WO2010033466A1

WO2010033466A1 - Macrocyclic inhibitors of hepatitis c protease

Info

Publication number: WO2010033466A1
Application number: PCT/US2009/056859
Authority: WO
Inventors: Juan Manuel Betancort; Michael E. Hepperle; David Alan Campbell; David T. Winn
Original assignee: Phenomix Corporation
Priority date: 2008-09-16
Filing date: 2009-09-14
Publication date: 2010-03-25

Abstract

The invention provides macrocyclic compounds inhibitory to the Hepatitis C viral protease, compositions and combinations including the compounds, methods of treatment of conditions wherein inhibition of the Hepatitis C viral protease is medically indicated, and methods of treatment of a Hepatitis C viral infection in a human patient.

Description

MACROCYCLIC INHIBITORS OF HEPATITIS C PROTEASE

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the priority of U.S. Ser. No. 61/097,414, filed September 16, 2008, the disclosure of which is incorporated herein by reference in its entirety.

BACKGROUND

Hepatitis C virus ("HCV") is the causative agent for hepatitis C, a chronic infection characterized by jaundice, fatigue, abdominal pain, loss of appetite, nausea, and darkening of the urine. HCV, belonging to the hepacivirus genus of the Flaviviriae family, is an enveloped, single-stranded positive-sense RNA-containing virus. The long-term effects of hepatitis C infection as a percentage of infected subjects include chronic infection (55-85%), chronic liver disease (70%), and death (1-5%). Furthermore, HCV is the leading indication for liver transplant. In chronic infection, there usually presents progressively worsening liver inflammation, which often leads to more severe disease states such as cirrhosis and hepatocellular carcinoma.

The HCV genome (Choo et al, Science 1989, 244, 359-362; Simmonds et al., Hepatology 1995, 21, 570-583) is a highly variable sequence exemplified by GenBank accession NC_004102 as a 9646 base single-stranded RNA comprising the following constituents at the parenthetically indicated positions: 5' NTR (i.e., non-transcribed region) (1-341); core protein (i.e., viral capsid protein involved in diverse processes including viral morphogenesis or regulation of host gene expression) (342-914); El protein (i.e., viral envelope) (915-1490); E2 protein (i.e., viral envelope) (1491-2579); p7 protein (2580- 2768); NS2 protein (i.e., non-structural protein 2) (2769-3419); NS3 protease (3420-5312); NS4a protein (5313-5474); NS4b protein (5475-6257); NS5a protein (6258-7601); NS5b RNA-dependent RNA polymerase (7602-9372); and 3' NTR (9375-9646). Additionally, a 17-kDalton -2/+1 frameshift protein, "protein F", comprising the joining of positions (342-369) with (371-828) may provide functionality originally ascribed to the core protein. The NS3 (i.e., non-structural protein 3) protein of HCV exhibits serine protease activity, the N-terminus of which is produced by the action of a NS2- NS3 metal-dependent protease, and the C-terminus of which is produced by auto-pro teoly sis. The HCV NS3 serine protease and its associated cofactor, NS4a, process all of the other non-structural viral proteins of HCV. Accordingly, the HCV NS3 protease is essential for viral replication.

Several compounds have been shown to inhibit the hepatitis C serine protease, but all of these have limitations in relation to the potency, stability, selectivity, toxicity, and/or pharmacodynamic properties. Such compounds have been disclosed, for example, in published U.S. Patent Application Nos. 2004/0266731, 2002/0032175, 2005/0137139, 2005/0119189, and 2004/0077600A1, and in published PCT patent applications WO 2005/037214 and WO 2005/035525. Macrocyclic inhibitors of HCV have been disclosed by the inventors herein in patent application U.S. Serial No. 60/883,946.

SUMMARY

The present invention is directed to compounds of Formula I, the compounds being adapted to inhibit the viral protease NS3 of the Hepatitis C Virus (HCV), to the use of compounds of compounds of Formula I in the treatment of malconditions for which inhibition of HCV protease is medically indicated, such as in the treatment of HCV infections, and to pharmaceutical compositions and combinations including a compound of Formula I as defined herein. The compounds of Formula I are adapted to bind to, and thus block the action of, an HCV-encoded protease enzyme that is required by the virus for the production of intact, mature, functional viral proteins from the viral polyprotein as translated from the viral RNA, and therefore for the formation of infectious particles, and ultimately for viral replication. The compounds of the invention are mimics or analogs of the peptide domain immediately N-terminal of the substrate site where the viral protease cleaves its native substrate viral polyprotein, and are believed to bind to and inhibit the protease by virtue of this mimicry or analogy. Various embodiments of the invention provide a compound of Formula I:

I

and stereoisomers, solvates, hydrates, tautomers, prodrugs, salts, pharmaceutically acceptable salts, and mixtures thereof, wherein: A is

R^c at each occurrence is independently H, or a substituted or unsubstituted alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, or heteroarylalkyl; wherein any carbon atom or nitrogen atom can be substituted with a J group; or two R^c groups together with a nitrogen atom to which they are bound form together with the nitrogen atom a 5- 11 membered mono- or bicyclic heterocyclic ring system that is unsubstituted or is substituted with 1-3 J groups;

B is CH₂;

R¹, R^la, R² and R^2a are independently H , halo, or alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J;

X is a bond, O, S, CH(R³) or N(R⁴);

Y is a bond, CH₂, C(O), C(O)C(O), S(O), S(O)₂ or S(O)(NR⁴); provided that when X and Y are both bonds, taken together they form a single bond: and

Z is hydrogen, alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, OR⁵, or N(R⁵)₂, wherein a heterocyclyl or heteroaryl group can be bonded by a carbon atom or by a heteroatom, wherein any carbon atom or nitrogen atom is unsubstituted or is substituted with J;

R³ is hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J;

R⁴ is independently at each occurrence hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J, or aralkanoyl, heteroaralkanoyl, C(O)R³ , SO ₂R³ or carboxamido, wherein any aralkanoyl or heteroaralkanoyl is substituted with 0-3 J groups;

R⁵ is independently at each occurrence hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, or heteroarylalkyl, wherein any alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, or heteroarylalkyl is substituted with 0-3 J, or two R groups which are bound to a nitrogen atom can form together with the nitrogen atom a 5-11 membered mono- or bicyclic heterocyclic ring system wherein the heterocyclic ring system is substituted with 0-3 J groups and can contain 0-3 additional heteroatoms selected from the group consisting of O, N, NR', S, S(O), and S(O)₂. D is CR'; m is 0, 1, 2, 3 or 4; n is O, 1, 2, 3 or 4; p is 1, 2, 3, or 4; L is O, S, C₂, C₂H₂ or C₂H₄;

M is O, S, S(O), S(O)₂;

J is halogen, R', OR', CN, CF₃, OCF₃, O, S, C(O), S(O), methylenedioxy, ethylenedioxy, (CH₂)₀-_pN(R')₂, (CH₂)₀-_pSR', (CH₂)₀-_pS(O)R', (CH₂V_pS(O)₂R', (CH₂)o-_pS(0)₂N(R')₂, (CH₂V_pSO₃R', (CH₂V_pC(O)R', (CH₂V_pC(O)C(O)R, (CH₂VpC(O)CH₂C(O)R, (CH₂ VpC(S)R', (CH₂ VpC(O)OR', (CH₂VpOC(O)R, (CH₂V_pC(O)N(R)₂, (CH₂V_pOC(O)N(R)₂, (CH₂V_pC(S)N(R)₂, (CH₂)₀-_pNH- C(O)R, (CH₂)o-_pN(R')N(R')C(0)R', (CH₂)₀-_pN(R)N(R')C(O)OR', (CH₂V _pN(R)N(R)CON(R')₂, (CH₂V_pN(R)SO₂R', (CH₂)₀-_pN(R')SO₂N(R)₂, (CH₂V pN(R)C(0)0R, (CH₂)o-_pN(R')C(0)R', (CH₂VpN(R)C(S)R, (CH₂V pN(R)C(0)N(R')₂, (CH₂)o-_pN(R')C(S)N(R)₂, (CH₂ V_pN(COR)COR, (CH₂V _pN(0R)R', (CH₂)o-_pC(=NH)N(R')₂, (CH₂)₀-_pC(O)N(OR)R', or (CH₂V pC(=N0R)R'; wherein, each R is independently at each occurrence hydrogen, (Ci-Ci₂)-alkyl, (C₂-Ci₂)-alkenyl, (C₂-Ci₂)-alkynyl, (C₃-Cio)-cycloalkyl, (C₃-Cio)-cycloalkenyl, [(C₃-Cio)cycloalkyl or (C₃-Ci₀)-cycloalkenyl]-[(Ci-Ci₂)-alkyl or (C₂-Ci₂)- alkenyl or (C₂-Ci₂)-alkynyl], (C₆-C₁₀)-aryl, (C₆-C₁₀)-aryl-[(C₁-C₁₂)-alkyl or (C₂- Ci₂)-alkenyl or (C₂-C i₂)-alkynyl], mono- or bicyclic 3-10 membered heterocyclyl, mono- or bicyclic 3-10 membered heterocyclyl-[(Ci-Ci₂)-alkyl or (C₂-Ci₂)-alkenyl or (C₂-Ci₂)-alkynyl], mono- or bicyclic 5-10 membered heteroaryl, or mono- or bicyclic 5-10 membered heteroaryl-[(Ci-Ci₂)-alkyl or (C₂-Ci₂)-alkenyl or (C₂-Ci₂)-alkynyl], wherein R' is substituted with 0-3 substituents selected independently from J; or, when two R' are bound to a nitrogen atom or to two adjacent nitrogen atoms, the two R groups together with the nitrogen atom or atoms to which they are bound can form a 3- to 8-membered monocyclic heterocyclic ring, or an 8- to 20-membered, bicyclic or tricyclic, heterocyclic ring system, wherein any ring or ring system can further contain 1 -3 additional heteroatoms selected from the group consisting of N, NR', O, S, S(O) and S(O)₂, wherein each ring is substituted with 0-3 substituents selected independently from J; wherein, in any bicyclic or tricyclic ring system, each ring is linearly fused, bridged, or spirocyclic, wherein each ring is either aromatic or nonaromatic, wherein each ring can be fused to a (Ce-Cio)aryl, mono- or bicyclic 5-10 membered heteroaryl, (C₃-Cio)cycloalkyl or mono- or bicyclic 3-10 membered heterocyclyl;

T is R⁶, alkyl-R⁶, alkenyl-R⁶, or alkynyl-R⁶; and

R⁶ is independently at each occurrence hydrogen, alkyl, alkoxy, aryl, aralkyl, cycloalkyl, cycloalkenyl, [cycloalkyl or cycloalkenyl]- [alkyl or alkenyl], heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any R⁶ except hydrogen is substituted with 0-3 J groups.

Various embodiments of the invention are directed to methods for synthesis of a compound of Formula I.

Various embodiments of the invention are further directed to pharmaceutical compositions comprising a compound of Formula I and a suitable excipient.

Various embodiments of the invention are further directed to pharmaceutical combinations comprising a compound of Formula I in a therapeutically effective amount and a second medicament in a therapeutically effective amount. A pharmaceutical combination of the invention may be formulated as a pharmaceutical composition of the invention.

Various embodiments of the invention are further directed to methods of treatment of a HCV infection in a patient in need thereof, or in a patient when inhibition of an HCV viral protease is medically indicated, comprising administering a therapeutically effective amount of a compound of Formula I to the patient.

Various embodiments of the invention are further directed to methods of treatment of a HCV infection in a patient in need thereof, or in a patient when inhibition of an HCV viral protease is medically indicated, comprising administering a therapeutically effective amount of a compound of Formula I to the patient in conjunction with administering an effective amount of a second medicament to the patient. The second medicament can include another antiviral agent, an antibiotic, or a medicament for alleviating symptoms of a Hepatitis C infection. Various embodiments of the invention are directed to use of an inventive compound in preparation of a medicament for the treatment of an HCV infection in a patient. In further embodiments, an effective amount of a second bioactive agent can be used in preparation of a medicament for treatment of an HCV infection in a patient.

Various embodiments of the invention provide an inventive compound for use in combination with an effective amount of a second medicament in treatment of an HCV infection in a patient.

DETAILED DESCRIPTION

The terms "HCV NS3 serine protease", "HCV NS3 protease", "NS3 serine protease", and "NS3 protease" denote all active forms of the serine protease encoded by the NS3 region of the hepatitis C virus, including all combinations thereof with other proteins in either covalent or non-covalent association. For example, other proteins in this context include without limitation the protein encoded by the NS4a region of the hepatitis C virus. Accordingly, the terms "NS3/4a" and "NS3/4a protease" denote the NS3 protease in combination with the HCV NS4a protein.

The term "other type(s) of therapeutic agents" as employed herein refers to one or more antiviral agents, other than HCV NS3 serine protease inhibitors of the invention.

"Subject" as used herein, includes mammals such as humans, non-human primates, rats, mice, dogs, cats, horses, cows and pigs.

The term "treatment" is defined as the management and care of a patient for the purpose of combating the disease, condition, or disorder and includes administering a compound of the present invention to prevent the onset of the symptoms or complications, or alleviating the symptoms or complications, or eliminating the disease, condition, or disorder.

"Treating" within the context of the instant invention means an alleviation of symptoms associated with a disorder or disease, or inhibition of further progression or worsening of those symptoms, or prevention or prophylaxis of the disease or disorder. Thus, treating a hepatitis C viral infection includes slowing, halting or reversing the growth of the virus and/or the control, alleviation or prevention of symptoms of the infection. Similarly, as used herein, an "effective amount" or a "therapeutically effective amount" of a compound of the invention refers to an amount of the compound that alleviates, in whole or in part, symptoms associated with the disorder or condition, or halts or slows further progression or worsening of those symptoms, or prevents or provides prophylaxis for the disorder or condition. In particular, a "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result by inhibition of HCV NS3 serine protease activity. A therapeutically effective amount is also one in which any toxic or detrimental effects of compounds of the invention are outweighed by the therapeutically beneficial effects. For example, in the context of treating HCV infection, a therapeutically effective amount of a HCV NS3 serine protease inhibitor of the invention is an amount sufficient to control HCV viral infection.

All chiral, diastereomeric, racemic forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated. Compounds used in the present invention include enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions. Both racemic and diastereomeric mixtures, as well as the individual optical isomers can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these are all within the scope of the invention.

By "chemically feasible" is meant a bonding arrangement or a compound where the generally understood rules of organic structure are not violated; for example a structure within a definition of a claim that would contain in certain situations a pentavalent carbon atom that would not exist in nature would be understood to not be within the claim.

When a substituent is specified to be an atom or atoms of specified identity, "or a bond", a configuration is referred to when the substituent is "a bond" that the groups that are immediately adjacent to the specified substituent are directly connected to each other by a chemically feasible bonding configuration.

The term "amino protecting group" or "N-protected" as used herein refers to those groups intended to protect an amino group against undesirable reactions during synthetic procedures and which can later be removed to reveal the amine. Commonly used amino protecting groups are disclosed in Protective Groups in Organic Synthesis, Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, New York, NY, (3rd Edition, 1999). Amino protecting groups include acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2- bromoacetyl, trifluoroacetyl, trichloroacetyl, o-nitrophenoxyacetyl, α- chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the like; alkoxy- or aryloxy-carbonyl groups (which form urethanes with the protected amine) such as benzyloxycarbonyl (Cbz), p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2- nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4- dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4- dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5- dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, l-(p- biphenylyl)-l-methylethoxycarbonyl, α,α-dimethyl-3,5- dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl, 2- trimethylsilylethyloxycarbonyl (Teoc), phenoxycarbonyl, 4- nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; aralkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and the like. Amine protecting groups also include cyclic amino protecting groups such as phthaloyl and dithiosuccinimidyl, which incorporate the amino nitrogen into a heterocycle. Typically, amino protecting groups include formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, Alloc, Teoc, benzyl, Fmoc, Boc and Cbz. It is well within the skill of the ordinary artisan to select and use the appropriate amino protecting group for the synthetic task at hand. The term "hydroxyl protecting group" or "O-protected" as used herein refers to those groups intended to protect an OH group against undesirable reactions during synthetic procedures and which can later be removed to reveal the amine. Commonly used hydroxyl protecting groups are disclosed in Protective Groups in Organic Synthesis, Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, New York, NY, (3rd Edition, 1999). Hydroxyl protecting groups include acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2- chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4- bromobenzoyl, 4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the like; acyloxy groups (which form urethanes with the protected amine) such as benzyloxycarbonyl (Cbz), p- chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p- nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4- dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5- dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, l-(p- biphenylyl)- 1 -methylethoxycarbonyl, α,α-dimethyl-3,5- dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl, 2- trimethylsilylethyloxycarbonyl (Teoc), phenoxycarbonyl, 4- nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; aralkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and the like. It is well within the skill of the ordinary artisan to select and use the appropriate hydroxyl protecting group for the synthetic task at hand.

