MXPA06006569A - Inhibitors of hepatitis c virus ns3/ns4a serine protease - Google Patents

Abstract

The present invention discloses novel compounds which have HCV protease inhibitory activity as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising such compounds as well as methods of using them to treat disorders associated with the HCV protease.

Description

INHIBITORS OF THE NS3 / NS4A SERINE PROTEASE OF HEPATITIS C VIRUS FIELD OF THE INVENTION The present invention relates to new inhibitors of hepatitis C virus proteases ("HCV"), pharmaceutical compositions containing one or more such inhibitors, methods for preparing such inhibitors and methods for using such inhibitors to treat hepatitis C. and related disorders. This invention also describes new macrocyclic compounds as inhibitors of the NS3 / NS4a serine protease of HCV. The present application claims the priority of the provisional patent application of E.U.A. Serial No. 60/528845 filed on December 11, 2003.

BACKGROUND OF THE INVENTION The hepatitis C virus (HCV) is a positive-sense single-stranded RNA virus that has been considered as the main causative agent of non-B non-B hepatitis (NANBH), particularly in NANBH associated with blood. (BB-NANBH) (see International Patent Application Publication No. WO 89/04669 and European Patent Application Publication No. EP 381 216). NANBH should be distinguished from other types of liver disease induced by viruses, such as hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis delta virus (HDV), cytomegalovirus (CMV) and the Epstein-Barr virus (EBV), as well as other forms of liver disease such as alcoholism and primary biliary cirrhosis. Recently, a HCV protease for polypeptide processing and viral replication has been identified, cloned and expressed; . { see, for example, the U.S. Patent. No. 5,712,145). This polyprotein of approximately 3000 amino acids contains, from the amino terminus to the carboxyl terminus, a nucleocapsid protein (C), envelope proteins (E1 and E2) and several non-structural proteins (NS1, 2, 3, 4a, 5a and 5b). NS3 is a protein of approximately 68 kda, encoded by approximately 1893 nucleotides of the HCV genome, and has two distinct domains: (a) a serine protease domain consisting of approximately 200 of the N-terminal amino acids and (b) a ATPase domain dependent on RNA at the C-terminal end of the protein. The NS3 protease is considered a member of the chymotrypsin family due to the similarities in the protein sequence, the overall three-dimensional structure and the mechanism of catalysis. Other enzymes of the chymotrypsin type are elastase, factor Xa, thrombin, trypsin, plasmin, urokinase, tPA and PSA. The NS3 serine protease of HCV is responsible for the proteolysis of the polypeptide (polyprotein) at the junctions NS3 / NS4a, NS4a / NS4b, NS4b / NS5a and NS5a / NS5b and thus is responsible for generating four viral proteins during viral replication. This has made NS3 serine protease an attractive target for antiviral chemotherapy. The compounds of the invention can inhibit such a protease. They can also modulate the processing of the hepatitis C virus (HCV) polypeptide. It has been determined that the NS4a protein, a polypeptide of about 6 kda, is a co-factor for the serine protease activity of NS3. The auto-rupture of the NS3 / NS4a junction by the NS3 / NS4a serine protease occurs intramolecularly (ie in cis) while the other cleavage sites are processed intramolecularly (i.e. in trans). The analysis of the natural break sites for the protease of HCV revealed the presence of cysteine in P1 and serine in P1 'and that these residues are conserved strictly in the NS4a / NS4b, NS4b / NS5a and NS5a / NS5b junctions. The NS3 / NS4a junction contains a threonine in P1 and a serine in P1 '. The Cys? Thr substitution in NS3 / NS4a is postulated to explain the requirement of cis processing rather than the trans in this binding. See, for example, Pizzi et al. (1994) Proc. Nati Acad. Sci (USA) 91: 888-892, Failla et al. (1996) Folding & Design 1: 35-42. The breaking site of NS3 / NS4a is also more tolerant to mutagenesis than the other sites. See, for example, Kollykhalov et al. (1994) J. Virol. 68: 7525-7533. It has also been found that acidic residues are required in the region upstream of the rupture site for efficient rupture. See, for example, Komoda et al. (1994) J. Virol. 68: 7351-7357. Inhibitors of the HCV protease that have been reported include antioxidants. { see, International Patent Application Publication No. WO 98/14181), certain peptides and peptide analogues. { see, International Patent Application Publication No. WO 98/17679, Landro et al. (1997) Biochem. 36: 9340-9348, Ingallinella et al. (1998) Biochem. 37: 8906-8914, Llinás-Brunet et al. (1998) Bioorg. Med. Chem. Lett. 8: 1713-1718), inhibitors based on the 70 amino acid eglin c polypeptide (Martin et al. (1998) Biochem. 37: 11459-11468, the affinity of the inhibitors selected from the secretory pancreatic trypsin inhibitor (hPSTI-C3) and minibody repertoires (MBip) (Dimasi et al (1997) J. Virol. 71: 7461-7469), cVHE2 (a "camelized" variable domain antibody fragment) (Martin et al. (1997) Protein Eng. 10: 607-614), and α1-antichymotrypsin (ACT) (Elzouki et al.) (1997) J. Hepat. 27: 42-28). Recently, a ribozyme designed to selectively destroy hepatitis C virus RNA has been described. { see, BioWorld Today 9 (217): 4 (November 10, 1998)). Reference is also made to PCT Publication No. WO 98/17679, published April 30, 1998 (Vértex Pharmaceuticals Incorporated); WO 98/22496, published May 28, 1998 (F. Hoffmann-La Roche AG); and WO 99/07734, published February 18, 1999 (Boehringer Ingelheim Canada Ltd.). HCV has been linked to cirrhosis of the liver and the induction of hepatocellular carcinoma. The prognosis for patients suffering from HCV infection is usually poor. HCV infection is more difficult to treat than other forms of hepatitis due to the lack of immunity or remission associated with HCV infection. Current data indicate a survival rate of less than 50% in the four years after the diagnosis of cirrhosis. Patients diagnosed with localized resectable hepatocellular carcinoma have a 5-year survival rate of 10% to 30%, while patients with unresectable localized hepatocellular carcinoma have a 5-year survival rate of less than 1%. Reference is made to WO 00/59929 (US 6,608,027, Assignee: Boehringer Ingelheim (Canada) Ltd .. Published October 12, 2000) which describes the peptide derivatives of the formula: Reference is made to A. Marchetti ei al, Synlett, S1_, 1000-1002 (1999), which describes the synthesis of bicyclic analogues of an inhibitor of the NS3 protease of HCV. A compound described therein has the formula: Reference is also made to W. Han eí al, Bioorganic & Medicinal Chem. Lett, (2000) 10, 711-713, which describes the preparation of certain α-ketoamides, α-keto esters and α-diketones containing allyl and ethyl functionalities. Reference is also made to WO 00/09558 (Assignee: Boehringer Ingelheim Limited; Published February 24, 2000), which describes peptide derivatives of the formula: ~ s where the various elements are defined. An illustrative compound of that series is: Reference is also made to WO 00/09543 (Assignee: Boehringer Ingelheim Limited, Published February 24, 2000), which describes peptide derivatives of the formula; where the various elements are defined. An illustrative compound of that series is: the American Application is also referenced 6,608,027 (Boehringer Ingelheim, Canada) describing NS3 protease inhibitors of the type: where the various radicals are defined. Current therapies for hepatitis C include interferon-a (INFa) and combination therapy with ribavirin and interferon. See, for example ^ Beremguer et al. (1998) Proc. Assoc. Am. Physicians 110 (2): 98-112. These therapies suffer from a low percentage of sustained response and frequent side effects. See, for example, Hoofnagle et al. (1997) N. Engl. J. Med? 336: 347. Currently, there is no vaccine available for HCV infection. Reference is also made to WO 01/74768 (Assignee: Vértex Pharmaceuticals Inc) published on October 11, 2001, which discloses certain compounds of the following general formula (R is defined therein) as inhibitors of the NS3-serine protease of the Hepatitis C virus: a specific compound described in WO 01/74768 mentioned above has the following formula: Reference is also made to WO02 / 18369 (Eli Lilly &Company), which describes protease inhibitors. An illustrative compound described therein has the structure: Reference is also made to WO03 / 006490 (Vértex Pharmaceuticals), which describes inhibitors of bicyclic proteases with bridging groups. An illustrative compound described therein has the structure: PCT Publications WO 01/77113, WO 01/081325, WO 02/08198, WO 02/08256, WO 02/08187, WO 02/08244, WO 02/48172, WO 02/08251 and the patent application of E.U.A. Serial No. 10 / 052,386, filed January 18, 2002, discloses various types of peptides and / or other compounds as inhibitors of NS3 / NS4a serine protease of hepatitis C virus. Moreover, pending applications of E.U.A. Serial No. 60 / 506,637 filed on September 26, 2003, 60 / 497,749 filed on August 26, 2003 and 60 / - (Case No. IN06122) filed on November 20, 2003 describe various types of protease inhibitors . Descriptions of those applications are incorporated herein by reference. There is a need for new treatments and therapies for HCV infection. There is a need for compounds useful in the treatment or prevention or alleviation of one or more symptoms of hepatitis C. There is a need for methods of treatment or prevention or alleviation of one or more symptoms of hepatitis C. There is a need for methods for modulate the activity of serine proteases, particularly the NS3 / NS4a serine protease of HCV, using the compounds provided therein. There is a need for methods to modulate the processing of the HCV polypeptide using the compounds provided herein.

