HK1101402B

HK1101402B - Inhibitors of serine proteases, particularly hcv ns3-ns4a protease

Info

Publication number: HK1101402B
Application number: HK07106097.2A
Authority: HK
Inventors: K．M．科特勒尔; J．J．科特; D．D．丹宁格; L．J．法默尔; J．皮特里克; R.B.珀尼
Original assignee: 沃泰克斯药物股份有限公司
Priority date: 2004-02-04
Filing date: 2005-02-04
Publication date: 2012-08-24

Description

Inhibitors of serine proteases, particularly HCV NS3-NS4A protease

Technical Field

The present invention relates to compounds that inhibit serine protease activity, particularly the activity of hepatitis c virus NS3-NS4A protease. Thus, they act by interfering with the life cycle of the hepatitis C virus and are useful as antiviral agents. The invention further relates to compositions comprising these compounds for ex vivo use or for administration to a patient suffering from HCV infection. The invention also relates to methods of treating HCV infected patients by administering pharmaceutical compositions comprising the compounds of the present invention.

Background

Hepatitis c virus ("HCV") infection is a human medical problem that needs to be addressed. HCV is recognized as the cause of most non-a, non-b Hepatitis and has been estimated to have a 3% prevalence in the global population (a. alberti et al, "Natural History of Hepatitis C",J.Hepatology，31.，(Suppl.1)，pp.17-24(1999)]. In The United States alone, nearly four million people may have been infected [ M.J. Alter et al, "The epidemic of viral Hepatitis in The United States,Gastroenterol.Clin.North Am.，23，pp.437-455(1994)；M.J.Alter“Hepatitis C VirusInfection in the United States，”J.Hepatology，31.，(Suppl.1)，pp.88-91(1999)]。

upon first exposure to HCV, only about 20% of infected individuals develop acute clinical hepatitis, while others appear to resolve the infection spontaneously. Is not limited toHowever, in almost 70% of cases, The virus establishes a Chronic infection lasting for decades [ S.Iward, "The Natural Courseof viral Hepatitis, ]"FEMS Microbiology Reviews，14，pp.201-204(1994)；D.Lavanchy，“Global Surveillance and Control ofHepatitis C，”J.Viral Hepatitis，6，pp.35-47(1999)]. This often leads to recurrent and progressive worsening liver inflammation, often leading to more severe disease states such as cirrhosis and hepatocellular Carcinoma [ m.c. kew, "hepatotis C and hepatocellular carcinosoma",FEMS Microbiology Reviews，14, pp.211-220 (1994); saito et al, "Patitis C Virus Infection associated with the Development of a Heatocellular Carcinoma"Proc.Natl.Acad.Sci.USA，87，pp.6547-6549(1990)]. Unfortunately, no generally effective treatment can impair the progression of chronic HCV.

The HCV genome encodes a 3010-3033 amino acid polyprotein [ Q.L.Choo, et al, "Genetic Organization and Diversity of the Hepatitis C Virus ].Proc.Natl.Acad.Sci.USA，88, pp.2451-2455 (1991); kato et al, "Molecular Cloning of the Human hepatites C Virus genome from Japan Patients with Non-A, Non-B hepatites"Proc.Natl. Acad.Sci.USA，87，pp.9524-9528(1990)；A.Takamizawa et.al.，“Structure and Organization of the Hepatitis C Virus GenomeIsolated From Human Carriers，”J.Virol.，65，pp.1105-1113(1991)]. The HCV Nonstructural (NS) proteins are postulated to provide the necessary catalytic mechanism for viral replication. NS proteins are derived from proteolytic Cleavage of polyproteins [ R.Bartenschlager et al, "Nonstructural Protein 3 of the Hepatitis C viruses encodings an Protein-Type Protein requiring for Cleavage at the NS3/4 and dNS 4/5 Junctions"J.Virol.，67，pp.3835-3844(1993)；A.Grakouiet.al.，“Characterization of the Hepatitis C Virus-EncodedSerine Proteinase：Determination of Proteinase-DependentPolyprotein Cleavage Sites，”J.Virol.，67，pp.2832-2843 (1993); grakoui et al, "Expression and Identification of Heapatitis C Virus Polyprotein Clearance Products"J.Virol.，67, pp.1385-1395 (1993); tomei et al, "NS 3 is a serine protease requiring for processing of hepatis C virus polyprotein",J. Virol.，67，pp.4017-4026(1993)]。

HCV NS protein 3(NS3) contains serine protease activity, which contributes to the processing of most viral enzymes, and is therefore considered essential for viral replication and infectivity. Mutations in the NS3 Protease of yellow Fever Virus are known to reduce the infectivity of the Virus [ Chambers, T.J., et al, "infectious fact that the N-tertiary Domain of non-structural Protein NS3 FromYe11ow Fever Virus a series Protease reactivity for site-Specific cleavage in the Viral Polyprotein",Proc.Natl. Acad.Sci.USA，87，pp.8898-8902(1990)]. The first 181 amino acids of NS3 (1027-1207 residues of the viral polyprotein) have been shown to contain the Serine protease domain of NS3 at all four downstream sites of the processing of the HCV polyprotein [ C.Lin et al, "Hepatitis C Virus NS3 spring protease: Trans-CleavageRequirements and Processing tools',J.Virol.，68，pp.8147-8157(1994)]。

the HCV NS3 serine protease and its related cofactor NS4A contribute to the processing of all viral enzymes and are therefore considered essential for viral replication. This processing appears to be similar to that performed by the human immunodeficiency virus aspartyl protease, which is also involved in viral enzyme processing. HIV protease inhibitors, which inhibit viral protein processing, are effective human antiviral agents, indicating that interfering with this stage of the viral life cycle makes available therapeutically active agents. Therefore, the HCV NS3 serine protease is an attractive target for drug development.

There is currently no satisfactory anti-HCV agent or treatment developed. Until recently, the only established therapy for HCV disease was interferon therapy. However, interferons have significant side effects [ m.a. wlaker et al, "Hepatitis C viruses：An Overview of CurrentApproaches and Progress，”DDT4, pp.518-29 (1999); mordapour et al, "Current and Evaling therapeutics for hepatis C,"Eur. J.Gastroenterol.Hepatol.，11, pp.1199-1202 (1999); janssen et al, "Suicide Associated with Alfa-interference therapy for Viral Hepatitis," L.A.Janssen et al "J.Hepatol.，21, pp.241-243 (1994); renault et al, "Side Effects of AlphaInterferon"Seminars in Liver Disease，9，pp.273-277.(1989)]And cause long-term remission in only a fraction (-25%) of cases o.weiland, "interference Therapy in Chronic Hepatitis C Virus Infection",FEMS Microbiol.Rev.，14，pp.279-288(1994)]. More recently, PEG conjugated forms of interferon(s) (II)And) Interferon conjugated with ribavirin and PEGThe introduction of the combination therapy of (a) results in only a small improvement in remission rate and only a partial reduction in side effects. Moreover, the prospects for effective anti-HCV vaccines remain uncertain.

Thus, there is a need for more effective anti-HCV therapies. Such inhibitors would have therapeutic potential as protease inhibitors, particularly as serine protease inhibitors, more particularly as HCV NS3 protease inhibitors. In particular, such compounds are useful as antiviral agents, particularly as anti-HCV agents.

Disclosure of Invention

The present invention provides compounds of formula I:

or a pharmaceutically acceptable salt or mixture thereof, wherein the variables are as defined herein.

The present invention also provides compounds of formula I-1:

The invention also relates to compositions comprising the above compounds and uses thereof. Such compositions can pre-treat invasive elements embedded in a patient, treat biological samples such as blood prior to administration to the patient, and be used for direct administration to the patient. In each case, the compositions are useful for inhibiting HCV replication and reducing the risk or severity of HCV infection.

The invention also relates to a process for the preparation of compounds of formula I.

Detailed description of the invention

The present invention provides compounds of formula I:

or a pharmaceutically acceptable salt or mixture thereof, wherein:

z is 0 or 1;

v is-C (O) -, -S (O) -, -C (R')₂-or-S (O)₂-；

R is-C (O) -, -S (O)-，-S(O)₂-，-N(R₈) -, -O-or a bond;

t is:

(C₆-C₁₀) -an aryl group,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group, a hydroxyl group, a carboxyl group,

(C₃-C₁₀) -a cycloalkyl group or a cycloalkenyl group,

[(C₃-C₁₀) -cycloalkyl or-cycloalkenyl]-(C₁-C₁₂) -an aliphatic radical,

(C₃-C₁₀) -a heterocyclic group,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical,

(C₅-C₁₀) Heteroaryl, or

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic group;

wherein the carbon atoms of up to 3 aliphatic groups in T may optionally be substituted by-S-, -S (O)₂-, -O-, -N-, or-N (H) -is replaced in a chemically stable arrangement;

wherein each T may be optionally substituted with up to 3J substituents;

wherein J is halogen, -OR ', -OC (O) N (R')₂，-NO₂，-CN，-CF₃，-OCF₃-R ', oxo, thioxo, 1, 2-methylenedioxy, 1, 2-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，-OC(O)N(R′)₂，-C(S)N(R′)₂，-(CH₂)_0-2NHC(O)R′，-N(R′)N(R′)COR′，-N(R′)N(R′)C(O)OR′，-N(R′)N(R′)CON(R′)₂，-N(R′)SO₂R′，-N(R′)SO₂N(R′)₂，-N(R′)C(O)OR′，-N(R′)C(O)R′，-N(R′)C(S)R′，-N(R′)C(O)N(R′)₂，-N(R′)C(S)N(R′)₂，-N(COR′)COR′，-N(OR′)R′，-C(＝NH)N(R′)₂，-C(O)N(OR′)R′，-C(＝NOR′)R′，-OP(O)(OR′)₂，-P(O)(R′)₂，-P(O)(OR′)₂OR-P (O) (H) (OR'), wherein;

two R' groups together with the nitrogen to which they are bound form a 3-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic heterocyclic ring system;

wherein in the bicyclic and tricyclic ring systems each ring is linearly fused, bridged or spirocyclic;

wherein each ring is aromatic or non-aromatic;

wherein each heteroatom in the heterocyclic ring system is selected from the group consisting of N, NH, O, S, SO and SO₂；

Wherein each ring is optionally substituted with (C)₆-C₁₀) Aryl group, (C)₅-C₁₀) Heteroaryl, (C)₃-C₁₀) Cycloalkyl or (C)₃-C₁₀) Heterocyclyl-fused; and is

Wherein each ring is optionally substituted with up to 3 independently selected from J₂Substituted with the substituent(s);

each R' is independently selected from:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