In general, "substituted" refers to an organic group as defined herein in which one or more bonds to a hydrogen atom contained therein are replaced by one or more bonds to a non-hydrogen atom such as, but not limited to, a halogen (i.e., F, Cl, Br, and I); an oxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboyxlate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxylamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR', OC(O)N(R')₂, CN, CF₃, OCF₃, R', O, S, C(O), S(O), methylenedioxy, ethylenedioxy, N(R')₂, SR', SOR, SO₂R, SO₂N(R)₂, SO₃R, C(O)R, C(O)C(O)R, C(O)CH₂C(O)R, C(S)R', C(O)OR, OC(O)R, C(O)N(R)₂, 0C(0)N(R')₂, C(S)N(R')₂, (CH₂V₂NHC(O)R, N(R)N(R)C(O)R, N(R')N(R')C(0)0R', N(R')N(R')CON(R)₂, N(R')S0₂R', N(R)SO₂N(R)₂, N(R')C(0)0R', N(R')C(O)R', N(R')C(S)R, N(R)C(0)N(R')₂, N(R')C(S)N(R)₂, N(COR)COR', N(0R')R, C(=NH)N(R')₂, C(O)N(OR)R', or C(=N0R)R' wherein R' can be hydrogen or a carbon-based moiety, and wherein the carbon-based moiety can itself be further substituted. When a substituent is monovalent, such as, for example, F or Cl, it is bonded to the atom it is substituting by a single bond. When a substituent is more than monovalent, such as O, which is divalent, it can be bonded to the atom it is substituting by more than one bond, i.e., a divalent substituent is bonded by a double bond; for example, a C substituted with O forms a carbonyl group, C=O, wherein the C and the O are double bonded. Alternatively, a divalent substituent such as O, S, C(O), S(O), or S(O)₂ can be connected by two single bonds to two different carbon atoms. For example, O, a divalent substituent, can be bonded to each of two adjacent carbon atoms to provide an epoxide group, or the O can form a bridging ether group between adjacent or non-adjacent carbon atoms, for example bridging the 1,4-carbons of a cyclohexyl group to form a [2.2.1]- oxabicyclo system. Further, any substituent can be bonded to a carbon or other atom by a linker, such as (CH₂)_n or (CR₂)_n wherein n is 1, 2, 3, or more, and each R' is independently selected.

Substituted alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl groups as well as other substituted groups also include groups in which one or more bonds to a hydrogen atom are replaced by one or more bonds, including double or triple bonds, to a carbon atom, or to a heteroatom such as, but not limited to, oxygen in carbonyl (oxo), carboxyl, ester, amide, imide, urethane, and urea groups; and nitrogen in imines, hydroxyimines, oximes, hydrazones, amidines, guanidines, and nitriles.

Substituted ring groups such as substituted cycloalkyl, aryl, heterocyclyl and heteroaryl groups also include rings and fused ring systems in which a bond to a hydrogen atom is replaced with a bond to a carbon atom. Therefore, substituted cycloalkyl, aryl, heterocyclyl and heteroaryl groups can also be substituted with alkyl, alkenyl, and alkynyl groups as defined herein.

By a "ring system" as the term is used herein is meant a moiety comprising one, two, three or more rings, which can be substituted with non-ring groups or with other ring systems, or both, which can be fully saturated, partially unsaturated, fully unsaturated, or aromatic, and when the ring system includes more than a single ring, the rings can be fused, bridging, or spirocyclic. By "spirocyclic" is meant the class of structures wherein two rings are fused at a single tetrahedral carbon atom, as is well known in the art. Alkyl groups include straight chain and branched alkyl groups and cycloalkyl groups having from 1 to about 20 carbon atoms, and typically from 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec -butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed above, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. Cycloalkyl groups are cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 5, 6, or 7. Cycloalkyl groups further include poly cyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined above. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or

2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term "cycloalkenyl" alone or in combination denotes a cyclic alkenyl group. The terms "carbocyclic" and "carbocycle" denote a ring structure wherein the atoms of the ring are carbon. In some embodiments, the carbocycle has 3 to 8 ring members, whereas in other embodiments the number of ring carbon atoms is 4, 5, 6, or 7. Unless specifically indicated to the contrary, the carbocyclic ring can be substituted with as many as N-I substituents wherein N is the size of the carbocyclic ring with, for example, alkyl, alkenyl, alkynyl, amino, aryl, hydroxy, cyano, carboxy, heteroaryl, heterocyclyl, nitro, thio, alkoxy, and halogen groups, or other groups as are listed above.

(Cycloalkyl)alkyl groups, also denoted cycloalkylalkyl, are alkyl groups as defined above in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a cycloalkyl group as defined above.

Alkenyl groups include straight and branched chain and cyclic alkyl groups as defined above, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=CH(CHs), -CH=C(CH₃)₂, -C(CH₃)=CH₂, -C(CH₃)=CH(CH₃),

-C(CH₂CH₃)=CH₂, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others. Cycloalkenyl groups include cycloalkyl groups having at least one double bond between 2 carbons. Thus for example, cycloalkenyl groups include but are not limited to cyclohexenyl, cyclopentenyl, and cyclohexadienyl groups. Cycloalkenyl groups can have from 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like, provided they include at least one double bond within a ring. Cycloalkenyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined above.

(Cycloalkenyl)alkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a cycloalkenyl group as defined above. Alkynyl groups include straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to -C≡CH, -OC(CH₃), -C≡C(CH₂CH₃), -CH₂C≡CH, -CH₂C≡C(CH₃), and -CH₂C≡C(CH₂CH₃) among others.

Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined above. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or 2-8 substituted naphthyl groups, which can be substituted with carbon or non-carbon groups such as those listed above.

Aralkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined above. Representative aralkyl groups include benzyl and phenylethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl.

Aralkenyl group are alkenyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined above.

Heterocyclyl groups include aromatic and non-aromatic ring compounds containing 3 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S. In some embodiments, heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members. Ring systems can be monocyclic, bicyclic, or tricyclic; for example a 10-membered heterocyclyl within the meaning herein can be composed of two fused six-membered rings, a least one of which must contain at least one heteroatom such as N, O, or S. A heterocyclyl group designated as a C₂-heterocyclyl can be a 5 -ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C₄- heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. A heterocyclyl ring can also include one or more double bonds. A heteroaryl ring is an embodiment of a heterocyclyl group. The phrase "heterocyclyl group" includes fused ring species including those comprising fused aromatic and non-aromatic groups. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups within the meaning herein. The phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. Heterocyclyl groups can be unsubstituted, or can be substituted as discussed above. Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Representative substituted heterocyclyl groups can be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6-substituted, or disubstituted with groups such as those listed above.

Heteroaryl groups are aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group designated as a C₂-heteroaryl can be a 5 -ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C₄-heteroaryl can be a 5 -ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Ring systems can be monocyclic, bicyclic, or tricyclic; for example a 10- membered heteroaryl within the meaning herein can be composed of two fused six-membered aromatic rings, a least one of which must contain at least one heteroatom such as N, O, or S. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed above. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed above.

Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N- hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1- anthracenyl, 2-anthracenyl, 3-anthracenyl), thiophenyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5- imidazolyl), triazolyl (1,2,3-triazol-l-yl, l,2,3-triazol-2-yl l,2,3-triazol-4-yl, l,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2- thiazolyl, 4-thiazolyl, 5 -thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4-pyridazinyl, 5-pyridazinyl), quinolyl (2- quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6- isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl, 6-benzo[b]furanyl, 7- benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro- benzo[b]furanyl), 3-(2,3-dihydro-benzo[b]furanyl), 4-(2,3-dihydro- benzo[b]furanyl), 5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro- benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl (2- benzo[b] thiophenyl, 3 -benzo[b] thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6-benzo[b] thiophenyl, 7-benzo[b]thiophenyl),

2,3-dihydro-benzo[b]thiophenyl, (2-(2,3-dihydro-benzo[b]thiophenyl), 3-(2,3- dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro-benzo[b]thiophenyl), 5-(2,3- dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro-benzo[b]thiophenyl), 7 -(2,3- dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2- benzoxazolyl), benzothiazolyl (1-benzothiazolyl, 2-benzothiazolyl, 4- benzothiazolyl, 5 -benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-l-yl, 5H-dibenz[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine- 5-yl), 10,1 l-dihydro-5H-dibenz[b,f]azepine (10,1 l-dihydro-5H- dibenz[b,f]azepine-l-yl, 10,1 l-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,11- dihydro-5H-dibenz[b,f]azepine-3-yl, 10,l l-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,1 l-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like.

Heterocyclylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group as defined above is replaced with a bond to a heterocyclyl group as defined above. Representative heterocyclyl alkyl groups include, but are not limited to, furan-2-yl methyl, furan-3-yl methyl, pyridine-3-yl methyl, tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl.

Heteroarylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined above.

The term "alkoxy" refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined above. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include one to about 12-20 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group is an alkoxy group within the meaning herein. A methoxyethoxy group is also an alkoxy group within the meaning herein. "Halo" as the term is used herein includes fluoro, chloro, bromo, and iodo. A "haloalkyl" group includes mono-halo alkyl groups, and poly-halo alkyl groups wherein all halo atoms can be the same or different. Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3- dibromo-3,3-difluoropropyl and the like.

The terms "aryloxy" and "arylalkoxy" refer to, respectively, an aryl group bonded to an oxygen atom and an aralkyl group bonded to the oxygen atom at the alkyl moeity. Examples include but are not limited to phenoxy, naphthyloxy, and benzyloxy. An "acyl" group as the term is used herein refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is also bonded to another carbon atom, which can be part of an alkyl, aryl, aralkyl cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group or the like. In the special case wherein the carbonyl carbon atom is bonded to a hydrogen, the group is a "formyl" group, an acyl group as the term is defined herein. An acyl group can include 0 to about 12-20 additional carbon atoms bonded to the carbonyl group. An acyl group can include double or triple bonds within the meaning herein. An acryloyl group is an example of an acyl group. An acyl group can also include heteroatoms within the meaning here. A nicotinoyl group (pyridyl-3 -carbonyl) group is an example of an acyl group within the meaning herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like. When the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a "haloacyl" group. An example is a trifluoroacetyl group.

The term "amine" includes primary, secondary, and tertiary amines having, e.g., the formula N(group)3 wherein each group can independently be H or non-H, such as alkyl, alkenyl, or alkynyl as defined herein, cycloalkyl or heterocyclyl as defined herein, aryl or heteroaryl as defined herein,, and the like. Amines include but are not limited to R-NH₂, for example, alkylamines, arylamines, alkylarylamines; R₂NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term "amine" also includes ammonium ions as used herein.

An "amino" group is a substituent of the form -NH₂, -NHR, -NR₂, -NR₃ ⁺, wherein each R is independently selected, and protonated forms of each. Accordingly, any compound substituted with an amino group can be viewed as an amine.

An "ammonium" ion includes the unsubstituted ammonium ion NH₄ ⁺, but unless otherwise specified, it also includes any protonated or quaternarized forms of amines. Thus, trimethylammonium hydrochloride and tetramethylammonium chloride are both ammonium ions, and amines, within the meaning herein.

The term "amide" (or "amido") includes C- and N-amide groups, i.e., -C(O)NR₂, and -NRC(O)R groups, respectively. Amide groups therefore include but are not limited to carbamoyl groups (-C(O)NH₂) and formamide groups (-NHC(O)H). A "carboxamido" group is a group of the formula C(O)NR₂, wherein R can be H, alkyl, aryl, etc.

The term "urethane" (or "carbamyl") includes N- and O-urethane groups, i.e., -NRC(O)OR and -OC(O)NR₂ groups, respectively.

The term "sulfonamide" (or "sulfonamido") includes S- and N- sulfonamide groups, i.e., -SO₂NR₂ and -NRSO₂R groups, respectively.

Sulfonamide groups therefore include but are not limited to sulfamoyl groups (- SO₂NH₂). An organosulfur structure represented by the formula -S(O)(NR)- is understood to refer to a sulfoximine, wherein both the oxygen and the nitrogen atoms are bonded to the sulfur atom, which is also bonded to two carbon atoms. The term "amidine" or "amidino" includes groups of the formula

-C(NR)NR₂. Typically, an amidino group is -C(NH)NH₂.

The term "guanidine" or "guanidino" includes groups of the formula -NRC(NR)NR₂. Typically, a guanidino group is -NHC(NH)NH₂.

A "salt" as is well known in the art includes an organic compound such as a carboxylic acid, a sulfonic acid, or an amine, in ionic form, in combination with a counterion. For example, acids in their anionic form can form salts with cations such as metal cations, for example sodium, potassium, and the like; with ammonium salts such as NH₄ ⁺ or the cations of various amines, including tetraalkyl ammonium salts such as tetramethylammonium, or other cations such as trimethylsulfonium, and the like. A "pharmaceutically acceptable" or "pharmacologically acceptable" salt is a salt formed from an ion that has been approved for human consumption and is generally non-toxic, such as a chloride salt or a sodium salt. A "zwitterion" is an internal salt such as can be formed in a molecule that has at least two ionizable groups, one forming an anion and the other a cation, which serve to balance each other. For example, amino acids such as glycine can exist in a zwitterionic form. A "zwitterion" is a salt within the meaning herein.

A "hydrate" is a compound that exists in a composition with water molecules. The composition can include water in stoichiometic quantities, such as a monohydrate or a dihydrate, or can include water in random amounts. A "solvate" is a similar composition except that a solvent other that water replaces the water. For example, methanol or ethanol can form an "alcoholate", which can again be stoichiometic or non-stoichiometric. "Tautomers" are two forms of a substance differing only by the position of a hydrogen atom in the molecular structures.

A "prodrug" as is well known in the art is a substance that can be administered to a patient where the substance is converted in vivo by the action of biochemicals within the patients body, such as enzymes, to the active pharmaceutical ingredient. Examples of prodrugs include esters of carboxylic acid groups, which can be hydrolyzed by endogenous esterases as are found in the bloodstream of humans and other mammals.

In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. For example, if X is described as selected from the group consisting of bromine, chlorine, and iodine, claims for X being bromine and claims for X being bromine and chlorine are fully described. Moreover, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any combination of individual members or subgroups of members of Markush groups. Thus, for example, if X is described as selected from the group consisting of bromine, chlorine, and iodine, and Y is described as selected from the group consisting of methyl, ethyl, and propyl, claims for X being bromine and Y being methyl are fully described.

In various embodiments, the compound or set of compounds, such as are used in the inventive methods, can be any one of any of the combinations and/or sub-combinations of the above-listed embodiments.

Without wishing to be bound by theory, the standard nomenclature of Schechter & Berger (Biochem. Biophys. Res. Comm., 1967, 27, 157-162) regarding the identification of residues in the polypeptide substrate of serine proteases will be employed herein unless other indicia of identification are specifically provided. Within the nomenclature of Schechter & Berger , the residues of the substrate, in the direction from the N-terminal toward the C- terminal, are labeled (Pi, ..., P3, P2, Pl, Pl', P2', Pr' ..., Pj), wherein cleavage is catalyzed between Pl and Pl '. Within the context of this nomenclature, compounds of Formulas I can be considered as mimics of at least the tripeptide P3-Pro-Pl, wherein the analog of Pl, as a moiety of the macrocyclic structure, is:

wherein the wavy lines indicate points of attachment within the macrocyclic ring, and those points of attachment are ultimately connected to each other via the macrocyclic ring as described below.

