HK1183030B

HK1183030B - Heterocyclic antiviral compounds

Info

Publication number: HK1183030B
Application number: HK13110430.2A
Authority: HK
Inventors: Leanna Renee Staben; Francisco Xavier Talamas; Ryan Craig Schoenfeld
Original assignee: F. Hoffmann-La Roche Ag
Priority date: 2010-07-07
Filing date: 2011-07-04
Publication date: 2016-10-28

Description

Heterocyclic antiviral compounds

The present invention provides non-nucleoside compounds of formula I and certain derivatives thereof, which inhibit HCV RNA-dependent RNA viral polymerase. These compounds are useful for treating RNA-dependent RNA viral infections. They are particularly useful as inhibitors of Hepatitis C Virus (HCV) NS5B polymerase, inhibitors of HCV replication, and for the treatment of hepatitis c infections.

Hepatitis C virus is the leading cause of chronic liver disease worldwide (Boyer, N. et al, J.hepatol.200032: 98-112). Patients infected with HCV are at risk of developing cirrhosis and subsequent hepatocellular carcinoma, and therefore HCV is the primary indication for liver transplantation.

HCV has been classified as a member of the Flaviviridae family of viruses, which includes the Flaviviridae (flaviviruses), pestiviruses (pestiviruses) and hapaceiviruses including hepatitis C virus (Rice, C.M., Flaviviridae: Thevirusand third edition in Fields virology, editors: B.N.fields, D.M.Knipe and P.M.Howley, Lippincott-Raven Press, Philadelphia, Pa., Chapter 30, 931-959, 1996). HCV is an enveloped virus containing a positive-sense single-stranded RNA genome of approximately 9.4 kb. The viral genome consists of a highly conserved 5 'untranslated region (UTR), a long open reading frame encoding a polyprotein precursor of approximately 3011 amino acids, and a short 3' UTR.

Genetic analysis of HCV has identified 6 major genotypes that differ in DNA sequence by more than 30%. More than 30 subtypes have been distinguished. In the united states, approximately 70% of infected individuals suffer from type 1a and type 1b infections. In asia, type 1b is the most prevalent subtype. (X.Forns and J.Bukh, ClinicsilverDisase 19993:693-716; J.Bukh et al, Semin.Liv.Dis.199515: 41-63). Unfortunately, type 1 infections are more resistant to treatment than type 2 or type 3 genotypes (N.N.Zein, Clin. Microbiol. Rev.,200013: 223-.

The virus structural proteins include a nucleocapsid core protein (C) and two envelope glycoproteins E1 and E2. HCV also encodes two proteases, a zinc-dependent metalloprotease encoded by the NS2-NS3 region and a serine protease encoded in the NS3 region. These proteases are required to cleave specific regions of the precursor polyprotein into the mature peptide. The carboxy portion of nonstructural protein 5(NS5B) contains an RNA-dependent RNA polymerase. The functions of the remaining nonstructural proteins NS4A and NS4B, as well as NS5A (the amino terminal portion of nonstructural protein 5) remain unknown. It is believed that most of the non-structural proteins encoded by the hcv RNA genome are involved in RNA replication.

The number of therapies currently approved for the treatment of HCV infection is limited. New and existing therapeutic approaches to treat HCV infection and inhibit HCV ns5B polymerase activity have been reviewed: r.g.gish, sim.livedis, 199919: 5; DiBesceglie, a.m. and Bacon, b.r., scientific american, 10 months 1999, 80-85; Lane-Bakaar, Current and FutureTherapyre for Chronic hepatitis CVirusLiverDisase, Current, drug Targ, InfectDis.20033(3): 247-253; hoffmann et al, Recentryttententotherapy for Patititis Cvirucinfunction (1999) 2002, exp. opin. Ther. Patents200313(11): 1707) 1723; walker et al, research Candidates for the molecular of chronologically antipathotics C, exp, Opin, investing, drugs200312(8): 1269-; L.Tan et al, hepatotiss CTherepeutics, CurrentStatus and EmergingStrategies, NatureRev. drug Discov.20021: 867-; J.Z.Wu and Z.hong, Targeting NS5 BRNA-dependentNapolymeans for anti-HCVCheheating, curr.DrugTafg. -infusion.Dis.20033 (3): 207-219.

Ribavirin (1- ((2R,3R,4S,5R) -3, 4-dihydroxy-5-hydroxymethyl-tetrahydro-furan-2-yl) -1H- [1,2,4]Triazole-3-carboxylic acid amide;) Is a synthesized non-interferon-induced broad-spectrum antiviral nucleoside analogue. Ribavirin has activity against several DNA and RNA viruses in vitro, including the flaviviridae family (gary l. davis. gastroenterology,2000,118:S104-S114). Although ribavirin reduces serum aminotransferase levels to normal in 40% of patients in monotherapy, it does not reduce serum levels of HCV-RNA. Ribavirin also exhibits significant toxicity, known to induce anemia. Viramidine (williamidine) is a ribavirin prodrug that is converted to ribavirin in hepatocytes by adenosine deaminase (J.Z.Wu, Antivir.chem.Chemothet 200617(1): 33-9).

Interferons (IFNs) have been used for the treatment of chronic hepatitis for nearly a decade. IFNs are glycoproteins produced by immune cells in response to viral infection. Two different types of interferons were identified: type 1 includes several interferon alphas and one interferon beta, type 2 includes interferon gamma. Type 1 interferons are produced primarily by infected cells, protecting neighboring cells from new infections. IFNs inhibit viral replication of a wide variety of viruses, including HCV, and when used alone to treat hepatitis c infection, IFNs inhibit serum HCV-RNA to undetectable levels. In addition, IFN normalizes serum aminotransferase levels. Unfortunately, the effects of IFN are transient. Discontinuation of therapy resulted in a recurrence rate of 70%, with only 10-15% showing a sustained virologic response and normal serum alanine transferase levels (Davis, Luke-Bakaar, supra).

One limitation of early IFN therapy was the rapid clearance of proteins from the blood. Chemical derivatization of IFNs with polyethylene glycol (PEG) has led to significant improvements in the pharmacokinetic properties of proteins.Is a conjugate of interferon α -2a and 40kD branched monomethoxy PEG, PEG-Is a conjugate of interferon α -2b and 12kD monomethoxy PEG (B.A. Luxon et al, Clin. Therap.200224(9):13631383; A.Kozlowski and J.M. Harris, J.Control. Release, 200172: 217-224).

Currently, ribavirin combined with interferon-alpha HCV therapy is the best HCV therapy. The combination of ribavirin and PEG-IFN (see below) produced a Sustained Virological Response (SVR) in 54-56% of HCV type I patients. For HCV types 2 and 3, the SVR reaches 80% (Walker, supra). Unfortunately, combination therapy also produces side effects that pose clinical challenges. Depression, influenza-like symptoms and skin reactions are associated with subcutaneous IFN- α, and hemolytic anemia is associated with the continuous treatment of ribavirin.

Several potential molecular targets for anti-HCV therapeutic drug development have now been identified,

including but not limited to NS2-NS3 self-protease (autoprotease), NS3 protease, NS3 helicase, and NS5B polymerase. RNA-dependent RNA polymerase is absolutely essential for replication of a single-stranded positive-sense RNA genome. This enzyme has attracted great interest to medicinal chemists.

The compounds of the present invention and their pharmaceutically acceptable salts are also useful for the treatment and prevention of viral infections, particularly hepatitis c infections and diseases, in living hosts when used in combination with each other and with other biologically active agents including, but not limited to, interferons, pegylated interferons, ribavirin, protease inhibitors, polymerase inhibitors, small compounds that interfere with RNA, antisense compounds, nucleotide analogs, nucleoside analogs, immunoglobulins, immunomodulators, hepatoprotectants, anti-inflammatory agents, antibiotics, antiviral agents and anti-infective compounds. Such combination therapy may also include the concurrent or sequential provision of a compound of the invention with other pharmaceutical drugs or potentiators, such as ribavirin and related compounds, amantadine and related compounds, various interferons such as interferon- α, interferon- β, interferon γ, and the like, as well as alternative forms of interferon such as pegylated interferon. Furthermore, the combination of ribavirin and interferon may also be administered as an additional combination therapy with at least one compound of the present invention.

