected to silica gel chromatography eluting with DCM / MeOH. The product-containing fractions were concentrated, and the resulting brown foam was then subjected to a second silica gel chromatography eluting with MTBE / acetone to give (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide (390 mg, 0.776 mmol, 40.9% yield) as a colorless foam. 1H NMR (chloroform-d) δ 9.82 (s, 1H), 9.52 (s, 1H), 8.42 (s, 1H), 7.28 (app d, J = 8.4 Hz, 1H), 7.21 (t, J = 7.9 Hz, 1H), 7.02 (s, 1H), 6.87-6.43 (m, 3H), 5.90 (s, 1H), 4.69 (s, 1H), 4.35 (s, 1H), 4.26 (t, J = 9.4 Hz, 1H), 3.82 (dd, J = 10.3, 4.6 Hz,1H),3.49(s,1H),3.28(m,2H),2.99(s,1H),2.87(s,1H),2.54(s,1H),2.34(m,1H),2.11-1.49(m,17H).ESI MS m / z=503.1 [M+H] + .

[0176] Example 6: Synthesis of (1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0177] The synthesis of Example 6 was similar in nature to the synthesis of Example 5, with the following changes: 1. In step 1, (1S,3aR,6aS)-2-(4-(methoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid was used in place of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid.

[0178] Characterization data for Example 6 was obtained: ESI MS m / z=464.1 [M+H] + .

[0179] Example 7: Synthesis of (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopiperidin-3-yl)ethyl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide. [ka]

[0180] Step 1: A flask was charged with dimethyl (tert-butoxycarbonyl)-L-glutamate (6.5 g) and THF (70 mL). The flask was cooled to -78 °C under a nitrogen atmosphere. LiHMDS (52 mL, 1 M in THF) was then added over 5 minutes. After 1 hour, 3-bromopropanenitrile (3 mL) was added dropwise. After 90 minutes, the reaction mixture was warmed to -55 °C and then quenched with aqueous NH4Cl. The reaction mixture was allowed to reach room temperature and then diluted with 20 mL of water. The product was extracted with MTBE and then concentrated. An additional 30 mL of MTBE was added, causing the formation of a precipitate. This was filtered off, and the filtrate was concentrated to give an orange oil, which was used directly in the next step.

[0181] Step 2: A flask was charged with cobalt(II) chloride hexahydrate (2.8 g). A solution of the product from Step 1 in THF (20 mL) was then transferred to this flask using MeOH rinses (140 mL). After cooling the flask to 0° C., sodium borohydride (3.6 g) was added over 20 minutes. The reaction was allowed to reach room temperature and stirred for 24 hours. Most of the volatiles were then removed under reduced pressure. EtOAc (100 mL) and 1 M HCl (40 mL) were added. The product was extracted with EtOAc, and the combined organic layers were washed with 1 M HCl, brine, and then concentrated. The residue was purified on silica gel to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-((S)-2-oxopiperidin-3-yl)propanoate (1.4 g, 20% over two steps). ESI MS m / z=301.1 [M+H] + .

[0182] Step 3: A flask was charged with methyl (S)-2-((tert-butoxycarbonyl)amino)-3-((S)-2-oxopiperidin-3-yl)propanoate (421 mg), followed by the addition of 4 M ammonia in MeOH (2.8 mL). The reaction mixture was stirred for 72 hours and then heated to 65° C. for 1.5 hours. The volatiles were removed, and the residue was purified on silica gel to give tert-butyl ((S)-1-amino-1-oxo-3-((S)-2-oxopiperidin-3-yl)propan-2-yl)carbamate (237 mg). This was added to a flask containing Pd(COCF) (28 mg) and MeCN (5 mL). Water (2 mL) and 2,2-dichloroacetonitrile (1.3 mL) were then added. After purging with nitrogen gas, the flask was heated to 60° C. for 2 hours. The reaction mixture was diluted with EtOAc and washed with water, then with brine. The organic extract was concentrated, and the residue was purified on silica gel to give tert-butyl ((S)-1-cyano-2-((S)-2-oxopiperidin-3-yl)ethyl)carbamate (88 mg).