BRIEF DESCRIPTION OF THE INVENTION The aforementioned WO 02/08244 widely discloses several compounds. Applicants found that a specific type of compound surprisingly exhibits good inhibitory activity of the NS3 / NS4a HCV serine protease. Thus, in its many embodiments, the present invention provides a new class of HCV protease inhibitors, pharmaceutical compositions containing one or more of the compounds, methods for preparing pharmaceutical formulations comprising one or more such compounds, and methods of treating or preventing HCV or alleviating one or more of the symptoms of hepatitis C using one or more such compounds or one or more of such formulations. Methods for modulating the interaction of a HCV polypeptide with the HCV protease are also provided. Among the compounds provided herein, compounds that inhibit the activity of the NS3 / NS4a serine protease of HCV are preferred. Thus, the present invention describes a compound, or enantiomers, stereoisomers, rotamers, tautomers, diastereomers, or racemates of said compound, or a pharmaceutically acceptable salt, solvate or ester of said compound, said compound having the general structure that is shows in Formula I: Formula I in which: R1 is H, OR8, NR9R10, or CHR9R10, where R8, R9 and R10 may be the same or different, each being independently selected from the group consisting of H, alkyl-, aryl-, heteroalkyl-, heteroaryl-, cycloalkyl-, cycloalkyl-, arylalkyl- and heteroarylalkyl; E and J can be equal or different, each being selected independently from the group consisting of R, OR, NHR, NRR7, SR, halo and S (02) R, or E and J can directly connect with each other to form a cycloalkyl of three to eight members, or a heterocyclyl radical of three to eight members; Z is N (H), N (R) or O, with the proviso that when Z is O, G is present or absent and if G is present when Z is O, then G is C (= 0); G may be present or absent, and if G is present, G is C (= 0) or S (02), and when G is absent, Z is directly connected to Y; And it is selected from the group consisting of: X = Q, S, NH X = 0, S, NH X = Q, S / NK M N - - •: '' \ - i '. ^ -, And ? - (MIR, R7, R2, R3, R4 and R5 can be identical or different, each being independently selected from the group consisting of H, alkyl-, alkenyl-, alkynyl-, cycloalkyl-, heteroalkyl-, heterocyclyl- , aryl-, heteroaryl-, (cycloalkyl) alkyl-, (heterocyclyl) alk-, aryl-alkyl-, and heteroaryl-alkyl-, wherein each of said heteroalkyl, heteroaryl and heterocyclyl has independently from one to six oxygen, nitrogen, sulfur or phosphorus atoms, wherein each of said alkyl, heteroalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl radicals may be unsubstituted or optionally substituted independently with one or more radicals selected from the integrated group by alkyl, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, heterocyclyl, halo, hydroxy, thio, alkoxy, aryloxy, alkylthio, arylthio, amino, amido, ester, carboxylic acid, carbamate, urea, ketone, aldehyde, cyano, nitro, sulf onamido, sulfoxide, sulfone, sulfonyl urea, hydrazide and hydroxamate. The compounds represented by Formula 1, by themselves or in combination with one or more additional suitable agents described herein, may be useful for the treatment of diseases such as HCV, HIV, (AIDS or Acquired Immunodeficiency Syndrome), and related disorders, as well as to modulate the activity of the hepatitis C virus protease (HCV), prevent HCV, or alleviate one or more symptoms of hepatitis C. Such modulation, treatment, prevention or relief can be performed with the compounds of the invention as well as pharmaceutical compositions or formulations comprising such compounds. Without being limited to theory, it is believed that the HCV protease may be the NS3 or NS4a protease. The compounds of the invention can inhibit such a protease. They can also modulate the processing of the hepatitis C virus (HCV) polypeptide.

DETAILED DESCRIPTION OF THE INVENTION In one embodiment, the present invention describes compounds that are represented by Formula 1 or its pharmaceutically acceptable salt, solvate or ester, wherein the various radicals are as defined above. In another embodiment, R1 is NR9R1 °, where R9 is H, R10 is H or R14, where R14 is alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl, alkyl-aryl-, alkyl-heteroaryl-, aryl-alkyl-, alkenyl, alkynyl or heteroaryl-alkyl-. In another embodiment, R14 is selected from the group consisting of: OMe OH MOH. -OMe, 1-3 ^ '1-3 In another embodiment, R2 is selected from the group consisting of the following radicals: In another embodiment, R3 is selected from the group consisting of: 'OH where C (0) N (H) tBu. In another modality, R is selected from the group consisting of: In another embodiment, the radical is selected from the group consisting of: where Y31 is selected from the group consisting of: OR, NHR and NRR7 and Y32 is selected from the group consisting of: in another embodiment, Z is NH. In another embodiment, Z is N (R). In another embodiment, Z is O; when Z is O, then G may be present or absent, and if G is present when Z is O, then G is 10 C (= 0). In another modality, G is present and G is C (= 0) or S (02). In another modality, G is absent. In another modality, Y is selected from the group consisting of: i5 X = Oß, NH X = 0, S NH X = Q, S, NH A = O, NH In another modality, the radical: it is selected from the group consisting of: In general, the symbols containing arrows in a radical represent the connecting points of the radical at and respective positions that are shown in the parent structure, for example in Formula I.

Yet another embodiment of the invention describes some of the compounds of the invention of Formula I in Table 1. Also described in Table 1 the inhibitory activity of NS3 / NS4a serine protease (Ki * expressed in nanoMolar) for some of the compounds of the invention.

TABLE 1 As used above and throughout this description, the following terms, unless otherwise indicated, will have the following meanings: "Patient" includes both humans and animals. "Mammal" means human beings and other mammalian animals. "Alkyl" means an aliphatic hydrocarbon group which may be straight or branched chain and comprises about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. "Lower alkyl" means a group having about 1 to about 6 carbon atoms in the chain which may be straight or branched. The term "substituted alkyl" means that the alkyl group may be substituted with one or more substituents which may be the same or different, each substituent is independently selected from the group consisting of halo, alkyl, aryl, cycloalkyl, cyano, hydroxy, alkoxy , alkylthio, amino, -NH (alkyl), -NH (cycloalkyl), -N (alkyl) 2, -N (alkyl) 2, carboxy and -C (O) O-alkik). Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl and t-butyl. "Alkenyl" means an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched and comprises about 2 to about 15 carbon atoms in the chain. Preferred alkenyl groups have from about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkenyl chain, "lower alkenyl" means about 2 to about 6 carbon atoms in the chain which may be straight or branched. The term "substituted alkenyl" means that the alkenyl group may be substituted with one or more substituents which may be the same or different, each substituent is independently selected from the group consisting of halo, alkyl, aryl, cycloalkyl, cyano, alkoxy and - S (alkyl). Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyl and decenyl. "Alkynyl" means an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and which may be straight or branched and comprises about 2 to about 15 carbon atoms in the chain. Preferred alkynyl groups have from about 2 to about 12 carbon atoms in. chain; and more preferably about 2 to about 4 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. "Lower alkynyl" means about 2 to about 6 carbon atoms in the chain which may be straight or branched. Non-limiting examples of suitable alkynyl groups include ethynyl, propynyl, 2-butynyl and 3-methylbutynyl. The term "substituted alkynyl" means that the alkynyl group may be substituted with one or more substituents which may be the same or different, each substituent is independently selected from the group consisting of alkyl, aryl and cycloalkyl. "Aryl" means an aromatic monocyclic or multicyclic ring system comprising about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl group may be optionally substituted with one or more "ring system substituents" which may be the same or different, and they are as defined in the present. Non-limiting examples of suitable aryl groups include phenyl and naphthyl. "Heteroaryl" means a monocyclic or multicyclic ring system comprising about 5 to about 14 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the ring atoms is a non-carbon element, for example nitrogen, oxygen or sulfur, alone or in combination. Preferred heteroaryls contain about 5 to about 6 ring atoms. The "heteroaryl" may be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above. The prefix aza, oxa or thia before the heteroaryl root name means that at least one nitrogen atom, oxygen or sulfur respectively, is present as a ring atom. A nitrogen atom of a heteroaryl can optionally be oxidized to the corresponding N-oxide. Non-limiting examples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones), isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl, 1,4- thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl, imidazo [1,2-a] pyridinyl, imidazo [2,1-b] thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzoimidazolyl, benzothienyl, quinolinyl, imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl, pyrrolopyridyl, midazopyridyl, isoquinolinyl, benzoazaindolyl, 1,4-triazinyl, benzothiazolyl and the like. The term "heteroaryl" further refers to partially saturated heteroaryl radicals, for example, tetrahydroisoquinolyl, tetrahydroquinolyl, and the like. "Aralkyl" or "arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl are as previously described. Preferred aralkyls comprise a lower alkyl group. Non-limiting examples of preferred aralkyl groups include benzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parent radical is through alkyl. "Alkylaryl" means an alkyl-aryl- group in which the alkyl and aryl are as previously described. Preferred alkynes comprise a lower alkyl group. A non-limiting example of a suitable alkylaryl group is tolyl. The link to the parent radical is through the aryl. "Cycloalkyl" means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7 ring atoms. The cycloalkyl may be optionally substituted with one or more "ring system substituents" which may be identical or different, and are as defined above. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1-decalinyl, norbornyl, adamantyl and the like, as well as partially saturated species such as, for example, indanyl, tetrahydronaphthyl and the like. "Halogen" or "halo" means fluorine, chlorine, bromine, or iodine. Fluorine, chlorine and bromine are preferred. "Ring system substituent" means a substituent attached to an aromatic or non-aromatic ring system that, for example, replaces a hydrogen available in the ring system. The substituents of the ring system can be the same or different, each being independently selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl, heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo, nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, alkylsulfoniium,. arylsulfonyl, heteroarylsulfonyl, alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio, cycloalkyl, heterocyclyl, -C (= N-CN) -NH2, -C (= NH) -NH2l -C (= NH) -NH (alkyl), Y1Y2N -, Y-, Y2N-alkylo-, Y ^ NCÍO) -, Y ^ NSOz- and -S02NY1Y2, where Y-, and Y2 may be the same or different and are independently selected from the group consisting of hydrogen, alkyl , aryl, cycloalkyl, and aralkyl. "Ring system substituent" may also mean a single radical that simultaneously replaces two available hydrogens on adjacent carbon atoms (one H on each atom) in a ring system. Examples of such radicals are methylenedioxy, ethylenedioxy, -C (CH3) 2- and the like forming radicals such as, for example: "Heterocyclyl" means a saturated non-aromatic monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and / or sulfur atoms present in the ring system. Preferred heterocyclyls contain about 5 to about 6 ring atoms. The prefix aza, oxa or tia before the heterocyclic root name means that at least one nitrogen, oxygen or sulfur atom respectively is present as a ring atom. Any -NH in a heterocyclyl ring may exist protected such as, for example, a group -N (Boc), -N (CBz), -N (Tos) and the like; such protections are further considered part of this invention. The heterocyclyl may be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein. The nitrogen or sulfur atom of the heterocyclyl may be optionally oxidized to the corresponding N-oxide, S-oxide or S, S-dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuran, tetrahydrothiophenyl, lactam, lactone, and the like. It should be noted that in the ring systems containing heteroatoms of this invention, there are no hydroxyl groups on the carbon atoms adjacent to an N, O or S, nor are there any N or S groups on the carbon adjacent to another heteroatom. In this way, for example, in the ring: there is no -OH attached directly to the carbons labeled 2 and 5. It should also be noted that the tautomeric forms such as, for example, the radicals: they are considered equivalent in certain embodiments of this invention. "Alkynylalkyl" means an alkynyl-alkyl- group in which the alkynyl and alkyl are as previously described. Preferred alkynylalkyls contain a lower alkynyl and a lower alkyl group. The bond to the parent radical is through alkyl. Non-limiting examples of suitable alkynylalkyl groups include propargylmethyl. "Heteroaralkyl" means a heteroaryl-alkyl- group in which the heteroaryl and alkyl are as previously described. Preferred heteroaralkyls contain a lower alkyl group. Non-limiting examples of aralkyl groups include pyridylmethyl, and quinolin-3-ylmethyl. The bond to the parent radical is through alkyl. "Hydroxyalkyl" means an HO-alkyl- group in which the alkyl is as previously defined. Preferred hydroxyalkyls contain lower alkyl. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl. "Acyl" means a group H-C (O) -, alkyl-C (O) - or cycloalkyl-C (O) -, in which the various groups are as previously described.