Other interferons currently under development include alb interferon- α -2b (albuferon), IFN-omega with DUROS, LOCTERON^TMAnd interferon- α -2bXL because of these and theirIt is marketed as interferon, so it is desirable to use them in combination therapy with the compounds of the present invention.

HCV polymerase inhibitors are another target for drug development, and compounds under development include R-1626, R-7128, IDX184/IDX102, PF-868554(Pfizer), VCH-759 (Virocchem), GS-9190(Gilead), A-837093 and A-848837(Abbot), MK-3281(Merck), GSK949614 and GSK625433(Glaxo), ANA598(Anadys), VBY708 (Viroay).

HCV ns3 protease inhibitors have also been identified as potentially useful for the treatment of HCV. Protease inhibitors in clinical trials include VX-950(Telaprevir, Vertex), SCH503034(Broceprevir, Schering), TMC435350(Tibotec/Medivir), and ITMN-191 (Intermune). Other protease inhibitors in the early stages of development include MK7009(Merck), BMS-790052(Bristol Myers Squibb), VBY-376(Virobay), IDXSCA/IDXSCB (Idenix), BI12202(Boehringer), VX-500(Vertex), PHX1766 Phenomix.

Other targets under development for anti-HCV therapy include cyclophilin inhibitors that inhibit RNA binding to NS5b, nitazoxanide, ciguatvir (Migenix), alpha-glucosidase-1 inhibitors, caspase inhibitors, Toll-like receptor agonists, and immune stimulants such as zadaxin (sciclone).

There is currently no prophylactic treatment of Hepatitis C Virus (HCV) and currently approved therapies that exist against HCV only are limited. The design and development of new pharmaceutical compounds is essential. The present invention provides compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R¹、R²、R³And X are as follows.

X is CH or N.

R¹Is selected from R^1a、R^1b、R^1c、R^1dAnd R^1e：

Wherein R is⁵Is hydrogen or C_1-3Alkyl, and R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy.

R²Is (a) aryl, (b) heteroaryl or (c) NR^aR^bWherein said aryl or said heteroaryl is optionally and independently substituted with one to three substituents selected from hydroxy, C_1-6Alkoxy radical, C_1-6Alkyl radical, C_1-6Hydroxyalkyl, halogen, (CH)₂)_nNR^cR^dCyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6An alkylsulfonyl group.

R^aAnd R^bTogether with the nitrogen to which they are attached are cyclic amines independently substituted by (CH) where n is 0 to 2₂)_nNR^cR^dSubstituted and additionally optionally substituted by one or two independently selected from C_1-6Alkyl or halogen.

R^cAnd R^dIndependently of each other is hydrogen, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Acyl radical, O₂SR⁴Wherein R is⁴Is C_1-6Alkyl radical, C_1-6Haloalkyl, C_3-7Cycloalkyl radical, C_3-7cycloalkyl-C_1-3Alkyl radical, C_1-6alkoxy-C_1-6Alkyl, -SO₂-NR^eR^f。

R^eAnd R^f(i) Independently of each other is hydrogen, C_1-3Alkyl or (CH)₂)_2-6NR^gR^hOr (ii) nitrogen-to which they are attachedIs (CH)₂)₂X⁵(CH₂)₂Wherein X is⁵Is O or NR^gAnd R is^gIs hydrogen, C_1-3Alkyl radical, C_1-3Acyl or C_1-3An alkylsulfonyl group.

R³Is CF₃、CH₂CF₃、C_3-5Cycloalkyl, halogen, C_1-6Alkoxy radical, C_1-3Haloalkoxy, CHR^4aR^4bOr CR^4aR^4bR^4cWherein:

(i)R^4a、R^4band R^4cIndependently selected from C_1-3Alkyl, CD₃、C_1-2Alkoxy radical, C_1-2Fluoroalkyl, C_1-3Hydroxyalkyl, cyano or hydroxy; or

(ii) When they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs hydrogen, C_1-3Alkyl, CD₃、C_1-2Alkoxy, halogen, C_1-3Hydroxyalkyl, cyano or C_1-2Fluoroalkyl or R^4aAnd R^4bTogether with the carbon to which they are attached is 3-oxetanyl or tetrahydrofuran-2-yl.

The invention also provides pharmaceutically acceptable salts of the compounds of formula I.

The present invention also provides a method of treating a disease, Hepatitis C Virus (HCV) viral infection, by administering to a patient in need thereof a therapeutically effective amount of a compound of formula I. These compounds may be administered alone or in combination with other antiviral compounds or immunomodulators.

The present invention also provides methods of inhibiting HCV replication in a cell by administering a compound of formula I in an amount effective to inhibit HCV.

The present invention also provides a pharmaceutical composition comprising a compound of formula I and at least one pharmaceutically acceptable carrier, diluent or excipient.

An entity as used herein means one (one) or more (a plurality) of such entities; for example, a compound means one or more compounds or at least one compound. Thus, the entities, the terms "entity(s)" and "at least one entity" are used interchangeably herein.

The phrase "as defined above" refers to the broadest definition of each group as provided in the summary of the invention or the broadest claim. In all other embodiments provided below, substituents that may be present in each embodiment and that are not explicitly defined remain with the broadest definition provided in the summary of the invention.

The terms "comprising" or "including" as used in this specification are to be interpreted as having an open-ended meaning, whether in transitional phrases or in the body of the claims. That is, the term is to be construed as synonymous with the phrases "having at least" or "including at least". When used in the context of a method, the term "comprising" means that the method includes at least the recited steps, but may include additional steps. The term "comprising" when used in the context of a compound or composition means that the compound or composition includes at least the recited features or components, but may also include additional features or components.

The term "independently" is used herein to indicate that a variable applies in any one instance regardless of whether a variable having the same or different definition is present within the same compound. Thus, in a compound where R "occurs twice and is defined as" independently carbon or nitrogen, "both R" may be carbon, both R "may be nitrogen, or one R" may be carbon and the other R "may be nitrogen.

When any variable (e.g. R)¹、R^4a、Ar、X¹Or Het) when occurring more than one time in any moiety or formula depicting and describing compounds employed or claimed in this invention, its definition on each occurrence is independent of its definition at every other occurrence. Likewise, only such compounds give rise to stable compounds, substituents and/orCombinations of variables are allowed.

The symbol "at the end of a bond" or "plotted across a bond" each refers to the point of attachment of a functional group or other chemical moiety to the rest of the molecule of which it is a part. Thus, for example:

MeC(＝O)OR⁴wherein

The bond drawn into the ring system (as opposed to being attached at a distinct vertex) indicates that the bond may be attached to any suitable ring atom.

The term "optionally" is used herein to indicate that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted" means that the optionally substituted moiety may have added hydrogen or a substituent.

The term "about" is used herein to mean approximately, on the left or right, roughly, or roughly. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the limits above and below the numerical values set forth. Generally, the term "about" is used herein to change a numerical value by a difference of 20% above and below the stated value.

As used herein, the recitation of a numerical range for a variable is intended to indicate that the invention can be practiced with the variable equaling any value within the range. Thus, for variables that are non-continuous in nature, the variable may equal any integer value of a range of values, including the endpoints of the range. Similarly, for a variable that is continuous in nature, the variable may be equal to any real value of a range of values, including the endpoints of the range. For example, a variable described as having a value of 0 to 2 may be 0,1, or 2 for a variable that is non-continuous in nature, and 0.0, 0.1, 0.01, 0.001, or any other real value for a variable that is continuous in nature.

The compounds of formula I exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer often yields mixtures whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. Common prototropic tautomers include keto/enolAmide/imide acid (And amidinesTautomers. The latter two are particularly common in heteroaryl and heterocyclic rings, and the present invention encompasses all tautomeric forms of the compounds.