[0183] Step 4: A vial was charged with tert-butyl ((S)-1-cyano-2-((S)-2-oxopiperidin-3-yl)ethyl)carbamate (88 mg) and DCM (1 mL). TFA (2 mL) was then added. After 1 h, the volatiles were removed and the product, (S)-2-amino-3-((S)-2-oxopiperidin-3-yl)propanenitrile 2,2,2-trifluoroacetate, was used without further purification.

[0184] Step 5: A vial was charged with (1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid (50 mg), (S)-2-amino-3-((S)-2-oxopiperidin-3-yl)propanenitrile 2,2,2-trifluoroacetate (20 mg), DMF (1 mL), and DIPEA (0.1 mL). HATU (40 mg) was then added. After 30 min, the reaction mixture was purified by RPHPLC to give the product (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopiperidin-3-yl)ethyl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (3 mg). ESI MS m / z=478.1 [M+H] + .

[0185] Example 8: Synthesis of (1S,3aR,6aS)-N-((S)-4-(cyclohexylamino)-3,4-dioxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0186] Step 1: A vial was charged with (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide (58 mg) and DCM (1 mL). The flask was cooled to 0° C. under a nitrogen atmosphere. Acetic acid (0.3 mL) was then added as a solution in DCM (1 mL). Isocyanocyclohexane (0.3 mL) was then added and the reaction was allowed to reach room temperature. After 2 h, the volatiles were removed. The residue was dissolved in MeOH (1 mL) and cooled to −40° C. Potassium carbonate (23 mg) was added and the reaction was warmed to 0° C. Water (0.2 mL) was then added, followed by 3 M aqueous HCl (0.5 mL). The product was extracted with ethyl acetate and concentrated. The residue was purified on silica gel to give the product (1S,3aR,6aS)-N-((2S)-4-(cyclohexylamino)-3-hydroxy-4-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (39 mg). ESI MS m / z=630.1 [M+H] + .

[0187] Step 2: A vial was charged with (1S,3aR,6aS)-N-((2S)-4-(cyclohexylamino)-3-hydroxy-4-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (39 mg), DCM (1 mL), and Hunig's base (0.033 mL) at 0° C. Py-SO (30 mg) was added as a solution in DMSO (1 mL). An additional portion of Py-SO (30 mg) in Hunig's base (0.033 mL) and DMSO (1 mL) was added. The reaction mixture was diluted with ethyl acetate and water. The organic layer was removed, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were concentrated, and the residue was crystallized twice from diethyl ether:THF to give the product (1S,3aR,6aS)-N-((S)-4-(cyclohexylamino)-3,4-dioxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (2 mg). ESI MS m / z=628.1 [M+H] + .

[0188] Example 9: Synthesis of (1S,3aR,6aS)-N-((S)-4-(cyclohexylamino)-3,4-dioxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0189] The synthesis of Example 9 was similar in nature to the synthesis of Example 8, with the following changes: 1. In step 1, (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide was used in place of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-N-((S)-1-oxo-3-((S)-2-oxopyrrolidin-3-yl)propan-2-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide.

[0190] Characterization data for Example 9 was obtained: ESI MS m / z=592.1 [M+H] + .

[0191] Example 10: Synthesis of (1S,3aR,6aS)-N-((S)-4-hydroxy-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0192] Step 1: A vial was charged with (1S,3aR,6aS)—N-((S)-4-chloro-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (99 mg), cesium fluoride (67 mg), 2-oxo-2-phenylacetic acid (38 mg), and DMF (2 mL). The flask was heated to 65° C. for 75 minutes. After cooling to room temperature, the reaction mixture was partitioned between ethyl acetate and water. The product was extracted with ethyl acetate, and the combined organics were concentrated. The product, (S)-3-((1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butyl 2-oxo-2-phenylacetate, was used in the next step without further purification.

[0193] Step 2: A vial was charged with (S)-3-((1S,3aR,6aS)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamido)-2-oxo-4-((S)-2-oxopyrrolidin-3-yl)butyl 2-oxo-2-phenylacetate (121 mg) and MeOH (3 mL). Potassium carbonate (27 mg) was then added. After 1 h, 1 M aqueous HCl was added (0.5 mL). The reaction mixture was filtered and concentrated. The residue was purified by RP-HPLC to give the product (1S,3aR,6aS)-N-((S)-4-hydroxy-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-methoxy-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide (3 mg). ESI MS m / z = 497.1 [M+H] + .