[(C₃-C₁₀) -cycloalkyl or-cycloalkenyl]-(C₁-C₁₂) -an aliphatic radical-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₁₂) An aliphatic group-,

(C₅-C₁₀) -heteroaryl-, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₁₂) -an aliphatic radical-,

wherein R' is optionally substituted with up to 3 independently selected from J₂Substituted with the substituent(s);

wherein J₂Is halogen, -OR ', -OC (O) N (R')₂，-NO₂，-CN，-CF₃，-OCF₃-R ', oxo, thioxo, 1, 2-methylenedioxy, -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，-OC(O)N(R′)₂，-C(S)N(R′)₂，-(CH₂)_0-2NHC(O)R′，-N(R′)N(R′)COR′，-N(R′)N(R′)C(O)OR′，-N(R′)N(R′)CON(R′)₂，-N(R′)SO₂R′，-N(R′)SO₂N(R′)₂，-N(R′)C(O)OR′，-N(R′)C(O)R′，-N(R′)C(S)R′，-N(R′)C(O)N(R′)₂，-N(R′)C(S)N(R′)₂，-N(COR′)COR′，-N(OR′)R′，-C(＝NH)N(R′)₂，-C(O)N(OR′)R′，-C(＝NOR′)R′，-OP(O)(OR′)₂，-P(O)(R′)₂，-P(O)(OR′)₂OR-P (O), (H), (OR'); or

T is:

wherein:

R₁₀the method comprises the following steps:

the presence of hydrogen in the presence of hydrogen,

(C₁-C₁₂) -an aliphatic radical,

(C₆-C₁₀) -an aryl group,

(C₃-C₁₀) -a cycloalkyl group or a cycloalkenyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical,

(C₅-C₁₀) -heteroaryl, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₁₂) -an aliphatic group;

k is a bond, (C)₁-C₁₂) -aliphatic radical, -O-, -S-, -NR₉-, - (O) -or-C (O) -NR₉-, wherein R₉Is hydrogen or (C)₁-C₁₂) -an aliphatic group;

n is 1 to 3; or

T is N (R)₁₇)₂；

Wherein each R₁₇Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl-or cycloalkenyl-,

[(C₃-C₁₀) -cycloalkyl-or cycloalkenyl]-(C₁-C₁₂) -an aliphatic radical-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical-,

(C₅-C₁₀) Heteroaryl-, or

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic radical-, or

Two R₁₇The groups together with the nitrogen to which they are bonded form a 3-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic heterocyclic ring system;

wherein each ring is aromatic or non-aromatic;

Wherein each ring is optionally substituted with up to 3 substituents independently selected from J;

w is:

wherein:

e is selected from N (R)₁₇) Or a bond;

wherein each ring is aromatic or non-aromatic;

R₅and R_5’Independently are:

the presence of hydrogen in the presence of hydrogen,

(C₁-C₁₂) -an aliphatic radical,

(C₆-C₁₀) Aryl, or

(C₅-C₁₀) (ii) a heteroaryl group, wherein,

wherein (C)₁-C₁₂) -any hydrogen in the aliphatic group is optionally replaced by halogen;

wherein any terminal carbon atom is optionally substituted with a mercapto or hydroxy group; while

Wherein up to two aliphatic carbon atoms may be selected from N, NH, O, S, SO or SO₂Heteroatom substitution of (a);

wherein any ring is optionally substituted with up to 3 substituents independently selected from J₂Substituted with the substituent(s); and is

Wherein each heteroatom in the heteroaryl ring system is selected from the group consisting of N, NH, O, S, SO, and SO₂(ii) a Or

R₅And R_5’Optionally together with the atoms to which they are bonded form a compound having up to 2 atoms selected from N, NH, O, S, SO or SO₂3-to 6-membered ring of the heteroatom(s); wherein the ring is optionally substituted with up to 2 substituents independently selected from J;

R₁(if present), R_1′(if present), R₁₁，R_11′，R₁₃And R_13’Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₁₂) An aliphatic group, a hydroxyl group, a carboxyl group,

(C₅-C₁₀) -heteroaryl-, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₁₂) -an aliphatic radical-,

wherein each R₁(if present), R_1′(if present), R₁₁、R_11′、R₁₃And R_13’Independently and optionally substituted with up to 3 substituents independently selected from J;

wherein any ring is optionally substituted with (C)₆-C₁₀) Aryl group, (C)₅-C₁₀) Heteroaryl, (C)₃-C₁₀) Cycloalkyl or (C)₃-C₁₀) Heterocyclyl-fused;

wherein each R₁(if present), R_1′(if present), R₁₁、R_11′、R₁₃And R_13′Up to 3 aliphatic carbon atoms in (A) may be selected from O, N, NH, S, SO or SO₂The heteroatoms of (a) are replaced in a chemically stable arrangement; or

R₁And R_1′Optionally formed with up to 2 atoms selected from N, NH, O, S, SO or SO, if any, bonded thereto₂3-to 6-membered ring of the heteroatom(s); wherein the ring system is optionally substituted with up to 2 substituents independently selected from J; or

R₁₁And R_11′Optionally together with the atoms to which they are bonded form a compound having up to 2 atoms selected from N, NH, O, S, SO or SO₂3-to 6-membered ring of the heteroatom(s); wherein the ring is optionally substituted with up to 2 substituents independently selected from J; or

R₁₃And R_13’Optionally together with the atoms to which they are bonded form a compound having up to 2 atoms selected from N, NH, O, S, SO or SO₂3-to 6-membered ring of the heteroatom(s); wherein the ring is optionally substituted with up to 2 substituents independently selected from J;

R₂，R₄，R₈(if present) and R₁₂Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₅-C₁₀) -heteroaryl-, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₁₂) -an aliphatic radical-,

wherein each R₂、R₄、R₈(if present) and R₁₂Independently and optionally substituted with up to 3 substituents independently selected from J;

wherein R is₂、R₄、R₈(if present) and R₁₂Up to two aliphatic carbon atoms in (B) may be selected from O, N, NH, S, SO or SO₂Is replaced by a heteroatom of (a); or

R₁₁And R₁₂Together with the atoms to which they are bonded form a 3-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic carbocyclic or heterocyclic ring system;

wherein each ring is aromatic or non-aromatic;

Wherein each ring is optionally substituted with up to 3 substituents independently selected from J; or

R₁₂And R₁₃Together with the atoms to which they are bonded form a 4-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic carbocyclic or heterocyclic ring system;

wherein each ring is aromatic or non-aromatic;

R₁₁And R₁₃Together with the atoms to which they are bonded form a 5-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic carbocyclic or heterocyclic ring system;

wherein each ring is aromatic or non-aromatic;

R₁₁、R₁₂And R₁₃Together with the atoms to which they are bonded form an 8-to 20-membered bicyclic or tricyclic carbocyclic or heterocyclic ring system;

wherein each ring is aromatic or non-aromatic;

R₁₃' and R₂Together with the atom to which they are bonded form a 3-to 20-membered monocyclic, 8-to 20-membered bicyclic or tricyclic ringA carbocyclic or heterocyclic ring system of rings;

wherein each ring is aromatic or non-aromatic;

R₅And R₁₃A carbocyclic or heterocyclic ring system which together with the atoms to which they are bonded forms an 18-to 23-membered monocyclic, 19-to 24-membered bicyclic, or 20-to 25-membered tricyclic ring;

wherein each ring is aromatic or non-aromatic;

Wherein each ring is optionally substituted with up to 6 substituents independently selected from J; or

R₁(if present) and R₁₂Together with the atoms to which they are bonded form an 18-to 23-membered monocyclic, 19-to 24-membered bicyclic, or 20-to 25-membered tricyclic carbocyclic or heterocyclic ring system；

wherein each ring is aromatic or non-aromatic;

Wherein each ring is optionally substituted with up to 6 substituents independently selected from J.

The present invention also provides compounds of formula I-1:

or a pharmaceutically acceptable salt or mixture thereof, wherein W, V, R, T, R₁、R_1′、R₂、R₄、R₅、R_5′、R₈、R₁₁、R_11′、R₁₂、R₁₃And R_13′Are as defined above for compounds of formula I.

Definition of

The term "aryl" as used herein refers to a monocyclic or bicyclic carbocyclic aromatic ring system. Phenyl is an example of a monocyclic aromatic ring system. Bicyclic aromatic ring systems include ring systems in which both rings are aromatic, such as naphthyl, and ring systems in which only one of the two rings is aromatic, such as tetrahydronaphthalene. It is to be understood that the term "(C) as used herein₆-C₁₀) -aryl- "includes C₆、C₇、C₈、C₉And C₁₀Monocyclic or bicyclic carbocycleAny of the aromatic ring systems.

The term "heterocyclyl" as used herein, means a monocyclic or bicyclic non-aromatic ring system having in each ring a chemically stable arrangement of 1 to 3 substituents selected from O, N, NH, S, SO or SO₂A heteroatom or heteroatom group of (a). In bicyclic non-aromatic ring system embodiments of "heterocyclyl", one or both rings may contain the heteroatom or heteroatom group. It is to be understood that the term "(C) as used herein₅-C₁₀) -heterocyclyl- "includes: a monocyclic or bicyclic non-aromatic ring system of any 5, 6, 7,8, 9 and 10 atoms having a chemically stable arrangement of from 1 to 3 heteroatoms or heteroatom groups selected from O, N, NH and S in each ring.

The term "heteroaryl" as used herein refers to a monocyclic or bicyclic aromatic ring system having in each ring a chemically stable arrangement of 1 to 3 heteroatoms or heteroatom groups selected from O, N, NH or S. In such a "heteroaryl" bicyclic aromatic ring system embodiment:

one or both rings may be aromatic; and is

One or both rings may contain said heteroatom or heteroatom group. It is to be understood that the term "(C) as used herein₅-C₁₀) -heteroaryl- "includes: a monocyclic or bicyclic aromatic ring system of any 5, 6, 7,8, 9 and 10 atoms having a chemically stable arrangement of from 1 to 3 heteroatoms or heteroatom groups selected from O, N, NH and S in each ring.

Heterocycles include, but are not limited to, 3-1H-benzimidazol-2-one, 3- (1-alkyl) -benzimidazol-2-one, 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-tetrahydropyrazinyl, 2-tetrahydropyrazinyl, 3-tetrahydropyrazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, dihydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiepin (benzodithiolane), benzodithiane, and 1, 3-dihydro-imidazol-2-one.