The compounds of formula I may contain an acidic or basic centre and suitable salts are formed from acids or bases which form non-toxic salts which have similar antiviral activity. Examples of salts of inorganic acids include hydrochloride, hydrobromide, hydroiodide, chloride, bromide, iodide, sulfate, bisulfate, nitrate, phosphate, and biphosphate. Examples of salts of organic acids include acetate, fumarate, pamoate, aspartate, benzenesulfonate, carbonate, bicarbonate, camphorsulfonate, D and L-lactate, D and L-tartrate, ethanesulfonate, methanesulfonate, malonate, orotate, glucoheptanoate, methylsulfate, stearate, glucuronate, 2-naphthalenesulfonate, toluenesulfonate, hydroxybenzoylbenzoate (hibenzate), nicotinate, isethionate, malate, maleate, citrate, gluconate, succinate, sucrose, benzoate, ethanesulfonate, and pamoate. For a review of suitable salts, see: berge et al, j.pharm.sci.,197766:1-19 and g.s.paulekuhn et al, j.med.chem.200750: 6665.

Technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention pertains, unless otherwise specified. Reference may be made herein to various methodologies and materials known to those skilled in the art. Standard reference works giving general principles of pharmacology include the pharmacologic basises of therapeutics, 10 th edition, mcgrawhillcompany inc, new york (2001), Goodman and Gilman. The starting materials and reagents used in the preparation of these compounds are generally available from commercial suppliers such as Aldrich chemical company, or prepared by methods known to those skilled in the art, according to the methods given in the literature. The materials, reagents, etc. mentioned in the following description and examples are available from commercial sources unless otherwise indicated. General synthetic methods have been described in the articles, for example: FieserandFieser' sRegigentsfororganic Synthesis, Wiley & Sons, New York, volumes 1-21; larock, comprehensive organic transformations, 2 nd edition, Wiley-VCH, New York 1999; comprehensive organic synthesis, b.trost and i.fleming (editor), volumes 1-9, Pergamon, oxford, 1991; comprehensive heterocyclic chemistry, a.r.katritzky and c.w.rees (editor), Pergamon, oxford, 1984, volumes 1-9; comprehensive heterocyclic chemistry ii, a.r.katritzky and c.w.rees (editor), Pergamon, oxford, 1996, volumes 1-11; and organic reactions, Wiley & Sons, New York, 1991, volumes 1-40, which are familiar to those skilled in the art.

In one embodiment of the invention, there is provided a compound of formula I, wherein R is¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein，R¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: whereinX is CH, R¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein X is N, R¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein X is CH, R¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR⁴aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein X is CH, R¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is a pyridyl radical, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR⁴aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4aAnd R^4bTogether are C₂Alkylene and R^4cIs C_1-3Alkyl, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical、C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is NR^aR^bAnd R is³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is NR^aR^bAnd R is³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is N1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is N- (S) -1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1aOptionally substituted by halogen, C_1-6Alkyl radical, C_1-3Haloalkyl, C_1-6Alkoxy or hydroxy substitution, R²Is N- (S) -1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3An alkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is (a) aryl or (b) heteroaryl, wherein said aryl or said heteroaryl is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CF₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present inventionIn one embodiment, there is provided a compound of formula I as defined below: wherein R is¹Is 2, 4-dioxo-tetrahydro-pyrimidin-1-yl, R²Is a pyridyl radical, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4aAnd R^4bTogether are C₂Alkylene and R^4cIs C_1-3Alkyl, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is NR^aR^bAnd R is³Is CF₃、CH₂CF₃、CHR^4aR^4bOr CR^4aR^4bR^4cWherein (i) R^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃Or (ii) when they are taken together, R^4aAnd R^4bTogether are C_2-4Alkylene and R^4cIs C_1-3Alkyl, halogen, cyano or C_1-2A fluoroalkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is NR^aR^bAnd R is³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d、C_1-6Alkoxy or hydroxy, R²Is N1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is N- (S) -1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3Alkyl or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1d，R²Is N- (S) -1-pyrrolidin-3-ylmethyl-methanesulfonamide, and R³Is CF₃Or CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIndependently selected from C_1-3An alkyl group.

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1b，R⁵Is hydrogen or methyl, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1c，R⁵Is hydrogen or methyl, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6Alkylsulfonyl, n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there are provided compounds of formula I as defined below: wherein R is¹Is R^1e，R⁵Is hydrogen or methyl, R²Is phenyl, is (CH)₂)_nNR^cR^dSubstituted and additionally optionally and independently substituted with one to two substituents selected from cyano, C_1-6Alkoxycarbonyl, carbamoyl, N-alkylcarbamoyl, N-dialkylcarbamoyl, (CH)₂)_0-2CO₂H、SO₂NH₂、C_1-6Alkylsulfinyl and C_1-6An alkyl sulfonyl group, a carboxyl group,n is 0, and R³Is CR^4aR^4bR^4cWherein R is^4a、R^4bAnd R^4cIs Me or CD₃。

In another embodiment of the present invention, there is provided a compound selected from table 1.

In another embodiment of the present invention, there is provided a method of treating HCV infection in a patient in need thereof, comprising administering a therapeutically effective amount of a compound of formula I, wherein R is¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

In another embodiment of the present invention, there is provided a method of treating an HCV infection in a patient in need thereof, comprising co-administering a therapeutically effective amount of a compound of formula I wherein R is a compound of formula I, wherein R is a compound of formula¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

In another embodiment of the present invention, there is provided a method of treating a disease caused by HCV in a patient in need thereof, the method comprising co-administering a therapeutically effective amount of a compound of formula I and at least one immune system modulator, wherein R is¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above, and the at least one immune system modulator is selected from an interferon, an interleukin, a tumor necrosis factor or a colony stimulating factor.

In bookIn another embodiment of the invention, there is provided a method of treating an HCV infection in a patient in need thereof, comprising co-administering a therapeutically effective amount of a compound of formula I and an interferon or chemically derivatized interferon, wherein R is¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

In another embodiment of the present invention, there is provided a method of treating HCV infection in a patient in need thereof, the method comprising co-administering a therapeutically effective amount of a compound of formula I wherein R is an antiviral compound¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above, said antiviral compound being selected from the group consisting of HCV protease inhibitors, other HCV polymerase inhibitors, HCV helicase inhibitors, HCV primase inhibitors, and HCV fusion inhibitors.

In another embodiment of the present invention, there is provided a method of inhibiting viral replication in a cell by delivering a therapeutically effective amount of a compound of formula I, wherein R is in admixture with at least one pharmaceutically acceptable carrier, diluent or excipient¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

In another embodiment of the present invention, there is provided a composition comprising a compound of formula I wherein R is¹、R²、R³、R⁴、R^4a、R^4b、R^4c、R⁵、R^a、R^b、R^c、R^d、R^e、R^f、R^gAnd n is as defined above.

The term "alkyl", as used herein, alone or in combination with other groups, without further limitation, means a straight or branched chain saturated monovalent hydrocarbon residue containing 1 to 10 carbon atoms. "C" as used herein_1-6Alkyl "refers to an alkyl group containing 1 to 6 carbons. Examples of alkyl groups include, but are not limited to, lower alkyl groups including methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, neopentyl, hexyl and octyl. Any carbon-hydrogen bond may be replaced by a carbon-deuterium bond without departing from the scope of the present invention.

The definitions described herein may be appended to form chemically relevant combinations, such as "heteroalkylaryl," "haloalkylheteroaryl," "arylalkyl heterocyclyl," "alkylcarbonyl," "alkoxyalkyl," and the like. When the term "alkyl" is used as a suffix following another term, as in "phenylalkyl" or "hydroxyalkyl", this is intended to refer to an alkyl group, as defined above, substituted with one to two substituents selected from the other specifically recited group. Thus, for example, "phenylalkyl" refers to an alkyl group having one to two phenyl substituents, thus including benzyl, phenylethyl, and biphenyl. "alkylaminoalkyl" is an alkyl group having one to two alkylamino substituents. "hydroxyalkyl" includes 2-hydroxyethyl, 2-hydroxypropyl, 1- (hydroxymethyl) -2-methylpropyl, 2-hydroxybutyl, 2, 3-dihydroxybutyl, 2- (hydroxymethyl), 3-hydroxypropyl and the like. Thus, the term "hydroxyalkyl" as used herein is used to define a subset of heteroalkyl groups defined below. The term- (ar) alkyl refers to unsubstituted alkyl or aralkyl. The term (hetero) aryl or (hetero) aryl refers to aryl or heteroaryl.