[0194] Example 11: Synthesis of (1S,2S,5R)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0195] The synthesis of Example 11 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 7, (1S,2S,5R)-3-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid was used in place of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid.

[0196] Characterization data for Example 11 was obtained: ESI MS m / z=500.1 [M+H] + .

[0197] Example 12: Synthesis of (S)—N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-1-(4-(difluoromethoxy)-1H-indole-2-carbonyl)pyrrolidine-2-carboxamide [ka]

[0198] The synthesis of Example 12 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 7, (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid was replaced with (4-(difluoromethoxy)-1H-indole-2-carbonyl)-L-proline.

[0199] Characterization data for Example 12 was obtained: ESI MS m / z=460.1 [M+H] + .

[0200] Example 13: Synthesis of (1S,3aR,6aS)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-2-(4,6-difluoro-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide. [ka]

[0201] The synthesis of Example 13 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 7, (1S,3aR,6aS)-2-(4,6-difluoro-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid was used in place of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid.

[0202] Characterization data for Example 13 was obtained: ESI MS m / z=470.1 [M+H] + .

[0203] Example 14: Synthesis of (1S,3aR,6aS)-2-(4-chloro-1H-indole-2-carbonyl)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)octahydrocyclopenta[c]pyrrole-1-carboxamide. [ka]

[0204] The synthesis of Example 14 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 7, (1S,3aR,6aS)-2-(4-chloro-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid was used in place of (1S,3aR,6aS)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxylic acid.

[0205] Characterization data for Example 14 was obtained: ESI MS m / z=469.1 [M+H] + .

[0206] Example 15: Synthesis of (1S,3aR,6aS)-N-((S)-4-(benzylamino)-3,4-dioxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)-2-(4-(difluoromethoxy)-1H-indole-2-carbonyl)octahydrocyclopenta[c]pyrrole-1-carboxamide [ka]

[0207] The synthesis of Example 15 was similar in nature to the synthesis of Example 8, with the following changes: 1. In step 1, (isocyanomethyl)benzene was used in place of isocyanocyclohexane.

[0208] Characterization data for Example 15 was obtained: ESI MS m / z=636.1 [M+H] + .

[0209] Example 16: Synthesis of (1R,2S,5S)—N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-(5,7-difluoro-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0210] The synthesis of Example 16 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 5,7-difluoro-1H-indole-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0211] Characterization data for Example 16 was obtained: ESI MS m / z=470.1 [M+H] + .

[0212] Example 17: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-(5,6-difluoro-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0213] The synthesis of Example 17 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 5,6-difluoro-1H-indole-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0214] Characterization data for Example 17 was obtained: ESI MS m / z=470.1 [M+H] + .

[0215] Example 18: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-(4,6-difluoro-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0216] The synthesis of Example 18 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 4,6-difluoro-1H-indole-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0217] Characterization data for Example 18 was obtained: ESI MS m / z=470.1 [M+H] + .

[0218] Example 19: Synthesis of (2S,4S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-1-(4-(difluoromethoxy)-1H-indole-2-carbonyl)-4-phenoxypyrrolidine-2-carboxamide [ka]

[0219] The synthesis of Example 19 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 5, ethyl (2S,4S)-4-phenoxypyrrolidine-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0220] Characterization data for Example 19 was obtained: ESI MS m / z=552.1 [M+H] + .

[0221] Example 20: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-(4,7-difluoro-1H-indole-2-carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0222] The synthesis of Example 20 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 4,7-difluoro-1H-indole-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0223] Characterization data for Example 20 was obtained: ESI MS m / z=470.1 [M+H] + .

[0224] Example 21: Synthesis of (1R,2S,5S)-3-(4-chloro-1H-indole-2-carbonyl)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0225] The synthesis of Example 21 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 4-chloro-1H-indole-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0226] Characterization data for Example 21 was obtained: ESI MS m / z=469.1 [M+H] + .

[0227] Example 22: Synthesis of (1R,2S,5S)-3-(4-chlorobenzofuran-2-carbonyl)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0228] The synthesis of Example 22 was similar in nature to the synthesis of Example 3, with the following changes: 1. In step 4, 4-chlorobenzofuran-2-carboxylic acid was used instead of 4-(difluoromethoxy)-1H-indole-2-carboxylic acid. 2. In step 5, ethyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was used instead of ethyl (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carboxylate hydrochloride.