The link to the parent radical is through the carbonyl. Preferred acyls contain a lower alkyl. Non-limiting examples of suitable acyl groups include formyl, acetyl and propanoyl. "Aroyl" means an aryl-C (O) - group in which the aryl group is as previously described. The link to the parent radical is through the carbonyl. Non-limiting examples of suitable groups include benzoyl and 1-naphthoyl. "Alkoxy" means an alkyl-O- group in which the alkyl group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The link to the parent radical is through the oxygen of the ether. . "Aryloxy" means an aryl-O- group in which the aryl group is as previously described. Non-limiting examples of suitable aryloxy groups include phenoxy and naphthoxy. The link to the parent radical is through the oxygen of the ether.

"Aralkyloxy" means an aralkyl-O- group in which the aralkyl group is as previously described. Non-limiting examples of suitable aralkyloxy groups include benzyloxy and 1- or 2-naphthalenemethoxy. The link to the parent radical is through the oxygen of the ether. "Alkylthio" means an alkyl-S- group in which the alkyl group is as previously described. Non-limiting examples of suitable alkylthio groups include methylthio and ethylthio. The link to the parent radical is through sulfur. "Arylthio" means an aryl-S- group in which the aryl group is as previously described. Non-limiting examples of suitable arylthio groups include phenylthio and naphthylthio. The link to the parent radical is through sulfur. "Aralkylthio" means an aralkyl-S- group in which the aralkyl group is as previously described. A non-limiting example of a suitable aralkylthio group is benzylthio. The link to the parent radical is through sulfur. "Alkoxycarbonyl" means an alkyl-O-C (O) - group. Non-limiting examples of alkoxycarbonyl groups include methoxycarbonyl and ethoxycarbonyl. The link to the parent radical is through the carbonyl. "Aryloxycarbonyl" means an aryl-O-C (O) - group. Non-limiting examples of suitable aryloxycarbonyl groups include phenoxycarbonyl and naphthoxycarbonyl. The link to the parent radical is through the carbonyl. "Aralkoxycarbonyl" means an aralkyl-O-C (O) - group. A non-limiting example of a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The link to the parent radical is through the carbonyl. "Alkylsulfonyl" means an alkyl-S (02) - group. Preferred groups are those in which the alkyl group is lower alkyl. The link to the parent radical is through the sulfonyl. "Arylsulfonyl" means an aryl-S (02) - group. The link to the parent radical is through the sulfonyl. The term "substituted" means that one or more hydrogens on the designated atom is replaced with a selection of the indicated group, provided that the normal valence of the designated atom under the existing circumstances is not exceeded, and that the substitution results in a stable compound . Combinations of substituents and / or variables are allowed only if such combinations result in stable compounds. By "stable compound" or "stable structure" is meant a compound that is strong enough to survive isolation to a useful degree of purity from a reaction mixture, and the formulation into an effective therapeutic agent. The term "one or more" or "at least one", when indicating the amount of substituents, compounds, combination agents and the like, refers to at least one, and up to the maximum amount of substituents, compounds, combination agents and similar allowed for chemical and physical use, which are present or added, depending on the context. Such techniques and knowledge are well known to those skilled in the art. The term "optionally substituted" means the optional substitution with the groups, radicals or radicals specified. The term "isolated" or "in isolated form" for a compound refers to the physical state of said compound after being isolated from a synthesis process or natural source or their combination. The term "purified" or "in purified form" for a compound refers to the physical state of said compound after having been obtained from a purification process or methods described herein or known to the person skilled in the art, in sufficient purity to be characterized by standard analytical techniques described herein or well known to those skilled in the art. It should be further noted that it is presumed that any heteroatom with valences not satisfied in the text, schemes, examples and tables in the present, has the hydrogen atom (s) to satisfy the valences. When a functional group in a compound is called "protected", this means that the group is in modified form to avoid unwanted side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those skilled in the art as well as by reference to standard textbooks such as, for example, T. W. Greene et al, Protective Groups in Organic Synthesis (1991), Wiley, New York. When any variable (for example, aryl, heterocycle, R2, etc.) occurs more than once in any constituent or in Formula 1, its definition in each occurrence is independent of its definition in another occurrence. As used herein, the term "composition" encompasses a product that comprises the specified ingredients in the specified amounts, as well as any product that results, directly or indirectly, from the combination of the specified ingredients in the specified amounts. Also contemplated herein are prodrugs and solvates of the compounds of the invention. The term "prodrug", as used herein, denotes a compound that is a drug precursor which, when administered to a subject, undergoes a chemical conversion by metabolic or chemical processes to give a compound of Formula 1 or its salt and / or solvate. A discussion on prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association & Pergamon Press, which are incorporated into the present in its entirety as a reference. "Solvate" means a physical association of a compound of this invention with one or more solvent molecules. This physical association "involves varying degrees of ionic and covalent binding, including hydrogen bonding.In certain circumstances the solvate may be isolated, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid." Solvate " It encompasses both insoluble solvates and solution phase solvents.Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. "Hydrate" is a solvate where the solvent molecule is H20. "Effective amount" or "therapeutically effective amount" is used. to describe an amount of a compound or a composition of the present invention effective to inhibit the CDK (s) and thereby produce the desired therapeutic, relieving, inhibiting or preventive effect The compounds of Formula 1 may form salts which are also within the scope of this invention Reference is made to a compound of formula 1 to include reference to its salts, unless Indicate otherwise. The term "salt (s)", as used herein, denotes acid salts formed with inorganic and / or organic acids, as well as basic salts formed with inorganic and / or organic bases. Further, when a compound of Formula 1 contains a basic radical, such as, but not limited to, a pyridine or imidazole, and an acidic radical, such as, but not limited to a carboxylic acid, zwitterions ("internal salts") can be formed. ) and are included within the term "salt (s)" as used herein. Pharmaceutically acceptable salts (ie non-toxic, acceptable for physiological use) are preferred, although other salts are also useful. Salts of the compounds of formula 1 can be formed, for example, by reacting a compound of formula 1 with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in a aqueous medium followed by lyophilization.

Examples of addition salts with acids include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, and hydroiods., lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartrates, thiocyanates, toluenesulfonates (also known as tosylates) and the like. Moreover, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are mentioned, for example, in P. Stahl et al, Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66 (1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson went to, The Practice of Medicinal. Chemistry (1996), Academic Press, New York; and in The Orange Book (Food &Drug Administration, Washington, D.C. on its website). These descriptions are incorporated herein by reference. Examples of basic salts include ammonium salts, alkali metal salts such as sodium, lithium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines. , t-butyl amines, and salts with amino acids such as arginine, lysine and the like. Basic groups containing nitrogen can be quatemized with agents such as lower alkyl halides (for example methyl, ethyl and butyl chlorides, bromides and iodides), dialkyl sulfates (for example dimethyl, diethyl and dibutyl sulphates), long chain halides (for example decyl, lauryl and stearyl chlorides, bromides and iodides), aralkyl halides (for example benzyl and phenethyl bromides), and others. All of these acid salts and base salts are pharmaceutically acceptable salts within the scope of the invention and all salts of acids and bases are considered equivalent to the free forms of the corresponding compounds for the purposes of the invention. The pharmaceutically acceptable esters of the present compounds include the following groups: (1) carboxylic acid esters obtained by esterification of the hydroxy groups, in which the non-carbonyl radical of the carboxylic acid moiety of the straight-chain alkyl ester group or branched (for example acetyl, n-propyl, t-butyl or n-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (for example benzyl), aryloxyalkyl (for example phenoxymethyl), aryl (for example phenyl optionally substituted with, for example, halogen, C- [alpha] alkyl, or C-α-4 alkoxy or amino); (2) sulfonate esters, such as those of alkyl or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters (e.g., L-valyl or L-isoleucyl); (4) phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate esters can be further esterified, for example, by a C-20 alcohol or a reactive derivative thereof, or by a 2,3-di (C6-2) acyl glycerol. The compounds of Formula 1, and their salts, solvates, esters and prodrugs, can exist in their tautomeric form (for example, as an amide or an ether). All tautomeric forms are contemplated herein as part of the present invention. All stereoisomers (for example geometric isomers, optical isomers and the like) of the present compounds (including those of the salts, solvates and prodrugs of the compounds as well as the salts and solvates of the prodrugs), such as those that may exist due to the asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropoisomers, and diastereomeric forms, are considered within the scope of the present invention, as are the positional isomers (such as as, for example, 4-pyridyl and 3-pyridyl). The individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be mixed, for example, as racemates or with other or other selected stereoisomers. The chiral centers of the present invention may have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms "salt", "solvate" "prodrug" and the like applies equally to the salt, solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates or prodrugs of the compounds of the invention. The compounds of Formula 1, and its salts, solvates and prodrugs, can exist in their polymorphic forms. The polymorphic forms of the compounds of Formula 1, and of the salts, solvates and prodrugs of the compounds of Formula I, are included in the present invention. All polymorphic forms or forms are contemplated herein as part of the present invention. It should be understood that the utility of the compounds of Formula 1 for the therapeutic applications mentioned herein is applicable to each compound by itself or to the combination or combinations of one or more compounds of Formula 1 as illustrated, for example, in the next immediate paragraph. The same understanding also applies to the pharmaceutical composition (s) comprising such and such compounds and method (s) of treatment involving such or such compounds. The compounds according to the invention may have pharmacological properties; in particular, the compounds of Formula 1 can be HCV protease inhibitors, each compound alone or one or more compounds of Formula 1 can be combined with one or more compounds selected from within Formula 1. The compound (s) may be useful to treat diseases such as, for example, HCV, HIV, (AIDS, Acquired Immune Deficiency Syndrome), and related disorders, as well as to modulate the activity of hepatitis C virus protease (HCV), prevent HCV, or alleviating one or more symptoms of hepatitis C. The compounds of Formula 1 can be used for the manufacture of a medicament for treating disorders associated with HCV protease, for example the method comprises intimately contacting a compound of Formula 1 with a acceptable vehicle from the pharmaceutical point of view. In another embodiment, this invention provides pharmaceutical compositions comprising the compound or compounds of the invention as an active ingredient. The pharmaceutical compositions generally comprise, in addition, at least one vehicle, excipient or pharmaceutically acceptable vehicle diluent (collectively referred to herein as carrier materials). Due to its HCV inhibitory activity, such pharmaceutical compositions possess utility in the treatment of hepatitis C and related disorders. In yet another embodiment, the present invention describes methods for preparing pharmaceutical compositions comprising the compounds of the invention as an active ingredient. In the pharmaceutical compositions and methods of the present invention, the active ingredients will normally be administered in admixture with appropriate carrier materials selected in an appropriate manner with respect to the intended administration form, ie oral tablets, capsules (either filled with solids). , filled with semisolids or filled with liquids), powders for constitution, oral gels, elixirs, dispersible granules, syrups, suspensions and the like, and consistent with conventional pharmaceutical practices. For example, for oral administration in the form of tablets or capsules, the active pharmacological component can be combined with any pharmaceutically acceptable inert non-toxic carrier, such as lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, ethyl alcohol (liquid forms) and the like. Moreover, when desired or needed, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. The powders and tablets may be formed from about 5 to about 95 percent of the composition of the invention. Suitable binders include starch, gelatin, natural sugars, corn-based sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol and waxes. Among the lubricants there may be mentioned for use in these dosage forms, boric acid, sodium benzoate, sodium acetate, sodium chloride, and the like. The disintegrants include starch, methylcellulose, guar gum and the like; Flavoring and sweetening agents and preservatives can also be included where appropriate. Some of the terms mentioned above, i.e., disintegrants, diluents, lubricants, binders and the like, are described below in greater detail. Additionally, the compositions of the present invention may be formulated in sustained release form to provide the rate of controlled release of any one or more of the components or active ingredients to optimize the therapeutic effects, ie, HCV inhibitory activity and the like. . Suitable dosage forms for prolonged release include layered tablets containing layers of different rates of disintegration or controlled release polymer matrices impregnated with the active ingredients and in the form of a tablet or capsules containing said impregnated or encapsulated porous polymer matrices. Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injections or the addition of sweeteners or opacifiers for oral solutions, suspensions and emulsions. Liquid form preparations may also include solutions for intranasal administration. Aerosol aerosol preparations may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier such as inert compressed gas, for example nitrogen. To prepare suppositories, a low melting point wax such as a mixture of fatty acid glycerides such as cocoa butter is first melted, and the active ingredient is dispersed homogeneously with stirring or similar mixing. The molten homogeneous mixture is then poured into molds of suitable size, allowed to cool and consequently solidified.