Heteroaryl rings include, but are not limited to: 2-furyl group, 3-furyl group, N-imidazolyl group, 2-imidazolyl group, 4-imidazolyl group, 5-imidazolyl group, benzimidazolyl group, 3-isoxazolyl group, 4-isoxazolyl group, 5-isoxazolyl group, 2-oxazolyl group, 4-oxazolyl group, 5-oxazolyl group, N-pyrrolyl group, 2-pyrrolyl group, 3-pyrrolyl group, 2-pyridyl group, 3-pyridyl group, 4-pyridyl group, 2-pyrimidyl group, 4-pyrimidyl group, 5-pyrimidyl group, pyridazinyl group (e.g., 3-pyridazinyl group), 2-thiazolyl group, 4-thiazolyl group, 5-thiazolyl group, tetrazolyl group (e.g., 5-tetrazolyl group), triazolyl group (e.g., 2-triazolyl group and 5-triazolyl group), 2-thienyl, 3-thienyl, benzofuranyl, benzothienyl, indolyl (e.g., 2-indolyl), pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1, 2, 3-oxadiazolyl, 1, 2, 5-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 3-triazolyl, 1, 2, 3-thiadiazolyl, 1, 3, 4-thiadiazolyl, 1, 2, 5-thiadiazolyl, purinyl, pyrazinyl, 1, 3, 5-triazinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).

The term "aliphatic group" as used herein refers to a straight or branched alkyl, alkenyl or alkynyl group. It is to be understood that the term "(C) as used herein₁-C₁₂) -aliphatic radical- "C comprising a carbon atom₁、C₂、C₃、C₄、C₅、C₆、C₇、C₈、C₉、C₁₀、C₁₁And C₁₂Any of a linear or branched alkyl series. It is understood that alkenyl or alkynyl embodiments need to have at least two carbon atoms in the aliphatic chain. The term "cycloalkyl or cycloalkenyl" refers to a monocyclic or fused or bridgedA bicyclic carbon ring system which is not an aromatic ring. The cycloalkenyl ring has one or more units of unsaturation. It is also understood that the term "(C) as used herein₃-C₁₀) -cycloalkyl-or-cycloalkenyl- "includes any C₃、C₄、C₅、C₆、C₇、C₈、C₉And C₁₀Monocyclic or fused or bridged bicyclic carbocycle. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, norbornyl (nornbornyl), adamantyl and decahydronaphthyl.

As used herein, carbon atom symbols can have the indicated integers and any intervening integers. For example, in (C)₁-C₄) The number of carbon atoms in the alkyl group is 1, 2, 3 or 4. It is to be understood that these symbols refer to the total number of atoms in the appropriate group. For example, in (C)₃-C₁₀) In the heterocyclyl group, the total number of carbon and heteroatoms is 3 (as in aziridine), 4, 5, 6 (as in morpholine), 7,8, 9 or 10.

The phrase "chemically stable arrangement" as used herein refers to a structure of a compound that confers stability to the compound sufficient to permit manufacture and administration to a mammal by methods known in the art. Typically, such compounds are stable for at least one week at temperatures of 40 ℃ or below in the absence of moisture or other chemical reaction conditions.

The term "interferon" as used herein refers to members of the highly homologous genus-specific protein family that inhibit viral replication and cellular proliferation, and modulate immune responses, such as interferon alpha, interferon beta, or interferon gamma. The Merck Index, entry 5015, 12 th edition.

Detailed description of the preferred embodiments

In one form of any embodiment of the present invention, there is provided a compound of formulae IIIA, IIIB, IIIC and IIID, wherein P is₁、P₂、P₃、P₄Denotes the residue of a serine protease inhibitor known to the person skilled in the art, E is selected from N (R)₁₇) Or a bond, V, R, T and R₁₇As defined in any one of the embodiments herein.

Thus, all have: 1) structural units of serine protease inhibitors; and 2) a diketohydrazide moiety (IIIA), or a diketohydroxamic acid moiety (IIIB), or a diketosulfonamide moiety (IIIC), or a diketosulfenamide moiety (IIID) are considered part of the present invention.

Compounds having structural units of serine protease inhibitors include, but are not limited to, compounds in the following publications: WO 97/43310, US 20020016294, WO 01/81325, WO 02/08198, WO 01/77113, WO 02/08187, WO 02/08256, WO 02/08244, WO 03/006490, WO 01/74768, WO 99/50230, WO 98/17679, WO 02/48157, US 20020177725, WO 02/060926, US 20030008828, WO 02/48116, WO01/64678, WO 01/07407, WO 98/46630, WO 00/59929, WO 99/07733, WO 00/09588, US 20020016442, WO 00/09543, WO 99/07734, US6,018,020, WO 98/22496, US 5,866,684, WO 02/079234, WO 00/31129, WO 99/38888, WO 99/64442, and WO 02/18369, which are incorporated herein by reference.

Thus, any of the compounds in the above publications can be modified to include a diketohydrazide moiety (IIIA), a diketohydroxamic acid moiety (IIIB), a diketosulfonamide moiety (IIIC), or a diketosulfenamide moiety (IIID), or derivatives thereof. Any such is part of the present invention. For example, compound a in WO 02/18369 (p.41):

may be modified to provide the following compounds of the invention:

wherein R is₁₇As defined in any one of the embodiments herein.

In other forms of any of the embodiments of the present invention, there is provided a compound of formulae IIIA-1, IIIB-1, IIIC-1, and IIID-1, wherein P₁、P₂And P₃Denotes the residue of a serine protease inhibitor known to the person skilled in the art, E is selected from N (R)₁₇) Or a bond, V, R, T and R₁₇As defined in any one of the embodiments herein.

In any of the other forms of the embodiments of the present invention,

R₁₁is H; and is

R₁₂Is that

(C₁-C₆) -an aliphatic radical,

(C₃-C₁₀) -a cycloalkyl group,

[(C₃-C₁₀) -cycloalkyl radical]-(C₁-C₁₂) -an alkyl group,

(C₆-C₁₀) -an aryl group,

(C₆-C₁₀) -aryl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₅-C₁₀) -heteroaryl, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₆) -an alkyl group.

In other forms of any of the embodiments of the present invention, R₁₂Is isobutyl, cyclohexyl, cyclohexylmethyl, benzyl or phenethyl.

In any of the other forms of the embodiments of the present invention,

R₁₁the method comprises the following steps:

(C₁-C₆) -an aliphatic radical,

(C₃-C₁₀) -a cycloalkyl group,

[(C₃-C₁₀) -cycloalkyl radical]-(C₁-C₁₂) -an alkyl group,

(C₆-C₁₀) -an aryl group,

(C₆-C₁₀) -aryl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₅-C₁₀) -heteroaryl, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₆) -an alkyl group; and is

R₁₂Is H.

In other forms of any of the embodiments of the present invention, R_11′And R₁₂Is H. In any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

wherein n is 0 or 1, and Z' are S or O.

In any of the other forms of the embodiments of the present invention,

the radicals are:

wherein each B independently forms a 3-to 20-membered carbocyclic or heterocyclic ring system;

wherein each ring B is aromatic or non-aromatic;

wherein each heteroatom in the heterocyclic ring system is N, NH, O, S, SO or SO₂；

Wherein each ring is optionally substituted with up to 3 substituents independently selected from J.

In any of the other forms of the embodiments of the present invention,

the radicals are:

wherein each ring C, D and E is as defined above for ring B, Z₃Is a carbon atom, -CHR₁₈-N-，-HN-CR₁₈-or-CHR₁₈-CHR₁₈-，-O-CHR₁₈-，-S-CHR₁₈-，-SO-CHR₁₈-，-SO₂-CHR₁₈-, or-N-. In another embodiment, R₁₈Is (C)₁-C₁₂) -aliphatic radical, (C)₆-C₁₀) -aryl, (C)₆-C₁₀) Aryl radical- (C)₁-C₁₂) -aliphatic radical or (C)₃-C₁₀) -a cycloalkyl group. In another embodiment, R₁₈Is (C)₁-C₆) -alkyl or (C)₃-C₇) -a cycloalkyl group.

In other forms of any of the embodiments of the invention, in

In the group, ring C is:

wherein R is₁₉The method comprises the following steps:

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

(C₆-C₁₀) -aryl-, or

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-.

In other forms of any of the embodiments of the invention, in

In the group, ring C is:

wherein R is₁₉The method comprises the following steps:

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

(C₆-C₁₀) -aryl-, or

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-.

In other forms of any of the embodiments of the invention, in

In which ring D is selected from:

wherein R is₁₉The method comprises the following steps:

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

(C₆-C₁₀) -aryl-, or

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-.

In other forms of any of the embodiments of the invention, in

In which ring D is selected from:

wherein R is₁₉The method comprises the following steps:

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-,

(C₆-C₁₀) -aryl-, or

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-.

In other forms of any of the embodiments of the present invention, rings a and B, together with the rings to which they are attached, include:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

wherein each ring B is aromatic or non-aromatic;

In any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

in any of the other forms of the embodiments of the present invention,

the radicals are:

wherein B forms a 4-to 20-membered carbocyclic or heterocyclic ring system;

wherein each ring B is aromatic or non-aromatic;

Wherein each ring is optionally substituted with (C)₆-C₁₀) Aryl group, (C)₅-C₁₀) Heteroaryl, (C)₃-C₁₀) Cycloalkyl or (C)₃-C₁₀) Heterocyclyl-fused;

wherein, in a carbocyclic or heterocyclic ring system, each ring is linearly fused, bridged or spirocyclic; and is

In any of the other forms of the embodiments of the present invention,

the radicals are:

in the above groups, it is understood that R₁₁The variable is H.

In other forms of any of the embodiments of the present invention, R₁₁And R₁₂Together with the atoms to which they are bonded form a 6-to 10-membered mono-or bicyclic carbocyclic or heterocyclic ring system;

wherein each heteroatom in the heterocyclic ring system is selected from the group consisting of N, NH, O, S, SO and SO₂(ii) a And is

Wherein the ring has up to 3 substituents independently selected from J.

Any of the ring systems may be substituted, as listed herein. In other forms of any of the embodiments of the present invention, the ring substituents are oxo, fluoro, difluoro (especially the adjacent difluoro) and hydroxy. In other forms of any of the embodiments of the present invention, the following ring systems:

optionally substituted with oxo, fluoro, difluoro (especially vicinal difluoro) and hydroxy; wherein ring B is a 5-membered carbocyclic ring, optionally having one unsaturated bond.

In other forms of any of the embodiments of the present invention, the heteroatom is selected from the group consisting of N, NH, O, SO, and SO₂。

In other forms of any of the embodiments of the present invention, R_5′Is H, R₅Is optionally substituted by 1 to 3 fluoro groups or 1-SH group₁-C₆) -an aliphatic radical, or R₅Is (C)₆-C₁₀) -an aryl group.