The term "alkylene" as used herein denotes a divalent saturated straight chain hydrocarbon group of 1 to 10 carbon atoms (e.g., (CH)₂)_n) Or a branched saturated divalent hydrocarbon group of 2 to 10 carbon atoms (e.g. -CHMe-or-CH)₂CH(i-Pr)CH₂-, unless otherwise indicated. C_0-4Alkylene means containing from 1 to 4 carbon atomsA linear or branched saturated divalent hydrocarbon radical, or in C₀In the case of (3), the alkylene group is omitted. Except that in the case of methylene, the open valences of the alkylene groups are not attached to the same atom. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, 2-methyl-propylene, 1-dimethyl-ethylene, butylene, 2-ethylbutylene.

The term "alkoxy" as used herein denotes-O-alkyl, wherein alkyl is as defined above, e.g. methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, including isomers thereof. "lower alkoxy" as used herein denotes alkoxy having "lower alkyl" as previously defined. "C" as used herein₁-₁₀Alkoxy "means wherein alkyl is C_1-10an-O-alkyl group of (a).

The term "haloalkyl" as used herein denotes a straight or branched chain alkyl group as defined above wherein 1,2,3 or more hydrogen atoms are substituted by halogen. Examples are 1-fluoromethyl, 1-chloromethyl, 1-bromomethyl, 1-iodomethyl, difluoromethyl, trifluoromethyl, trichloromethyl, 1-fluoroethyl, 1-chloroethyl, 12-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2, 2-dichloroethyl, 3-bromopropyl or 2,2, 2-trifluoroethyl. The term "fluoroalkyl" as used herein refers to a haloalkyl moiety wherein the halogen is fluorine.

The term "acyl" (or "alkanoyl") as used herein refers to a group of formula-C (= O) R, wherein R is hydrogen or lower alkyl as defined herein. The term "alkylcarbonyl" as used herein refers to a group of formula C (= O) R wherein R is alkyl as defined herein. Term C_1-6Acyl or "alkanoyl" refers to a group-C (= O) R containing 1 to 6 carbon atoms. C₁Acyl is formyl wherein R = H, C₆Acyl refers to hexanoyl when the alkyl chain is unbranched. The term "arylcarbonyl" or "aroyl" as used herein refers to a group of formula C (= O) R wherein R is aryl; the term "benzoyl" as used herein refers to an "arylcarbonyl" or "aroyl" group wherein R is phenyl.

The terms "alkylsulfonyl" and "arylsulfonyl" as used herein represent the formula-S (= O)₂R, wherein R is independently alkyl or aryl and alkyl and aryl are as defined herein. The term C as used herein_1-3Alkylsulfonylamino refers to the group RSO₂NH-wherein R is C as defined herein_1-3An alkyl group. Term C_1-6Haloalkylsulfonyl group, C_3-7Cycloalkylsulfonyl radical, C_3-7cycloalkyl-C_1-3Alkyl-sulfonyl or C_1-6alkoxy-C_1-6Alkylsulfonyl refers to the radical S (= O)₂R, wherein R is each C_1-6Haloalkyl, C_3-7Cycloalkyl radical, C_3-7cycloalkyl-C_1-3Alkyl and C_1-6alkoxy-C_1-6An alkyl group.

The terms "alkylsulfonylamino" and "arylsulfonylamino" as used herein represent the formula-NR' S (= O)₂R, wherein R is independently alkyl or aryl, R' is hydrogen or C_1-3Alkyl, and alkyl and aryl are as defined herein. The term "sulfonamido" may be used as a prefix, while "sulfonamide" is the corresponding suffix.

The term "cycloalkyl" as used herein denotes a saturated carbocyclic ring containing from 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. "C" as used herein_3-7Cycloalkyl "refers to cycloalkyl groups containing from 3 to 7 carbons in the carbocyclic ring.

The term "cycloalkylalkyl" as used herein refers to the group R 'R "-, wherein R' is cycloalkyl as defined herein and R" is alkylene as defined herein, it being understood that the point of attachment of the cycloalkylalkyl moiety will be on the alkylene. Examples of cycloalkylalkyl groups include, but are not limited to, cyclopropylmethyl, cyclohexylmethyl, cyclopentylethyl. C_3-7cycloalkyl-C_1-3Alkyl refers to the group R 'R ", wherein R' is C as defined herein_3-7Cycloalkyl and R' is C as defined herein_1-3An alkylene group.

The term "halogen" or "halo" as used herein means fluorine, chlorine, bromine or iodine.

The terms "hydroxyalkyl" and "alkoxyalkyl" as used herein denote alkyl groups as defined herein in which 1 to 3 hydrogen atoms on different carbon atoms are replaced by hydroxy or alkoxy groups, respectively. C_1-3alkoxy-C_1-6Alkyl moiety means a compound in which 1 to 3 hydrogen atoms are replaced by C_1-3C with alkoxy radicals replaced and the point of attachment of the alkoxy radicals being an oxygen atom_1-6An alkyl substituent.

The terms "alkoxycarbonyl" and "aryloxycarbonyl" as used herein, represent groups of the formula-C (= O) OR, wherein R is alkyl OR aryl, respectively, and alkyl and aryl are as defined herein.

The term "cyano" as used herein refers to a carbon attached to nitrogen through a triple bond, i.e., -C ≡ N. The term "nitro" as used herein refers to the group-NO₂. The term "carboxy" as used herein refers to the group-CO₂H。

The term oxo refers to double-bonded oxygen (= O), i.e. carbonyl.

The term "acyl" (or "alkanoyl") as used herein refers to a group of formula-C (= O) R, wherein R is hydrogen or lower alkyl as defined herein. The term "alkylcarbonyl" as used herein refers to a group of formula C (= O) R wherein R is alkyl as defined herein. Term C_1-6Acyl or "alkanoyl" refers to a group-C (= O) R containing 1 to 6 carbon atoms. C₁Acyl is formyl wherein R = H, C₆Acyl refers to hexanoyl when the alkyl chain is unbranched. The term "arylcarbonyl" or "aroyl" as used herein denotes a group of formula C (= O) R wherein R is aryl; the term "benzoyl" as used herein refers to an "arylcarbonyl" or "aroyl" group wherein R is phenyl.

The term "heteroaryl" as used herein, without further definition or limitation, refers to the "pyridyl", "pyrazinyl" and "pyridazinyl" rings. The term "pyridine" ("pyridyl") refers to a 6-membered heteroaromatic ring having one nitrogen atom. The terms "pyrimidine" (pyrimidinyl), "pyrazine" ("pyrazinyl") and "pyridazine" ("pyridazinyl") refer to 6-membered non-fused heteroaromatic rings having two nitrogen atoms arranged in a 1,3, 1,4 and 1,2 relationship, respectively. The respective radical names are in brackets.

The term "sulfamoyl" as used herein refers to the group-S (O)₂NH₂. The terms "N-alkylsulfamoyl" and "N, N-dialkylsulfamoyl" as used herein refer to the group-S (O)₂NR ' R ", wherein R ' and R" are hydrogen and lower alkyl and R ' and R "are independently lower alkyl, respectively. Examples of N-alkylsulfamoyl substituents include, but are not limited to, methylaminosulfonyl, isopropylaminosulfonyl. Examples of N, N-dialkylsulfamoyl substituents include, but are not limited to, dimethylaminosulfonyl, isopropyl-methylaminosulfonyl.

The term "carbamoyl" as used herein denotes the group-CONH₂. The prefixes "N-alkylcarbamoyl" and "N, N-dialkylcarbamoyl" denote the groups CONHR ' or CONR ' R ", respectively, wherein the R ' and R" groups are independently alkyl as defined herein. The prefix "N-arylcarbamoyl" denotes a group CONHR 'wherein R' is aryl as defined herein.

The terms "alkylsulfinyl" and "arylsulfinyl" as used herein, denote groups of the formula-S (= O) R, wherein R is alkyl or aryl, respectively, and alkyl and aryl are as defined herein.