Also included are preparations in solid form which are intended to be converted, immediately before use, into liquid form preparations for either oral or parenteral administration. Said liquid forms include solutions, suspensions and emulsions. The compounds of the invention can also be administered transdermally. The transdermal compositions can take the form of creams, lotions, aerosols and / or emulsions and can be included in a transdermal patch of the matrix or reservoir type which are conventional in the art for this purpose. The compounds of the invention can also be administered orally, intravenously, intranasally or subcutaneously. The compounds of the invention may further comprise preparations that are in a unit dosage form. In such form, the preparation is subdivided into unit doses of appropriate size containing appropriate amounts of the active ingredients, for example, an effective amount to achieve the desired purpose. The amount of the active composition of the invention in a unit dose preparation can be varied or adjusted generally from about 1.0 milligram to about 1,000 milligrams, preferably from about 1.0 to about 950 milligrams, more preferably from about 1.0 to about 500 milligrams, and usually from about 1 to about 250 milligrams, according to the particular application. The actual dose used may vary depending on the age, sex, weight of the patient and the severity of the condition treated. Said techniques are known to those with experience in the art. Generally, the oral dosage form for humans containing the active ingredients can be administered 1 or 2 times per day. The amount and frequency of administration will be regulated according to the criteria of the attending physician. A generally recommended regimen of daily dosage for oral administration may vary from about 1.0 milligram to about 1,000 milligrams per day, in one or several. dose. Some useful terms are described below: Capsule - refers to a special container or container made with methyl cellulose, polyvinyl alcohols or denatured gelatins or starch to retain or contain the compositions comprising the active ingredients. The hard coating capsules are generally made with mixtures of pig skin gelatins and relatively high gel strength bone. The capsules themselves may contain small amounts of dyes, opacifying agents, plasticizers and preservatives. "Compressed" - refers to a compressed or molded solid dosage form containing the active ingredients with appropriate diluents. The tablet can be prepared by compression of mixtures or granulations obtained by wet granulation, dry granulation or by compaction. Oral gel - refers to active ingredients dispersed or solubilized in a semi-solid hydrophilic matrix. Powder for constitution refers to mixtures of powders containing the active ingredients and suitable diluents that can be suspended in water or juice. Diluent - refers to substances that usually make up the largest portion of the composition or dosage form. Suitable diluents include sugars such as lactose, sucrose, mannitol and sorbitol; starches derived from corn, wheat, rice and potatoes; and celluloses such as microcrystalline cellulose. The amount of diluent in the composition can vary from about 10 to about 90% by weight of the total composition, preferably from about 25 to about 75%, more preferably from about 30 to about 60% by weight, even more preferably from about 12 to about 60%. Disintegrant - refers to materials added to the composition to help break up (disintegrate) and release medications. Suitable disintegrants include starches; Modified "cold water soluble" starches such as sodium carboxymethyl starch; natural or synthetic gums such as acacia, carayá, guar, tragacanth and agar; cellulose derivatives such as methylcellulose and sodium carboxymethylcellulose; microcrystalline celluloses and cross-linked microcrystalline celluloses such as croscarmellose sodium; alginates such as alginic acid and sodium alginate; clays such as bentonites; and effervescent mixtures. The proportion of disintegrant in the composition can vary from about 2 to about 15% by weight of the composition, more preferably from about 4 to about 10% by weight. Binder - refers to substances that agglutinate or "stick" the powders and make them cohesive to form granules, thus serving as the "adhesive" of the formulation. The binders add cohesive strength already available in the diluting agent or binder. Suitable binders include sugars such as sucrose; starches derived from corn, wheat, rice and potatoes; natural gums such as acacia, gelatin and tragacanth; marine algae derivatives such as alginic acid, sodium alginate and calcium and ammonium alginat; cellulose-containing materials such as methylcellulose and sodium carboxymethylcellulose and hydroxypropylmethylcellulose; polyvinyl pyrrolidone; and inorganics such as magnesium aluminum silicate. The proportion of binder in the composition can vary from about 2 to about 20% by weight of the composition, with. greater preference from about 3 to about 10% by weight, even more preferably from about 3 to about 6% by weight. Lubricant - refers to one. substance added to the dosage form to allow the tablet, granules, etc. After they have been compressed, they are released from the mold or matrix reducing friction or friction. Suitable lubricants include metal stearates such as magnesium stearate, calcium stearate or potassium stearate; stearic acid; high melting point waxes; and water soluble lubricants such as sodium chloride, sodium benzoate, sodium acetate, sodium oleate, polyethylene glycols and d'l-leucine. Lubricants are usually added in the last step before compression, since they must be present on the surfaces of the granules and between them and the parts of the tablet press. The proportion of lubricant in the composition can vary from about 0.2 to about 5% by weight of the composition, preferably from about 0.5 to about 2%, more preferably from about 0.3 to about 1.5% by weight. Sliding - material that prevents agglomeration and improves the flow characteristics of the granulations, so that the flow is smooth and uniform. Suitable glidants include silicon dioxide and talc. The proportion of glidant in the composition can vary from about 0.1% to about 5% by weight of the total composition, preferably from about 0.5 to about 2% by weight. Coloring agents - excipients that provide coloration to the composition or dosage form. Such excipients may include food grade dyes or food grade dyes adsorbed onto a suitable adsorbent such as clay or aluminum oxide. The proportion of coloring agent may vary from about 0.1 to about 5% by weight of the composition, preferably from about 0.1 to about 1%. Bioavailability - refers to the rate and extent to which the active pharmacological ingredient or therapeutic radical is absorbed into the systemic circulation from a dosage form administered, as compared to a standard or control. Conventional methods for preparing tablets are known. Such methods include dry methods such as direct compression and granulation compression produced by compaction, or wet methods or other special procedures. Conventional methods for performing other forms of administration such as, for example, capsules, suppositories and the like are likewise known. Another embodiment of the invention describes the use of the compounds of the invention or pharmaceutical compositions described above for the treatment of diseases such as, for example, hepatitis C and the like. The method comprises administering a therapeutically effective amount of compound or pharmaceutical composition of the invention to a patient having said disease or diseases and in need of such treatment. Even in another embodiment, the compounds of the invention can be used for the treatment of HCV in humans in monotherapy modality or in a combination therapy modality (eg combination). dual, triple combination, etc.) such as, for example, in combination with antiviral agents and / or immunomodulators. Some examples of such antiviral and / or immunomodulatory agents include Ribavirin (from Schering-Plow Corporation, Madison, New Jersey) and Levovirin ™. (from ICN Pharmaceuticals, Costa Mesa, California), VP 50406 ™ (from Viropharma, Incorporated, Exton, Pennsylvania), ISIS 14803 ™ (from ISIS Pharmaceuticals, Carlsbad, California), Heptazyme ™ (from Ribozyme Pharmaceuticals, Boulder, Colorado), VX 497 ™ (from Vértex Pharmaceutlcals, Cambridge, Massachusetts), Thymosin ™ (from SciClone P.harmaceuticals, San Mateo, California), Maxamine ™ (Maxim Pharmaceuticals, San Diego, California), mycophenolate mofetil (from Hoffman-LaRoche, Nutley, New Jersey), interferon (such as, for example, interferon-alpha, PEG-interferon alpha conjugates) and the like. The "PEG-interferon alpha conjugates" are interferon alpha molecules covalently linked to a PEG molecule. Illustrative PEG-interferon alpha conjugates include interferon alfa-2a (Roferon ™, from Hoffman La-Roche, Nutley, New Jersey) in the form of pegylated interferon alfa-2a (eg, marketed under the Pegasys ™ brand), interferon alpha-2b (Intron ™, Schering-Plow Corporation) in the form of pegylated interferon alpha-2b (eg, marketed under the trade name PEG-Intron ™), interferon alpha-2c (Berofor Alfa ™, from Boehringer Ingelheim, Ingelheim, Germany) or consensus interferon as defined by determination of a consensus sequence of natural alpha interferons (Infergen ™, from Amgen, Thousand Oaks, California).

As indicated above, the invention also includes tautomers, rotamers, enantiomers and other stereoisomers of the compounds of the invention. Thus, as will be appreciated by one of ordinary skill in the art, some of the compounds of the invention may exist in appropriate isomeric forms. Said variations are considered within the scope of the invention. Another embodiment of the invention describes a method of preparing the compounds described herein. The compounds can be prepared by various techniques known in the art. The illustrative procedures are defined in the following reaction schemes. The illustrations should not be considered as limiting the scope of the invention defined in the appended claims. Mechanistic pathways and alternative analogous structures will be obvious to those skilled in the art. It should be understood that although the following illustrative schemes describe the preparation of some representative compounds of the invention, appropriate substitution of either the natural or non-natural amino acids will result in the formation of the desired compounds based on said substitution. Such variations are considered within the scope of the invention. For the procedures described below, the following abbreviations are used: THF: Tetrahydrofuran DMF: N, N-Dimethylformamide EtOAc: Ethyl acetate AcOH: Acetic acid HOOBt: 3-Hydroxy-1, 2,3-benzotriazin-4 (3H) -one EDCl: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride NMM: N-Methylmorpholine ADDP: 1,1 '- (Azodicarbobil) dipiperidine DEAD: Diethylazodicarboxylate MeOH: Methanol EtOH: Ethanol Et20: Diethyl ether DMSO: Dimethyl sulfoxide HOBt : N-Hydroxybenzotriazole PyBrOP: Bromo-tris-pyrrolidinephosphonium hexafluorophosphate DCM: Dichloromethane DCC: 1,3-Dicyclohexylcarbodiimide TEMPO: 2,2,6,6-Tetramethyl-1-piperidinyloxy Phg: Phenylglycine Chg: Cyclohexylglycine Bn: Benzyl Bzl: Benzyl Et : Ethyl Ph: Phenyl, Boc: isobutoxycarbonyl, Pr: isopropyl, Bu or Bu1: tert-Butyl, Boc: tert-Butyloxycarbonyl, Cbz: Benzyloxycarbonyl, Cp: Cyclopentyldienyl, Ts: p-toluenesulfonyl, Me: Methyl, HATU: 0- (7-azabenzotriazole, hexafluorophosphate -1-il) -1, 1, 3,3- tetrameth luronium DMAP: 4-N, N-Dimethylaminopyridine BOP: Benzotriazol-1-yl-oxy-tris (dimethylamino) hexafluorophosphate PCC: Pyridinium chlorochromate KHMDS: Potassium hexamethyldisilazide or potassium bis (trimethylallylamide) NaHMDS: Hexamethyldisilazide Sodium or sodium bis (trimethylsilylamide) LiHMDS: Lithium hexamethyldisilazide or lithium bis (trimethylsilylamide) 10% Pd / C: 10% Palladium on carbon (by weight).