[0229] Characterization data for Example 22 was obtained: ESI MS m / z=470.1 [M+H] + .

[0230] Example 23: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-2-(cyclopentanecarboxamido)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0231] Step 1: A vial was charged with Boc-L-tert-leucine (1.55 g), DCM (20 mL), and DMF (5 mL). The vial was cooled to 0° C., and then HATU (2.2 g) was added. Hunig's base (1.8 g) was then added. The yellow suspension was stirred for 2 minutes, after which methyl (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate hydrochloride was added. Hunig's base (1.8 g) was then added, and the reaction was allowed to reach room temperature. After 2 hours, the reaction mixture was diluted with ethyl acetate and partitioned with water. The product was extracted with ethyl acetate. The combined organic layers were concentrated, and the residue was purified on silica gel to give the product, methyl (1R,2S,5S)-3-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate (1.5 g). ESI MS m / z=383.1 [M+H] + .

[0232] Step 2: A vial was charged with methyl (1R,2S,5S)-3-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylate (300 mg) and MeOH (4 mL). The vial was cooled to 0 °C, and then 2.5 M aqueous LiOH (2 mL) was added. After 3.5 h, the reaction mixture was concentrated. Next, 4 M HCl in dioxane (5.5 mL) was added, and the reaction mixture was stirred for 1 h. The reaction mixture was then concentrated to provide the product (1R,2S,5S)-3-((S)-2-amino-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid, which was used crude without further purification. ESI MS m / z=269.1 [M+H] + .

[0233] Step 3: A vial was charged with (1R,2S,5S)-3-((S)-2-amino-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid (304 mg), triethylamine (0.42 mL), and MeCN (6 mL). The vial was stirred at 0°C, and then cyclopentanecarbonyl chloride (147 mg) was added dropwise under a nitrogen atmosphere. After 1 h, DCM (25 mL) was added. The reaction mixture was partitioned with water (25 mL) and stirred at room temperature. After 10 min, the organic layer was removed. The aqueous layer was acidified to pH 5 with 1 M aqueous HCl. The product was extracted with ethyl acetate, dried over sodium sulfate, filtered, and concentrated. The product (1R,2S,5S)-3-((S)-2-(cyclopentanecarboxamido)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid was used in crude form without further purification. ESI MS m / z=365.1 [M+H] + .

[0234] Step 4: A vial was charged with (1R,2S,5S)-3-((S)-2-(cyclopentanecarboxamido)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid (130 mg) and DMF (5 mL). DIEA (139 mg) was then added. HATU (274 mg) was then added at room temperature, and the reaction mixture was stirred for 10 minutes. tert-Butyl ((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)carbamate (155 mg) was then added. After 1 hour, the reaction mixture was poured into water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by RP-HPLC to give the product (1R,2S,5S)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-2-(cyclopentanecarboxamido)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide (45 mg). ESI MS m / z=500.1 [M+H] + .1H NMR(300 MHz,DMSO-d6)δ 8.96(d,J=8.5 Hz,1H),7.76(d,J=9.2 Hz,1H),7.66(s,1H),5.04-4.89(m,1H),4.38(d,J=9.2 Hz,1H),4.12(s,1H),3.91-3.76(m,2H),3.20-2.98(m,2H),2.82-2.66(m,1 H),2.47-2.37(m,1H),2.23-2.01(m,2H),1.76-1.42(m,11H),1.28(d,J=7.6 Hz,1H),1.02(s,3H),0.94(s,9H),0.82(s,3H).

[0235] Example 24: Synthesis of (1R,2S,5S)—N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-3,3-dimethyl-2-(2-phenylacetamido)butanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0236] The synthesis of Example 24 was similar in nature to that of Example 23, with the following changes: 1. In step 3, 2-phenylacetyl chloride was used instead of cyclopentanecarbonyl chloride.