In another form of any embodiment of the present invention, (C)₁-C₆) -the aliphatic group is substituted with 1 to 3 fluoro groups.

In other forms of any of the embodiments of the present invention, R₅And R_5′Independently hydrogen or:

in other forms of any of the embodiments of the present invention, R_5′Is H, and R₅The method comprises the following steps:

in other forms of any of the embodiments of the present invention, R₅And R_5′The method comprises the following steps:

in other forms of any of the embodiments of the present invention, R_13′Is hydrogen, and

R₁₃the method comprises the following steps:

(C₁-C₆) -an aliphatic radical,

(C₃-C₁₀) -a cycloalkyl group,

[(C₃-C₁₀) -cycloalkyl radical]-(C₁-C₁₂) -an alkyl group,

(C₆-C₁₀) -an aryl group,

(C₆-C₁₀) -aryl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₅-C₁₀) -heteroaryl, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₆) -an alkyl group;

wherein R is₁₃Optionally substituted with up to 3 substituents independently selected from J; and is

Wherein R is₁₃Up to 3 aliphatic carbon atoms in (A) may be selected from O, NH, SO or SO₂Are replaced in a chemically stable arrangement.

In other forms of any of the embodiments of the present invention, R_13′Is hydrogen, R₁₃The method comprises the following steps:

in any of the other forms of the embodiments of the present invention,

R₁if present, is:

(C₁-C₆) -an aliphatic radical,

(C₃-C₁₀) -a cycloalkyl group,

[(C₃-C₁₀) -cycloalkyl radical]-(C₁-C₁₂) -an alkyl group,

(C₆-C₁₀) -an aryl group,

(C₆-C₁₀) -aryl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₆-C₁₀) -heterocyclyl- (C)₁-C₆) An alkyl group, a carboxyl group,

(C₅-C₁₀) -heteroaryl, or

(C₅-C₁₀) -heteroaryl- (C)₁-C₆) -an alkyl group;

wherein R is₁Optionally substituted with up to 3 substituents independently selected from J; and is

Wherein R is₁Up to 3 aliphatic carbon atoms in (A) may be selected from O, NH, S, SO or SO₂In a chemically stable arrangement。

In other forms of any of the embodiments of the present invention, R_1′If present, is hydrogen, R₁If present, is:

in other forms of any embodiment of the present invention, T is selected from:

(C₆-C₁₀) -an aryl group,

(C₃-C₁₀) -a cycloalkyl group or a cycloalkenyl group,

(C₃-C₁₀) -a heterocyclic group,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical,

(C₅-C₁₀) Heteroaryl, or

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic radical,

wherein each T is optionally substituted with up to 3J substituents.

In other forms of any of the embodiments of the present invention, T is (C)₅-C₁₀) Heteroaryl, wherein T is optionally substituted with up to 3J substituents.

In other forms of any embodiment of the present invention, T is:

in other forms of any embodiment of the present invention, T is:

or

In other forms of any of the embodiments of the present invention, T contains at least one substituent selected from the group consisting of-NH₂Hydrogen bond donor moieties of-NH-, -OH and-SH.

In other forms of any embodiment of the present invention, T is:

wherein:

t is optionally substituted with up to 4J substituents, wherein J is as defined herein;

z is independently O, S, NR₁₀、C(R₁₀)₂；

n is independently 1 or 2; and is

Independently a single or double bond.

In other forms of any embodiment of the present invention, T is:

wherein:

z is independently O, S, NR₁₀、C(R₁₀)₂、SO、SO₂；

n is independently 1 or 2; and is

Independently a single or double bond.

In other forms of any embodiment of the present invention, T is:

wherein:

t is optionally substituted with up to 4J substituents, wherein J is as defined herein; and is

Z is independently O, S, NR₁₀、C(R₁₀)₂、SO、SO₂。

In other forms of any embodiment of the present invention, T is:

in other forms of any embodiment of the present invention, V-R-T is selected from:

wherein R is₁₂And each R₁₇As defined herein.

In other forms of any embodiment of the present invention, V-R-T is:

wherein:

a R₁₇Is hydrogen; and is

A R₁₇The method comprises the following steps:

(C₁-C₁₂) -an aliphatic group-;

(C₁-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-, or

(C₃-C₁₀) -cycloalkyl or-cycloalkenyl-;

wherein R is₁₇Up to 3 aliphatic carbon atoms in (A) may be selected from O, N, NH, S, SO or SO₂The heteroatoms of (a) are replaced in a chemically stable arrangement; and is

Wherein R is₁₇Optionally substituted with up to 3 substituents independently selected from J.

In other forms of any embodiment of the present invention, V-R-T is:

in other forms of any of the embodiments of the present invention, R₂And R₄Each is independently H or (C)₁-C₃) -alkyl, R₈If presentIs H or (C)₁-C₃) -an alkyl group.

In other forms of any of the embodiments of the present invention, R₂And R₄Each is H, R₈If present, is H.

In other forms of any of the embodiments of the present invention, R₈If present, is hydrogen, V is-C (O) -, R is a bond, and T is as defined in any of the embodiments herein.

In another form of any embodiment of the invention, W is:

wherein:

e is selected from N (R)₁₇) Or a bond;

each R₁₇Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl-or cycloalkenyl-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) Aliphatic radicals

(C₅-C₁₀) A heteroaryl-,

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic radical-, or

wherein each ring is aromatic or non-aromatic;

Wherein each ring optionally has up to 3 substituents independently selected from J.

In another form of any embodiment of the invention, W is:

wherein

Each R₁₇Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl-or cycloalkenyl-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical-,

(C₅-C₁₀) A heteroaryl-,

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic radical-, or

wherein each ring is aromatic or non-aromatic;

In another form of any embodiment of the invention, W is:

wherein

Each R₁₇Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -cycloalkyl-or cycloalkenyl-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic group-,

(C₃-C₁₀) -a heterocyclic radical-,

(C₃-C₁₀) -heterocyclyl- (C)₁-C₁₂) -an aliphatic radical-,

(C₅-C₁₀) A heteroaryl-,

(C₅-C₁₀) Heteroaryl- (C)₁-C₁₂) -an aliphatic radical-, or

wherein each ring is aromatic or non-aromatic;

In other forms of any of the embodiments of the present invention, J is halogen, -OR', -NO₂，-CF₃，-OCF₃-R ', oxo, 1, 2-methylenedioxy, -N (R')₂，-SR′，-SOR′，-SO₂R′，-C(O)R′，-COOR′-CON(R′)₂-N (R ') COR', -N (COR ') COR', -CN, or-SO₂N(R′)₂。

In other forms of any of the embodiments of the present invention, J₂Is halogen, -OR', -NO₂，-CF₃，-OCF₃-R ', oxo, 1, 2-methylenedioxy, -N (R')₂，-SR′，-SOR′，-SO₂R′，-C(O)R′，-COOR′-CON(R′)₂-N (R ') COR', -N (COR ') COR', -CN, or-SO₂N(R′)₂。

In other forms of any of the embodiments of the present invention, in J and J₂Wherein halogen is chlorine or fluorine. In other forms of any of the embodiments of the present invention, the halogen is fluorine.

In other forms of any of the embodiments of the present invention, R_1′If present, is H.

In other forms of any of the embodiments of the present invention, R_13′Is H.

In other forms of any of the embodiments of the present invention, R_11′Is H.

In other forms of any of the embodiments of the present invention, R₁₂Is H.

In another form of any embodiment of the present invention, there is provided a method of making a compound of the present invention. These methods are described in the schemes and examples.

In other forms of any embodiment of the invention, the compound is:

the compounds of the present invention may contain one or more asymmetric carbon atoms and may therefore occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. It is clear that all such isomeric forms of these compounds are included within the scope of the present invention. Each stereoisomeric carbon may be in the R or S configuration.

In another embodiment, the compounds of the present invention have the structures and stereochemistry described in compounds 1-8.

It should be appreciated that any of the above-listed embodiments, including those within the above categories, may be combined to produce another embodiment of the invention.

Abbreviations subsequently used in the schemes, preparations and examples are:

THF: tetrahydrofuran (THF)

DMF: n, N-dimethylformamide

DMA: dimethylacetamide

EtOAc: ethyl acetate

AcOH: acetic acid

NMM: n-methylmorpholine

NMP: n-methyl pyrrolidone

EtOH: ethanol

t-BuOH: tert-butyl alcohol

Et₂O: ether (A)

DMSO, DMSO: dimethyl sulfoxide

DCCA: dichloroacetic acid

DIEA: diisopropylethylamine

MeCN: acetonitrile

TFA: trifluoroacetic acid

DBU: 1, 8-diazabicyclo [5.4.0] undec-7-ene

DEAD: azanedicarboxylic acid diethyl ester

HOBt: 1-hydroxybenzotriazole hydrate

HOAt: 1-hydroxy-7-azabenzotriazoles

EDC: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

Boc: tert-butyloxycarbonyl radical

Boc₂O: di-tert-butyl dicarbonate

Cbz: benzyloxycarbonyl group

Cbz-Cl: benzyl chloroformate

Fmoc: 9-fluorenylmethoxycarbonyl

Chg: cyclohexylglycine

t-BG: tert-butyl glycine

mBPA: 3-Chloroperbenzoic acid

IBX: o-iodoxybenzoic acid

DAST: (diethylamino) sulfur trifluoride

TEMPO: 2, 2,6, 6-tetramethyl-1-piperidinyloxy, free radical

PyBROP: tris (pyrrolidinyl) phosphonium bromide hexafluorophosphate

TBTU or HATU: 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate

DMAP: 4-dimethylaminopyridine

AIBN: 2, 2' -azobisisobutyronitrile

DMEM: dulbecco's minimum essential Medium

PBS: phosphate buffered saline

RT or RT: at room temperature

ON: overnight

ND: not determined

MS: mass spectrometry

LC: liquid chromatography

Xaa: commercially available amino acids or synthetically prepared from commercially available intermediates and reagents

General synthetic methods:

in general, the compounds of the present invention may be prepared by methods known to those skilled in the art. Schemes 1-22 and 19a-20a below illustrate synthetic routes for the compounds of the present invention. The various molecular moieties may be synthesized using other equivalent schemes that will be apparent to the ordinarily skilled organic chemical workers, as exemplified by the following general schemes and the preparation examples that follow.