The terms "alkylsulfonyl" and "arylsulfonyl" as used herein refer to the formula-S (= O)₂R groups, wherein R is independently alkyl or aryl, wherein alkyl and aryl are as defined herein.

The term "benzyl" as used herein refers to C₆H₅CH₂Wherein the phenyl ring may be optionally substituted with one or more, preferably one or three substituents independently selected from: hydroxy, thio, cyano, alkyl, alkoxy, lower haloalkoxy, alkylthio, halogen, haloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, aminoAlkylamino, dialkylamino, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl, alkylsulfonyl, arylsulfinyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, carbamoyl, alkylcarbamoyl and dialkylcarbamoyl, arylcarbamoyl, alkylcarbonylamino, arylcarbonylamino, unless otherwise stated.

The terms "oxetane" (oxetanyl), "tetrahydrofuran" (tetrahydrofuryl) and "tetrahydropyran" (tetrahydropyranyl) refer to 4,5 and 6-membered non-fused heterocycles, respectively, each containing one oxygen atom.

The term "aryl" as used herein refers to phenyl.

The term "cyclic amine" refers to a saturated carbocyclic ring containing 3 to 6 carbon atoms as defined above and in which at least one carbon atom is replaced by a heteroatom selected from N, O and S, such as piperidine, piperazine, morpholine, thiomorpholine, dioxo-thiomorpholine, pyrrolidine, pyrazoline, imidazolidine, azetidine, wherein a ring carbon atom is optionally substituted by one or more substituents selected from halogen, hydroxy, phenyl, lower alkyl, lower alkoxy, or 2 hydrogen atoms on carbon are each replaced by oxo (= O). When the cyclic amine is piperazine, one nitrogen atom may optionally be replaced by C_1-6Alkyl radical, C_1-6Acyl radical, C_1-6Alkylsulfonyl.

The term (i)2, 4-dioxo-1, 2,3, 4-tetrahydro-pyrimidin-5-yl (when R is⁵Is H), ()2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl,(iii)2, 4-dioxo-1, 2,3, 4-tetrahydro-2H-pyrimidin-1-yl or (iv) 3-methyl-5-oxo-1, 5-dihydro- [1,2,4]Triazol-4-yl refers to the following moieties:

3-oxo-3, 4-dihydro-pyrazin-2-yl, (ii) 3-oxo-2, 3-dihydro-pyridazin-4-yl, (iii) 2-oxo-1, 2-dihydro-pyrimidin-4-one-5-yl, (iv) 2-oxo-1, 2-dihydro-pyridin-3-yl, (v) 6-oxo-1, 6-dihydro- [1,2,4] triazin-5-yl and (vi)

Commonly used abbreviations include: acetyl (Ac), aqueous or aqueous (aq.), atmospheric (Atm), 2 '-bis (diphenylphosphino) -1,1' -Binaphthyl (BINAP), tert-butyloxycarbonyl (Boc), di-tert-butyldicarbonate or Boc anhydride (BOC)₂O), benzyl (Bn), butyl (Bu), chemical abstracts accession number (CASRN), benzyloxycarbonyl (CBZ or Z), Carbonyldiimidazole (CDI), 1, 5-diazabicyclo [4.3.0 ]]Non-5-ene (DBN), 1, 8-diazabicyclo [5.4.0]Undec-7-ene (DBU), N' -Dicyclohexylcarbodiimide (DCC), 1, 2-Dichloroethane (DCE), Dichloromethane (DCM), diethyl azodicarboxylate (DEAD), diisopropyl azodicarboxylate (DIAD), diisobutylaluminum hydride (DIBAL or DIBAL-H), Diisopropylethylamine (DIPEA), N-Dimethylacetamide (DMA), 4-N, N-Dimethylaminopyridine (DMAP), N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), ethyl (Et), ethyl acetate (EtOAc), ethanol (EtOH), 2-ethoxy-2H-quinoline-1-carboxylic acid Ethyl Ester (EEDQ), Diethyl ether (Et)₂O), O- (7-azabenzotriazol-1-yl) -N, N, N 'N' -tetramethylurea hexafluorophosphateAcetic acid (HATU), acetic acid (HOAc), 1-N-hydroxybenzotriazole (HOBt), High Pressure Liquid Chromatography (HPLC), Isopropanol (IPA), methanol (MeOH), melting point (mp), MeSO₂- (methylsulfonyl or Ms), methyl (Me), acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA), mass spectrometry (Ms), methyl tert-butyl ether (MTBE), N-methylmorpholine (NMM), N-methylmorpholinePyrrolidone (NMP), phenyl (Ph), propyl (Pr), isopropyl (i-Pr), pounds per square inch (psi), pyridine (pyr), room temperature (RT or RT), satd. (saturated), tert-butyldimethylsilyl, or t-BuMe₂Si (TBDMS), triethylamine (TEA or Et)₃N), trifluoromethanesulfonate or CF₃SO₂- (Tf), trifluoroacetic acid (TFA), O-benzotriazol-1-yl-N, N, N ', N' -tetramethylureaTetrafluoroborate (TBTU), Thin Layer Chromatography (TLC), Tetrahydrofuran (THF), Tetramethylethylenediamine (TMEDA), trimethylsilyl or Me₃Si (TMS), p-toluenesulfonic acid monohydrate (TsOH or pTsOH), 4-Me-C₆H₄SO₂-or tosyl (Ts), N-urethane-N-carboxy anhydride (UNCA). When used with alkyl moieties, the conventional nomenclature including the prefixes n- (n-), iso (i-), secondary (sec-), tertiary (tert-), and neo- "have their usual meaning. (J.Rigaudy and D.P.Klesney, nomenclature in organic chemistry (Nomelantereinorganic chemistry), IUPAC1979Pergamon Press, Oxford.)

The following table provides examples of representative compounds encompassed by and within the scope of the present invention. The following examples and preparations are provided to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.

Generally, the nomenclature used herein is based on AUTONOM^TMv.4.0, a beilstein institute computer system used to generate IUPAC systematic nomenclature. The depicted structure is used to control if there is a discrepancy between the depicted structure and the name given to the structure. In addition, if the stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or the portion of the structure is to be interpreted as encompassing all stereoisomers of it.

The compounds of the present invention can be prepared by a variety of methods depicted in the illustrative synthetic reaction schemes shown and described below. The starting materials and reagents for preparing these compounds are generally available from commercial suppliers (e.g., Aldrich chemical CO.), or may be prepared by methods known to those skilled in the art according to procedures described in the literature, e.g., FieserandFieser's research for organic Synthesis, Wiley & Sons: New York, volumes 1-21; larock, comparative organic transformations, 2 nd edition, Wiley-VCH, New York 1999, comparative organic Synthesis, B.Trost and I.Fleming, Vol.1-9, Pergamon, Oxford City, 1991, comparative heterocyclic chemistry, A.R.Katritzky and C.W.Rees (eds), Pergamon, Oxford City, 1984, Vol.1-9, comparative heterocyclic chemistry II, A.R.Katritzky and C.W.Rees (eds), Pergamon, Oxford City, 1996, Vol.1-11, and organic reactions, Wiley & Sons, New York, 1991, Vol.1-40. The following synthetic reaction schemes are merely illustrative of some of the methods by which the compounds of the present invention can be synthesized, and various modifications can be made to these synthetic reaction schemes and will be suggested to one skilled in the art having referred to the disclosure contained herein.

The starting materials and intermediates of the synthetic reaction schemes may be isolated and purified as appropriate using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography and the like. These substances can be characterized by conventional means, including physical constants and spectral data.

Unless indicated to the contrary, the reactions described herein are preferably carried out under an inert atmosphere at atmospheric pressure, with reaction temperatures ranging from about-78 ℃ to about 150 ℃, more preferably from about 0 ℃ to about 125 ℃, most preferably and conveniently at about room (or ambient) temperature, for example about 20 ℃.

Some of the compounds in the following schemes are depicted with generalized substituents; however, those skilled in the art will immediately appreciate that the nature of the R groups may be varied to provide a variety of compounds of interest for the present invention. Moreover, the reaction conditions are exemplary and alternative conditions are well known. The order of reaction in the following examples is not meant to limit the scope of the invention described in the claims.