[0237] Characterization data for Example 24 was obtained: ESI MS m / z=522.1 [M+H] + . 1H NMR(300 MHz,DMSO-d6)δ 8.97(d,J=8.6 Hz,1H),8.13(d,J=9.0 Hz,1H),7.66(s,1H),7.30-7.14(m,6H),5.03-4.88(m,1H),4.36(d,J=6.0 Hz,1H),4.11(s,1H),3.90-3.81(m,1H),3.74(d,J=10.4 Hz,1H),3.56-3.41(m,2H),3.19-2.99(m,2H),2.27-1.99(m,3H),1.77-1.63(m,2H),1.54-1.48(m,1H),1.27(d,J=7.6 Hz,1H),1.01(s,3H),0.91(s,9H),0.74(s,3H).

[0238] Example 25: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-2-(cyclohexanecarboxamido)-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0239] The synthesis of Example 25 was similar in nature to that of Example 23, with the following changes: 1. In step 3, cyclohexanecarbonyl chloride was used instead of cyclopentanecarbonyl chloride.

[0240] Characterization data for Example 25 was obtained: ESI MS m / z=514.1 [M+H] + . 1H NMR(300 MHz,DMSO-d6)δ 8.95(d,J=8.5 Hz,1H),7.73-7.62(m,2H),5.00-4.90(m,1H),4.35(d,J=9.2 Hz,1H),4.11(s,1H),3.90-3.77(m,4H),3.19-2.99(m,2H),2.45-2.24(m,2H),2.22-2. 00(m,2H),1.75-1.47(m,8H),1.24-1.10(m,4H),1.01(s,3H),0.93(s,9H),0.82(s,3H).

[0241] Example 26: Synthesis of (1R,2S,5S)—N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-3,3-dimethyl-2-propionamidobutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0242] The synthesis of Example 26 was similar in nature to that of Example 23, with the following changes: 1. In step 3, propionyl chloride was used instead of cyclopentanecarbonyl chloride.

[0243] Characterization data for Example 26 was obtained: ESI MS m / z=460.1 [M+H] +. 1H NMR(300 MHz,DMSO-d6)δ 8.96(d,J=8.6 Hz,1H),7.79(d,J=8.9 Hz,1H),7.65(s,1H),5.01-4.90(m,1H),4.34(d,J=9.0 Hz,1H),4.11(s,1H),3.89-3.76(m,2H),3.08(dq,J=16.4,9.4 Hz,2H),2.45-2.35(m,1H),2.19-2.01(m,4H),1.78-1.63(m,2H),1.58-1.50(m,1H),1.28(d,J=7.7 Hz,1H),1.02(s,3H),0.94(t,J=6.0 Hz,1H),0.92(s,9H),0.84(s,3H).

[0244] Example 27: Synthesis of (1R,2S,5S)—N—((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-2-(isobutyramido-3,3-dimethylbutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide. [ka]

[0245] The synthesis of Example 27 was similar in nature to that of Example 23, with the following changes: 1. In step 3, isobutyryl chloride was used instead of cyclopentanecarbonyl chloride.

[0246] Characterization data for Example 27 was obtained: ESI MS m / z=474.1 [M+H] +. 1H NMR(300 MHz,DMSO-d6)δ 8.95(d,J=8.5 Hz,1H),7.76(d,J=9.2 Hz,1H),7.65(s,1H),5.01-4.89(m,1H),4.39(d,J=9.2 Hz,1H),4.12(s,1H),3.91-3.75(m,2H),3.20-2.97(m,2H),2.58(dd,J=13.6,6.8 Hz,1H),2.46-2.36(m,1H),2.23-2.02(m,2H),1.78-1.63(m,2H),1.56-1.49(m,1H),1.28(d,J=7.6 Hz,1H),1.02(s,3H),1.00-0.85(m,15H),0.82(s,3H).

[0247] Example 28: Synthesis of (1R,2S,5S)—N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-3-((S)-3,3-dimethyl-2-pivalamidobutanoyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0248] The synthesis of Example 28 was similar in nature to that of Example 23, with the following changes: 1. In step 3, pivaloyl chloride was used instead of cyclopentanecarbonyl chloride.

[0249] Characterization data for Example 28 was obtained: ESI MS m / z=488.1 [M+H] +. 1H NMR(300 MHz,DMSO-d6)δ 8.95(d,J=8.5 Hz,1H),7.67(s,1H),6.88(d,J=7.3 Hz,1H),5.02-4.89(m,1H),4.46(s,1H),4.13(s,1H),3.93-3.81(m,1H),3.73(d,J=10.3 Hz,1H),3.21-2.98(m,2H),2.45-2.36(m,1H),2.19-2.04(m,2H),1.79-1.63(m,2H),1.58-1.49(m,1H),1.29(d,J=7.6 Hz,1H),1.08(s,9H),1.02(s,3H),0.92(s,9H),0.81(s,3H).