EXAMPLE OF PREPARATION 1 Step A A solution of pyrazinecarboxylic acid 1a (3 g) in 150 ml of anhydrous dichloromethane and 150 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 6.03 g). L-cyclohexylglycine 1b hydrochloride (1.2 eq, 6.03 g) was added in small portions. Then, it was added. N-methylmorpholine (4 eq, 10 ml, d 0.920) dropwise. The reaction mixture was gradually warmed to room temperature and stirred for 20 h. All volatiles were removed in vacuo and the residue was dissolved in 500 ml of ethyl acetate. The organic phase was washed with water (100 ml), aqueous 1 N HCl (100 ml), saturated aqueous sodium bicarbonate solution (100 ml), and brine (100 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 5:95 to 3: 7) to obtain the product as a white solid.

Step B A solution of methyl ester 1c (6.5 g) in 270 ml of a 1: 1: 1 mixture of THF / MeOH / water was cooled to 0 ° C and treated with lithium hydroxide monohydrate (2.5 eq, 2.45 g). The mixture was stirred and monitored by TLC (acetone / hexanes: 2: 8). Once all of the starting material had been consumed, the reaction mixture was treated with 100 ml of 1N aqueous HCl and the mixture was concentrated on the rotary evaporator. Dichloromethane (250 ml) was added and the phases were separated. The aqueous phase was extracted with dichloromethane (3 x 80 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to obtain the product as a white solid.

Step C HX C-H3 (C02CH3 A H.HCI you Aminoester 1e was prepared following the method of R. Zhang and JS Madalengoitia (J. Org. Chem. 1999, 64, 330), with the exception that the Boc group was cleaved by reaction of the Boc-protected amino acid with methanolic HCl ( 4M HCl in dioxane was also used for deprotection). (Note: In a variation of the aforementioned synthesis, the sulfonium ylide was replaced with the corresponding phosphonium ylide).

A solution of Boc-ter-Leu 1f (Fluka, 5.0 g, 21.6 mmol) in anhydrous CH2Cl2 / DMF (50 mL, 1: 1) was cooled to 0 ° C and treated with amine hydrochloride 1f (5.3 g, 25.7 mmol), NMM (6.5 g, 64.8 mmol) and BOP reagent (11.6 g, 25.7 mmol). The reaction was stirred at room temperature for 24 h, diluted with aqueous HCl (1 M) and extracted with CH2Cl2. The combined organic phases were washed with 1 M aqueous HCl, saturated NaHCO 3, brine, dried (MgSO 4), filtered and concentrated in vacuo and purified by chromatography (SiO 2, Acetone / Hexane 1: 5) to obtain 1 g as a colorless solid.

Step E A solution of methyl ester 1g (4.0 g, 10.46 mmol) was dissolved in 4M HCl in dioxane and stirred at room temperature for 3 h. The reaction mixture was concentrated in vacuo to obtain the 1h amine hydrochloride salt, which was used without purification.

Step F A solution of 1d acid (100 mg) in 5 ml of anhydrous dichloromethane and 5 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 202 mg). Amine hydrochloride 1h (1.2 eq, 146 mg) was added. Then, N-methylmorpholine (4 eq, 0.17 ml, d 0.920) was also added. The reaction mixture was stirred at 0CC overnight. All volatiles were removed in vacuo and the residue was dissolved in 80 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 1: 9 to 4: 6) to obtain product 1i as a white solid.

Step G A solution of methyl ester 1i (180 mg) in 9 ml of a 1: 1: 1 mixture of THF / MeOH / water was cooled to 0 ° C and treated with lithium hydroxide monohydrate (2.5 eq, 35 mg). The mixture was stirred and monitored by TLC (acetone / hexanes; 3: 7). Once all of the starting material had been consumed, the reaction mixture was treated with 50 ml of 1 N aqueous HCl and the mixture was concentrated on the rotary evaporator. Dichloromethane (80 ml) was added and the phases were separated. The aqueous phase was extracted with dichloromethane (3 x 50 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to obtain product 1j as a white solid.

Step H A solution of 1k acid (2 g) in 100 ml of anhydrous dichloromethane and 5 ml of DMF was treated with N, 0-dimethylhydroxylamine hydrochloride (1.1 eq, 986 mg), BOP reagent (1.1 eq, 4.47 g), and N -methylmorpholine (3.3 eq, 3.3 ml, d 0.920) in that order. The mixture was heated to 50 ° C overnight. The reaction mixture was concentrated to half its volume and diluted with 400 ml of ethyl acetate. The organic phase was washed with water (80 ml), 1 M aqueous HCl (80 ml), saturated aqueous sodium bicarbonate solution (80 ml), and brine (80 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated. at reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 5:95 to 3: 7) to obtain product 11 as a translucent oil.

He passed A solution of amide 11 (2.2 g) in 100 ml of anhydrous THF was cooled to ° C. Lithium aluminum double hydride solution (1.3 eq) was added dropwise. The cooling bath was removed after 5 min and the mixture was allowed to reach room temperature. Analysis by TLC (ethyl acetate / hexanes: 2: 8) showed that all the initial material had been consumed. The excess LAH was cautiously tempered with the addition of saturated aqueous sodium hydrogen sulfate drops. The mixture was diluted with 200 ml of ether and saturated aqueous sodium hydrogen sulfate was added in small portions until a white solid precipitated. The mixture was filtered through celite and the filtrate was washed with 50 ml of brine. The organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: ethyl acetate / hexanes: 5:95 to 4: 6) to obtain the aldehyde product 1m as a colorless oil.

Step J A solution of the aldehyde 1m (1.8 g) in 100 ml of anhydrous dichloromethane was treated with isonitrile (1.1 eq, 680 mg) and acetic acid (2 eq, 1.02 ml, d 1.0149). The mixture was stirred overnight. All volatiles were removed in vacuo and the residue was chromatographed on silica gel (gradient: ethyl acetate / hexanes; 2: 8 to 6: 4) to obtain product 1n as a white solid.

Step K A solution of acetate 1o (1.6 g) in 60 ml of a 1: 1: 1 mixture of THF / MeOH / water was treated with lithium hydroxide monohydrate and stirred for about 1 h until all of it had been consumed. initial material as determined by TLC analysis (ethyl acetate / hexanes: 1: 1). The volatiles were removed on a rotary evaporator and the residue was diluted with dichloromethane (150 ml). The phases were separated and the aqueous phase was diluted with 30 ml of saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (3 x 80 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated to obtain the 1p product as a white solid.

Step L The protected N-Boc 1p amine (1.5 g) was dissolved in 20 ml of 4M HCl in dioxane. The reaction mixture was stirred for about 1 h until all the starting material had been consumed. All volatiles were removed in vacuo to obtain the 1q product as a white solid.

Step M A solution of the acid 1j (50 mg) in 2 ml of anhydrous dichloromethane and 2 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 52 mg). 1q amine hydrochloride (1.2 eq, 26 mg) was added. Then, N-methylmorpholine (4 eq, 0.042 ml, d 0.920) was also added. The reaction mixture was stirred at 0 ° C overnight. All volatiles were removed in vacuo and the residue was dissolved in 80 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The product 1r was used without further purification.

Step N A solution of the alcohol 1r (65 mg) in 5 ml of anhydrous dichloromethane was treated with Dess-Martin periodinan (3 eq., 121 mg). The reaction mixture was stirred at room temperature for 45 min. The mixture was treated with 1M aqueous sodium thiosulfate solution (10 ml) and saturated aqueous sodium bicarbonate solution (10 ml) and stirred for 15 min. The mixture was extracted with dichloromethane (3 x 20 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes; 2: 8 to 5: 5) to obtain product 1 as a white solid. HRMS (FAB) calculated for C36H54N7? 6 [M + H] 680.4136; found 680.4131.

EXAMPLE PE PREPARATION 2 Step A u To a solution of 4-pentin-1-ol, 2a (4.15 g, Aldrich) was added Dess-Martin periodinana (30.25g, Aldrich) and the resulting mixture was stirred for 45 minutes before the addition of (tert-butoxycarbonylmethylene) triphenylphosphorane (26.75 g, Aldrich). The resulting dark reaction was stirred overnight, diluted with ethyl acetate), washed with aqueous sodium sulfite, saturated aqueous sodium bicarbonate, water, brine and dried. Volatiles were removed under reduced pressure and the residue was purified by silica gel column chromatography using 1% ethyl acetate in hexanes as eluent to give the desired compound 2b (3.92g). Some impure fractions were also obtained but they were left aside at this time.

Step B Using the alkene 2b (1.9 g) in r > -propanol (20ml, Aldrich)), benzyl carbamate (4.95g, Aldrich) in n-propanol (40ml), NaOH (1.29g) in water (79ml), tere-butyl hypochlorite (3.7ml), (DHQ) 2PHAL (0.423g; Aldrich)) in n-propanol (37.5ml), and potassium osmiate: dihydrate (0.1544g; Aldrich) and the procedure set forth in Angew. Chem. Int. Ed. Engl (1998), 35, (23/24), pp. 2813-7, gave a crude product which was purified by silica gel column chromatography using EtOAc: Hexanes (1: 5) to give the desired amino alcohol 2c (1.37 g, 37%) as a white solid.

Step C To ester 2c (0.700g) was added 4M HCl in dioxane (20ml, Aldrich) and the resulting mixture was allowed to stand at room temperature overnight. The volatiles were removed under reduced pressure to give the acid 2d (0.621 g) as a white solid.

Step D BOP reagent (3.65 g, Sigma) followed by triethylamine (3.45 ml) was added to a dichloromethane solution (20 ml) of 2d carboxylic acid (2.00 g) and allyl amine. (0.616 ml) at room temperature and the resulting mixture was stirred overnight. The reaction mixture was partitioned between EtOAc and 10% aqueous HCl. The organic phase was separated, washed with saturated aqueous sodium bicarbonate, water, dried (magnesium sulfate). The crude reaction product was purified by silica gel column chromatography using (EtOAc: Hexanes: 70:30) as eluent to give the desired amide 2e (1.73 g) as a yellow viscous oil. .