Procedure A

Quinazoline derivatives within the scope of the present invention were prepared from 7-bromo-4-hydroxy-2-tert-butyl-quinazoline (22 a). Preparation of 22a was accomplished by cyclization of 20b, which was prepared by acetylation of 5-bromo-anthranilamide. The 4-methanesulfonamide substituent was introduced by a palladium catalyzed coupling reaction of 4-methylsulfonylamino-phenylboronic acid with 22a using the Suzuki (Suzuki) protocol.

Suzuki reaction is boric acid (R-B (OH)₂) (wherein R is aryl or vinyl) with aryl or vinyl halides or triflates (R 'Y, wherein R' is aryl or vinyl; Y = halogen or-OSO)₂CF₃) Palladium catalyzed coupling is carried out to give the compound R-R'. Typical catalysts include Pd (PPh)₃)₃、Pd(OAc)₂And PdCl₂(dppf). Using PdCl₂(dppf) it is possible to couple primary alkyl borane compounds with aryl or vinyl halides or triflates without β -elimination catalysts have been identified (see, for example, J.P.Wolfe et al, J.Am.chem.Soc.1999121(41):9550-9561 and A.F.Littke et al, J.Am.chem.Soc2000122(17): 4020-4028.) the reaction can be carried out in a variety of organic solvents including toluene, THF, bis (toluene)Alkane, 1, 2-dichloroethane, DMF, PhMe, MeOH, DMSO and acetonitrile, aqueous solvent and under biphasic conditions. The reaction is usually carried out inFrom about room temperature to about 150 ℃. Additives (e.g., CsF, KF, TlOH, NaOEt, and KOH) often accelerate coupling. There are a number of parameters in the Suzuki reaction, including palladium source, ligand, additives and temperature, and optimal conditions sometimes require parameter optimization for a given pair of reactants. Littke et al (ibid) discloses methods for using Pd₂(dba)₃/P (tert-bu)₃Conditions for Suzuki Cross-coupling with arylboronic acids at high yields at RT and for use with Pd (OAc)₂/P(C₆H₁₁)₃Conditions for cross-coupling of aryl-and vinyl-triflates at room temperature. Wolf et al (supra) disclose methods for using Pd (OAc)₂Effective conditions for Suzuki cross-coupling with o- (di-tert-butylphosphino) biphenyl or o- (dicyclohexylphosphino) biphenyl. The optimal conditions can be determined by those skilled in the art without undue experimentation.

Amine substituent NR^aR^bThe palladium catalyzed amination is performed. The replacement of suitable leaving groups such as chloro, bromo, iodo, mesylate or triflate substituents on aryl or heteroaryl rings with amines has become a well established procedure (e.g., Buchwald-Hartwig coupling; see (a) J.P.Wolfe, S.Wagaw and S.L.Buchwald J.Am.Chem.Soc.1996,118,7215-7216; (b) J.P.Wolfe and S.L.Buchwald tetrahedron Lett.1997,38,6359-6362; (c) J.P.Wolfe, S.Wagaw, J.F.Marcoux and S.L.Buchwald Acc.Res.1998, 31,805-818; (d) B.H.Yang and S.L.Buchwald J.Organomet.146. IV.125. 125. Cheg.576, 125-ChegJ.7. EgW.C.; (c.7, 15-7, J.W). Amination of (hetero) aryl halides or sulfonates with palladium catalysts such as tris- (dibenzylideneacetone) dipalladium (0) (Pd)₂(dba)₃) Or Pd (OAc)₂Phosphine ligands such as triphenylphosphine, rac-2, 2' -bis (diphenylphosphino) -1,1' -binaphthyl (rac-BINAP), dicyclohexyl- (2',4',6' -triisopropyl-biphenyl-2-yl) -phosphine (X-Phos), (R) - (-) -1- [ (S) -2- (dicyclohexylphosphino) ferrocenyl]Ethyl di-tert-butylphosphine (Josiphos; see Q.Shen, S.Shekhar, J.P.Stambuli and J.F.HartwiggAngew.Chem.Int.Ed.2005, 44,1371-₆H₁₁)₃、P(ortho-Tol)₃Or P (tert-Bu)₃And (3) catalyzing. In general, in solvents such as toluene, EtOH, DME, bisAlkaline additives such as Cs in alkanes or water or mixtures thereof₂CO₃、K₃PO₄Or KO-tert-Bu. C-N formation can be carried out at room temperature or at elevated temperature, where heating can be accomplished conventionally or by microwave irradiation (see also Palladium (0) complex organic chemistry, organometallics synthesis (m.schlosser editions), chapter 4, 2 nd edition, 2002, john wiley&Sons, Ltd, Chichester, UK and D.Prim et al, Tetrahedron200258: 2041-.

Followed by treatment with POCl₃The heteroaryl substituent is introduced by converting the 4-hydroxy group to the corresponding chloride. Optionally substituted 2-methoxy-pyridin-3-ylboronic acids (or corresponding borates) can be suzuki coupled with 24 to give 26, which is demethylated with HBr/HOAc to give the desired product.

Boronic acids useful in preparing compounds of the present invention include, but are not limited to, 2-methoxy-pyridin-3-yl boronic acid (CASRN163105-90-6), 2-benzyloxy-3-pyridineboronic acid, 2-oxo-1, 2-dihydropyridine-3-boronic acid (CASRN951655-49-5), 5-fluoro-2-methoxy-3-pyridineboronic acid (CASRN957120-32-0), 2-methoxy-6-methyl-pyridin-3-yl boronic acid (CASRN1000802-75-4), 5-chloro-2-methoxy-pyridin-3-yl boronic acid (CASRN943153-22-8), 2, 6-dimethoxy-pyridin-3-yl boronic acid (CASRN221006-70-8), or 2, 4-dioxo-1, 2,3, 4-tetrahydropyrimidin-5-ylboronic acid (CASRN 70523-22-7). One skilled in the art will recognize that boronic acids and boronic esters such as 4,4,5, 5-tetramethyl- [1,3,2] dioxaborolan-2-yl may be used interchangeably in suzuki couplings.

Alternatively, the2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl group can be introduced by copper catalyzed aryl amination, replacing the aryl halide with uracil. A number of methods for CuI-catalyzed aryl amination have been reported (r.wagner et al, WO2009/039127 discloses CuI catalysis with uracil instead of aryl halides). Dibromide 42 (via 3, 3-dimethyl-2, 3-dihydro-benzene)And sequential monobromination of furan) was first performed with 29 to afford 44 and the isomeric coupled products. Separating the isomers, all using uracil, CuI, (2-cyano-phenyl) -pyridine-2-carboxamide and Cs₂CO₃Amination is carried out to obtain I-1 and I-4.

The analog pyrido [3,2-d ] pyrimidine was prepared similarly except that 3-amino-5-bromo-pyridin-2-ylcarboxamide was used instead of 20 a. The 4-and 7-substituents may be introduced as described above.

Wherein R is¹Is R^1bAnd R is⁵Is C_1-3Alkyl compounds can be prepared as follows: palladium catalyzed coupling of 2, 4-dimethoxy-pyrimidin-5-ylboronic acid (or ester thereof) followed by treatment of the adduct with methyl iodide (or equivalent) affords a 4-methoxy-1-methyl-2-oxo-1H-pyrimidin-5-yl derivative which can be treated with HBr/HOAc as described herein to afford a 1-methyl-2, 4-dioxo-1H-pyrimidin-5-yl analog. Wherein R is¹Is R^1eThe compounds of (3) are prepared from the corresponding 4-amino derivatives, which can be prepared as described above. The triadimefon is obtained by treatment of the amine with triethyl orthoacetate and methyl hydrazinoformate under mildly acidic conditions and subsequent cyclization with an alkali metal alkoxide.