Scheme 1:

scheme 1 above provides a synthetic route to the intermediate ester hydrochloride 14 from intermediate acid 9. Intermediate acid 9 was prepared according to the procedure described in Harbeson, S. et al, J.Med.chem., Vol.37, No.18, pp.2918-2929 (1994).

And (2) a flow scheme:

scheme 2 above provides intermediates 21 in the preparation of amino esters from commercially available amino esters, amino esters prepared according to any of the schemes herein, or amino esters prepared according to any of the issued patents or published patent applications incorporated herein by reference (wherein V is-C (O) -, and R is a bond, -N (R)₁₂)CH(R₁₁) The C (O) -group being a bicyclic proline derivative, T, R₁、R₁₃And ring B is as defined in any of the embodiments herein). It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to scheme 1 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (3) a flow path: wherein W is C (O) NHN (R)₁₇)₂Synthesis of compounds of formula I:

schemes 1 and 2 in combination with scheme 3 above provide general routes for the preparation of compounds of formula I wherein V is-C (O) -, R is a bond, -N (R)₁₂)CH(R₁₁) The C (O) -group being a bicyclic proline derivative, W being C (O) NHN (R)₁₇)₂And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. The coupling process to convert intermediate 21 to amide 22, followed by oxidation with Dess Martin periodinane (periodinane) to final product 23 is accomplished according to the procedures detailed in the examples described herein. Wherein W is C (O) NHN (R)₁₇)₂And P is₂The preparation of compounds of formula I other than bicyclic prolines can also be accomplished by the general route provided in schemes 1 and 2, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1 and 2, wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (4) a flow chart: wherein W is C (O) NHOR₁₇Synthesis of compounds of formula I:

schemes 1 and 2 in conjunction with scheme 4 above provide for the preparation of R from intermediate acid 21 and commercially available₁₇General route to the preparation of compounds of formula I from hydroxyamines, wherein V is-C (O) -, R is a bond, -N (R)₁₂)CH(R₁₁) The C (O) -group being a bicyclic proline derivative, W being C (O) NHOR₁₇And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. The coupling process to convert intermediate 21 to amide 24, followed by periodoxylation to final product 25 with Dess Martin was carried out according to the procedures detailed in the examples described herein. Wherein P is₂Is not bicyclic proline and W is C (O) NHOR₁₇Can also be prepared by the general route provided in schemes 1, 2 and 4, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1, 2 and 4 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (5) a flow chart: wherein W is C (O) NHSO₂N(R₁₇)₂Synthesis of compounds of formula I:

scheme 5 above provides for the preparation of intermediate acids 21 and commercially available R₁₇General route to amino sulfonamides 26 for the preparation of compounds of formula I, wherein V is-C (O) -, R is a bond, -N (R)₁₂)CH(R₁₁) The C (O) -group being a bicyclic proline derivative, W being C (O) NHSO₂N(R₁₇)₂And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. Coupling of intermediate 21 with aminosulfonamide 26 to produce ketoamide 27 is accomplished according to the following procedure: rossier, s. et al, biorg.med.chem.lett., 12, p.2523(2002) and Pelletier, j. et al, Synlett, 11, p.1141 (1995). The final oxidation of intermediate 27 to a compound of formula I (shown as compound 28) is accomplished according to known procedures using desssmatin periodinane as the oxidant. Wherein W is C (O) NHSO₂N(R₁₇)₂And P is₂The preparation of compounds of formula I other than bicyclic prolines can also be accomplished by the general route provided in schemes 1, 2 and 5, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1, 2 and 5 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (6) a flow path: wherein W is C (O) NHSO₂R₁₇Synthesis of compounds of formula I:

the above flowScheme 6 provides for the preparation of R from intermediate acid 21 and commercially available R₁₇General route to sulfonamide 29 Compounds of formula I, wherein V is-C (O) -, R is a bond, -N (R)₁₂)CH(R₁₁) The C (O) -group being a bicyclic proline derivative, W being C (O) NHSO₂R₁₇And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. Coupling of intermediate 21 with sulfonamide 29 to make ketoamide 30 is accomplished according to the following procedure: rossier, s. et al, biorg.med.chem.lett., 12, p.2523(2002) and Pelletier, j. et al, Synlett, 11, p.1141 (1995). Oxidation of ketol 30 to the diketoamide 31 was accomplished using Dess Martin periodinane according to procedures known in the art. Wherein W is C (O) NHSO₂R₁₇And P is₂The preparation of compounds of formula I other than bicyclic prolines can also be accomplished by the general route provided in schemes 1, 2 and 6, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1, 2 and 6 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 7: wherein W is C (O) NHS (O) N (R)₁₇)₂Synthesis of compounds of formula I:

scheme 7 above provides a general route for the preparation of compounds of formula I, wherein W is C (O) NHS (O) N (R) R (R) H, from intermediate acids 21 and aminosulfinamide 32 (either commercially available or prepared according to the procedures of Schipper, E. et al, J.org.chem., 34, p.2397 (1969)), wherein W is C (O)₁₇)₂And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. Oxidation of the intermediate alcohol 33 to the final product 34 with IBX (o-iodoxybenzoic acid) as oxidant according to Wu, Y, et al, Organic Letters, 4, p.2141 (2002). Wherein W is C (O) NHS (O) N (R)₁₇)₂And P is₂The preparation of compounds of formula I other than bicyclic prolines can also be accomplished by the general route provided in schemes 1, 2 and 7, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1, 2 and 7 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (3) a process 8: wherein W is C (O) NHS (O) R₁₇Synthesis of compounds of formula I:

scheme 8 above provides a general route for the preparation of compounds of formula I, wherein W is C (O) NHS (O) R₁₇And T, R₁、R₁₃、R₁₇And ring B is as defined in any of the embodiments herein. The oxidation of the intermediate alcohol 36 to the final product 37 is carried out using IBX (o-iodoxybenzoic acid) as oxidant according to the procedure of Wu, Y.et al, Organic Letters, 4, p.2141 (2002). Wherein W is C (O) NHS (O) R₁₇And P is₂The preparation of compounds of formula I other than bicyclic prolines can also be accomplished by the general route provided in schemes 1, 2 and 8, starting from the appropriately protected P₂The amino ester is started. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1, 2 and 8 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (3) a process 9:

schemes 1-8 in combination with scheme 9 above provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-9 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

A process 10:

schemes 1-8 in combination with scheme 10 above provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 10 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 11:

schemes 1-8 in combination with scheme 11 above provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 11 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (3) a process 12:

schemes 1-8 in combination with scheme 12 above provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 12 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 13:

schemes 1-8 in combination with scheme 13 above provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 13,

wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

The process 14:

starting from commercially available CBz 4-ketoprolinesAmino methyl esters, and utilizing schemes 1-8 in conjunction with scheme 14 above, provide another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 14 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 15:

starting with the commercially available CBz 4-ketoproline methyl ester and utilizing schemes 1-8 in conjunction with scheme 15 above, another general method of preparation of compounds of formula I is provided, wherein-N (R)₁₂)CH(R₁₁) The C (O) -group is as shown above, and wherein n is 0 or 1. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 15 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

And (3) a process 16:

schemes 1-8 in conjunction with scheme 16 above, another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The c (o) -group is as shown above, and wherein R' is as defined in any embodiment herein. The allyl esters can be cleaved according to procedures known to those skilled in the art. For example, allyl ester can be substituted with Pd (Ph)₃)₄With pyrrolidine in CH₂Cl₂And CH₃Cracking in CN to obtain free acid. The acid is coupled to intermediate 14 followed by the general procedure of schemes 1-8 to give compounds of formula I, wherein-N(R₁₂)CH(R₁₁) The C (O) -group is as shown above. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 16 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 17:

schemes 1-8 in conjunction with scheme 17 above, another general method for the preparation of compounds of formula I, wherein-N (R)₁₂)CH(R₁₁) The c (o) -group is as shown above, and wherein R' is as defined in any embodiment herein. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 17 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

The process 18:

scheme 18 above provides a general route for the preparation of compounds wherein V-R-T is as shown above, R₁₇As described in any of the embodiments herein. Wherein a commercially available amine and sulfonyl chloride are condensed and then hydrolyzed under basic conditions to give an intermediate acid. This acid is then further converted to the compound of formula I using the procedures described in schemes 1-8. It will also be appreciated by those skilled in the art that compounds of formula I wherein z is zero can be prepared according to schemes 1-8 and 18 wherein a smaller CBz-Xaa-OH group is coupled during the preparation of the compound.

Scheme 19:

scheme 19 above provides a general route for the preparation of compounds wherein V-R-T is as depicted above, R₁₇、R₁And R_1′As described in any of the embodiments herein. Wherein commercially available amino acid esters are converted to the corresponding N-chlorosulfonyl esters according to the process described by Kempf, D.J. et al, J.Med.chem., pp.320-330 (1993). Coupling of sulfonyl chlorides with the relevant hydrazines (either commercially available or prepared according to procedures well known in the art) followed by basic hydrolysis affords the intermediate acids. The acid intermediate is then converted to the compound of formula I-1 using the procedures described in schemes 1-8 above.

Scheme 19 a:

scheme 19a above provides a general route for the preparation of compounds wherein V-R-T is as depicted above, R₁₇、R₁₃And R_13′As described in any of the embodiments herein. Wherein commercially available amino acid esters are converted to the corresponding N-chlorosulfonyl esters according to the process described by Kempf, D.J. et al, J.Med.chem., pp.320-330 (1993). Coupling of sulfonyl chlorides with the relevant hydrazines (either commercially available or prepared according to procedures well known in the art) followed by basic hydrolysis affords the intermediate acids. The acid intermediate is then converted to the compound of formula I, wherein z is zero, using the procedures described in schemes 1-8 above.

A process 20:

scheme 20 above provides a general route for the preparation of compounds whereinV-R-T is as described above, R₁₇、R₁And R_1′As described in any of the embodiments herein. The chloroacid ester 19b was prepared according to the procedure described in j.org.chem., pp.2624-2629 (1979). Coupling of commercially available amino-tert-butyl ester 19a with chloride 19b affords sulfonamide 19 c. Basic hydrolysis of the mixed ester 19c followed by coupling with a commercially available amine 19d affords intermediate ester 19 e. Oxidation with one equivalent of mBPA to give the sulfoxide, where V is-S (O)₁-. Alternatively, oxidation with two equivalents of mBPA gives the sulfone, where V is-S (O)₂-. Acidic hydrolysis of tert-butyl ester 19e affords acid 19f, which is then further processed to the compound of formula I-1 according to the procedures described in schemes 1-8 above.