Various embodiments of the invention provide a compound of Formula I:

I

B is CH₂;

X is a bond, O, S, CH(R³) or N(R⁴);

M is O, S, S(O), S(O)₂;

T is R⁶, alkyl-R⁶, alkenyl-R⁶, or alkynyl-R⁶; and

In various embodiment, a compound of the invention is as shown in Table 1, or a compound as shown in Table 2, below.

In various embodiments of the invention, M is O. In various embodiments of the invention, D is CH or C(CH₃).

In various embodiments of the invention, X-Y is O, thus forming an ether linkage between Z and the pyrrolidine ring.

In various embodiments of the invention, T is R⁶ and R⁶ is alkyl.

In various embodiments of the invention, R⁶ is methyl. In various embodiments T and the CR of D form a spirocyclic ring comprising 3-6 atoms including 0-2 heteroatoms selected from the set consisting of O, NR, S, S(O), and S(O)₂.

In various embodiments, a compound of the invention can have the formula

wherein A, T, X, Y, and Z are as defined herein. In various embodiments, the spirocyclic ring formed by T and the CR' of D is a 3, 4, or 5 membered ring that is carbocyclic or that contains a single oxygen atom.

In various embodiments of the invention, B is CH₂. In various embodiments, B is absent, such that a single bond exists between the carbon atom bearing A and the adjacent carbon atom.

In various embodiments of the invention, a carbon atom bearing T is of an R absolute configuration, or is of an S absolute configuration, or is a mixture thereof. In various embodiments of the invention, A is a group of formula

In various embodiments of the invention, A is a group of formula

In various embodiments of the invention, X is O, or X is a bond. In various embodiments of the invention, Y is a bond, or C(O).

In various embodiments of the invention, both X and Y are bonds, taken together forming a single bond.

In various embodiments of the invention, Z can be an unsubstituted heteroaryl group or is a heteroaryl group mono- or independently pluri- substituted with J.

In various embodiments of the invention, Z can be a substituted quinolyl group, a substituted triazolyl group, a substituted tetrazolyl group, or a substituted isoindolidinyl group, wherein any group is mono- or independently pluri-substituted with J. In various embodiments of the invention, Z can be a thiazolyl-substituted quinolyl group, or a pyrazolyl-substituted quinolyl group, wherein any group is mono- or independently pluri-substituted with J. In various embodiments of the invention, X-Y-Z comprises a group of the formula

In various embodiments of the invention, m is O- 1.

In various embodiments of the invention, n is 1-4.

In various embodiments of the invention, Z is a group of formula

wherein a wavy line indicates a point of attachment.

In various embodiments, L is C₂H₂. More specifically, the C₂H₂ group of L can be in a cis configuration in the macrocyclic ring.

In various embodiments of the invention the compound of formula I is any of the compounds shown below in Table 1 or Table 2, or any Example herein, provided that the definition of A falls within the limitations of claim 1, or any stereoisomer, solvate, hydrate, tautomer, prodrug, salt, or pharmaceutically acceptable salt, or mixture thereof.

In various embodiments the invention provides a method of preparation of a compound of formula I of claim 1. More specifically, in various embodiments the invention provides a method of preparation of a compound of formula (Ha) wherein L is C₂H₂, comprising contacting a compound of formula II

wherein A' is -C(=O)OR^C and m = 0, and an olefin metathesis catalyst under conditions sufficient to bring about formation of the compound of formula Ha

wherein L is C₂H₂ and m = 0. In various embodiments the invention further provides a method of converting the compound of formula (Ha) to the compound of formula (I) of claim 1, comprising converting the -C(=O)OR^c group of A' of the compound of formula (Ha) to either of the

groups of A of the compound of formula (I), and then, optionally, contacting the compound of formula I wherein L is C₂H₂ and hydrogen in the presence of a catalyst to provide a compound of formula I wherein L is C2H4. The hydrogenation catalyst can be platinum or palladium.

The olefin metathesis catalyst can be dichloro(o- isopropoxyphenylmethylene) (tricyclohexylphosphine)ruthenium(II), also known as Hoveyda-Grubbs 1st generation catalyst.

Methods of Use

In one aspect, the invention provides methods of inhibiting HCV NS3 protease. The methods include contacting the hepatitis C viral serine protease with a compound as described herein. In other embodiments, the methods of inhibiting HCV NS3 protease include administering a compound as described herein to a subject infected with hepatitis C virus.

In another aspect, the invention provides methods for treating hepatitis C viral infection. The methods include administering to a subject in need of such treatment an effective amount of a compound of the invention as described herein. As used herein, "a compound" can refer to a single compound or a plurality of compounds. In some embodiments, the methods for treating hepatitis C viral infection include administering to a subject in need of such treatment an effective amount of a composition comprising a compound of the invention and a pharmaceutically acceptable carrier.

In another embodiment, the invention provides methods for treating hepatitis C viral infection comprising administering to a subject in need of such treatment an effective amount of a compound of the invention in combination with another medicament, such as another anti-viral agent. The term "anti-viral agent" as used herein denotes a compound which interferes with any stage of the viral life cycle to slow or prevent HCV reproduction. Representative anti-viral agents include, without limitation, INTRON-A, (interferon alfa-2b available from Schering Corporation, Kenilworth, N.J.), PEG-INTRON (peginteferon alfa-2b, available from Schering Corporation, Kenilworth, N.J.), ROFERON-A (recombinant interferon alfa-2a available Hoffmann-La Roche, Nutley, N.J.), PEGASYS (peginterferon alfa-2a available Hoffmann-La Roche, Nutley, N.J.), INFERGEN A (Schering Plough, inteferon-alpha 2B+Ribavirin), WELLFERON (interferon alpha-nl), cyclophilin inhibitors, nucleoside analogues, IRES inhibitors, El inhibitors, E2 inhibitors, IMPDH inhibitors, NS5B polymerase inhibitors such as R- 1626, R-7128, MK-0608, A837093, GS9190 and PF- 868554, NS4B inhibitors, NS5A inhibitors, and/or NTPase/helicase inhibitors. In certain embodiments, the methods of treating HCV infection include administering to a subject in need of such treatment an effective amount of a compound of the invention in combination with another NS3/4A inhibitor. Examples of other NS3/4A inhibitors which can be administered in combination with compounds of the present invention include, without limitation, telaprevir (VX950), boceprevir (SCH503034), ITMN191, TMC 435350, MK7009, and PHXl 766.

Still other antiviral agents that may be used in conjunction with inventive compounds for the treatment of HCV infection include, but are not limited to, ribavirin (l-beta-D-ribofuranosyl-lH-l,2,- 4-triazole-3-carboxamide, available from ICN Pharmaceuticals, Inc., Costa Mesa, Calif.; described in the Merck Index, entry 8365, Twelfth Edition); REBETROL.RTM. (Schering Corporation, Kenilworth, N.J.), COPEGASUS. RTM. (Hoffmann-La Roche, Nutley, N.J.); BEREFOR.RTM. (interferon alfa 2 available from Boehringer Ingelheim Pharmaceutical, Inc., Ridgefield, Conn.); SUMIFERON.RTM. (a purified blend of natural alpha interferons such as Sumiferon available from Sumitomo, Japan); ALFERON.RTM. (a mixture of natural alpha interferons made by Interferon Sciences, and available from Purdue Frederick Co., CT); .alpha.-interferon; natural alpha interferon 2a; natural alpha interferon 2b; pegylated alpha interferon 2a or 2b; consensus alpha interferon (Amgen, Inc., Newbury Park, Calif); VIRAFERON.RTM.; INFERGEN.RTM.; REBETRON.RTM. (Schering Plough, Inteferon-alpha 2B+Ribavirin); pegylated interferon alpha (Reddy, K. R. et al. "Efficacy and Safety of Pegylated (40-kd) Interferon alpha-2a Compared with Interferon alpha-2a in Noncirrhotic Patients with Chronic Hepatitis C (Hepatology, 33, pp. 433-438 (2001); consensus interferon (Kao, J. H., et al., " Efficacy of Consensus Interferon in the Treatment of Chronic Hepatitis" J. Gastroenterol. Hepatol. 15, pp. 1418-1423 (2000); lymphoblastoid or "natural" interferon; interferon tau (Clayette, P. et al., "IFN- tau, A New Interferon Type I with Antiretroviral activity" Pathol. Biol. (Paris) 47, pp. 553-559 (1999); interleukin 2 (Davis, G. L. et al., "Future Options for the Management of Hepatitis C." Seminars in Liver Disease, 19, pp. 103-112 (1999); Interleukin 6 (Davis et al. "Future Options for the Management of Hepatitis C." Seminars in Liver Disease 19, pp. 103-112 (1999); interleukin 12 (Davis, G. L. et al., "Future Options for the Management of Hepatitis C." Seminars in Liver Disease, 19, pp. 103-112 (1999); and compounds that enhance the development of type 1 helper T cell response (Davis et al., "Future Options for the Management of hepatitis C." Seminars in Liver Disease, 19, pp. 103-112 (1999)). Also included are compounds that stimulate the synthesis of interferon in cells (Tazulakhova, E. B. et al., "Russian Experience in Screening, analysis, and Clinical Application of Novel Interferon Inducers" J. Interferon Cytokine Res., 21 pp. 65-73) including, but are not limited to, double stranded RNA, alone or in combination with tobramycin, and Imiquimod (3M Pharmaceuticals; Sauder, D. N. "Immunomodulatory and Pharmacologic Properties of Imiquimod" J. Am. Acad. Dermatol, 43 pp. S6-11 (2000)

In another embodiment, the invention provides a method for treating hepatitis C viral infection, comprising administering to a subject in need of such treatment an effective amount of a compound of the invention in combination with an anti-proliferative agent. The term "antiproliferative agent" as used herein denotes a compound which inhibits cellular proliferation. Cellular proliferation can occur, for example without limitation, during carcinogenesis, metastasis, and immune responses. Representative anti-proliferative agents include, without limitation, 5-fluorouracil, daunomycin, mitomycin, bleomycin, dexamethasone, methotrexate, cytarabine, and mercaptopurine.

In another embodiment, the invention provides a method for treating hepatitis C viral infection, comprising administering to a subject in need of such treatment an effective amount of a compound of the invention in combination with an immune modulator. The term "immune modulator" as used herein denotes a compound or composition comprising a plurality of compounds which changes any aspect of the functioning of the immune system. In this context, immune modulator includes without limitation anti-inflammatory agents and immune suppressants. Representative immune modulator include without limitation steroids, non-steroidal antiinflammatories, COX2 inhibitors, anti- TNF compounds, anti-IL- 1 compounds, methotrexate, leflunomide, cyclosporin, FK506 and combinations of any two or more thereof. Representative steroids in this context include without limitation prednisone, prednisolone, and dexamethasone. Representative non-steroidal anti-inflammatory agents in this context include without limitation ibuprofen, naproxen, diclofenac, and indomethacin. Representative COX2 inhibitors in this context include without limitation rofecoxib and celecoxib. Representative Anti-TNF compounds in this context include without limitation enbrel, infliximab, and adalumimab.

Representative anti-IL-1 compounds in this context include without limitation anakinra. Representative immune suppressants include without limitation cyclosporin and FK506.

In another embodiment, the invention provides a method for treating hepatitis C viral infection, comprising administering to a subject in need of such treatment an effective amount of a compound of the invention in combination with an antibiotic. As used herein, an antibiotic includes an anti-bacterial or anti-fungal agent. Agents of this type are useful in combating, for example, secondary bacterial or fungal infections that may accompany a hepatitis C infection. Examples include beta-lactam antibiotics, macrocyclic antibiotics, triazole anti-fungal agents, and the like.

Compounds of the invention include mixtures of stereoisomers such as mixtures of diastereomers and/or enantiomers. In some embodiments, the compound, e.g. of Formula I, is 90 weight percent (wt %) or greater of a single diastereomer of enantiomer. In other embodiments, the compound is 92, 94, 96, 98 or even 99 wt % or more of a single diastereomer or single enantiomer.

A variety of uses of the invention compounds are possible along the lines of the various methods of treating a subject as described above. Exemplary uses of the invention methods include, without limitation, use of a compound of the invention in a medicament or for the manufacture of a medicament for treating a condition that is regulated or normalized via inhibition of the HCV NS3 serine protease.

An embodiment of the invention is directed to use of an inventive compound in preparation of a medicament for the treatment of an HCV infection in a patient. In further embodiments, an effective amount of a second bioactive agent, such as described above, can be used in preparation of a medicament for treatment of an HCV infection in a patient. An embodiment of the invention provides an inventive compound for use in combination with an effective amount of a second medicament, such as described above, in treatment of an HCV infection in a patient.

Biochemical methods

Fluorescence resonance energy transfer (FRET; see e.g., Heim et al, (1996) Curr. Biol. 6: 178-182; Mitra et al., (1996) Gene 173: 13-17; and Selvin et al., (1995) Meth. Enzymol. 246:300-345) is an exquisitely sensitive method for detecting energy transfer between two fluorophoric probes. As known in the art, such probes are given the designations "donor" and "acceptor" depending on the relative positions of the maxima in the absorption and emission spectra characterizing the probes. If the emission spectrum of the acceptor overlaps the absorption spectrum of the donor, energy transfer can occur. Because of the known and highly non-linear relationship of energy transfer and distance between fluorophores, approximated by an inverse sixth power dependence on distance, FRET measurements correlate with distance. For example, when the probes are in proximity, such as when the probes are attached to the N- and C- termini of a peptide substrate, and the sample is illuminated in a spectrofluorometer, resonance energy can be transferred from one excited probe to the other resulting in observable signal. Upon scission of the peptide linking the probes, the average distance between probes increases such that energy transfer between donor and accept probe is not observed. As a result, the degree of hydrolysis of the peptide substrate, and the level of activity of the protease catalyzing hydrolysis of the peptide substrate, can be quantitated. Accordingly, using methods known in the arts of chemical and biochemical kinetics and equilibria, the effect of inhibitor on protease activity can be quantitated.

Compounds of the invention will be found to have activity in this assay when employed to evaluate the inhibition of the HCV NS3 protease. Compositions and Combination Treatments

A. Compositions.

Another aspect of the invention provides compositions of the compounds of the invention, alone or in combination with another NS3 protease inhibitor or another type of antiviral agent and/or another type of therapeutic agent. As set forth herein, compounds of the invention include stereoisomers, tautomers, solvates, prodrugs, pharmaceutically acceptable salts and mixtures thereof. Compositions containing a compound of the invention may be prepared by conventional techniques, e.g. as described in Remington: The Science and Practice of Pharmacy, 21st Ed., (2005). The compositions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications.

Typical compositions include a compound of the invention which inhibits the enzymatic activity of the HCV NS3 protease, and a pharmaceutically acceptable excipient which may be a carrier or a diluent. For example, the active compound will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet, paper, or other container. When the active compound is mixed with a carrier, or when the carrier serves as a diluent, it may be solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The active compound can be adsorbed on a granular solid carrier, for example contained in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.

The formulations can be mixed with auxiliary agents which do not deleteriously react with the active compounds. Such additives can include wetting agents, emulsifying and suspending agents, salt for influencing osmotic pressure, buffers and/or coloring substances preserving agents, sweetening agents or flavoring agents. The compositions can also be sterilized if desired.

The route of administration may be any route which effectively transports the active compound of the invention which inhibits the enzymatic activity of the HCV NS3 protease to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal or parenteral, e.g., rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment, the oral route being preferred. If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution. Injectable dosage forms generally include aqueous suspensions or oil suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent Injectable forms may be in solution phase or in the form of a suspension, which is prepared with a solvent or diluent. Acceptable solvents or vehicles include sterilized water, Ringer's solution, or an isotonic aqueous saline solution. Alternatively, sterile oils may be employed as solvents or suspending agents. Preferably, the oil or fatty acid is non-volatile, including natural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the formulation may also be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates. For injection, the formulations may optionally contain stabilizers, pH modifiers, surfactants, bioavailability modifiers and combinations of these. The compounds may be formulated for parenteral administration by injection such as by bolus injection or continuous infusion. A unit dosage form for injection may be in ampoules or in multi-dose containers.