[0250] Example 29: Synthesis of (1R,2S,5S)-3-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-((S)-1-cyano-2-((S)-2-oxopyrrolidin-3-yl)ethyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxamide [ka]

[0251] The synthesis of Example 29 was similar in nature to that of Example 23, with the following changes: 1. In step 3, acetyl chloride was used instead of cyclopentanecarbonyl chloride.

[0252] Characterization data for Example 29 was obtained: ESI MS m / z=446.1 [M+H] +. 1H NMR(300 MHz,DMSO-d6)δ 8.97(d,J=8.6 Hz,1H),7.87(d,J=8.7 Hz,1H),7.65(brs,1H),4.99-4.92(m,1H),4.30(d,J=8.7 Hz,1H),4.11(s,1H),3.90-3.75(m,2H),3.18-2.99(m,2H),2.45-2.35(m,1H),2. 20-1.98(m,2H),1.83(s,3H),1.79-1.62(m,2H),1.57-1.50(m,1H),1.28(d,J=7.7 Hz,1H),1.02(s,3H),0.94(s,9H),0.86(s,3H).

[0253] biological activity SARS-CoV-2 3C-like (3CL) protease fluorescence assay (FRET): Recombinant SARS-CoV-2 3CL protease was expressed and purified. The TAMRA-SITSAVLQSGFRKMK-Dabcyl-OH peptide 3CL pro substrate was synthesized. Black, small-volume, round-bottom, 384-well microplates were used. In a typical assay, 0.85 μL of test compound was dissolved in DMSO and then incubated with SARS-CoV-2 3CL protease (10 nM) in 10 μL of assay buffer (50 mM HEPES [pH 7.5], 1 mM DTT, 0.01% BSA, 0.01% Triton-X 100) for 30 minutes at room temperature. Next, 10 μL of 3CL protease substrate (40 μM) in assay buffer was added, and the assay was monitored continuously for 1 h on an Envision multimode plate reader operating in fluorescence kinetic mode with excitation at 540 nm and emission at 580 nm at room temperature. Compound (DMSO only) and enzyme controls were routinely included on each plate. All experiments were performed in duplicate.

[0254] Data Analysis: SARS-CoV-2 3CL-protease enzyme activity was measured as the initial velocity of the linear phase (RFU / s) and normalized to control samples DMSO (100% activity) and no enzyme (0% activity) to determine percent residual activity at various concentrations of test compound (0-10 μM). Data were fitted to a normalized activity (variable slope) versus concentration fit in GraphPad Prism 7 to calculate IC 50 All experiments were performed in duplicate and IC 50 Ranges are reported as follows: A<0.1 μM; B 0.1-1 μM; C>1 μM. [Table 7]

[0255] While the present invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

1. A compound represented by formula (VI-6a), or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 In the above formula, X is —CN; and A is selected from: 1) optionally substituted -C １ -C ８ Alkyl; 2) optionally substituted -C ３ -C １２ cycloalkyl; 3) optionally substituted 3- to 12-membered heterocycloalkyl; 4) optionally substituted aryl; and 5) optionally substituted heteroaryl.

2. The compound of claim 1, wherein A is optionally substituted -C 1 -C 8 alkyl.

3. The compound of claim 1, wherein A is an optionally substituted heteroaryl.

4. The compound of claim 1, wherein A is selected from the group consisting of: 【Chemistry 2】 、 However, each is optionally substituted.

5. The compound of claim 1, which is a compound represented by formula (X-e), or a pharmaceutically acceptable salt thereof: 【Chemistry 2】 In the above formula, X is —CN and A is selected from the group consisting of: 【Transformation 3】 。

6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

7. The pharmaceutical composition according to claim 6 for use in treating or preventing coronavirus infection.

8. The pharmaceutical composition described in claim 7, wherein the coronavirus is 229E, NL63, OC43, HKU1, SARS-CoV or MERS coronavirus.

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