Scheme 20 a:

scheme 20a above provides a general route for the preparation of compounds wherein V-R-T is as depicted above, R₁₇、R₁₃And R_13′As described in any of the embodiments herein. The chloroacid ester 19b was prepared according to the procedure described in j.org.chem., pp.2624-2629 (1979). Coupling of commercially available amino tert-butyl ester 19g with chloride 19b gave sulfonamide 19 h. Basic hydrolysis of the mixed ester 19h followed by coupling with a commercially available amine 19d affords intermediate ester 19 i. Oxidation with one equivalent of mBPA to give the sulfoxide, where V is-S (O)₁-. Alternatively, oxidation with two equivalents of mBPA gives the sulfone, where V is-S (O)₂-. Acidic hydrolysis of tert-butyl ester 19I affords acid 19j, which is then further processed to compounds of formula I, where z is zero, according to the procedures described in schemes 1-8 above.

Scheme 21:

scheme 21 above provides a route to compound 5 of the present invention from intermediate 32 a. Intermediate 32a was prepared according to the general procedure outlined in schemes 1 and 2 from commercially available starting materials. The experimental procedure for the preparation of compound 5 from intermediate 32a and intermediate 14 is described in detail in the examples section provided herein.

The process 22:

scheme 22 above provides a route to compound 6 of the present invention from intermediate 39. Intermediate 39 was prepared from commercially available starting materials according to the general procedures outlined in schemes 1, 2 and 21. The experimental procedure for the preparation of compound 6 from intermediate 39 is described in detail in the examples section provided herein.

The preparation of compounds of formula I by the preparation of various other optionally substituted polycyclic azaheterocyclyl intermediates of schemes 1-8 above may be accomplished by the methods described in PCT publication WO 02/18369 and the references cited therein.

Various can be used as P₂Some of the 3, 4 and 5-substituted proline analogs are either commercially available or can be prepared according to known literature methods. For example, certain 3-substituted proline analogs of interest can be prepared according to the method of Holladay, m.w. et al, j.med.chem., 34, pp.457-461 (1991). In addition, various 3, 4-disubstituted proline analogues can be prepared according to the method of Kanamasa, S. et al, J.org.chem, 56, pp.2875-2883 (1991). In relation to 3, 4 or 5-substituted prolines or 3, 4-disubstitutedIn each of the syntheses of proline, the intermediates may be further processed by the routes defined above in schemes 1-8 for the preparation of the compounds of the invention.

While certain embodiments are depicted and described below, it will be appreciated that the compounds of the invention may be prepared according to the methods generally described above, using starting materials that are generally available to those of ordinary skill.

In other forms of any of the embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt or mixture of salts thereof. In other forms of any of the embodiments of the present invention, the compounds of the present invention are present in an amount effective to reduce the amount of virus loaded into a sample or in a patient, wherein the virus encodes a serine protease essential to the viral life cycle, and a pharmaceutically acceptable carrier.

If pharmaceutically acceptable salts of the compounds of the present invention are used in these compositions, it is preferred that those salts are derived from inorganic or organic acids and bases. Included among such acid salts are the following: ethylhydrochloric acid, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentane-propionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate (pectinate), persulfate, 3-phenyl-propionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and the like.

Similarly, basic nitrogen-containing groups may be quaternized with lower alkyl halide-based agents such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl, diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromine and iodine, aralkyl halides such as benzyl and phenethyl bromides and others. Thereby obtaining water or oil-soluble or dispersible products.

The compounds used in the compositions and methods of the invention may also be modified by the addition of suitable functional groups, which may add selective biological properties. Such modifications are known in the art and include those that enhance biological penetration of the biological system being administered (e.g., blood, lymphatic system, central nervous system), enhance oral efficacy, enhance solubility for administration by injection, alter metabolism, and alter rate of excretion.

Pharmaceutically acceptable carriers that may be used in these compositions include, but are not limited to: ion exchange resins, aluminum oxide, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, paraffin wax, polyethylene-polyoxypropylene-block copolymers, polyethylene glycol and wool fat.

In other forms of any of the embodiments of the present invention, the compositions of the present invention are formulated for pharmaceutical administration to a mammal. In other forms of any of the embodiments of the present invention, the compositions of the present invention are formulated for pharmaceutical administration to a human.

Such pharmaceutical compositions of the invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an embedded reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the composition is administered orally or intravenously.

Sterile injectable forms of the compositions of the present invention may be aqueous or oily suspensions. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Acceptable excipients and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, for example natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. The oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant, for example carboxymethyl cellulose or similar dispersing agents, which are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants such as Tweens, Spans, and other emulsifying agents or bioavailability enhancers, which are commonly used to prepare pharmaceutically acceptable solid, liquid, or other dosage forms, may also be used for formulation purposes.

Dosage levels of the protease inhibitor compounds described herein of between about 0.01 and about 100mg/kg body weight per day are effective for use in monotherapy for the prophylaxis and treatment of viruses, particularly against HCV-mediated diseases. In other forms of any of the embodiments of the present invention, a dosage level of the protease inhibitor compounds described herein of between about 0.5mg/kg and about 75mg/kg body weight per day is effective for use in monotherapy for the prophylaxis and treatment of viral diseases, particularly against HCV mediated diseases. Typically, the pharmaceutical compositions of the present invention are administered from about 1 to about 5 times per day, or as a continuous infusion. Such administration may be for chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Typical formulations will contain from about 5% to about 95% active compound (w/w). Another such formulation contains from about 20% to about 80% of the active compound.

When the compositions of the present invention comprise a combination of a compound of formula I and one or more additional therapeutic or prophylactic agents, the dosage level of both the compound and the additional agent should be between about 10% and 100% of the dosage normally administered in a monotherapy regimen. In other forms of any of the embodiments of the invention, the additional agent should be present at a dosage level of between about 10 to 80% of the dosage normally administered in a monotherapy regimen.

The pharmaceutical compositions of the present invention may be administered orally in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in the form of capsules, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of the present invention may be administered in the form of suppositories for rectal administration. These compositions may be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of the present invention may also be administered topically, particularly when the target site of treatment includes sites or organs accessible by topical application, including ocular, dermatological, or lower intestinal disorders. Suitable topical formulations for each of these sites or organs can be readily prepared.

Topical administration for the lower intestinal tract may be carried out in the form of rectal suppository formulations (see above) or suitable enema formulations. Topical-transdermal patches may also be used.

For topical administration, the pharmaceutical compositions may be formulated in a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions may be formulated in a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical composition may be formulated as a micronized suspension in isotonic, pH adjusted, sterile saline, preferably as a solution in isotonic, pH adjusted, sterile saline, whether or not a preservative such as benzalkonium chloride is present. Alternatively, for ophthalmic use, the pharmaceutical composition may be formulated in an ointment such as paraffin oil.

The pharmaceutical compositions of the present invention may also be administered by means of nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in physiological saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents.

In any other form of the embodiment of the present invention, the pharmaceutical composition is formulated for oral administration.

In other forms of any of the embodiments of the present invention, the compositions of the present invention additionally comprise another antiviral agent, preferably an anti-HCV agent. Such antiviral agents include, but are not limited to: immunomodulators, such as alpha-, beta-, and gamma-interferons, pegylated-derived interferon-alpha compounds, and thymosins; other antiviral agents, such as ribavirin, tricyclodecylamine, and telbivudine (telbivudine); inhibitors of other hepatitis C proteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors); inhibitors of other targets in the HCV life cycle, including helicase and polymerase inhibitors; an internal ribosome channel inhibitor; broadspectrum viral inhibitors, such as IMPDH inhibitors (e.g., the compounds of U.S. Pat. Nos. 5,807,876, 6,498,178, 6,344,465, 6,054,472, WO 97/40028, WO 98/40381, WO 00/56331, and mycophenolic acid and derivatives thereof, including but not limited to VX-497, VX-148, and/or VX-944); or a combination of any of the above. See w.markland et al,Antimicrobial & Antiviral Chemotherapy44, p.859(2000) and us6,541,496.

The following definitions are used herein (by the filing date of this application, trademarks on products exist).

"Peg-Intron" meansPegionterferon α -2b available from Schering Corporation, Kenilworth, NJ;

"Intron" meansInterferon alpha-2 b available from schering corporation, Kenilworth, NJ;

"ribavirin" refers to ribavirin (1- β -D-ribofuranosyl-1H-1, 2, 4-triazole-3-carboxamide, available from ICN Pharmaceuticals, Inc., Costa Mesa, Calif.; described in Merck Index, entry 8365, Twelfth Edition; and optionallyObtained from Schering Corporation, Kenilworth, NJ, orTrademarks of (D) are available from Hoffmann-La Roche, Nutley, NJ;

"Pagasys" meansPEG-IFN α -2a from Hoffmann-La Roche, Nutley, NJ;

by "Roferon" is meantRecombinant interferon alpha-2 a, available from Hoffmann-La Roche, Nutley, NJ;

by "Berefor" is meantInterferon alpha 2, available from boehringer ingelheim Pharmaceutical, inc, ridgfield, CT;

a pure mixture of natural alpha interferons such as Sumiferon, available from Sumitomo, Japan;

interferon alfa n1, available from Glaxo-Wellcome ltd, GreatBritain;

a mixture of natural alpha interferons prepared by Interferon Sciences, available from Purdue Frederick co., CT;

In other forms of any of the embodiments of the present invention, the interferon is interferon-alpha. In any other form of the embodiment of the present invention, the therapeutic combination of the present invention employs natural interferon alpha 2 a. Alternatively, the therapeutic combination of the present invention employs natural interferon-alpha 2 b. In any other form of the embodiment of the invention, the therapeutic combination therapy of the invention employs recombinant interferon-alpha 2a or 2 b. In other forms of any of the embodiments of the present invention, the interferon is a pegylated interferon alfa 2a or 2 b. Interferons suitable for the present invention include:

(a) intron (interferon-. alpha.2B, Schering Plough),

(b)Peg-Intron，

(c)Pegasys，

(d)Roferon，

(e)Berofor，

(f)Sumiferon，

(g)Wellferon，

(h) consensus interferon-alpha, available from Amgen, inc., Newbury Park, CA,

(i)Alferon；

(j)Viraferon；

(k)Infergen。

as the skilled artisan will recognize, oral administration of protease inhibitors is preferred. Interferons cannot generally be administered orally. However, there is no limitation to any particular dosage form or mode of the methods or compositions of the invention herein. Thus, each of the components of the combination according to the invention may be administered separately, together or in any combination thereof.