The formulations of the invention may be designed to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art. Thus, the formulations may also be formulated for controlled release or for slow release.

Compositions contemplated by the present invention may comprise, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and/or delivery effect. Therefore, the formulations may be compressed into pellets or cylinders and implanted intramuscularly or subcutaneous Iy as depot injections or as implants such as stents. Such implants may employ known inert materials such as silicones and biodegradable polymers, e.g., polylactide-polyglycolide. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides).

For nasal administration, the preparation may contain a compound of the invention which inhibits the enzymatic activity of the HCV NS3 protease, dissolved or suspended in a liquid carrier, preferably an aqueous carrier, for aerosol application. The carrier may contain additives such as solubilizing agents, e.g., propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabens.

For parenteral application, particularly suitable are injectable solutions or suspensions, preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.

Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application. Preferable carriers for tablets, dragees, or capsules include lactose, corn starch, and/or potato starch. A syrup or elixir can be used in cases where a sweetened vehicle can be employed.

A typical tablet that may be prepared by conventional tabletting techniques may contain:

Core: Active compound (as free compound or salt thereof) 250 mg

Colloidal silicon dioxide (Aerosil)® 1.5 mg Cellulose, microcryst. (Avicel)® 70 mg Modified cellulose gum (Ac-Di-Sol)® 7.5 mg Magnesium stearate Ad. Coating:

HPMC approx. 9 mg *Mywacett 9-40 T approx. 0.9 mg

*Acylated monoglyceride used as plasticizer for film coating. A typical capsule for oral administration contains, for example, compounds of the invention (250 mg), lactose (75 mg) and magnesium stearate (15 mg). The mixture is passed through a 60 mesh sieve and packed into a No. 1 gelatin capsule. A typical injectable preparation is produced by aseptically placing, for example, 250 mg of compounds of the invention into a vial, aseptically freeze-drying and sealing. For use, the contents of the vial are mixed with 2 mL of sterile physiological saline, to produce an injectable preparation.

The compounds of the invention may be administered to a mammal, especially a human in need of such treatment, prevention, elimination, alleviation or amelioration of the various diseases as mentioned above, e.g., HCV infection. Such mammals include also animals, both domestic animals, e.g. household pets, farm animals, and non-domestic animals such as wildlife.

The compounds of the invention are effective over a wide dosage range. For example, in the treatment of adult humans, daily dosages ranging from about 1 to about 3000 mg may be used. Doses of about 1 mg to about 1000 mg can be provided to the patient up to three times per day. In choosing a regimen for patients it may frequently be necessary to begin with a higher dosage and when the condition is under control to reduce the dosage. The exact dosage will depend upon the activity of the compound, mode of administration, on the therapy desired, form in which administered, the subject to be treated and the body weight of the subject to be treated, and the preference and experience of the physician or veterinarian in charge. HCV NS3 protease inhibitor activity of the compounds of the invention may be determined by use of an in vitro assay system which measures the potentiation of inhibition of the HCV NS3 protease. Inhibition constants (i.e., K₁ or IC50 values as known in the art) for the HCV NS3 protease inhibitors of the invention may be determined by the method described in the Examples.

The invention also encompasses prodrugs of a compound of the invention which on administration undergo chemical conversion by metabolic or other physiological processes before becoming active pharmacological substances. Conversion by metabolic or other physiological processes includes without limitation enzymatic (e.g, specific enzymatically catalyzed) and non- enzymatic (e.g., general or specific acid or base induced) chemical transformation of the prodrug into the active pharmacological substance. In general, such prodrugs will be functional derivatives of a compound of the invention which are readily convertible in vivo into a compound of the invention. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985. In another aspect, there are provided methods of making a composition of a compound described herein comprising formulating a compound of the invention with a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutically acceptable carrier or diluent is suitable for oral administration. In some such embodiments, the methods may further comprise the step of formulating the composition into a tablet or capsule. In other embodiments, the pharmaceutically acceptable carrier or diluent is suitable for parenteral administration. In some such embodiments, the methods further comprise the step of lyophilizing the composition to form a lyophilized preparation. B. Combinations.

The compounds of the invention may be used in combination with i) one or more other NS3 protease inhibitors and/or ii) one or more other types of antiviral agents (employed to treat viral infection and related diseases) and/or one or more other types of therapeutic agents which may be administered orally in the same dosage form, in a separate oral dosage form (e.g., sequentially or non-sequentially) or by injection together or separately (e.g., sequentially or non- sequentially).

Accordingly, in another aspect the invention provides combinations, comprising: a) a compound of the invention as described herein; and b) one or more compounds comprising: i) other compounds of the present invention ii) anti-viral agents including, but not limited to, other NS3 protease inhibitors iii) anti-proliferative agents iv) immune modulators v) antibiotics.

Combinations of the invention include mixtures of compounds from (a) and (b) in a single formulation and compounds from (a) and (b) as separate formulations. Some combinations of the invention may be packaged as separate formulations in a kit. In some embodiments, two or more compounds from (b) are formulated together while a compound of the invention is formulated separately. Combinations of the invention can further comprise a pharmaceutically acceptable carrier. In some embodiments, the compound of the invention is 90 wt % or more of a single diastereomer or single enantiomer. Alternatively, the compound of the invention can be 91, 92, 93, 94, 95, 96, 97, 98, or 99 wt % or more of a single diastereomer or single enantiomer. The dosages and formulations for the other antiviral agent to be employed, where applicable, will be as set out in the latest edition of the

Physicians ' Desk Reference.

In carrying out the methods of the invention, a composition may be employed containing the compounds of the invention, with or without another antiviral agent and/or other type therapeutic agent, in association with a pharmaceutical vehicle or diluent. The composition can be formulated employing conventional solid or liquid vehicles or diluents and pharmaceutical additives of a type appropriate to the mode of desired administration. The compounds can be administered to mammalian species including humans, monkeys, dogs, etc. by an oral route, for example, in the form of tablets, capsules, granules or powders, or they can be administered by a parenteral route in the form of injectable preparations. The dose for adult humans is preferably between 1 and 3,000 mg per day, which can be administered in a single dose or in the form of individual doses of about 1 mg to about 1,000 mg up to 3 times per day.

The compounds and processes of the present invention will be better understood in connection with the following examples, which are intended as an illustration only and not to limit the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications including, without limitation, those relating to the chemical structures, substituents, derivatives, formulations and/or methods of the invention may be made without departing from the spirit of the invention and the scope of the appended claims.

Examples

The following abbreviations are used throughout this document. BOP Benzotriazol-l-yl-oxy-tris-(dimethylamino)phosphonium hexafluorophosphate CDI Carbonyl diimidazole

DBU Diazabicycloundecane

DCM Dichloromethane

DIPEA, ¹Pr₂EtN N,N-Diisopropylethylamine

DMAP 4-(Ν,Ν-dimethylamino)pyridine DMF N,N-Dimethylformamide

DMSO Dimethylsulfoxide

EDAC 1 -Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride eq Equivalents Et₂O Diethyl ether

EtOAc Ethyl acetate h Hours

HATU O-(7-Azabenzotriazole-l-yl)-N,N,N'N'- tetramethyluronium hexafluorophosphate

HCl Hydrochloric acid

HOAT Hydroxyazabenztriazole

HOBT Hydroxybenzotriazole

LiHDMS Lithium hexamethyldisilazide

LiOH Lithium hydroxide mg Milligrams min Minutes mL Milliliters μL Microliters mmole Millimoles

MS Mass spectroscopy

MeOH Methanol

NaBH₃CN Sodium cyanoborohydride

NaH Sodium hydride

NaIO₄ Sodium periodate

NMM N-Methylmorpholine rb Round-bottom

RT Room temperature sat. Saturated

TEA Triethylamine

TFA Trifluoroacetic acid

THF Tetrahydrofuran to (range, e.g., X-Y = X to Y)

Compound 1.

Compound 1 was prepared according to the following protocol.

Synthesis of compound E2.

E^{1 E2}

To a solution of methyl 2-hydroxyacetate (2.8 g, 31.1 mmol) in dry THF (60 mL) was slowly added NaH (2.23 g, 93.3 mmol) and the resulting solution was stirred in an ice-bath for 15 min. 6-bromo-l-hexene (5.54 g, 34.2 mmol) was added and the reaction mixture was warmed up to room temperature overnight. Then the reaction mixture was acidified with 1 N HCl and diluted with H₂O (300 mL). The mixture was extracted with ethyl acetate (200 mL x 3) and the organic layer was dried and concentrated to afford 3.0 g of crude compound El. The crude product was dissolved in THF (15 mL) and 1 N LiOH (15 mL) was added. The reaction mixture was stirred at room temperature for 4 h. TLC indicated the complete consumption of starting material. The mixture was acidified to pH 3 and then diluted with H₂O (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic layer was dried over Na₂SO₄, filtered and concentrated under reduced pressure to afford 2.7 g of compound E2 in a total yield of 55 %.

Synthesis of compound E3.

E3

To a solution of the protected cis-hydroxyproline shown (4.0 g, 16 mmol) in THF (190 mL) was added 1 N LiOH (190 mL) and the reaction mixture was stirred at room temperature for 4.5 h. After TLC indicated the consumption of starting material, the mixture was acidified to pH 2 and extracted with ethyl acetate (100 mL x 3). The organic layer was dried and concentrated to afford 3.65 g of compound E3 which was used in the next step directly. Synthesis of compound E4.

To a solution of compound E3 (500 mg, 2.16 mmol) in dried DCM (20 mL) in an ice-bath was added HATU (905 mg, 2.38 mmol) and the mixture was stirred for 15 min. Then l-amino-2-vinyl-cyclopropanecarboxylic acid methyl ester tosylate salt (778 mg, 2.38 mmol) and DIPEA (836 mg, 6.48 mmol) were added and the reaction mixture was slowly warmed up to room temperature over a period of 3 h. Then the reaction solution was washed with 1 N HCl (30 mL) and saturated aqueous NaHCCh (30 mL), dried, filtered and concentrated to afford 1.1 g of a crude product. After purification by column chromatography, 460 mg of compound E4 were isolated in a yield of 60 %.

Synthesis of compound E5.

To a solution of compound E4 (460 mg, 1.30 mmol) in dried DCM (2 mL) was added TFA (2 mL) and the reaction solution was stirred at room temperature for 3 h. After TLC analysis indicated the consumption of starting material, the reaction solution was concentrated under reduced pressure to give compound E5 as a crude product which was used in the next step directly. Synthesis of compound E6.

To a solution of compound E2 (223 mg, 1.30 mmol) in dried DCM (20 mL) cooled with an ice-bath was added HATU (741 mg, 1.95 mmol) and the mixture was stirred for 15 min. Then crude compound E5 from the previous step and DIPEA (840 mg, 6.5 mmol) were added and the reaction mixture was slowly warmed up to room temperature over 3 h. Then the reaction solution was washed with 1 N HCl (10 mL) and saturated aqueous NaHCO₃ (10 mL), dried, filtered and concentrated to afford 430 mg of a crude product. After purification by column chromatography, 210 mg of compound E6 were isolated in a yield of 41 %.

Synthesis of compound E7.

metathesis

E6 E7 A solution of compound E6 (210 mg, 0.55 mmol) in dry DCM (40 mL) was deoxygenated by bubbling argon for 30 min through the solution. Hoveyda- Grubbs 1^st generation catalyst (17 mg, 6 mol %) was then added as a solid and the reaction was refluxed under argon for 48 h. The red-orange solution was evaporated to yield a brown residue. After the purification by column chromatography (EtOAc / DCM), 90 mg of compound E7 were obtained in a yield of 46%. Synthesis of compound E8.

E7 E8

To a solution of compound E7 (220 mg, 0.60 mmol) and DIPEA (155 mg, 2 eq.) in DCM (10 mL) was added slowly mesylate chloride (102 mg, 1.5eq.) at 0 ⁰C. The reaction was kept for 3 hours at 0 ⁰C. After letting the reaction come to room temperature, ethyl acetate (30 mL) was added followed by washing with aqueous 5% citric acid (10 mL), water (10 mL), IM NaHCCh (10 mL) and brine respectively. The organic phase was separated, dried over Na₂SO₄, filtered, and evaporated, producing 240 mg of crude compound E8 which was used in the next step without further purification. Mass (ESI, positive): 445 m/z [M+l]⁺.

Synthesis of compound ElO.

E8 E10 To a solution of compound E8 (240 mg, 0.54 mmol) and 2-(4- isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol E9 (203 mg, 1.1 eq) in dry DMF (5 mL) was added cesium carbonate (1.06 g, 6 eq). The reaction mixture was stirred at 70 ⁰C for 12 hours, cooled to room temperature and then diluted with ethyl acetate. It was then washed with a sodium carbonate solution and water, dried over Na₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography using petroleum ether/ethyl acetate mixtures to give 110 mg of compound ElO in a 30% yield. Mass (ESI, positive): 663 m/z [M+H]⁺. Synthesis of compound Ell.

E10 E11

A IN LiOH solution in water (0.34 mL, 2 eq.) was added to a solution of compound ElO (110 mg, 0.17 mmol) in THF (2 mL) and the reaction mixture was allowed to stir at room temperature for 12 hours. THF was removed under reduced pressure and DCM was added. The pH was adjusted to ~1, the DCM layer was separated and the water layer extracted with ethyl acetate three times. The organic layers were combined, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give 97 mg of compound Ell in a 90% yield. Mass (ESI, positive): 649 m/z [M+H]⁺

Synthesis of compound E13 (Compound 1)

E11 E13

To a solution of compound Ell (97 mg, 0.15 mmol) in DMF (10 mL) under N₂ was added CDI (45 mg, 2 eq) and the mixture was stirred at 45 ⁰C overnight. It was then cooled down to room temperature. Cyclopropanesulfonamide E12 (84 mg, 4 eq) was added followed by the addition of DBU (83 mg, 4 eq). The resulting mixture was stirred at 55 ⁰C overnight. DMF was removed under reduced pressure. The residue was diluted with ethyl acetate and the pH adjusted to ~6. The organic phase was washed with saturated NaCl one time, dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by column chromatography using petroleum ether/ethyl acetate mixtures to give 10 mg of the desired compound E13 in a 9% yield. Mass (ESI, positive): 752 m/z [M+H]⁺

Compound 2.

Compound 2 was prepared according to the following protocol.

Synthesis of compound E14.

E14 To a solution of 6-bromo- 1 -hexene (5 g, 31 mmol) in acetone (50 mL) was added NaI (6.9 g, 46 mmol). The mixture was refluxed for 4 hours (progress of the reaction was monitored by GC). Solid NaBr was removed by filtration, and the acetone was evaporated under reduced pressure. The residue was dissolved in DCM (100 mL), washed with water (50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to afford the corresponding iodide (5.2 g, 80% yield) as a yellow liquid.

Silver oxide (835 mg, 3.6 mmol) was added to a mixture of (+)-methyl D-lactate (500 mg, 4.8 mmol) and 6-iodo-l -hexene (1.5 g, 7.2 mmol) in ether. The reaction mixture was stirred for 2 days. Then the reaction was diluted with ether (10 mL) and filtered. Solvents were removed under reduced pressure. The crude product was purified by column chromatography with petroleum ether/ethyl acetate mixtures to afford compound E14 (500 mg, 56% yield).

Synthesis of compound E15.

E14 E15

To a solution of compound E14 (500 mg, 2.7 mmol) in THF was added a IN aqueous solution of LiOH (4 mL). The reaction mixture was stirred for 3 hours at room temperature. Then, most of the THF was removed under reduced pressure and the pH adjusted to 3 with aqueous IN HCl. The solution was extracted with DCM, dried over anhydrous Na₂SO₄ and filtered. Solvents were removed under reduced pressure to yield compound E15 (370 mg, 80% yield).

Alternative method for the synthesis of compound E15.