The activity of the compounds of the present invention as inhibitors of HCV activity can be determined by any suitable method known to those skilled in the art, including in vivo and in vitro assays. For example, the HCV NS5B inhibitory activity of the compounds of formula I can be determined using standard assays described in Behrens et al, EMBOJ.199615:12-22, Lohmann et al, Virology1998249:108-118 and Ranjith-Kumar et al, J.virology200175: 8615-8623. Compounds of the invention have been demonstrated to have in vitro hcv ns5B inhibitory activity in such standard assays, unless otherwise indicated. Example 8 describes HCV polymerase assay conditions for compounds of the present invention. Cell-based replicon systems for HCV have been developed in which nonstructural proteins stably replicate subgenomic viral RNA in Huh7 cells (V.Lohmann et al, Science1999285:110 and K.J.Blight et al, Science2000290: 1972). Example 4 describes cell-based replicon assay conditions for compounds of the invention. In the absence of purified functional HCV replicase, composed of viral non-structural and host proteins, our understanding of flaviviridae RNA synthesis comes from studies with active recombinant RNA-dependent RNA-polymerases and validation of these studies in the HCV replicon system. Inhibition of recombinantly purified HCV polymerase in vitro biochemical assays can be demonstrated using the replicon system by which the polymerase is present in a replicase complex with appropriate stoichiometry of other viral and cellular polypeptides. Demonstration of cell-based inhibition of HCV replication may be more predictive of in vivo function than demonstration of HCV ns5B inhibitory activity in an in vitro biochemical assay.

The compounds of the present invention may be formulated in a variety of oral administration dosage forms and carriers. Oral administration may be in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions, syrups or suspensions. In other routes of administration, the compounds of the invention are effective when administered by other routes of administration, including continuous (intravenous drip) topical parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer), buccal, nasal, inhalation, and suppository administration. The preferred mode of administration is generally oral using a convenient daily regimen which may be adjusted according to the degree of involvement and the response of the patient to the active ingredient.

One or more compounds of the present invention and their pharmaceutically acceptable salts, together with one or more conventional excipients, carriers or diluents, can be formulated into pharmaceutical compositions and unit dosage forms. The pharmaceutical compositions and unit dosage forms can contain conventional ingredients in conventional proportions, with or without additional active compounds or ingredients, and the unit dosage forms can contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed. The pharmaceutical composition may be employed in the form of: solid such as tablets or filled capsules, semi-solid, powder, sustained release formulations or liquid such as solutions, suspensions, emulsions, elixirs or filled capsules for oral use; or suppositories for rectal or vaginal administration; or a sterile injectable solution for parenteral use. Typical formulations will contain from about 5% to about 95% active compound (w/w). The terms "formulation" or "dosage form" are intended to include both solid and liquid formulations of the active compound, and those skilled in the art will appreciate that the active ingredient may be present in different formulations, depending on the target organ or tissue and the desired dosage and pharmacokinetic parameters.

The term "excipient", as used herein, means a compound that is useful in preparing a pharmaceutical composition, is generally safe, non-toxic, neither biologically nor otherwise undesirable, and includes excipients that are acceptable for veterinary use as well as human pharmaceutical use. The compounds of the present invention may be administered alone, but may generally be administered together with one or more suitable pharmaceutical excipients, diluents and carriers selected with regard to the intended route of administration and standard pharmaceutical practice.

"pharmaceutically acceptable" means useful in preparing a pharmaceutical composition that is generally safe, non-toxic, neither biologically nor otherwise undesirable, including those that are acceptable for human pharmaceutical use.

The "pharmaceutically acceptable salt" form of the active ingredient may also initially impart to the active ingredient the desired pharmacokinetic properties not present for the non-salt form and may even positively influence the pharmacodynamics of the active ingredient with respect to its therapeutic activity in vivo. The phrase "pharmaceutically acceptable salt" of a compound refers to a salt that is pharmaceutically acceptable and has the intended pharmacological activity of the parent compound. Such salts include: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with organic acids; examples of the organic acid include acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, laurylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) a salt formed when the acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion; or a salt formed when coordinated with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, or the like.

Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances that also can act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is generally a finely divided solid which is in admixture with the finely divided active component. In tablets, the active ingredient is usually mixed with a carrier having the required binding capacity in suitable proportions and compacted in the shape and size desired. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low melting wax, cocoa butter, and the like. Solid form preparations may contain, in addition to the active ingredient, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

Liquid preparations are also suitable for oral administration, and include emulsions, syrups, elixirs, aqueous solutions, and aqueous suspensions. They include solid form preparations which are intended to be converted, shortly before use, to liquid form preparations. Emulsions may be prepared in solution, for example, in aqueous propylene glycol, or may contain emulsifying agents, for example, lecithin, sorbitan monooleate, or acacia. Aqueous solutions can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavors, stabilizers, and thickening agents. Aqueous suspensions may be prepared by dispersing the finely divided active component in water containing viscous material, for example, natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

The compounds of the invention may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion) and may be presented in unit dosage form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, for example, solutions in aqueous polyethylene glycol. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil) and injectable organic esters (e.g., ethyl oleate), which may contain formulatory agents such as preservatives, wetting, emulsifying or suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic packaging of the sterilized solid or by freeze-drying of the solution, for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

The compounds of the invention may be formulated as ointments, creams or lotions or as a transdermal patch for topical application to the epidermis. For example, ointments and creams may be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. Formulations suitable for oral topical administration include: lozenges comprising the active agent in a flavored base (usually sucrose and acacia or tragacanth); pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The compounds of the present invention may be prepared for administration in the form of suppositories. A low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active ingredient is dispersed homogeneously, for example by stirring. The molten homogeneous mixture is then poured into a suitably sized mold, cooled and solidified.

The compounds of the invention may be prepared in a form for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate. The compounds of the present invention may be formulated for nasal administration. The solution or suspension is applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or nebulizer. These formulations may be provided in single or multiple dose forms. When in a dropper or pipette multi-dose form, administration may be by administering to the patient an appropriate predetermined volume of solution or suspension. When a nebulizer is employed, administration can be carried out, for example, by a metered dose nebulizer pump.

The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract and including intranasal administration. The compounds typically have small particle sizes, for example on the order of 5 microns or less. The particle size may be obtained by methods known in the art, such as micronization. The active ingredient may be provided in pressurized packs with a suitable propellant, for example a chlorofluorocarbon (CFC) such as dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, carbon dioxide or other suitable gas. The aerosol may also conveniently contain a surfactant such as lecithin. The dose of medicament may be controlled by a metering valve. Alternatively, the active ingredient may be provided in the form of a dry powder, for example a powder mix of the compound with a suitable powder base such as lactose, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP). The powder carrier can form a gel in the nasal cavity. The powder compositions may be presented in unit dosage form, for example gelatin or blister packs of capsules or cartridges, from which the powder may be administered by inhalation.

When desired, the formulations may be prepared with enteric coatings suitable for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention may be formulated in a transdermal or subcutaneous drug delivery device. These delivery systems are advantageous when sustained release of the compound is necessary and when patient compliance with the treatment regimen is critical. The compounds in transdermal delivery systems are typically attached to a skin-adherent solid support. The compound of interest may also be added with a penetration enhancer such as azone (1-dodecaazacycloheptan-2-one). The sustained release delivery system may be implanted subcutaneously in the subcutaneous layer by surgery or injection. Subcutaneous implants encapsulate the compound in a lipid-soluble membrane such as silicone rubber or a biodegradable polymer such as polylactic acid.

Suitable formulations and pharmaceutical carriers, diluents and excipients are described in Remington: the science and practice of pharmacy (1995.e.w. martin, edited by mark (Mack) publishing company, 19 th edition, easton, pa). The skilled formulation scientist is able to modify the formulation within the teachings of the specification to provide a wide variety of formulations for specific routes of administration without destabilizing the compositions of the invention or otherwise compromising their therapeutic activity.

Modifications of the compounds of the invention to render them more soluble in water or other vehicles can be readily accomplished, for example, by small modifications (salt formation, esterification, etc.), which are well within the ordinary skill of the art. It is also well within the ordinary skill in the art to modify the route of administration and dosage regimen of a particular compound in order to manage the pharmacokinetics of the compounds of the present invention to achieve maximal beneficial effects in the patient.