In any other form of this embodiment of the invention, the protease inhibitor and interferon are administered in separate dosage forms. In other forms of any of the embodiments of the present invention, any additional agent is administered as part of a single dosage form with the protease inhibitor, or as a separate dosage form. Because the present invention relates to the combined use of compounds, the specific amount of each compound may depend on the specific amount of each other compound in the composition. As recognized by the skilled artisan, the dose of interferon is typically measured in IU's (e.g., about 4 million IU to about 12 million IU).

Accordingly, agents that may be used with the compounds of the present invention in compositions (whether acting as immunomodulators or otherwise) include, but are not limited to: interferon- α 2B (Intron, Schering Plough); rebatron (Schering Plough, interferon-. alpha.2B + ribavirin); pegylated interferon alpha (Red, K.R. et al, "efficiency and Safety of Pegylated (40-kd) interactive alpha-2 accessed with interactive alpha-2 a in noncirrhotic moieties with a cytotoxic moiety CHepatology33, pp.433-438 (2001); consensus Interferon (Kao, J.H., et al, "effectiveness of Consensus Interferon therapy of viral HepatitisJ.Gastroenterol.Hepatol.15，pp.1418-1423(2000)，interferon-alpha 2A(Roferon A；Roche)，lymphoblastoid or″natural″interferon；interferon tau (Clayette, p. et al, "IFN-tau, A New Interferon Type I with Antiretroviral activityPathol.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，19Pp.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 Disease19, pp.103-112 (1999); ribavirin; and increase the progression of type 1 helper T cell responses (Davis et al, "Future Options for the Management of hepatitis C.")Seminars in Liver Disease19, pp.103-112 (1999). Interferons can ameliorate viral infections by producing a direct antiviral effect and/or by modulating the immune response to the infection. The antiviral effects of interferons are often mediated by inhibiting viral penetration or uncoating, viral RNA synthesis, translation of viral proteins, and/or viral assembly and release.

Compounds which promote the synthesis of interferons in cells (Tazulakhova, E.B. et al, Russian experiment in Screening, analysis, and clinical application of Novel interference indicatorsJ.Interferon Cytokine Res.21pp.65-73) include but are not limited to: double-stranded RNA, alone or in combination with tobramycin and Imiquimod (3M Pharmaceuticals; Sauder, D.N. "immunomodulating and pharmacological Properties of ImiquimodJ. Am.Acad.Dermatol.，43 pp.S6-11(2000)。

Other non-immunomodulatory or immunomodulatory compounds that may be used in combination with the compounds of the invention include, but are not limited to: those described in detail in WO 02/18369, which is incorporated herein by reference (see, e.g., pages 273, 9-22 and lines 274, 4 to 276, 11).

The invention may also include administration of a cytochrome P450 monooxygenase inhibitor. CYP inhibitors can be effective to increase the liver concentration of CYP-inhibited compounds and/or increase blood levels.

If embodiments of the invention include a CYP inhibitor, any CYP inhibitor that enhances the pharmacokinetics of the protease related NS3/4A may be used in the methods of the invention. Such CYP inhibitors include, but are not limited to, ritonavir (WO 94/14436), ketoconazole, oleandomycin, 4-methylpyrazole, cyclosporin, clomerthiazol, cimetidine, itraconazole, fluconazole, miconazole, fluvoxamine, fluoxetine, nefazodone, sertraline, indinavir, nelfinavir, amprenavir, fosamprenavir, saquinavir, lopinavir, delavirdine, erythromycin, VX-944, and VX-497. Preferred CYP inhibitors include ritonavir, ketoconazole, triacetyl oleandomycin, 4-methylpyrazole, cyclosporin, and clomeprazole. For a preferred dosage form of ritonavir, see U.S. Pat. No. 6,037,157 and the following references: U.S. Pat. No. 5,484,801, U.S. application 08/402,690, and international applications WO 95/07696 and WO 95/09614).

Methods for determining the ability of a compound to inhibit cytochrome P50 monooxygenase activity are known (see US6,037,157 and Yun, et al,Drug Metabolism & Disposition，vol.21，pp.403-407(1993)。

once the patient's condition is improved, a maintenance dose of a compound, composition or combination of the invention can be administered, if desired. Subsequently, the dose or number of administrations, or both, as a parameter of the condition may be reduced to a level that maintains an improved condition, and the treatment should be discontinued when the condition has been alleviated to the desired level. However, patients may require intermittent therapy in a long-term manner in the event of recurrence of disease symptoms.

It will also be understood that the specific dose and treatment regimen for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease undergoing therapy. The amount of active ingredient also depends on the presence and performance of the specifically described compounds, and additional antiviral agents in the composition.

The present invention provides a method for treating a patient infected with a virus characterized by a virally-encoded serine protease essential to the viral life cycle by administering to said patient a pharmaceutically acceptable composition of the present invention. In other forms of any of the embodiments of the present invention, the methods of the present invention are used to treat a patient having an HCV infection. Such treatment may completely eradicate the viral infection or reduce its severity. In any other form of the embodiment of the present invention, the patient is a human.

In other forms of any of the embodiments of the present invention, the methods of the present invention further comprise the step of administering to said patient an antiviral agent, preferably an anti-HCV agent. Such antiviral agents include, but are not limited to: immunomodulators, such as alpha-, beta-, and gamma-interferons, pegylated-derived interferon-alpha compounds, and thymosins; other antiviral agents, such as ribavirin, tricyclodecylamine, and telbivudine; inhibitors of other hepatitis C proteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors); inhibitors of other targets in the HCV life cycle, including but not limited to helicase and polymerase inhibitors; an internal ribosome channel inhibitor; broadspectrum viral inhibitors, such as IMPDH inhibitors (e.g., VX-497 and other IMPDH inhibitors disclosed in U.S. patent nos. 5,807,876 and 6,498,178, mycophenolic acid and derivatives thereof); an inhibitor of cytochrome P-450, such as ritonavir, or a combination of any of the foregoing.

Such additional agents may be administered to the patient as part of a single dosage form comprising both the compound of the present invention and another antiviral agent. Alternatively, another agent may be administered separately from the compound of the invention as part of a multiple dosage form, wherein the other agent is administered prior to, together with, or after administration of a composition comprising the compound of the invention.

In other forms of any of the embodiments herein, the invention provides a method of pretreating a biological substance designed for administration to a patient, comprising the step of contacting the biological substance with a pharmaceutically acceptable composition comprising a compound of the invention. Such biological substances include, but are not limited to: blood and its components such as plasma, platelets, subpopulations of blood cells, and the like; organs such as kidney, liver, heart, lung, etc.; semen and egg cells; bone marrow and its components, and other fluids injected into a patient, such as saline, glucose, and the like.

In other forms of any of the embodiments of the present invention, the present invention provides methods of treating materials that can potentially be exposed to viruses characterized by virus-encoded serine proteases essential to the viral life cycle. The method comprises the step of contacting the substance with a compound according to the invention. Such materials include, but are not limited to: surgical instruments and clothing (e.g., coats, gloves, aprons, gowns, masks, lenses, footwear, etc.); laboratory instruments and clothing (e.g., coats, gloves, aprons, gowns, masks, lenses, footwear, etc.); blood collection devices and materials; and implanting components such as bypass tubes, stents, etc.

In any other form of this embodiment of the invention, the compounds of the invention may be used as a laboratory tool to aid in the isolation of virally encoded serine proteases. The method comprises the following steps: providing a compound of the invention attached to a solid support; contacting the solid support with a sample containing a viral serine protease under conditions that cause binding of the protease to the solid support; and eluting the serine protease from the solid support. In one embodiment, the viral serine protease isolated by this method is HCV NS3-NS4A protease.

In order that the invention may be more fully understood, the following preparation and test examples are set forth. These examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention in any way.

Examples

Recording at 500MHz using a Bruker AMX 500 instrument¹H-NMR spectrum. Mass spectral samples were analyzed on a MicroMass ZQ or QuattroII mass spectrometer with electrospray ionization operating in single MS mode. The sample is introduced into the mass spectrometer using Fluid Injection (FIA) or chromatography. All mobile phases for mass spectrometry consist of acetonitrile-water mixtures containing 0.2% formic acid as modifier.

The term "R" as used herein_t(min) "means the HPLC retention time in minutes, which is related to the compound. The HPLC retention times listed can be obtained either from mass spectral data or using the following methods:

the instrument comprises the following steps: hewlett Packard HP-1050;

column: YMC C₁₈(Cat.No.326289C46)；

Gradient/gradient time: 10-90% CH₃CN/H₂O9 min, then 100% CH₃CN 2 minutes;

flow rate: 0.8 ml/min;

detector wavelength: 215nM and 245 nM.

Using Cambridge Soft corporation ChemDrawThe nomenclature provided by version7.0.1 accomplishes the chemical nomenclature of the compounds selected herein.

Example 1:

3-amino-2-hydroxy-ethyl hexanoate hydrochloride(14)Preparation of

To carboxylic acids prepared according to Harbeson, S. et al, J.Med.chem.37, 18, pp.2918-2929(1994)9(4.5g，24.5mmol) of ethanol (50mL) is added 6mL of a saturated solution of hydrochloric acid in ethyl acetate, followed by a catalytic amount of concentrated sulfuric acid. The reaction was stirred at ambient temperature overnight. The solvent was removed and the residue was dried under vacuum. Based on TLC analysis, the amino acid was completely converted to the amino ester. The product was used without further purification.

Example 2: 3- { [1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carbonyl]-amino } -2-hydroxy-hexanoic acid ethyl ester(32)Preparation of

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino ] -2-carbonyl ] -amino-1-methyl-formamide in a dry flask with a 1: 1 mixture of dichloromethane and dimethylformamide (100mL)]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid32a(2.0g, 4.28mmol), PyBOP (2.5g, 4.7mmol) and N-methyl-morpholine (2.4ml, 21.4mmol) and the reaction was placed under a nitrogen atmosphere. Amine was placed in a 100mL addition funnel14(1.0g, 4.7mmol) with N-methyl-morpholine (0.60mL, 5.35mmol) in a 1: 1 mixture of dichloromethane-dimethylformamide (10 mL). The reaction was cooled to 0 ℃ and the amine was added while maintaining the temperature at 0 ℃. The reaction was allowed to warm to ambient temperature with stirring overnight. HPLC analysis showed the reaction was complete, then the reaction was diluted to 500mL with water. After stirring, the precipitate was filtered and dried to give 2.4g (82% yield)32It is a beige solid. The product was used in the next step without further purification. HPLC RT 5.66min, (10-90% acetonitrile-water over 7 min); LC/MS: retention time 3.94min (10-90% acetonitrile-water over 5 min); m + H⁺＝685.5。

Example 3:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino-]Acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid [1- (N '-methyl-N' -phenyl-hydrazinooxalyl) -butyl]-amides of(6)Preparation of

To acid33(100mg, 0.152mmol) in DMA (1mL) was added PyBrop (110mg, 0.213mmol), N-methylmorpholine (250. mu.L, 2.27mmol) andN-methyl-N-phenylhydrazine (122mg, 1.0mmol), the reaction was stirred until the starting acid was consumed according to HPLC analysis. The reaction was diluted with water and the product precipitated from solution. The solid was filtered and dried to give 70mg (60%) of the compound36Having consistent mass spectral data (M + H)⁺761). The material was used without further purification.