The sodium salt of D-lactic acid (1.26 g, 11.2 mmol) was dissolved in DMSO (40 mL) with some sonication. The reaction flask was placed in a water bath at 15-20 ⁰C and tBuOK (1.26 g, 11.2 mmol) was added in one portion. The reaction was stirred under argon for 90 minutes. Then 6-bromo-l-hexene (1.66 mL, 11.76 mmol) was added dropwise and the reaction stirred for 21 hours at room temperature. The contents of the flask were added portionwise to distilled water cooled to 0 ⁰C with vigorous stirring. Et₂O (~ 50 mL) was added to the same Erlenmeyer flask. The organic layer was separated and the aqueous one extracted with Et₂O (2 x 50 mL). The aqueous layer was cooled to 0 ⁰C and acidify to pH = 1-2 with aqueous IN HCl. The aqueous layer was extracted with Et₂O (2 x 50 mL). The combined ether layers were washed with brine and dried over sodium sulfate. After filtration, solvents were removed under reduced pressure to yield E15 (325 mg, 17%) which was used in a later step without further purification. Mass (ES negative): 171 [M-H]^".

Alternative method for the synthesis of compound E15.

L-Alanine (10 g, 112.25 mmol) was dissolved in a mixture of 48% HBr (116 mL) and H₂O (200 mL). To the solution was added cracked ice (-400 g). The mixture was also externally cooled with an ice bath. NaNO₂ (20.86 g, 302.32 mmol) was added to the above mixture in small portions with vigorous stirring. Na₂SO₄ (140 g, 985.64 mmol) was then added to the reaction in portions. The stirring was continued until the solution temperature reached 15°C. The resulting solution was decanted from the flask and extracted with Et₂O (100 mL x 5). The organics were combined, dried over MgSO₄, filtered and concentrated. The crude product was purified via vacuum distillation (25 torr, 104 - 108⁰C) to afford the desired (S)-2-bromopropanoic acid as a colorless liquid (14.08 g, 92.04 mmol, 82% yield).

To a suspension of NaH (95%, 347 mg, 13.74 mmol) in DMF (20 mL) at room temperature was added 5-hexen-l-ol (0.87 mL, 7.19 mmol) dropwise. The mixture was stirred at room temperature for 0.5 h before it was cooled to 0⁰C. A solution of the α-bromo acid obtained above (1 g, 6.54 mmol) in DMF (5 mL) was added to the mixture via cannula. The reaction was stirred at 0⁰C for 6 h. The reaction was quenched with dropwise addition of IN HCl until pH = 1-2. The resulting mixture was extracted with Et₂O (20 mL x 3). The organics were combined, dried over MgSO₄, filtered and concentrated. The crude was purified via flash column chromatography (25 g silica, MeOH/DCM = 0-5%) to afford the desired product as a colorless liquid (1.00 g, 5.82 mmol, 89% yield). R_f = 0.2 (MeOH/DCM = 1 :9).

Synthesis of compound E17.

To a solution of N-Boc-c/s-4-hydroxy-L-proline (4.9 g, 21 mmol), l-amino-2- vinyl-cyclopropanecarboxylic acid ethyl ester tosylate salt E16 (7.55 g, 23.3 mmol) and HATU (8.4 g, 22 mmol) in DMF (70 mL) under argon cooled to 0 ⁰C was added ¹Pr₂NEt (0.94 mL, 5.7 mmol) via syringe pump over 7 minutes. The reaction was stirred for 45 minutes at 0 ⁰C. It was diluted with Et₂O and quenched with IN HCl. The organic phase was separated and washed a second time with IN HCl. The combined ether layers were washed with a saturated aqueous solution of NaHCθ3, dried over MgSO₄ and filtered through a small plug of silica gel. Solvents were removed under reduced pressure and the residue was triturated with Et₂O. The solids were filtered off to afford compound E17 (6.1 g, 78%). Mass (ESI positive): 369 [M+H]⁺. Synthesis of compound E18.

Compound E17 (1 g, 2.7 mmol) was dissolved in 4N HCl dioxane (15 mL) and stir at room temperature for 1 hour. At this time, LCMS showed complete conversion to the desired product. Solvents were removed under reduced pressure to afford crude E18 that was used without further purification in the next step. Mass (ESI positive): 269 [M+H]⁺.

Synthesis of compound E19.

To a solution of E15 (2.7 mmol), E18 (crude from previous step, 2.7 mmol) and HATU (1.1 g, 2.8 mmol) in DMF (27 mL) cooled to 0°C was added ¹Pr₂NEt (0.94 mL, 5.7 mmol) dropwise over 8 minutes. The reaction was stirred for 1 hour at 0 ⁰C. Precooled IN HCl (20 mL) was added and the mixture vigorously stirred for 5 minutes. A 10% EtOAc/Et₂O solution (30 mL) was added with continuous stirring. The organic phase was separated and the aqueous one extracted with 10% EtOAc/Et₂O (8 x 25 mL). The combined organic layers were washed with brine, dried over Na₂SO₄ and filtered. Solvents were removed under reduced pressure. The residue was purified by column chromatography (100 g Biotage silica gel cartridge) by eluting with a gradient of methanol in dichloromethane (0% - 5%). Appropriate fractions were pooled and solvents removed under reduced pressure to afford compound E19 (816 mg, 71%). Mass (ESI positive): 423 [M+H]⁺. Synthesis of compound E20.

E19 E20

A two-neck round bottom flask was charged with a solution of compound E19 (923 mg, 2.2 mmol) in 1,2 dichloroethane (1 L). After the flask was fitted with a reflux condenser, argon gas was bubbled through the solution for 15 min at room temperature. Then the flask was placed in a 80 ⁰C oil bath and stirred for 15 min with a continuous flow of argon through the solution. Hoveyda-Grubbs 1^st generation catalyst (40 mg, 3 mol %) was added to the reaction in one portion. Subsequently, a solution of catalyst (92 mg, 7 mol%) in dichloroethane (15 mL) was added to the reaction over 7 hours by use of a syringe pump. Argon is bubbled through the solution until the starting material is consumed. After LCMS analysis indicated that compound E 19 was totally consumed, the reaction mixture was cooled to 50 ⁰C and treated with 2-mercaptonicotinic acid (180 mg). After stirring at 50 ⁰C overnight, the mixture was cooled to room temperature and concentrated to 1/5 of the original volume under reduced pressure. The reaction mixture was washed with aqueous saturated sodium bicarbonate, IN HCl and brine. The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (25 g Biotage silica gel cartridge) by eluting with a gradient of methanol in dichloromethane. After the fractions were checked by TLC, the clean fractions were combined, filtered through a fine fritted filter, and concentrated under reduced pressure to afford compound E20 (738 mg, 85%) as a white solid. Mass (ESI positive): 395 [M+H]⁺.

Synthesis of compound E21.

E20 E21 A solution of brosyl chloride (54 mg, 0.21 mmol) in CH₂Cl₂ (0.8 mL) was added to a solution of E20 (63 mg, 0.16 mmol), triethylamine (67 μL, 0.48 mmol), and DMAP (1 mg) in toluene (250 μL) at ambient temperature. The reaction mixture was stirred at ambient temperature overnight (15 h). HPLC indicated formation of the desired product. An additional portion of brosyl chloride (25 mg) was added to the reaction mixture. After stirring an additional 4 hr, the reaction mixture was diluted with CH₂Cl₂ (5 mL), washed with aqueous 0.5 N HCl (12 mL) and 5% NaHCCh (15 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to afford crude brosylate E21 (106 mg). This crude product was used directly for the next step.

E22

In a 25 mL sealed tube, a solution of brosylate E21 (106 mg, 0.17 mmol), 2-(4- isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol E9 (46 mg, 0.15 mmol), and Cs₂CCb (64 mg, 0.45 mmol) in NMP (0.5 mL) was submerged in 50 ⁰C oil bath and stirred overnight (15 hr). HPLC analysis indicated that the reaction was complete. After cooling to room temperature, EtOAc (12 mL) was added, and the suspension was washed with 5% NaHCθ3 (2 x 10 mL). The combined aqueous layers were backwashed with ethyl acetate (15 mL). The combined organic layers were washed with saturated NaHCθ3 (2 x 10 mL), 0.2 N HCl (20 mL) and brine (20 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to afford crude E22. The crude solid was purified by column chromatography (4 g silica gel, ISCO) by eluting with a 0% - 4% methanol gradient in CH₂Cl₂. Appropriate fractions were combined and concentrated under reduced pressure to afford compound E22 (75 mg, 68% for two steps). Mass (ESI positive): 691 [M+H]⁺.

E22 E23

A solution of lithium hydroxide (282 mg, 8.1 mmol) in water (17 mL) was added dropwise to a stirred solution of compound E22 (1.15 g, 1.7 mmol) in a mixture of tetrahydrofuran (34 mL) and methanol (17 mL) at room temperature. Subsequently, the flask was fitted with a reflux condenser and was submerged in a 40 ⁰C oil bath. After 4 hr, an additional portion of lithium hydroxide (60 mg) was added as a solid in one portion. After stirring for 90 additional minutes, LCMS analysis showed complete conversion to the desired product. After cooling to room temperature, the reaction mixture was submerged and stirred vigorously in an ice bath; aqueous 3N H₂SO₄ (~ 2.5 mL) was added slowly until the stirred solution reached a pH of around 4 to 5. After volatiles were removed under reduced pressure, the aqueous suspension was diluted with water (100 mL) and extracted with ethyl acetate (150 mL). The aqueous layer was extracted with additional ethyl acetate (2 x 40 mL). The combined organic layers were washed with water (70 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to afford compound E23 as a white solid. This product was used directly in the next step without any further purification.

E23 E24

Compound E23 (1.1 g, 1.6 mmol) was diluted with dry THF (14 mL) and stirred under argon at room temperature until it dissolves. CDI (530 mg, 3.2 mmol) was added in one portion. After stirring at room temperature for 5 min, the flask was submerged in a 65 ⁰C oil bath and was stirred under argon for 5 h. LCMS analysis showed conversion to the azalactone intermediate. After cooling the reaction to near room temperature, cyclopropylsulfonamide (446 mg, 3.5 mmol) was added in one portion, followed by dropwise addition of DBU (521 μL, 3.4 mmol). The reaction flask was submerged in a 50 ⁰C oil bath. After stirring under argon for 2 h, LCMS analysis showed complete conversion of the azalactone intermediate to the desired product. After cooling to room temperature, the reaction mixture was treated with 70 mL of an aqueous solution (pH ~ 5) of citric acid, extracted with ethyl acetate (4 x 60 mL) and washed with water (70 mL) and brine (70 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (50 g Biotage silica gel cartridge) by eluting with a gradient of methanol in dichloromethane (0 to 2%). Appropriate fractions were combined, filtered through a fine fritted filter, and concentrated under reduced pressure to afford compound E24 (748 mg) as an off white solid. The material arising from the combined mixed fractions (~ 100 mg) was applied to a 10 g silica gel column and eluted with a gradient of ethyl acetate in hexanes (30 - 50%) to afford an additional 70 mg of compound E24 (total of 818 mg, 64% yield). Mass (ESI positive): 766 [M+H]⁺. Compound 3.

Compound 3 was prepared according to the following protocol.

Synthesis of compound E25.

E²⁵

To a solution of 5-bromo-l-pentene (3.0 g, 20.1 mmol) in 30 ml of acetone was added NaI (4.53 g, 30.2 mmol). The resulting reaction mixture was refluxed for 5 hours. GC analysis showed that 5-bromo-l-pentene was completely consumed. After cooling to room temperature, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure to afford 5-iodo-l-pentene E25 (2.0 g, 51% yield) which was used in the next step without further purification.

Synthesis of compound E27.

E26 E27 Sliver oxide (1.4 g, 6.2 mmol) was added over 2 hours to a mixture of methyl D- lactate E26 (0.85 g, 8.1 mmol) and 5-iodo-l-pentene E25 (2.0 g, 10.3 mmol). The reaction mixture was stirred at room temperature for 3 days. GC analysis showed that no starting material was left. The reaction mixture was diluted with ether (50 mL) and filtered. The filtrate was concentrated under reduced pressure to give compound E27 (730 mg, 52% yield) that was used in the next step without further purification.

Synthesis of compound E28.

E27 E28 To a solution of E27 (0.73 g, 4.2 mmol) in THF (6.4 mL) was added a 1 M LiOH aqueous solution (6.4 mL). The reaction mixture was stirred at room temperature for 4 hours. TLC showed compound E27 was completely consumed. The reaction mixture was acidified to pH = 2 with a IN HCl aqueous solution and extracted with EtOAc (2 x 50 mL). The combined organic layers were dried over anhydrous MgSO₄ and concentrated under reduced pressure to afford E28 (650 mg, 97% yield) that was used in a later step without further purification.

Synthesis of compound E29.

To a mixture of N-Boc-c/s-4-hydroxy-L-proline (900 mg, 3.9 mmol) and 1- amino-2-vinyl-cyclopropanecarboxylic acid methyl ester tosylate salt (1.4 g, 4.3 mmol) in dichloromethane (20 ml) cooled in an ice/water bath was added HATU (1.6 g, 4.3 mmol). Then a solution of DIPEA (1.5 g, 11.7 mmol) in dichloromethane was added dropwise over 30 minutes. The reaction mixture was stirred at room temperature for 2.5 hours. The reaction was diluted with dichloromethane (20 mL) and sequentially washed with IN HCl (15 mL), aqueous saturated ΝaHCθ3 (15 mL) and brine (15 mL). The organic layer was dried over Na₂SO₄ and filtered. Solvents were removed under reduced pressure and the residue purified by silica gel column chromatography using a MeOH/CH₂Cl₂ solvent system to afford compound E29 (841mg, 61% yield). Mass (ESI positive): 355 [M+H]⁺

Synthesis of compound E30.

E29 E30

To a solution of compound E29 (1.6 g, 4.5 mmol) in dichloromethane (5 mL) was added TFA (5 mL). The reaction mixture was stirred at room temperature for 3h. The reaction mixture was then concentrated under reduced pressure to give compound E30 which was used in the next step directly. Synthesis of compound E31.

To a mixture of compound E28 (0.65 g, 4.1 mmol), HATU (1.72 g, 4.5 mmol) and compound E30 (4.5 mmol) in dichloromethane (30 mL) was added DIPEA 5 (1.59 g, 12.3 mmol) dropwise over 1 h. The reaction mixture was stirred for an additional 2 hours. LCMS analysis showed the starting material was completely consumed. The reaction solution was diluted with dichloromethane (50 mL) and washed with aqueous IN HCl, aqueous saturated NaHCθ3 and brine. The organic layer was dried over anhydrous Na₂SO₄, filtered and concentrated under 10 reduced pressure. The residue was purified by silica gel column chromatography using a MeOH/CH₂Cl₂ solvent system to give compound E31 (800 mg, 45% yield). Mass (ESI positive): 395 [M+H]⁺.

Synthesis of compound E32.

I₅ E31 E32

To a solution of compound E31 (800 mg, 1.96 mmol) in dichloromethane (10 mL) was added dropwise a solution of methanesulfonyl chloride (280 mg, 2.4 mmol) in dichloromethane (10 mL) at 0⁰C .The reaction was stirred for 2 hours. TLC showed that no starting material was left. The reaction mixture was 0 washed with saturated aqueous NaHCCh (2 x 5 mL) and brine. The organic layer was dried, filtered and concentrated under reduced pressure to give compound E32 (776 mg, 81% yield) that was used in the next step without further purification.

A mixture of mesylate E32 (776 mg, 1.6 mmol), compound E9 (515 mg, 1.64 mmol) and Cs₂CO₃ (1.08 g, 3.3 mmol) in DMF (5 mL) was heated at 70⁰C overnight. LCMS showed compound E32 was completely consumed. The reaction was cooled to room temperature and solvents were removed under reduced pressure. The residue was purified by silica gel column chromatography using a MeOHZCH₂Cl₂ solvent system to give compound E33 (658 mg, 58% yield). Mass (ESI positive): 691 [M+H]⁺.

E33 E34

A solution of compound E33 (658 mg, 0.95 mmol) in DCE (437 mL) was purged with N₂ for 30 min and then heated to 75 C. A solution of Hoveyda-Grubbs 1^st generation catalyst (57 mg, 0.095 mmol) in DCE (20 mL) was added dropwise over 2 hours. After 7 hours, TLC indicated that no starting material was left. The solvent was removed under reduced pressure and the residue purified by silica gel column chromatography using a MeOH/CH₂Cl₂ solvent system to give compound E34 (354 mg, 56% yield). Mass (ESI positive): 663 [M+H]⁺. ynthesis of compound E35.