Step E A solution of N-Cbz amine 2e (85.8 mg) in 5 ml of a 4: 1 mixture of trifluoroacetic acid / methyl sulfide was stirred at room temperature for about 3 h. All volatiles were removed under reduced pressure. The product 2f was subjected to high vacuum for about 3 h and was used without further purification.

Step F A solution of the acid 1j (50 mg) in 2 ml of anhydrous dichloromethane and 2 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 72 mg). The amine salt 2f (1.3 eq, 72 mg) was added in dichloromethane. Then, N-methylmorpholine (4 eq, 0.042 ml, d 0.920) was also added. The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 80 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The 2g product was used without further purification.

Step G A solution of the hydroxyamide 2g (67 mg) in 5 ml of anhydrous dichloromethane was treated with Dess-Martin periodinano (3 eq, 123 mg). The reaction mixture was stirred at room temperature for 45 min.

The mixture was treated with 1 M aqueous solution of sodium thiosulfate (10 ml) and saturated aqueous sodium bicarbonate (10 ml) and stirred for 15 min. The mixture was extracted with dichloromethane (3 x 20 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 4: 6) to obtain product 2 as a white solid. HRMS (FAB) calculated for C 34 H 52 N 7 6 6 [M + H] 690.3979; found 690.3995.

EXAMPLE OF PREPARATION 3 Step A The protected N-Boc amine 3a (3 g) was dissolved in 60 ml of 4M solution of HCl in dioxanes. The mixture was stirred at room temperature until all the starting material had been consumed according to the determination made by TLC (ethyl acetate / hexanes: 6: 4). After 2 h, all volatiles were removed under reduced pressure to obtain product 3b (2.4 g, 98%) as a white solid which was used without further purification.

Step B A solution of the acid 1j (150 mg) in 3 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 155 mg). Amine hydrochloride 3b (88 mg) was added followed by N-. methylmorpholine (4 eq, 0.13 ml, d 0.920). The mixture of. The reaction was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (.10 ml), and brine (8 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 3c (210 mg) was used without further purification.

Step C A solution of the hydroxyamide 3c (214 mg) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (3 eq, 371 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 45:55) to obtain product 3 as a semi-solid which was dissolved in 2 ml of dichloromethane and 10 ml of hexane, the solvent was removed at reduced pressure to give product 3 as a white solid (150 mg, 70% for two steps). HRMS (FAB) calculated for C36H5iF3N706 [M + H] 734.3853; found 734.3850.

EXAMPLE PE PREPARATION 4 Step A A solution of the acid 1j (150 mg) in 3 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 155 mg). Amine hydrochloride 4a (71 mg) was added followed by N-methylmorpholine (4 eq, 0.13 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of. ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate (10 ml), and brine (8 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 4b (190 mg) was used without further purification.

Step B A solution of hydroxyamide 4b (199 mg) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (3 eq, 371 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 4: 6) to obtain product 4 as a semi-solid which was dissolved in 2 ml of dichloromethane and 10 ml of hexane, the solvent was removed at reduced pressure to give product 4 (150 mg, 76% for two steps) as a white solid. HRMS (FAB) calculated for C36H54N706 [M + H] 680.4136; .. found 680.4165.

EXAMPLE PE PREPARATION 5 Step A A solution of the acid 1j (150 mg) in 3 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 155 mg). Amine hydrochloride 5a (76 mg) was added followed by N-methylmorpholine (4 eq, 0.13 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate (10 ml), and brine (8 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 5b (200 mg) was used without further purification.

Step B A solution of the hydroxyamide 5b (202 mg) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinan (3 eq, 371 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 45:55) to obtain product 5 as semi-solid which was dissolved in 2 ml of dichloromethane and 10 ml of hexane, the solvent was removed at reduced pressure to give product 5 (170 mg, 84% for two steps) as a white solid. HRMS (FAB) calculated for C 37 H 56 N 706 [M + H] 694.4292; found 694.4294.

EXAMPLE OF PREPARATION 6 Step A A solution of the acid 1j (150 mg) in 3 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 155 mg). Amine hydrochloride 6a (80 mg) was added followed by N-methylmorpholine (4 eq, 0.13 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate (10 ml), and brine (8 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 6b (205 mg) was used without further purification.

Step B A solution of the hydroxyamide 6b (206 mg) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (3 eq, 371 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate (10 ml) was added as well and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 45:55) to obtain product 6 as a semi-solid which was dissolved in 2 ml of dichloromethane and 10 ml of hexane, the solvent was removed at reduced pressure to give product 6 (169 mg, 82% for two steps) as a white solid. HRMS (FAB) calculated for C38H56N706 [M + H] 706.4292; Found 706.4280.

EXAMPLE PE PREPARATION 7 Step A A solution of acid 1j (80 mg) in 3 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 83 mg). Amine hydrochloride 7a (1.1 eq, 40 mg) was added followed by N-methylmorpholine (4 eq, 0.07 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (10 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (8 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 7b (105 mg) was used without further purification.

Step B A solution of the hydroxyamide 7b (108 mg) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinane (3 eq, 198 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 45:55) to obtain product 7 as semi-solid which was dissolved in 2 ml of dichloromethane and 10 ml of hexane. The solvent was removed under reduced pressure to give the product 7 (86 mg, 80% for two passages) as a white solid. HRMS (FAB) calculated for C 37 H 54 N 706 [M + H] 692.4136; found 692.4145.

EXAMPLE PE PREPARATION 8 8 A solution of the picolinic acid 8a (1.0 g) in 50 ml of anhydrous DMF and 50 ml of anhydrous dichloromethane was stirred at 0 ° C and treated with HATU (1.4 eq, 4.3 g). Methyl ether cyclohexylglycine hydrochloride, (1.1 eq, 1.85 g) followed by N-methylmorpholine (4 eq, 3.6 ml, d 0.920) was added. The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 500 ml of ethyl acetate. The organic phase was washed with water (100 ml), aqueous 1 N HCl (100 ml), saturated aqueous sodium bicarbonate solution (100 ml), and brine (100 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 5:95 to 35:65) to obtain the product 8c (1.9 g, 85%) as a translucent semi-solid.

Step B A solution of the methyl ester 8c (1.9 g) in THF / MeOH / H20 (100: 100: 50) was treated with lithium hydroxide monohydrate (2.5 eq, 2.82 g) at 0 ° C. The reaction mixture was stirred until all the starting material had been consumed as determined by TLC analysis (acetone / hexanes: 15:85). The reaction mixture was treated with 100 ml of aqueous 1N HCl (pH of the mixture was about 1) and all volatiles were removed under reduced pressure. The residue was extracted with dichloromethane (3 x 100 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The crude product 8d (1.6 g, 90%) was used without further purification.

Step C A solution of the acid 8d (235 mg) in 10 ml of anhydrous DMF and 10 ml of dichloromethane was stirred at 0 ° C and treated with HATU (1.4 eq, 480 mg).

The 1h amine salt (1.1 eq, 300 mg) was added followed by N-methylmorpholine (4 eq, 0.4 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue dissolved in 100 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (20 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 5:95 to 4: 6) to obtain product 8e (440 mg, 93%).

Step D A solution of the methyl ester 8e (440 mg) in 30 ml of a mixture of THF / MeOH / H20 (1: 1: 1) was treated with lithium hydroxide monohydrate (2.5 eq, 88 mg) at 0 ° C. The reaction mixture was stirred until all the starting material had been consumed according to the determination made by TLC (acetone / hexanes: 3: 7). The reaction mixture was treated with 20 ml of aqueous HCl N (pH of the mixture was about 1) and all volatiles were removed under reduced pressure. The residue was extracted with dichloromethane (3 x 60 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The crude product 8f (419 mg, 98%) was used without further purification.

Step E A solution of the acid 8f (80 mg) in 2 ml of anhydrous dichloromethane and 1 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 83 mg). The 1 g amine salt (1.1 eq, 38 mg) was added followed by N-methylmorpholine (4 eq, 0.07 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 8g (105 mg) was used without further purification.

Step F A solution of the hydroxyamide 8g (0.156 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinnan (2.3 eq, 152 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with aqueous solution 1 of sodium thiosulfate (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried. on magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 1: 9 to 45:55) to obtain product 8 as a solid which was dissolved in 0.5 ml of dichloromethane and 5 ml of hexane, the solvent at reduced pressure to give the product 8 (59 mg, 56% for two steps) as a white solid. HRMS (FAB) calculated for C 37 H 55 N 6 6 6 [M + H] 679.4183; found 679.4191.

EXAMPLE PE PREPARATION 9 Step A A solution of the acid 8f (80 mg) in 2 ml of anhydrous dichloromethane and 1 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 83 mg). Amine salt 4a (1.1 eq, 38 mg) was added followed by N-methylmorpholine (4 eq, 0.07 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), 1 N aqueous HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 9a (105 mg) was used without further purification.

Step B A solution of hydroxyamide 9a (0.156 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.3 eq, 152 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 1: 9 to 4: 6) to obtain product 9 as a solid which was dissolved in 0.5 ml of dichloromethane and 5 ml of hexane, the solvent at reduced pressure to give product 9 (68 mg, 64% for two steps) as a white solid. HRMS (FAB) calculated for C 37 H 55 N 6 6 6 [M + H]: 679.4183; found 679.4181.

EXAMPLE PE PREPARATION 10 Step A A solution of the acid 8f (80 mg) in 2 ml of anhydrous dichloromethane and 1 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 83 mg). Amine salt 5a (1.1 eq, 41 mg) was added followed by N-methylmorpholine (4 eq, 0.07 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 10a (105 mg) was used without further purification.

Step B A solution of hydroxyamide 10a (0.156 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.3 eq, 152 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and. they concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 1: 9 to 4: 6) to obtain the product 10 as a solid which was dissolved in 0.5 ml of dichloromethane and 5 ml of hexane, the solvent at reduced pressure to give the product 10 (68 mg, 63% for two steps) as a white solid. HRMS (FAB) calculated for C38H57N606 [M + H] 693.4340; found 693.4310.

EXAMPLE PE PREPARATION 11 eleven Step A A solution of N-Boc-cHex-Glycine 11a (916 mg) in 20 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 1.89 g). The 1h amine salt (1.1 eq, 1.2 g) was added in 30 ml of anhydrous dichloromethane followed by N-methylmorpholine (4 eq, 1.55 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 250 ml of ethyl acetate. The organic phase was washed with water (100 ml), aqueous 1 N HCl (50 ml), saturated aqueous sodium bicarbonate solution (50 ml), and brine (50 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 5:95 to 25:75) to obtain product 11b (1.65 g, 89%) as a white solid.