To hydroxy-hydrazides36(70mg, 0.092mmol) of CH₂Cl₂(2mL) solution Dess-Martin periodinane (100mg, 0.236mmol) and 100. mu.L t-butanol were added and the reaction was stirred at RT for 1 hour. The solvent was removed and the product was purified by flash chromatography on silica gel (50% EtOAc-hexanes) to give the product as a colorless glass. Dissolving the material in 50% acetonitrile-water, and lyophilizing to obtain 23mg of the compound6As a colorless solid with consistent mass spectral data (LC/MS; RT 4.30min, 10-90% acetonitrile-water over 7 min; M + H)⁺＝759.1)。

As hereinbefore described for the compounds6The process also prepares the compound7And8。

example 4:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid (1-phenoxyaminooxalyl-butyl) -amide(5)Preparation of

To acid33(200mg, 0.304mmol) in DMA (1mL) was added PyBrop (200mg, 0.43mmol), N-methylmorpholine (400. mu.L, 3.62mmol) and O-phenyl-hydroxylamine (100mg, 0.682mmol) and the reaction stirred until the starting acid was consumed according to HPLC analysis. The reaction was diluted with water and the product precipitated from solution. The solid was filtered and dried to yield 78mg (34%) of the compound34And used in the next step without further purification.

To hydroxy-amides34(78mg, 0.10mmol) of CH₂Cl₂(2mL) solution Dess-Martin periodinane (68mg, 0.16mmol) and 100. mu.L t-butanol were added and the reaction was stirred at RT for 1 hour. Removing solvent, and subjecting the product to preparative silica gel thin layer colorSpectrum purification (5% isopropanol-EtOAc) to give 5.8mg (6%) of the compound5As a colorless glass with consistent mass spectral data (LC/MS; RT 4.50min, (10-90% acetonitrile-water over 7 min); M + H⁺＝746.1)。

As hereinbefore described for the compounds5The process also prepares the compound1To4。

Example 5:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid (1-isobutoxyaminooxalyl-butyl) -amide(1)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 4 and 21. The product was isolated as a solid with consistent mass spectral data (LC/MS; retention time 4.2, M + H)⁺726 observation).

Example 6:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid (1-tert-butoxyaminooxalyl-butyl) -amide(2)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 4 and 21. The product was isolated as a solid with consistent mass spectral data (LC/MS; retention time 4.1, M + H)⁺726.3 observations).

Example 7:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid (1-benzyloxyaminooxalyl-butyl) -amide(3)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 4 and 21. The product was isolated as a solid with consistent mass spectral data (LC/MS; retention time 4.2, M + H)⁺760 observed values).

Example 8:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid (1-allyloxyaminooxalyl-butyl) -amide(4)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 4 and 21. The product was isolated as a solid with consistent mass spectral data (LC/MS; retention time 3.8, M + H)⁺710 observations).

Example 9:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid [1- (morpholin-4-yl) aminooxalyl-butyl]-amides of(7)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 3 and 22. The product was isolated as a colorless solid with consistent mass spectral data (LC/MS; retention time 3.7, M + H)⁺739.4 observations).

Example 10:

1- (2- { 2-cyclohexyl-2- [ (pyrazine-2-carbonyl) -amino group]-acetylamino } -3, 3-dimethyl-butyryl) -octahydro-indole-2-carboxylic acid [1- (N ', N' -dibenzyl-hydrazino) oxalyl-butyl]-amides of(8)Preparation of

This compound was prepared using procedures similar to schemes 1, 2, 3 and 22. The product was isolated as a colorless solid with consistent mass spectral data (LC/MS; retention time 4.94, M + H)⁺849 observations).

Example 11:

HCV Ki assay protocol:

HPLC Microbore method for separating 5AB substrate and product

Substrate:

NH₂-Glu-Asp-Val-Val-(α)Abu-Cys-Ser-Met-Ser-Tyr-COOH

stock solutions of 20mM 5AB (or concentration of you chose) were prepared in DMSO w/0.2M DTT. Stored in aliquots at-20 ℃.

Buffering agent: 50mM HEPES, pH 7.8; 20% of glycerol; the total test volume of 100mM NaCl was 100. mu.L

Reagent	X1(μL)	Concentration in assay
			Buffer solution	86.5	See above
5mM KK4A	0.5	25μL
			1M DTT	0.5	5mM
DMSO or inhibitors	2.5	2.5％v/v
			50μM tNS3	0.05	25nM
250 μ M5 AB (original)	20	25μM

Combining buffer, KK4A, DTT and tNS 3; each 78 μ L of this solution was dispensed into each well of a 96-well plate. Incubate at 30 ℃ for about 5-10 minutes.

To each well was added 2.5 μ L of a DMSO solution of the appropriate concentration of test compound (DMSO alone for control) and incubated at room temperature for 15 minutes.

The reaction was initiated by adding 20. mu.l of 250. mu.L of 5AB substrate (25. mu.M concentration equal to or slightly below Km of 5 AB).

Incubate at 30 ℃ for 20 minutes.

The reaction was stopped by adding 25. mu.L of 10% TFA.

Transfer 120 μ l aliquots to HPLC vials.

The SMSY product was isolated from the substrate and KK4A by the following method:

microbore separation method:

the instrument comprises the following steps: agilent 1100

Degassing device G1322A

Binary pump G1312A

Autosampler G1313A

Column thermostatic chamber G1316A

Diode matrix detector G1315A

Column:

phenomenex Jupiter; 5micron C18; 300 angstroms; 150x2 mm; P/O00F-4053-B0

Keeping the column constant temperature: 40 deg.C

Injection volume: 100 μ L

Solvent a ═ HPLC grade water + 0.1% TFA

Solvent B ═ HPLC grade acetonitrile + 0.1% TFA

Time (min)	％B	Flow rate (ml/min)	Maximum pressure
				0	5	0.2	400
12	60	0.2	400
				13	100	0.2	400
16	100	0.2	400
				17	5	0.2	400

Stopping time: 17min

Time after operation: for 10 min.

Table 1 below depicts Ki data for certain compounds of the present invention. Compounds with Ki below 1.5 μ M were designated as a. Compounds with Ki's ranging from 1.5. mu.M to 3. mu.M were designated B. Compounds with Ki higher than 3 μ M were designated C.

Table 1:

compound (I)	Ki(μM)
		1	A
2	A
		3	B
4	C
		5	B
6	A
		7	C
8	C

Claims

1. A compound of formula I:

or a pharmaceutically acceptable salt or mixture thereof, wherein:

z is 1;

v is-C (O) -;

r is a bond;

t is (C)₅-C₁₀) A heteroaryl group;

w is:

wherein:

each R₁₇Independently are:

hydrogen-,

(C₁-C₁₂) -an aliphatic radical-,

(C₃-C₁₀) -a cycloalkyl group-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-, or

(C₃-C₁₀) -heterocyclyl-;

R₅and R_5’Independently are: hydrogen or (C)₁-C₁₂) -an aliphatic group;

R₁，R_1′，R₁₁，R_11′，R₁₃and R_13’Independently are: hydrogen-, (C)₁-C₁₂) -an aliphatic radical-, or (C)₃-C₁₀) -a cycloalkyl group;

R₂，R₄ ，R₈and R₁₂Independently are: hydrogen-;

or R₁₁And R₁₂A carbocyclic or heterocyclic ring system which together with the atoms to which they are bonded forms an 8-to 20-membered bicyclic ring; wherein, in the bicyclic ring system, each ring is linearly fused; wherein each ring is aromatic or non-aromatic; wherein each heteroatom in the heterocyclic ring system is N or NH;

in the above definitions the aliphatic group means a straight or branched alkyl, alkenyl or alkynyl group.

2. A compound according to claim 1, wherein:

the radicals are:

3. a compound according to claim 2, wherein:

the radicals are:

4. a compound according to claim 1, wherein R₅And R_5′Independently hydrogen or:

5. a compound according to claim 1, wherein R_13′Is hydrogen, and R₁₃The method comprises the following steps: (C)₁-C₆) -an aliphatic group.

6. A compound according to claim 5, wherein R_13′Is hydrogen, and R₁₃The method comprises the following steps:

7. a compound according to claim 1, wherein R_1′Is hydrogen, and R₁The method comprises the following steps: (C)₃-C₁₀) -a cycloalkyl group.

8. A compound according to claim 7, wherein R_1′Is hydrogen, and R₁The method comprises the following steps:

9. the compound according to claim 1, wherein T is:

10. a compound according to claim 1, wherein W is:

wherein

Each R₁₇Independently are:

hydrogen-,

(C₆-C₁₀) -an aryl-group-,

(C₆-C₁₀) -aryl- (C)₁-C₁₂) An aliphatic radical-, or

(C₃-C₁₀) -heterocyclyl-.

11. A compound selected from the group consisting of:

12. a pharmaceutical composition comprising a serine protease inhibiting amount of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or adjuvant.

13. The pharmaceutical composition according to claim 12, wherein the adjuvant is an excipient.

14. The pharmaceutical composition according to claim 12, wherein the composition is formulated for administration to a patient.

15. Use of a compound according to any one of claims 1 to 11 in the manufacture of a medicament for inhibiting serine protease activity.

16. The use according to claim 15, wherein said serine protease is HCV NS3 protease.

17. Use of a compound according to any one of claims 1-11 in the manufacture of a medicament for treating HCV infection in a patient.

18. A method for eliminating or reducing HCV contamination of a biological sample or of a medical or laboratory device for non-diagnostic and therapeutic purposes comprising the step of contacting said biological sample or medical or laboratory device with a composition according to claim 12.

19. The method according to claim 18, wherein the biological sample is either a medical or laboratory device selected from the group consisting of biological tissue, surgical instruments, surgical gowns, laboratory instruments, lab coats, blood or other body fluid collection devices, blood or other body fluid storage materials.

20. The method according to claim 18, wherein the biological sample is blood or other body fluid.