E34 E35

A solution of lithium hydroxide monohydrate (520 mg, 12.7 mmol) in water (5 mL) was added to a stirred solution of methyl ester E34 (200 mg, 0.3 mmol) in a mixture of THF (10 mL) and of MeOH (7 mL). After 5 hours at room temperature, TLC showed that no starting material was left. The reaction mixture was quenched by addition of a saturated aqueous NH₄Cl solution. Solvents were partially removed under reduced pressure. The resulting solution was acidified to pH = 3 with 1 N HCl and extracted with dichloromethane. The organic phases were dried, filtered and evaporated under reduced pressure to give acid E35 (190 mg, 97% yield). Mass (ESI positive): 649 [M+H]⁺.

E35 E36 To a solution of acid E35 (190 mg, 0.3 mmol) in THF (10 mL) was added CDI (105 mg, 0.65 mmol). The reaction solution was refluxed under N₂ for 2 hours. After being cooled to room temperature, cyclopropylsulfonamide (135 mg, 1.1 mmol) and DBU (96 mg, 0.64 mmol) were added. The solution was heated at 50⁰C overnight. Then the reaction mixture was cooled down to room temperature and concentrated under reduced pressure. The residue was partitioned between dichloromethane and IN aqueous HCl. The organic layer was washed with brine, dried, filtered and evaporated under reduced pressure. Purification by column chromatography using a MeOH/Cf^C^ solvent system afforded compound E36 (82 mg, 37% yield). Mass (ESI positive): 752 [M+H]⁺.

Compound 4.

E37 Compound 4

Compound E37 (63 mg, 0.1 mmol) was dissolved in anhydrous dichloromethane (1 mL). EDAC (23 mg, 0.12 mmol) was added in one portion. The reaction was stirred at room temperature for 90 minutes. LCMS indicated complete conversion to the azalactone intermediate. Cyclopropyl sulfonamide (18 mg, 0.15 mmol) was added in one portion followed by dropwise addition of DBU (37 uL, 0.24 mmol). The reaction was heated to 35 ⁰C and stirred for 2-3 hours. LCMS showed there was complete reaction. The reaction was diluted with

CH₂CI₂ (3 mL) and washed with 5% citric acid and brine. The organic layer was dried over Na₂SO₄, filtered and concentrated under reduced pressure. The resulting mixture was purified in a Biotage instrument with a 4 gram silica gel column using a MeOH/CH₂Cl₂ solvent system. Appropriate fractions were combined and evaporated under reduced pressure to yield compound 4 as a white solid (50 mg, 67% yield). Mass (ESI positive): 749 [M+H]⁺.

Compound E37 was prepared following the procedures described in the synthesis of Compound 2 using 7-methoxy-8-methyl-2-(3-isopropyl-lH-pyrazol- l-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol. Compound 5.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(5- methylisoxazol-3-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7- methoxy-8-methylquinolin-4-ol. 7-Methoxy-8-methyl-2-(5-methylisoxazol-3- yl)quinolin-4-ol E39 was synthesized according to the following procedures.

Synthesis of compound E38.

E38 To a solution of S-methylisoxazole-S-carboxylic acid (1 g, 7.9 mmol) in DCM (15 mL) was sequentially added DMF (0.5 mL) and (COCl)₂ (0.8 mL, 9.5 mmol). Upon completion of the reaction as indicated by TLC, solvents were removed under reduced pressure. The crude was used directly in the next step without further purification. To a solution of crude 5-methylisoxazole-3-carbonyl chloride (6.8 mmol) in

DCM (7.5 mL) under nitrogen was added a solution of l-(2-amino-4-methoxy-3- methylphenyl)ethanone (1 g, 5.5 mmol) in DCM (7.5 mL). The reaction was stirred overnight and then solvents were removed under reduced pressure. The residue was sequentially washed with aqueous IN HCl and saturated NaCl solution. The organic layer was dried over Na₂SO₄, filtered and evaporated. The residue was used directly in the next step without further purification.

Synthesis of compound E39.

A mixture of E38 (1.6 g, 5.5 mmol) and t-BuOK (1.5 g, 13.1 mmol) in t-BuOH (15 mL) was refluxed overnight. Then solvents were removed under reduced pressure. The residue was dissolved in water (10 mL) and treated with IN HCl to adjust the pH to 3-4. The crude product precipitated out. It was then purified by silica gel column chromatography using a MeOH/DCM solvent system to afford compound E39 (0.5 g, 33% yield). Mass (ESI positive): 271 [M+H]⁺.

Compound 6.

Compound 6 was prepared according to the following protocols.

Synthesis of compound E40.

E40

To a solution of (-)-Methyl L-lactate (2 g, 19.2 mmol) in DMF (190 mL) cooled to 0 ⁰C was added 60% NaH (845 mg, 21.1 mmol) in three portions. The reaction was stirred at 0 ⁰C for 25 minutes and at room temperature for 15 minutes. 6-Bromo- 1 -hexene (3.2 mL, 23 mmol) was added and the reaction was allowed to stir overnight. The reaction was cooled down to 0 ⁰C and a precooled solution of saturated NH₄Cl (100 mL) was added dropwise. Et₂O (100 mL) was slowly added with vigorous stirring. A precipitate forms that dissolves upon addition of water (200 mL). The ether layer was separated and washed with water (3 x 100 mL). The organic layer was separated, dried, filtered and evaporated under reduced pressure. The residue was purified by silica gel column chromatography using a C^CVhexane solvent system to give compound E40 (1.2 g, 35% yield).

Synthesis of compound E41.

E40 E41

Compound E40 was saponified according to the procedure described above for the synthesis of Compound 2. Synthesis of compound E42.

Compound E41 was coupled to E18 according to the procedure described above for the synthesis of Compound 2.

Synthesis of compound E44.

E42 E43 E44

To a solution of compound E42 (1.05 g, 2.5 mmol) in DMF (10 mL) at room temperature was sequentially added imidazole (511 mg, 7.5 mmol) and triisopropylsilyl chloride (1.12 mL, 5.3 mmol). The reaction was stirred overnight. A second portion of imidazole (0.5 eq) and triisopropylsilyl chloride (0.5 eq) was added and the reaction was stirred for an additional 5 hours. A l N solution of HCl (10 mL) and a 4: 1 mixture of ether/ethyl acetate were added. The organic phase was washed with water (3 x 10 mL), dried, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/hexanes mixtures (10 - 20%) as solvent system. The fractions containing the compound with the higher Rf on TLC of the two stereoisomers were combined and evaporated to afford compound E44 (510 mg, 35% yield). Mass (ESI positive): 579 [M+H]⁺. Synthesis of compound E45.

E44 E45

Compound E45 was prepared following the procedures described in the synthesis of Compound 2 using compound E44 in place of compound E19. Mass (ESI positive): 551 [M+H]⁺.

Synthesis of compound E46.

E45 E46

A solution of compound E45 (330 mg, 0.59 mmol) in a 2: 1: 1 mixture of aqueous 3N HCl/EtOH/THF (10 mL) was stirred at room temperature for 3 hours. It was then diluted with an aqueous saturated NaHCθ3 solution followed by removal of the volatiles under reduced pressure. The resulting mixture was extracted with EtOAc (20 mL). The organic layer was washed with water (10 mL), brine, dried over Na₂SO₄, filtered and evaporated under reduced pressure. The resulting residue was purified by column chromatography (1O g Biotage silica gel cartridge) by eluting with a gradient of methanol in dichloromethane to afford compound E46. Mass (ESI positive): 395 [M+H]⁺.

Synthesis of compound E47 (Compound 6).

E47

Compound E47 was prepared following the procedures described in the synthesis of Compound 2 using compound E46 in place of compound E20. Mass (ESI positive): 766 [M+H]⁺.

Compound 7.

The title compound was prepared following the procedures described above using intermediate E49. E49 was synthesized according to the following procedures.

Synthesis of compound E48.

To a solution of ?rα«s-N-(?er?-butoxycarbonyl)-4-hydroxy-L-proline (2 g, 8.6 mmol), l-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester tosylate salt E16 (3.1 g, 9.5 mmol) and HATU (3.5 g, 9 mmol) in DMF (29 mL) under argon cooled to 0°C was slowly added ¹Pr₂NEt (4.7 mL, 26.7 mmol). The reaction was stirred for 45 minutes at 0°C and allowed to warm up to room temperature over 30 minutes. It was quenched with IN HCl and extracted three times with EtOAc. The combined organic layers were washed with a saturated aqueous solution of NaHCθ3, dried over Na₂SO₄ and filtered. Solvents were removed under reduced pressure and the residue purified by column chromatography (50 g Biotage silica gel cartridge) by eluting with a gradient of EtOAc in dichloromethane. Appropriate fractions were combined and washed with a saturated aqueous solution of NaHCθ3, dried over Na₂SO₄ and filtered. Solvents were removed under reduced pressure to afford compound E48 (2.84 g, 89 % yield). Mass (ESI positive): 369 [M+H]⁺.

To a solution of compound E48 (614 mg, 1.7 mmol) in dichloromethane (13 mL) was added CDI (252 mg, 2 mmol) in one portion. After stirring for 1 hour, there is still some E48 left so a second portion of CDI (63 mg, 0.5 mmol) was added. After stirring for an additional hour, isoindoline (568 μL, 5 mmol) was added dropwise over four minutes. The reaction mixture was stirred overnight. It was then cooled with an ice/water bath to 0 ⁰C, diluted with dichloromethane (20 mL) and washed with aqueous IN HCl (20 mL). The organic phase was separated, dried over Na₂SO₄ and filtered. The organic solvents were removed under reduced pressure and the residue was purified by silica gel column chromatography using an EtOAc/hexanes solvent system to afford compound E49 (690 mg, 80% yield). Mass (ESI positive): 514 [M+H]⁺.

Compound 10.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 2-(hex-5-enyloxy)-2-methylpropanoic acid in place of intermediate E15. The 2-(hex-5-enyloxy)-2-methylpropanoic acid was prepared according to the following procedure. Synthesis of E50.

E50

A mixture of 5-hexen-l-ol (5 g, 50 mmol) and potassium hydroxide (11.2 g, 200 mmol) in acetone (24 mL) was stirred for 30 minutes. It was then cooled to 0 ⁰C and CHCI3 (16 mL, 200 mmol) was added. The reaction was stirred for 2 hours at 0 ⁰C and for an additional 2 hours at reflux. It was then cooled down and filtered. The filtrate volume was reduced in half by evaporation under reduced pressure. 20% H₂SO₄ was added and the reaction mixture was stirred at room temperature overnight. It was then extracted with ethyl acetate. The organic layer was dried, filtered and evaporated under reduced pressure. The residue was purified by column chromatography using hexane/ethyl acetate mixtures to afford compound E50 (5.5 g, 59% yield).

Compound 13. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using 1-methyl-cyclopropanesulfonamide in place of the cyclopropanesulfonamide.

Compound 14. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using benzylsulfonamide in place of the cyclopropanesulfonamide.

Compound 15. The title compound was prepared following the procedures described in the synthesis of Compound 2 using (R)-(-)-mandelic acid in place of the (+)-Methyl D-lactate.

Compound 17. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using phenylsulfonamide in place of the cyclopropanesulfonamide. Compound 18.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 2-(4-isopropylthiazol-2-yl)-7-methoxyquinolin- 4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4- ol.

Compound 19.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 7-methoxy-2-(3-methyl-lH-pyrazol-l- yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol. The 7-methoxy-2-(3-methyl-lH-pyrazol-l-yl)quinolin-4-ol was prepared as described in WO 2000/059929 Al

Compound 20. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(3-methyl-lH- pyrazol-l-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy- 8-methylquinolin-4-ol.

Compound 21.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-2-(4-methyl-lH-pyrazol-l- yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol.

Compound 22.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 2-ethoxy-7-methoxy-8-methylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol. The 2-ethoxy-7-methoxy-8-methylquinolin-4-ol was prepared as described in WO 2008/059046 Al. Compound 23.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(3 -methyl- lH-pyrazol-1 -yl)quinolin-4- ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 24.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 7-methoxy-2-phenylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 25.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using 6-fluoro-2-phenylquinolin-4-ol in place of the 2- (4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 26.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using phenethylamine in place of the cyclopropanesulfonamide.

Compound 27.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using 1-pyrrolidinesulfonamide in place of the cyclopropanesulfonamide.

Compound 28.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using N,N-dimethylsulfamide in place of the cyclopropanesulfonamide.

Compound 29.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(lH-pyrazol-l- yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol. Compound 30.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(5-(4-methoxyphenyl)-2H-l,2,3- triazol-4-yl)thiazole in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol. The 2-(5-(4-methoxyphenyl)-2H-l,2,3-triazol-4-yl)thiazole was prepared according to WO 2008/021871 A2.

Compound 31.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 6-methoxy-2-phenylpyrimidin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol. The 6-methoxy-2-phenylpyrimidin-4-ol was prepared as described in WO 2008/095999 Al.

Compound 32.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 8-methyl-2-phenylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 33.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(6- methylpyridin-2-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7- methoxy-8-methylquinolin-4-ol. The 7-methoxy-8-methyl-2-(6-methylpyridin-2- yl)quinolin-4-ol was prepared as described in WO 2007/014926 Al.

Compound 34.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-phenylquinolin- 4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4- ol. Compound 35.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 6-fluoro-3-phenylquinolin-2-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 36.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using methanesulfonamide in place of the cyclopropanesulfonamide.

Compound 37.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using 7-bromo- 1 -heptene for the alkylation of the (+)-methyl D-lactate.

Compound 38.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 3 using 2-propanesulfonamide in place of the cyclopropanesulfonamide.

Compound 39.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-phenylquinolin-4-ol in place of the 2- (4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 40.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 8-chloro-2-methylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 41.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-2-propylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol. Compound 42.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 8-chloro-2-phenylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8-methylquinolin-4-ol.

Compound 43.

The title compound was prepared following the procedures described in the synthesis of Compound 20 using 1-methyl-cyclopropanesulfonamide in place of the cyclopropanesulfonamide.

Compound 44.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(6-isopropylpyridin-2-yl)-7-methoxy- 8-methylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol. The 2-(6-isopropylpyridin-2-yl)-7-methoxy-8- methylquinolin-4-ol was prepared as described in WO 2007/014926 Al.

Compound 45.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(4-isopropylthiazol-2-yl)-7- methoxyquinazolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol.

Compound 46. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(l -methyl- IH- pyrazol-3-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy- 8-methylquinolin-4-ol.

Compound 47.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(l,3-dimethyl-lH-pyrazol-5-yl)-7- methoxy-8-methylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7- methoxy-8-methylquinolin-4-ol. Compound 48.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 2-(2-isopropyl-5-methyloxazol-4-yl)-7- methoxy-8-methylquinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7- methoxy-8-methylquinolin-4-ol.

Compound 49.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(4-methylthiazol- 2-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol.

Compound 50.

The title compound was prepared following the procedures described in the synthesis of Compound 49 using 1-methyl-cyclopropanesulfonamide in place of the cyclopropanesulfonamide.

Compound 51.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(l -methyl- IH- pyrazol-5-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy- 8-methylquinolin-4-ol.

Compound 52. The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using 7-methoxy-8-methyl-2-(2-methyloxazol- 4-yl)quinolin-4-ol in place of the 2-(4-isopropylthiazol-2-yl)-7-methoxy-8- methylquinolin-4-ol.

Compound 53.

The title compound was prepared following the procedures described in the synthesis of Compounds 4 and 19 using 1-methyl-cyclopropanesulfonamide in place of the cyclopropanesulfonamide. Compound 54.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using (R)-3-hydroxy-2-methylpropanoic acid ester in place of the (+)-methyl D-lactate.

Compound 55.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using (R)-2-hydroxy-4-phenylbutanoic acid ethyl ester in place of the (+)-methyl D-lactate.