Step B A solution of methyl ester 11b (1.64 mg) in 60 ml of a mixture of THF / MeOH / H20 (1: 1: 1) was treated with lithium hydroxide, monohydrate (2.5 eq, 330 mg) at 0 ° C. The cold bath was removed and the reaction mixture was stirred until all the starting material had been consumed according to the determination made by TLC (acetone / hexanes: 3: 7). The reaction mixture was treated with 50 ml of 1 N aqueous HCl (pH of the mixture was about 1) and all volatiles were removed under reduced pressure. The residue was extracted with dichloromethane (3 x 100 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The product 11c to obtain a white solid (1.61 g, 98%) and was used without further purification.

Step C A solution of acid 11c (248 mg) in 10 ml of anhydrous dichloromethane and 5 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 120 mg). The 1 g amine salt (1.1 eq, 120 mg) was added followed by N-methylmorpholine (4 eq, 0.22 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 150 ml of ethyl acetate. The organic phase was washed with water (40 ml), aqueous 1 N HCl (20 ml), saturated aqueous sodium bicarbonate solution (20 ml), and brine (20 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 11d (330 mg) was used without further purification.

Step D A solution of hydroxyamide 11d (0.489 mmol) in 20 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.3 eq, 152 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (10 ml) was added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 1: 9 to 4: 6) to obtain product 11e as a solid which was dissolved in 1 ml of dichloromethane and 8 ml of hexane, the solvent at reduced pressure to give the product 11e ("280 mg, 85% for two passages) as a white solid HRMS (FAB) calculated for C36H6oN507 [M + H]: 674.4492; found 674.4507.

EXAMPLE OF PREPARATION 12 12 Step A The protected N-Boc amine 11 (80 mg) was dissolved in 5 ml of formic acid. The resulting solution was stirred at room temperature until it had been consumed. all the initial material according to the determination made by TLC (acetone / hexanes; 3: 7). After 4 h, the volatiles were removed under reduced pressure and the residue was placed under high vacuum. No further purification was performed for the product 12a (70 mg, 98%).

Step B The amine salt 12a (0.118 mmol) was dissolved in 5 ml of anhydrous dichloromethane and cooled to 0 ° C. N-methylmorpholine (2.5 eq, 0.032 ml, d 0.920) was added followed by pivalic anhydride (1.2 eq, 0.028 ml, d 0. 910) in 2 ml of anhydrous dichloromethane. The mixture was stirred overnight (temp 0 to 25 ° C). The reaction mixture was diluted with 50 ml of dichloromethane.

The solution was washed with 10 ml of 1 M aqueous HCl, 10 ml of saturated aqueous sodium bicarbonate solution, and 10 ml of brine. The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was analyzed by chromatography on silica gel (gradient: acetone / hexanes: 1: 9 to 1: 1) to obtain product 12 (28 mg, 36%) as a white solid. HRMS (FAB) calculated for C36H6oN506 [M + H] 658.4543; experimental 658.4558.

EXAMPLE OF PREPARATION 13 13 Step A A solution of 1d acid (90 mg) in 5 ml of anhydrous dichloromethane and 3 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 180 mg). Amine hydrochloride 13a (1.0 eq, 128 mg) was added followed by N-methylmorpholine (4 eq, 0.15 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 80 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (15 ml), saturated aqueous sodium bicarbonate solution (15 ml), and brine (15 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 1: 9 to 4: 6) to give product 13b (160 mg, 80%) as a white solid.

Step B The methyl ester 13b (160 mg) was dissolved in 15 ml of a 1: 1: 1 mixture of THF / methanol / water and treated with lithium hydroxide monohydrate (2.5 eq, 28 mg) at 0 ° C. The reaction mixture was gradually warmed to room temperature and stirred for 2 h until all the starting material had been consumed. Aqueous HCl 1 (30 ml) was added and all volatiles were removed on a rotary evaporator. The residue was extracted with dichloromethane (3 x 30 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The crude product 13c (150 mg, 98%) was used without further purification.

Step C A solution of 13c acid (75 mg) in 4 ml of anhydrous dichloromethane and 2 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 69 mg). Amine hydrochloride 7a (1.2 eq, 37 mg) was added followed by N-methylmorpholine (4 eq, 0.06 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 13d was used without further purification.

Step D A solution of hydroxyamide 13d (0.130 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.0 eq, 110 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (20 ml) was also added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 2: 8 to 5: 5) to obtain product 13 (69 mg, 70% for two passages) as a white solid. HRMS (FAB) calculated for C42H54N706 [M + H]: 752.4136; found 752.4122.

EXAMPLE OF PREPARATION 14 14 Step A A solution of 13c acid (75 mg) in 4 ml of anhydrous dichloromethane and 2 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 69 mg). Amine hydrochloride 4a (1.2 eq, 35 mg) was added followed by N-methylmorpholine (4 eq, 0.06 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 14a was used without further purification.

Step B A solution of hydroxyamide 14a (0.130 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.0 eq, 110 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for 5 min. Saturated aqueous sodium bicarbonate solution (20 ml) was also added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes, 2: 8 to 5: 5) to obtain product 14 (66 mg, 69% for two passages) as a white solid.

HRMS (FAB) calculated for C4? H54N706 [M + H] 740.4136; experimental 740.4146.

EXAMPLE OF PREPARATION 15 fifteen Step A The protected N-Boc amine 11 was dissolved in 5 ml of 4M solution of HCl in dioxanes. The resulting solution was stirred at room temperature for about 45 min. All volatiles were removed under reduced pressure to obtain product 15a (60 mg, 98%) as a white solid.

No further purification was carried out for the product.

Step B A solution of nicotinic acid 15b (12 mg) in 1 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 54 mg). Amine hydrochloride 15a (1.0 eq, 62 mg) was added in 3 ml of anhydrous dichloromethane followed by N-methylmorpholine (4 eq, 0.05 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate solution (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (gradient: acetone / hexanes: 2: 8 to 1: 1) to give product 15 (16 mg, 23%) as a white solid. HRMS (FAB) calculated for C 37 H 55 N 606 [M + H] 679.4183; Experimental 679.4193.

EXAMPLE OF PREPARATION 16 16 Step A A solution of acid 1j (80 mg) in 2 ml of anhydrous dichloromethane and 2 ml of anhydrous DMF was stirred at 0 ° C and treated with HATU (1.4 eq, 83 mg). Amine hydrochloride 16a (1.2 eq, 40 mg) was added followed by N-methylmorpholine (4 eq, 0.07 ml, d 0.920). The reaction mixture was stirred overnight (temp 0 at 25 ° C). All volatiles were removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The organic phase was washed with water (20 ml), aqueous 1 N HCl (10 ml), saturated aqueous sodium bicarbonate (10 ml), and brine (10 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product 16b was used without further purification.

Step B A solution of hydroxyamide 16b (0.155 mmol) in 10 ml of anhydrous dichloromethane was treated with Dess-Martin periodinone (2.0 eq, 131 mg). The reaction mixture was stirred at room temperature for 30 min. The mixture was treated with 1 M aqueous sodium thiosulfate solution (10 ml) and stirred for .5 min. Saturated aqueous sodium bicarbonate (20 ml) was also added and stirring was continued for an additional 10 min. The mixture was extracted with dichloromethane (3 x 30 ml). The combined organic phases were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed on silica gel (gradient: acetone / hexanes; 2: 8 to 5: 5) to obtain product 16 (55 mg, 51%) as a white solid. HRMS (FAB) calculated for C 37 H 56 N 7 6 6 [M + H] 694.4292; found 694.4310. The present invention relates to new inhibitors of the HCV protease. This utility may be manifested in its ability to inhibit NS2 / NS4a serine protease of HCV. A general procedure used for said manifestation is detailed through the in vitro assay that follows.

Assay to determine the inhibitory activity of the HCV protease Spectrophotometric assay The spectrophotometric assay for HCV serine protease was performed on the compounds of the invention following the procedure described by R, Zhang er al, Analytical Biochemistry, 270 (1999) 268-275, the disclosure of which is incorporated herein by reference . The assay, based on the proteolysis of chromogenic substrates with ester groups, is suitable for continuous monitoring of the activity of NS3 protease of HCV. The substrates were derived from the P side of the NS5A-NS5B binding sequence (Ac-DTEDVVX (Nva), where X = A or P) whose C-terminal carboxyl groups were esterified with one of four different chromophoric alcohols (3- or 4-) nitrophenol, 7-hydroxy-4-methyl-coumarin or 4-phenylazophenol). The synthesis, characterization and application of these new spectrophotometric substrates with ester groups to the high performance systematic selection and the detailed kinetic evaluation of the inhibitors of the HCV NS3 protease are illustrated below.

Materials and Methods: Materials: Chemical reagents for pH regulators related to the assay were obtained from Sigma Chemical Company (St. Louis, Missouri). Reagents for peptide synthesis were from Aldrich Chemicals, Novabiochem (San Diego, California), Applied Biosystems (Foster City, California) and Perseptive Biosystems (Framingham, Massachusetts). The peptides were synthesized manually or on an automated ABI model 431 A synthesizer (from Applied Biosystems). The UV? IS Spectrometer model LAMBDA 12 was from Perkin Elmer (Norwalk, Connecticut) and 96-well UV plates were obtained from Corning (Corning, New York). The preheat block was from USA Scientific (Ocala, Florida) and the vortex motion instrument for the 96-well plate is from Labline Instruments (Melrose Park, Illinois). A Spectramax Plus microtitre plate reader was obtained with a Molecular Devices monochromator (Sunnyvale, California).

Enzyme Preparation: NS3 / NS4A recombinant heterodimeric HCV protease (strain 1a) was prepared using previously published procedures (D. L. Sali et al, Biochemistry, 37 (1998) 3392-3401). Protein concentrations were determined by the Biorad dye method using recombinant protease of HCV standards previously quantified by amino acid analysis. Before starting the assay, the storage pH regulator (50 mM sodium phosphate pH 8.0, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, and 10 mM DTT) was exchanged for the test pH regulator (MOPS) 25 mM pH 6.5, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5 μM DTT) using a pre-packed Biorad Bio-Spin P-6 column.

Substrate synthesis and purification The synthesis of the substrates was carried out as reported by R. Zhang et al,. { ibid.) and is started by anchoring Fmoc-Nva-OH to 2-chlorotryril chloride resin using a standard protocol (K. Barios et al., Int. J. Pept. Protein.