Compound 56.

The title compound was prepared following the procedures described in the synthesis of Compound 2 using (R)-2-hydroxy-3-phenylpropanoic acid ethyl ester in place of the (+)-methyl D-lactate. The (R)-2-hydroxy-3-phenylpropanoic acid ethyl ester was prepared according to the following procedure.

Synthesis of compound E51.

(R)-2-Amino-3-phenylpropionic acid (5.3 g, 32 mmol) was placed into a three- necked flask, and water (24 mL) was added. The flask was fitted with two addition funnels. 2N H₂SO₄ (18 mL) was placed in one addition funnel and 2N aqueous NaNO₂ (18 mL) was placed in the other one. The reaction vessel was cooled to 0°C, and the acid was added dropwise with stirring. After the initial amino acid was dissolved, dropwise addition of the NaNO₂ solution started. Upon completion of the addition, the reaction was stirred at 0°C for 3 hours and then allowed to stir at room temperature for 2 days. Then the reaction mixture was extracted several times with ethyl acetate. The combined organic layers were dried, filtered and concentrated under reduced pressure to give E51 (3.6 g, 68% yield) as a white solid. Synthesis of compound E52.

E51 E52

Triethylsilylchloride (0.2 ml, 1.5 mmol) was added to a solution of acid E51 (2.5 g, 15.0 mmol) in diethoxyethane (24 mL) and EtOH (6 mL). The mixture was stirred for 16 hours at room temperature and the solvent removed under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate mixtures to give E52 (2.3 g, 79% yield) as a white solid. Mass (ESI positive): 195 [M+H]⁺

Compound 57.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using (R)-2-hydroxy-4-methylpentanoic acid ethyl ester in place of the (+)-methyl D-lactate.

Compound 58.

The title compound was prepared following the procedures described in the synthesis of Compounds 2 and 4 using (5)-3-hydroxy-2-methylpropanoic acid methyl ester in place of the (+)-methyl D-lactate.

The following compounds can also be prepared using the methods described above:

Table 2: Further Examples

Biochemical analysis of compounds against HCV genotypes Ib, Ia, 2a, and 3a

All biochemical assays were conducted using the buffer and substrate conditions specified in the Enzolyte 520 HCV Protease Assay Kit from Anaspec (#71145). HCV NS3/4A Genotype Ib from strain Con-1, Genotype Ia from strain H77, Genotype 2a from strain HC- J6, and Genotype 3a from strain N2L1 were custom prepared by Protein One (Bethesda, MD). The NS3 protease (amino acids 3 - 181) was expressed with an NS4A fragment (His-NS4A₂i-3₂- GSGS-NS3-181) as a single peptide in bacterial cells and purified using immobilized metal ion chromatography followed by additional FPLC methods. Purified HCV proteases were analyzed by Nu-PAGE and confirmed by western blot analysis. Phenomix protease inhibitors were prepared as 10 mM stocks in DMSO and stored at -20⁰C.

HCV protease inhibitors were screened using a fluorescence resonance energy transfer (FRET) based assay. The FRET peptide was derived from the natural cleavage site at the NS4A/NS4B junction of the HCV polypeptide, and dual labeled with 5-carboxyfluorescein (5-FAM) and QXL™520 dyes. 5-FAM fluorescence is quenched by QXL™520 when the peptide is intact, only to be recovered upon protease driven cleavage of the substrate. Protease enzymes were diluted in reaction buffer to provide a signal to noise (SfN) ratio above 4 after 10 minutes of substrate turnover at room temperature. Compounds were assayed in an 8 dose format using half-log dilutions ranging from 2 to 6250 nM. For compounds showing potency < 2 nM, assay sensitivities allowed for repeat testing at compound concentrations from 0.2 to 625 nM to quantitate activity.

Test wells in 96-well plates contained 45 uL of diluted protease and 5 uL of compound titrated in DMSO. Enzyme only (max) wells contained 45 uL of diluted protease and 5 uL of DMSO. No enzyme (min) wells contained 45 uL of reaction buffer and 5 uL of DMSO. All compounds were assayed in duplicate. The assay plate was incubated at room temperature for 20 minutes post compound addition to allow for equilibration of test compounds with the enzyme. 50 uL of FRET-substrate diluted in reaction buffer was then added to all wells followed by incubation at room temperature for 10 minutes.

Fluorescence in each well was measured at settings of Ex/Em= 485/535 nm using a Wallac Victor2 1420 plate reader (Perkin Elmer). The percent inhibition of protease activity was determined using the fluorescence value (FV) at each concentration of a given test compound. Percent inhibition was defined as: 100% X (max- FV) / (max- min)

IC50/90 calculations were performed from the calculated percent inhibition data by non-linear regression analysis using Prism software (GraphPad).

The following Table 3 lists compounds that were prepared according to any one of the above procedures and their activities in the assays described above. The compound numbers correspond to the compound numbers in Table 1.

Wherein:

A indicates IC50 of less than 10 nM B indicates 10 nM < IC50 <100 nM C indicates 100 nM < IC50 < 1 μM D indicates IC50 > 1 μM

Determination of the effective concentration in the cell based replicon assay.

Huh-luc/neo-ET cells expressing the persistent replicon I^luc-ubi- neo/NS3-37ET were purchased from Reblikon. Cells were cultured in DMEM media (Invitrogen) containing 10% FBS (Biowhittaker), IX Antibiotic- Antimycotic (Invitrogen), IX nonessential amino acids (Invitrogen), and 1 mg/mL G418 (Invitrogen). Cells were housed in a 37⁰C incubator with 5% CO2 and were passaged regularly. Huh-luc/neo-ET cells (5,000 cells per well) were seeded in a 96-well plate in DMEM media (without phenol red or G418) and were incubated at 37⁰C overnight. Compounds were serial diluted in the same media plus 1% BSA (Invitrogen) and 0.5% DMSO (Sigma) in a BSA pre-coated plate (1% BSA incubated for 24 hours at 37⁰C). Compounds were added to the Huh-luc/neo-ET cells and incubated for 72 hours. Bioluminecence was measured using the

BriteLite Ultra-High Sensitivity Reporter Assay System (PerkinElmer) and read in a Wallac Microbeta TriLux luminometer (PerkinElmer). EC50 values were calculated from a non-linear regression using Prism software (Graph Pad).

Table 4 shows the activity of compounds of this invention in the assay described above. The compound numbers correspond to the compound numbers in Table 1.

Wherein:

A indicates IC50 of less than 10 nM B indicates 10 nM < IC50 <100 nM C indicates 100 nM < IC50 < 1 μM D indicates IC50 > 1 μM Biochemical analysis of the compounds against mutant HCV proteases

The activity of compounds of this invention against clinically relevant HCV NS3/4A mutants was evaluated. All biochemical assays were conducted using the buffer and substrate conditions specified in the Enzolyte 520 HCV Protease Assay Kit from Anaspec (#71145). Mutant Proteases A156S/T/V, R155K, V36M, T54A, and D168A/V created from a genotype Ib strain Con-1 background were custom ordered from BioEnza (Mountain View, CA). The NS3 protease (amino acids 3 - 181) was expressed with an NS4A fragment (His- NS4A₂₁-3₂-GSGS-NS3-₁8₁) as a single peptide in bacterial cells and purified using immobilized metal ion chromatography followed by additional FPLC methods. Purified HCV proteases were analyzed by Nu-PAGE and confirmed by western blot analysis. Phenomix protease inhibitors were prepared as 10 mM stocks in DMSO and stored at -20⁰C. HCV protease inhibitors were screened using a fluorescence resonance energy transfer (FRET) based assay. The FRET peptide was derived from the natural cleavage site at the NS4A/NS4B junction of the HCV polypeptide, and dual labeled with 5-carboxyfluorescein (5-FAM) and QXL™520 dyes. 5-FAM fluorescence is quenched by QXL™520 when the peptide is intact, only to be recovered upon protease driven cleavage of the substrate. Protease enzymes were diluted in reaction buffer to provide a signal to noise (SfN) ratio above 4 after 10 minutes of substrate turnover at room temperature. Compounds were assayed in an 8 dose format using half-log dilutions. Test wells in 96-well plates contained 45 uL of diluted protease and 5 uL of compound titrated in DMSO. Enzyme only (max) wells contained 45 uL of diluted protease and 5 uL of DMSO. No enzyme (min) wells contained 45 uL of reaction buffer and 5 uL of DMSO. All compounds were assayed in duplicate. The assay plate was incubated at room temperature for 20 minutes post compound addition to allow for equilibration of test compounds with the enzyme. 50 uL of FRET-substrate diluted in reaction buffer was then added to all wells followed by incubation at room temperature for 10 minutes. Fluorescence in each well was measured at settings of Ex/Em= 485/535 nm using a Wallac Victor2 1420 plate reader (Perkin Elmer). The percent inhibition of protease activity was determined using the fluorescence value (FV) at each concentration of a given test compound. Percent inhibition was defined as: 100% X (max- FV) / (max- min)

Table 5 shows the activity of compounds of this invention in the assays described above. The compound numbers correspond to the compound numbers in Table 1.

Wherein:

Specificity Assays.

Although the HCV NS3/4A protease is of viral origin, it was identified as a chymotrypsin-like protease with some similarity to human proteases.

Therefore targeting NS3/4A requires screening against human proteases to assure selectivity. Compounds of the present invention were screened against five human proteases: chymotrypsin, human neutrophil elastase (HNE), cathepsin G, cathepsin B, and chymase. Chymotrypsin (#230900), human neutrophil elastase (HNE) (#324681), cathepsin B (#219362), and cathepsin G (#219373) were purchased from Calbiochem. Chymase (#SE-281) was from Biomol. Suc-Ala-Ala-Pro-Phe-AMC (#230914), MeOSuc-Ala-Ala-Pro-Val-AMC (#324740), and Z-Arg-Arg-AMC (#219392) fluorogenic substrates were purchased from Calbiochem. The reference compounds chymostatin (#230790) and cathepsin G Inhibitor I

(#219372) were also from Calbiochem. E-64 (#E3132) and MeOSuc-Ala-Ala- Pro-Val-chloromethyl ketone (#M0398) were available from Sigma. Phenomix protease inhibitors and reference compounds were prepared as 10 mM stocks in DMSO and stored at -20⁰C. HCV protease inhibitors were screened for selectivity using AMC- labeled fluorogenic substrates. Protease driven cleavage of AMC from the peptide substrate leads to detectable fluorescence, allowing for quantitation of protease activity. The chymotrypsin assay was run in 100 mM Hepes pH 7.5, 100 mM NaCl, 20 mM CaCl₂, and 0.125 mg/mL BSA using 20 uM of the Suc- Ala-Ala-Pro-Phe-AMC substrate. The HNE assay was run in 62.5 mM Hepes pH 7.8, 625 mM NaCl, and 1.25 mg/mL BSA using 200 uM MeOSuc-Ala-Ala- Pro-Val-AMC. The cathepsin B assay was run in 100 mM sodium/potassium phosphate buffer pH 6.8, 1 mM EDTA, and 2 mM DTT using 200 uM of the Z- Arg-Arg-AMC substrate. The cathepsin G assay was run in 100 mM Hepes pH 7.5, 500 mM NaCl, 1 mM DTT, and 0.125 mg/mL BSA using 200 uM Suc-Ala- Ala-Pro-Phe-AMC. The chymase assay was run in 200 mM Hepes pH 8.0, 2 M NaCl, and 0.01% Triton X-100 using 20 uM of the Sue-Ala- Ala-Pro-Phe- AMC substrate. Protease enzymes were diluted in the appropriate assay buffer to provide a signal to noise (SfN) ratio allowing for a sensitive and reproducible assay. Chymotrypsin, HNE and cathepsin B were optimized for a S/N ratio above 6 after 10 minutes of substrate turnover at room temperature. Due to the lower activity of cathepsin G and chymase, these enzymes required 25 to 30 minutes of substrate turnover to obtain a S/N ratio greater than 3. Compounds were assayed in an 8 dose format using half-log dilutions.

Test wells in 96-well plates contained 45 uL of diluted protease and 5 uL of compound titrated in DMSO. Enzyme only (max) wells contained 45 uL of diluted protease and 5 uL of DMSO. No enzyme (min) wells contained 45 uL of reaction buffer and 5 uL of DMSO. All compounds were assayed in duplicate. The assay plate was incubated at room temperature for 20 minutes post compound addition to allow for equilibration of test compounds with the enzyme. 50 uL of AMC-substrate diluted in reaction buffer was then added to all wells followed by incubation at room temperature for 10 to 30 minutes. Fluorescence in each well was measured at settings of Ex/Em= 380/460 nm using a Wallac Victor2 1420 plate reader (Perkin Elmer). The percent inhibition of protease activity was determined using the fluorescence value (FV) at each concentration of a given test compound. Percent inhibition was defined as: 100% X (max- FV) / (max- min) IC50/90 calculations were performed from the calculated percent inhibition data by non-linear regression analysis using Prism software (GraphPad).

Compounds 1, 2, 6 and 10 had IC50 values greater than 1 μM against all five human proteases. Single dose pharmacokinetic analysis of Compound 2 in solution in Sprague Dawley Rats.

The plasma pharmacokinetics and liver concentrations of Compound 2 after a single administration of 10 mg/kg in solution (60% PEG 400, 10% ethanol and 30% water) by oral gavage to three male Sprague Dawley rats were determined. Eight time points of plasma and 24 hr liver samples were taken from two rats, and seven time points of plasma and 12 hr liver samples were taken from the third animal for plasma and liver concentration determinations.

The PK parameters of Compound 2 were as follows: the mean T_max was 1.8 hr; the mean C_max was 629 ng/ml; the mean AUCo-mf value was 3590 hr*ng/ml; the mean terminal half-life was 3.7 hr. The mean plasma concentrations of Compound 2 were sustained more than 24 hr above its replicon EC₉0 (0.19 ng/ml). These data indicated that Compound 2 was orally available, with a bioavailability of 54% (an intravenous PK study served as the reference).

The rat 12 hr liver concentration was 216 ng/g of liver, which corresponds to 3.4-fold of the plasma concentration from the same animal at that timepoint. The 12 hr liver concentration was 1137-fold of the replicon EC₉0 of Compound 2. The mean 24 hr liver concentration was 3.6 ng/g of liver, which corresponds to 5.8-fold of the plasma concentration from the same animal at that timepoint. The 24 hr liver concentration was 19-fold of the replicon EC₉0 of Compound 2.

These data indicate that a 10 mg/kg dose of Compound 2 administered to rat achieved pharmacologically relevant concentrations exceeding the replicon EC₉0 in both plasma and liver.

When administered at 10 mg/kg as a single dose in solution, Compound 2 did not cause any observed adverse effects.

Single dose pharmacokinetic analysis of Compounds 18, 19, 20 and 43 in solution in Sprague Dawley Rats.

The plasma pharmacokinetics and liver concentrations of Compounds 18, 19, 20 and 43 after a single administration of 10 mg/kg in solution by oral gavage to male Sprague Dawley rats were determined. The PK parameters were as follows:

Single dose pharmacokinetic analysis of Compound 2 in solution in Beagle Dogs.

The plasma pharmacokinetics of Compound 2 after a single administration of 6 mg/kg in solution (60% PEG 400, 10% ethanol and 30% water) by oral gavage to 3 male Beagle dogs was determined. Ten time points of plasma samples were taken from each animal for plasma concentration determination.

The PK parameters of Compound 2 were as follows: the mean T_max was 1.3 hr; the mean C_max was 284 ng/ml; the mean AUCo_-mf value was 665 hr*ng/ml; the mean terminal half-life was 3.5 hr. The mean plasma concentrations of Compound 2 were sustained for over 23 hours above its replicon EC₉0 (0.19 ng/ml). These data indicated that Compound 2 was orally available, with a bioavailability of 26% (an intravenous PK study served as the reference).

These data indicate that a 6 mg/kg dose of Compound 2 administered to dog achieved pharmacologically relevant concentrations exceeding the replicon EC₉0 in plasma.

When administered at 6 mg/kg in a single dose in solution, Compound 2 did not cause any observed adverse effects.