Res., 37 (1991), 513-520). Peptides were subsequently assembled, using Fmoc chemistry, either manually or in an automatic ABI model 431 peptide synthesizer. N-acetylated and fully protected peptide fragments were dissociated from the resin by either acetic acid or % (HOAc) and 10% trifluoroethanol (TFE) in dichloromethane (DCM) during min, or by 2% trifluoroacetic acid (TFA) in DCM for 10 min. The combined filtrate and washing DCM was azeotropically evaporated (or repeatedly extracted with aqueous Na 2 CO 3 solution) to remove the acid used in the break. The DCM phase was dried over Na2SO4 and evaporated. The ester substrates were assembled using standard acid-alcohol coupling procedures (K. Holmber er al, Acta Chem. Scand., B33 (1979) 410-412). The peptide fragments were dissolved in anhydrous pyridine (30-60 mg / ml) to which 10 molar equivalents of chromophore and a catalytic amount (0.1 eq.) Of para-toluenesulfonic acid (pTSA) were added. Dicyclohexylcarbodiimide (DCC) was added, 3 eq.) To initiate coupling reactions. The product formation was monitored by HPLC and found to be complete after 12-72 hours of reaction at room temperature. The pyridine solvent was evaporated in vacuo and further removed by azeotropic evaporation with toluene. The peptide ester was deprotonated with 95% TFA in DCM for two hours and extracted three times with anhydrous ethyl ether to remove excess chromophore. The deprotected substrate was purified by reverse phase HPLC on a C3 or C8 column with a gradient of acetonitrile from 30% to 60% (using six column volumes). The overall yield after purification by HPLC was about 20-30%. The molecular mass can be confirmed by mass spectroscopy by ionization with electrospray. The substrates were stored as a dry powder with drying.

Spectra of Substrates and Products The spectra of the substrates and the corresponding chromophore products were obtained in the pH regulator of the pH 6.5 assay. The extinction coefficients were determined at the optimal wavelength of the main peak in 1 cm cuvettes (340 nm for 3-Np and HMC, 370 nm for PAP and 400 nm for 4-Np) using multiple dilutions. The optimal length of the main peak was defined as the wavelength that produced the maximum fractional difference in absorbance between the substrate and the product (OD of the product - OD of the substrate) / OD of the substrate).

Protease Assay HCV protease assays were performed at 30 ° C using a 200 μl reaction mixture in a 96-well microtiter plate. The conditions of the assay pH regulator (25 mM MOPS pH 6.5, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5 μM DTT) were optimized for the NS3 / NS4A heterodimer (DL Sali er al, ibid.)). Normally, mixtures of 150 μl pH, substrate and inhibitor buffer were placed in the wells (final concentration of DMSO <4% v / v) and left to incubate at 30 ° C for about 3 minutes. Then, a microtiter of preheated protease (12 nM, 30 ° C) was used in the assay pH buffer to initiate the reaction (final volume 200 μl). The plates were monitored for the duration of the assay (60 minutes) to determine the change in absorbance at the appropriate wavelength (340 nm for 3-Np and HMC, 370 nm for PAP, and 400 nm for 4-Np) using a Spectromax Plus microtitre plate equipped with a monochromator (acceptable results were obtained with plate readers that use cut-off filters). The proteolytic cleavage of the ester bond between the Nva and the chromophore at the appropriate wavelength against a target without enzymes was monitored as a control for nonenzymatic hydrolysis. The evaluation of the kinetic parameters of the substrate was performed on a 30-factor substrate concentration scale (-6-200 μM). The initial velocities were determined using linear regression and the kinetic constants were obtained by adapting the data to the Michaelis-Menten equation using non-linear regression analysis (Mac Curve Fit 1, 1, K. Raner). The resulting numbers (/ ccat) were calculated assuming that the enzyme is fully active.

Evaluation of Inhibitors and Inactivators The inhibition constants were determined experimentally (K) for the competitive inhibitors Ac-D- (D-Gla) -Ll- (Cha) -C-OH (27), Ac-DTEDVVA (Nva) -OH and Ac-DTEDVVP (Nva) -OH at concentrations fixed enzymes and substrate plotting a graph of vo / v¡ vs. inhibitor concentration ([l] o) according to the Michaelis-Menten rearranged equation for competitive inhibition kinetics: Vo / v¡ = 1 + [l] o / (Kj (1 + [S] o / Km) ), where Vo is the initial non-inhibited velocity, v, is the initial velocity in the presence of an inhibitor at a given inhibitor concentration ([l] 0) and [S] or is the substrate concentration used. The resulting data were adjusted using linear regression and the resulting slope, 1 / (K, (1+ [S] o / Km)> was used to calculate the value of Ki * The values thus obtained from Ki * for some of The compounds of the invention are shown in Table 2 and Table 3.

TABLE 2 Scale A < 100 nM; B > 100 nM < 1000 nM; C > 1000 nM.

TABLE 3 Although the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and other variants thereof will be apparent to those skilled in the art. All these alternatives, modifications and variants should fall within the spirit and scope of the present invention.

Claims (36)

NOVELTY OF THE INVENTION CLAIMS

1. - A compound, or enantiomers, stereoisomers, rotamers, tautomers, diastereomers or racemates of said compound, or a pharmaceutically acceptable salt, solvate or ester of said compound, said compound having the general structure shown in Formula I: Formula I wherein: R1 is H, OR8, NR9R10 or CHR9R10, where R8, R9 and R10 may be the same or different, each being independently selected from the group consisting of H, alkyl-, aryl-, heteroalkyl-, heteroaryl -, cycloalkyl-, cycloalkyl-, arylalkyl-, and heteroarylalkyl; E and J can be the same or different, each being selected independently from the group consisting of R, OR, NHR, NRR7, SR, halo and S (02) R, or E and J can directly connect with each other to form either a cycloalkyl of three to eight members, or a heterocyclyl moiety of three to eight members; Z is N (H), N (R) or O, with the proviso that when Z is O, G is present or absent and if G is present when Z is O, then G is C (= 0); G may be present or absent, and if G is present, G is C (= 0) or S (02), and when G is absent, Z is directly connected to Y; And it is selected from the group consisting of: X- < XS, NH X-Q.S. NH XX S, HH x = o, s, K? ? = O, NM R, R7, R2, R3, R4 and R5 can be the same or different, each being independently selected from the group consisting of H, alkyl-, alkenyl-, alkynyl-, cycloalkyl-, heteroalkyl-, heterocyclic- , aryl-, heteroaryl-, (cycloalkyl) alkyl-, (heterocyclic) alkyl-, aryl-alkyl- and heteroaryl-alkyl-, wherein each of said heteroalkyl, heteroaryl and heterocyclyl independently has from one to six oxygen atoms , nitrogen, sulfur, or phosphorus; wherein each of said alkyl, heteroalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl radicals may be unsubstituted or optionally substituted independently with one or more radicals selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, aralkyl , cycloalkyl, heterocyclyl, halo, hydroxy, thio, alkoxy, aryloxy, alkylthio, arylthio, amino, amido, ester, carboxylic acid, carbamate, urea, ketone, aldehyde, cyano, nitro, sulfonamido, sulfoxide, sulfone, sulfonyl urea, hydrazide and hydroxamate.

2. The compound according to claim 1, further characterized in that R1 is NR9R10 and R9 is H, R10 is H or R14, wherein R14 is alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl, alkyl-aryl-, alkyl- heteroaryl-, aryl-alkyl-, alkenyl, alkynyl or heteroaryl-alkyl.

3. The compound according to claim 2, further characterized in that R14 is selected from the group consisting of: OMe OH i- OH. l-OMe, 1-3 1-3 . Me ~ < \ 1-4 1-4

4. The compound according to claim 1, further characterized in that R2 is selected from the group consisting of the following radicals:

5. - The compound according to claim 1, further characterized in that R3 is selected from the group consisting of: wherein R31 is OH or O-alkyl; and R32 is H, C (0) CH3, C (0) OtBu or C (0) N (H) tBu.

6. The compound according to claim 5, further characterized in that R3 is selected from the group consisting of:

7. - The compound according to claim 1, further characterized in that R5: XY Í "íl 'o is selected from the group consisting of: where Y31 is selected from the group Integrated by: OR, NHR and NRR7 and Y32 is selected from the group consisting of:

8. - The compound according to claim 1, further characterized in that Z is NH.

9. The compound according to claim 1, further characterized in that Z is N (R).

10. The compound according to claim 1, further characterized in that Z is O, G is present or absent and if G is present, G is C (= 0).

11. The compound according to claim 1, further characterized in that G is present and is C (= 0) or S (02).

12. The compound according to claim 1, further characterized in that Y is selected from the group consisting of: XO., S, NH X = OrS, NH «S, HH X = OY, NH A = O, NH

13. - The compound according to claim 1, further characterized by: it is selected from the group consisting of:

14. - The compound according to claim 1, further characterized in that G is absent.

15. A pharmaceutical composition comprising as an active ingredient at least one compound as defined in claim 1.

16. The pharmaceutical composition according to claim 15, for use in the treatment of disorders associated with HCV.

17. - The pharmaceutical composition according to claim 15, further characterized in that it comprises at least one pharmaceutically acceptable carrier.

18. The pharmaceutical composition according to claim 17, further characterized in that it contains at least one antiviral agent.

19. The pharmaceutical composition according to claim 18, further characterized in that it still contains at least one interferon.

20. The pharmaceutical composition according to claim 19, further characterized in that at least one antiviral agent is ribavirin and said at least one interferon is a-interferon or pegylated interferon.

21. The use of a compound defined in claim 1, for the manufacture of a medicament for treating disorders associated with HCV.

22. The use claimed in claim 21, wherein said medicament is administrable orally or subcutaneously.

23- A method for preparing a pharmaceutical composition for treating disorders associated with HCV, said method comprising intimately contacting at least one compound as defined in claim 1 and at least one pharmaceutically acceptable carrier.

24. A compound exhibiting HCV protease inhibitory activity, or enantiomers, stereoisomers, rotamers, tautomers, diastereomers or racemates of said compound, or a pharmaceutically acceptable salt, solvate or ester of said compound, said compound being selected from the compounds of the structures listed below:

25. - A compound, or enantiomers, stereoisomers, rotamers, tautomers, diastereomers or racemates of said compound, or a pharmaceutically acceptable salt, solvate or ester of said compound, said compound being selected from the compounds of the structures listed below:

26. - A pharmaceutical composition for treating disorders associated with HCV, said composition comprising a therapeutically effective amount of one or more compounds as defined in claim 25 and a pharmaceutically acceptable carrier.

27. The pharmaceutical composition according to claim 26, further characterized in that it contains at least one antiviral agent.

28. The pharmaceutical composition according to claim 27, further characterized in that it still contains at least one interferon or conjugate PEG-interferon alpha.

29. The pharmaceutical composition according to claim 28, further characterized in that at least one antiviral agent is ribavirin and said at least one interferon is a-interferon or pegylated interferon.

30. The use of one or more compounds as defined in claim 25, for the preparation of a medicament for the treatment of a disorder associated with the hepatitis O virus.

31.- A method for modulating the activity of the protease. of hepatitis C virus (HCV), which comprises contacting the HCV protease with one or more compounds as defined in claim 25.

32.- The use of one or more compounds as defined in claim 25, for the preparation of a medication to treat, prevent or alleviate one or more symptoms of hepatitis C (HCV).

33. The use claimed in claim 32, wherein the HCV protease is the NS3 / NS4a protease.

34. The use claimed in claim 32, wherein the compound or compounds inhibit the NS3 / NS4a protease of HCV.

35.- A method for modulating the processing of the hepatitis C virus (HCV) polypeptide, which comprises contacting a composition containing the HCV polypeptide under conditions in which said polypeptide is processed with one or more compounds that are defined in claim 25.

36. The compound according to claim 1, in purified form.