Single dose pharmacokinetic analysis of Compound 2 in solution in Cynomolgus Monkeys.

The plasma pharmacokinetics and the liver concentrations of Compound 2 after a single administration of 6 mg/kg in solution (60% PEG 400, 10% ethanol and 30% water) by oral gavage to 4 male Cynomolgus monkeys were determined. Seven time points of plasma and one 12 hr liver sample from each of two animals, and eight time points of plasma and one 24 hr liver sample from each of the other two animals were taken for plasma and liver concentration determinations.

The PK parameters of Compound 2 were as follows: the mean T_max was 2.4 hr; the mean C_max was 603 ng/ml; the mean AUCo_-mf value was 4397 hr*ng/ml; the mean terminal half-life was 5.1 hr. The mean plasma concentrations of Compound 2 were sustained for over 24 hours above its replicon EC₉0 (0.19 ng/ml). The data indicated that Compound 2 was orally available, with a bioavailability of 57% (an intravenous PK study served as the reference).

The mean 12 hr liver concentration was 393 ng/g of liver, which corresponds to 2-fold of the plasma concentration from the same animals at that timepoint. The 12 hr liver concentration was 2066-fold of the replicon EC₉0 of Compound 2. The mean 24 hr liver concentration was 39 ng/g of liver, which corresponds to 3-fold of the plasma concentration from the same animals at that timepoint. The 24 hr liver concentration was 205-fold of the replicon EC₉0 of Compound 2.

These data indicate that a 6 mg/kg dose of Compound 2 administered to monkey achieved pharmacologically relevant concentrations exceeding the replicon EC₉₀ in both plasma and liver.

It is within ordinary skill to evaluate any compound disclosed and claimed herein for effectiveness in inhibition of HCV protease and in the various cellular assays using the procedures described above. Accordingly, the person of ordinary skill can prepare and evaluate any of the claimed compounds without undue experimentation.

Any compound found to be an effective inhibitor of HCV protease can likewise be tested in animal models and in human clinical studies using the skill and experience of the investigator to guide the selection of dosages and treatment regimens.

While the invention has been described and exemplified in sufficient detail for those skilled in this art to make and use it, various alternatives, modifications, and improvements will be apparent to those skilled in the art without departing from the spirit and scope of the claims.

All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

Claims

CLAIMSWhat is claimed is:

1. A compound of Formula I:

I

B is CH₂; R¹, R^la, R² and R^2a are independently H , halo, or alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J;

X is a bond, O, S, CH(R³) or N(R⁴);

Y is a bond, CH₂, C(O), C(O)C(O), S(O), S(O)₂ or S(O)(NR⁴); provided that when X and Y are both bonds, taken together they form a single bond: and Z is hydrogen, alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, OR⁵, or N(R⁵)₂, wherein a heterocyclyl or heteroaryl group can be bonded by a carbon atom or by a heteroatom, wherein any carbon atom or nitrogen atom is unsubstituted or is substituted with J; R³ is hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J; R⁴ is independently at each occurrence hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylalkenyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any carbon atom or nitrogen atom can be substituted with J, or aralkanoyl, heteroaralkanoyl, C(O)R³ , SO ₂R³ or carboxamido, wherein any aralkanoyl or heteroaralkanoyl is substituted with 0-3 J groups;

R⁵ is independently at each occurrence hydrogen, alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, or heteroarylalkyl, wherein any alkyl, alkenyl, aryl, aralkyl, aralkenyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, or heteroarylalkyl is substituted with 0-3 J, or two R⁵ groups which are bound to a nitrogen atom can form together with the nitrogen atom a 5-11 membered mono- or bicyclic heterocyclic ring system wherein the heterocyclic ring system is substituted with 0-3 J groups and can contain 0-3 additional heteroatoms selected from the group consisting of O, N, NR', S, S(O), and S(O)₂.

D is CR'; m is 0, 1, 2, 3 or 4; n is O, 1, 2, 3 or 4; p is 1, 2, 3, or 4; L is O, S, C₂, C₂H₂ or C₂H₄; M is O, S, S(O), S(O)₂; J is halogen, R', OR', CN, CF₃, OCF₃, O, S, C(O), S(O), methylenedioxy, ethylenedioxy, (CH₂)₀-_pN(R')₂, (CH₂)₀-_pSR', (CH₂)₀-_pS(O)R', (CH₂V_pS(O)₂R', (CH₂)o-_pS(0)₂N(R')₂, (CH₂VpSO₃R', (CH₂VpC(O)R', (CH₂VpC(O)C(O)R¹, (CH₂V_pC(O)CH₂C(O)R', (CH₂V_pC(S)R', (CH₂V_pC(O)OR', (CH₂)₀-_pOC(O)R', (CH₂V_pC(O)N(R)₂, (CH₂V_pOC(O)N(R)₂, (CH₂V_pC(S)N(R)₂, (CH₂)₀-_pNH- C(O)R, (CH₂)o-_pN(R')N(R')C(0)R', (CH₂)₀-_pN(R)N(R')C(O)OR', (CH₂V _pN(R)N(R)C0N(R')₂, (CH₂V_pN(R)SO₂R', (CH₂)₀-_pN(R')SO₂N(R')₂, (CH₂V pN(R)C(0)0R, (CH₂)o-_PN(R')C(0)R', (CH₂VpN(R)C(S)R, (CH₂V pN(R)C(0)N(R')₂, (CH₂)o-_pN(R')C(S)N(R)₂, (CH₂)o-_pN(COR)COR, (CH₂V pN(0R)R', (CH₂)o-_PC(=NH)N(R')₂, (CH₂VpC(O)N(OR)R', or (CH₂V _pC(=N0R)R'; wherein, each R is independently at each occurrence hydrogen, (Ci-Ci₂)-alkyl, (C₂-Ci₂)-alkenyl, (C₂-Ci₂)-alkynyl, (C₃-Cio)-cycloalkyl, (C₃-Ci₀)-cycloalkenyl, [(C₃-Cio)cycloalkyl or (C₃-Ci₀)-cycloalkenyl]-[(Ci-Ci₂)-alkyl or (C₂-C₁₂)- alkenyl or (C₂-C₁₂)-alkynyl], (C₆-C₁₀)-aryl, (C₆-Ci₀)-aryl-[(Ci-Ci₂)-alkyl or (C₂- Ci₂)-alkenyl or (C₂-C i₂)-alkynyl], mono- or bicyclic 3-10 membered heterocyclyl, mono- or bicyclic 3-10 membered heterocyclyl-[(Ci-Ci₂)-alkyl or (C₂-Ci₂)-alkenyl or (C₂-Ci₂)-alkynyl], mono- or bicyclic 5-10 membered heteroaryl, or mono- or bicyclic 5-10 membered heteroaryl-[(Ci-Ci₂)-alkyl or (C₂-Ci₂)-alkenyl or (C₂-Ci₂)-alkynyl], wherein R' is substituted with 0-3 substituents selected independently from J; or, when two R' are bound to a nitrogen atom or to two adjacent nitrogen atoms, the two R groups together with the nitrogen atom or atoms to which they are bound can form a 3- to 8-membered monocyclic heterocyclic ring, or an 8- to 20-membered, bicyclic or tricyclic, heterocyclic ring system, wherein any ring or ring system can further contain 1-3 additional heteroatoms selected from the group consisting of N, NR', O, S, S(O) and S(0)2, wherein each ring is substituted with 0-3 substituents selected independently from J; wherein, in any bicyclic or tricyclic ring system, each ring is linearly fused, bridged, or spirocyclic, wherein each ring is either aromatic or nonaromatic, wherein each ring can be fused to a (Ce-Cio)aryl, mono- or bicyclic 5-10 membered heteroaryl, (C₃-Cio)cycloalkyl or mono- or bicyclic 3-10 membered heterocyclyl;

T is R⁶, alkyl-R⁶, alkenyl-R⁶, or alkynyl-R⁶; and R⁶ is independently at each occurrence hydrogen, alkyl, alkoxy, aryl, aralkyl, cycloalkyl, cycloalkenyl, [cycloalkyl or cycloalkenyl]- [alkyl or alkenyl], heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, heteroaryl, heteroarylalkyl, or heteroarylalkenyl, wherein any R⁶ except hydrogen is substituted with 0-3 J groups.

2. The compound of claim 1, wherein M is O.

3. The compound of claim 1 wherein D is CH or C(CH₃).

4. The compound of claim 1, wherein T is R⁶ and R⁶ is alkyl.

5. The compound of claim 4, wherein R⁶ is methyl.

6. The compound of claim 1, wherein X-Y is O.

7. The compound of claim 1, wherein a carbon atom bearing T is of an R absolute configuration, or is of an S absolute configuration, or is a mixture thereof. The compound of claim 7 wherein the compound of formula (I) is:

wherein n, A, T, X, Y, and Z are as defined in claim 1.

9. The compound of claim 1, wherein A is a group of formula

10. The compound of claim 9, wherein A is a group of formula

11. The compound of claim 1 , wherein X is O.

12. The compound of claim 1, wherein Y is a bond, or Y is C(O).

13. The compound of claim 1, wherein X and Y are both bonds, taken together forming a single bond.

14. The compound of claim 1, wherein Z is an unsubstituted heteroaryl group or is a heteroaryl group mono- or independently pluri-substituted with J.

15. The compound of claim 14, wherein Z is a substituted quinolyl group, a substituted triazolyl group, a substituted tetrazolyl group, or a substituted isoindolinyl group, wherein any group is mono- or independently pluri- substituted with J.

16. The compound of claim 14, wherein Z is a thiazolyl-substituted quinolyl group, or a pyrazolyl-substituted quinolyl group, wherein any group is mono- or independently pluri-substituted with J.

17. The compound of claim 1, wherein m is 0-1, or n is 1-4, or both..

18. The compound of claim 1 wherein T and the CR' of D forms a spirocyclic ring comprising 3, 4, or 5 atoms wherein the spirocyclic ring atoms are all carbon atoms or are all carbon atoms except for a single oxygen atom.

19. The compound of claim 1, wherein Z is a group of formula

wherein a wavy line indicates a point of attachment.

20. The compound of claim 1, wherein L is C₂H₂.

21. The compound of claim 20, wherein the C₂H₂ is in a cis configuration.

22. The compound of claim 1, wherein the compound of formula I is any of the following:

or any salt, pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, or prodrug thereof.

23. A pharmaceutical composition comprising a compound of any one of claims 1-22 and a pharmaceutically suitable excipient.

24. A pharmaceutical combination comprising a compound of any one of claims 1-22 in a therapeutically effective dose and a second medicament in a therapeutically effective dose.

25. A pharmaceutical composition comprising the combination of claim 24 and a pharmaceutically suitable excipient.

26. A method of inhibition of a hepatitis C viral protease, comprising contacting the viral protease with an effective amount of the compound of any one of claims 1-22 or the composition of claim 23.

27. The method of claim 26 wherein the viral protease is present in a body of a living mammal.

28. A method of treatment of a hepatitis C viral infection in a patient, comprising administering to the patient the compound of any one of claims 1-22 or the composition of claim 23 in a therapeutically effective amount.

29. The method of claim 28 further comprising administering to the patient an effective amount of a second medicament.

30. The method of claim 29 wherein the second medicament comprises another compound of claim 1, an other HCV protease inhibitor, interferon alfa- 2b, peginteferon alfa-2b, recombinant interferon alfa-2a, peginterferon alfa-2a, inteferon-alpha 2B+Ribavirin, interferon alpha-nl, a nucleoside analog, an IRES inhibitor, an El inhibitor, an E2 inhibitor, an IMPDH inhibitor, anNS3 inhibitor, an NS5B inhibitor, an NS4B inhibitor, an NS5A inhibitor or an NTPase/helicase inhibitor, or any combination thereof.

31. The method of claim 30 wherein the other HCV protease inhibitor comprises telaprevir (VX950), boceprevir (SCH503034), ITMN191, TMC 435350, MK7009, and PHX1766.

32. The method of claim 29 wherein the second medicament comprises an anti-proliferative agent.

33. The method of claim 32 wherein the anti-proliferative agent comprises 5- fluorouracil, daunomycin, mitomycin, bleomycin, dexamethasone, methotrexate, cytarabine, or mercaptopurine.

34. The method of claim 29 wherein the second medicament comprises an immune modulator, preferably a steroid, preferably prednisone, prednisolone, or dexamethasone; a non-steroidal anti-inflammatory, preferably ibuprofen, naproxen, diclofenac, or indomethacin; a COX2 inhibitor, preferably rofecoxib or celecoxib; an anti-TNF compound, preferably enbrel, infliximab, or adaumimab; an anti-ILl compound, preferably anakinra; an interferon, preferably interferon alfa-2b, peginteferon alfa-2b, recombinant interferon alfa- 2a, peginterferon alfa-2a, or interferon alpha-nl; methotrexate; leflunomide; cyclosporin; FK506; or a combination of any two or more thereof.

35. The combination of claim 24, the composition of claim 25, or the method of claim 29, wherein the second medicament comprises an anti-viral agent other than a compound of formula I as defined herein; or comprises an anti- proliferative agent, an immune modulator, an antibiotic; or any combination thereof.

36. The use of the compound of any one of claim 1-22, the composition of claim 23, the combination of claim 24, or the composition of claim 25 in the preparation of a medicament for treatment of a hepatitis C viral infection or for treatment of a malcondition wherein inhibition of hepatitis C viral protease is medically indicated.

37. A method of preparation of a compound of formula I of claim 1 wherein L is C₂H₂ and m = 0, comprising contacting a compound of formula II

II wherein A' is -C(=O)OR^C and m = 0, and an olefin metathesis catalyst under conditions sufficient to brin^ about formation of the compound of formula Ha

Z

Ha wherein L is C₂H₂ and m = 0.

38. The method of claim 37 wherein the olefin metathesis catalyst comprises dichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II).

39. The method of claim 37 further comprising converting the compound of formula (Ha) to the compound of formula (I) wherein L = C₂H₂ and m = 0, comprising converting the -C(=O)OR^C group of A' of the compound of formula

(Ha) to either of the

groups of A of the compound of formula (I), and then, optionally, contacting the compound of formula I wherein L is C₂H₂ and hydrogen in the presence of a catalyst to provide a compound of formula I wherein L is C₂H₄.

40. Use of the compound of any one of claims 1 -22 in the preparation of a medicament for the treatment of Hepatitis C.

41. The use of claim 40 further comprising use of an additional bioactive agent or a plurality of additional bioactive agents for preparation of a medicament for the treatment of Hepatitis C.

42. The use of claim 41 wherein the additional bioactive agent or plurality of additional bioactive agents comprises another compound of claim 1, another HCV protease inhibitor, interferon alfa-2b, peginteferon alfa-2b, recombinant interferon alfa-2a, peginterferon alfa-2a, inteferon-alpha 2B+Ribavirin, interferon alpha-nl, a nucleoside analog, an IRES inhibitor, an El inhibitor, an E2 inhibitor, an IMPDH inhibitor, an NS3 inhibitor, an NS5B inhibitor, an NS4B inhibitor, an NS5A inhibitor or an NTPase/helicase inhibitor, or any combination thereof.

43. The use of claim 41 wherein the additional bioactive agent or plurality of additional bioactive agents comprises an anti-proliferative agent.

44. The use of claim 43 wherein the anti-proliferative agent comprises 5- fluorouracil, daunomycin, mitomycin, bleomycin, dexamethasone, methotrexate, cytarabine, or mercaptopurine.

45. The use of claim 41 wherein the additional bioactive agent or plurality of additional bioactive agents comprises an immune modulator, preferably a steroid, preferably prednisone, prednisolone, or dexamethasone; a non-steroidal anti-inflammatory, preferably ibuprofen, naproxen, diclofenac, or indomethacin; a COX2 inhibitor, preferably rofecoxib or celecoxib; an anti-TNF compound, preferably enbrel, infliximab, or adaumimab; an anti-ILl compound, preferably anakinra; an interferon, preferably interferon alfa-2b, peginteferon alfa-2b, recombinant interferon alfa-2a, peginterferon alfa-2a, or interferon alpha-nl; methotrexate; leflunomide; cyclosporin; FK506; or a combination of any two or more thereof.