The term "therapeutically effective amount" as used herein means the amount required to reduce the symptoms of a disease in an individual. The dosage will be adjusted to the individual needs in each particular case. The dosage may vary within wide limits depending on various factors, such as the severity of the disease being treated, the age and general health of the patient, other medications being used to treat the patient, the route and manner of administration, and the preferences and experience of the attending medical personnel. For oral administration, a daily dose of about 0.01 to about 1000mg/kg body weight/day should be appropriate in monotherapy and/or in combination therapy. Preferred daily dosages are from about 0.1 to about 500mg/kg body weight/day, more preferably from 0.1 to about 100mg/kg body weight/day, and most preferably from 1.0 to about 10mg/kg body weight/day. Thus, for administration to a 70kg human, the dose will range from about 7mg to 0.7 g/day. The daily dose may be administered as a single dose or in divided doses, typically 1 to 5 doses per day. Generally, treatment is initiated at smaller doses than the optimal dose of the compound. The dosage is then gradually increased in smaller increments until the optimum effect is achieved for the individual patient. The ordinarily skilled artisan, without undue experimentation in treating the diseases described herein, will be able to determine, depending on the individual knowledge, experience, and disclosure of the present application, a therapeutically effective amount of a compound of the invention for a given disease and patient.

In an embodiment of the invention, the active compound or salt may be administered in combination with other antiviral agents such as ribavirin, nucleoside HCV polymerase inhibitors, other HCV non-nucleoside polymerase inhibitors, or HCV protease inhibitors. When the active compound or derivative or salt thereof is administered in combination with other antiviral agents, the activity may be enhanced as compared to the parent compound. When the treatment is a combination therapy, such administration may be concurrent or sequential with respect to the administration of the nucleoside derivative. As used herein, "concurrently administering" thus includes administering each drug at the same time or at different times. Administration of two or more drugs at the same time may be achieved by a single formulation containing two or more active ingredients or by substantially simultaneous administration of two or more dosage forms containing a single active ingredient.

It will be appreciated that reference herein to treatment extends to prophylaxis as well as treatment of existing conditions. Furthermore, the term "treatment" of HCV infection as used herein also includes the treatment or prevention of a disease or disorder associated with or mediated by HCV infection, or the clinical symptoms thereof.

A therapeutically effective amount of a compound of the present invention, and optionally one or more additional antiviral agents, is an amount effective to reduce viral load or achieve a sustained virological response to treatment. In addition to viral load, indications useful for a sustained response include, but are not limited to, liver fibrosis, elevated serum transaminase levels, and necrotic inflammatory activity in the liver. An illustrative, but non-limiting, common example of a marker is serum alanine Aminotransferase (ALT), which is determined by standard clinical assays. In some embodiments of the invention, an effective treatment regimen is a regimen that reduces ALT levels to below about 45IU/mL serum.

The following examples illustrate the preparation and biological evaluation of compounds within the scope of the present invention. The following examples and preparations are provided so that those skilled in the art can more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.

Example 1

N- {4- [ 2-tert-butyl-4- (6-methyl-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (I-1; scheme A)

Step 1: to 20a (3g,14mmol, CASRN112253-70-0) in DCM cooled to 0 deg.C and maintained at N₂To the suspension under atmosphere was added TEA and the suspension was stirred for 15 minutes. Pivaloyl chloride (1.68g,14mmol) was slowly added to the suspension and the solution was stirred at room temperature overnight. The solution was cooled again to 0 ℃, a second aliquot of pivaloyl chloride (300 μ L) was added, and the resulting mixture was stirred at 0 ℃ for 2 hours and then at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give 20b, which was used without additional purification.

Step 2: to a suspension of 20b (4.2g,14mmol) and EtOH (28mL) was added aqueous NaOH (2.8mL,28mmol,10M solution) and the resulting mixture was placed in N₂The mixture was heated under reflux for 1.5 hours under an atmosphere. The solution was cooled to RT, poured onto ice and neutralized with 1n hcl. The resulting mixture was extracted twice with EtOAc. The combined extracts were washed with brine and dried (MgSO)₄) Filtered and evaporated to give 2.07g of 22a as a yellow solid.

Step 3: to a microwave vial was added 22a (0.5g,1.78mmol), 4-methanesulfonylamino-phenylboronic acid (0.421g,1.96mmol, CASRN380430-57-9), Pd (PPh)₃)₄(0.206g,0.178mmol)、Na₂CO₃(0.566g,5.34mmol), MeOH (3mL), and toluene (1.5 mL). Charging Ar into the small bottle for 5 minutes, sealing, and microwave synthesizingIrradiation was carried out at 115 ℃ in the apparatus. The reaction mixture was cooled and concentrated. The insoluble material was washed with Et₂Trituration with O gave 0.55g (84.3%) of 22b as a brown solid.

Step 4: to a suspension of 22b (0.1g,0.27mmol) in benzene (0.5mL) was added diethylaniline (73.0. mu.L, 0.45mmol) and POCl in that order₃(14.8. mu.L). The mixture was heated at reflux for 6 h, cooled and diluted with EtOAc. The resulting solution was treated sequentially with 1N HCl, H₂O, saturated NaHCO₃Aqueous solution, H₂O and brine wash. The solution was dried (MgSO₄) Filtered and evaporated to give 24.

Step 5: to a vial was added 24(0.107g,0.274mmol), 6-methyl-2-methoxy-pyridin-3-ylboronic acid (0.055g,0.33mmol), PdCl₂(dppf).CH₂Cl₂(0.010g,0.014mmol)、Cs₂CO₃(0.822g,0.268mmol), bisAlkane (1mL) and H₂O (0.25mL), cleaned with Ar for 10 minutes, sealed and heated for 1 hour. The solution was cooled to room temperature, diluted with EtOAc and successively with H₂O and brine were extracted twice. The resulting solution was dried (MgSO)₄) Filtering and vacuum concentrating. Passing the crude product through SiO₂Purification by chromatography, eluting with a gradient of EtOAc/hexanes (10 to 30% EtOAc) afforded 0.08g 26.

Step 6: a mixture of 26(0.08g,0.169mmol), 48% aqueous HBr (52. mu.L) and HOAc was heated in a sealed tube at 60 ℃ for 3 hours. The tube was cooled and the mixture was diluted with EtOAc and saturated NaHCO₃The aqueous solution was neutralized and stirred at room temperature overnight. Evaporation of EtOAc and filtration of the remaining yellow solid using EtOAc and H₂And O washing. The solid was dried in a vacuum oven at 70 deg.C overnight to give I-1.

Example 2

N- {4- [ 2-tert-butyl-4- (6-methyl-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (I-2)

At 0 deg.CN-pyrrolidin-3-ylmethyl-methanesulfonamide (28) -TEA (1.05mL,7.5mmol) was added to a solution of (R) -3- (aminomethyl) -1-N-BOC-pyrrolidine (1g,5mmol) in DCM (25 mL). Methanesulfonyl chloride (0.43mL,5.5mmol) was then added. After stirring at 0 ℃ for 2 hours, the reaction mixture was diluted with water. The organic phase was separated and dried (MgSO)₄) Filtering, and concentrating. The crude material was treated with a 1M solution of HCl in MeOH (25mL) at room temperature and stirred at room temperature for 20 hours. Volatiles were removed under reduced pressure to give 0.95g28 as a white solid.

Step 1To a vial was added 22a (0.154g0.55mmol), 28(0.130g, 0.6mmol), Pd (OAc)₂(6.17mg,0.03mmol), tert-butyl-di-1-adamantylphosphine (19.7mg,0.06mmol,321921-71-5), NaOAc (0.211g) and toluene (2mL), purged with argon for 10 minutes, sealed and irradiated in a microwave synthesizer at 120 ℃ for 20 minutes. The walls of the tubes were covered with a black film, the mixture was diluted with dry DMSO (0.7mL), the vials were sealed, and irradiated at 120 ℃ for an additional 20 minutes. The solution was cooled, diluted with EtOAc and washed with H₂And O washing. The organic extracts were dried, filtered and evaporated. Passing the residue through SiO₂Purification by chromatography, eluting with a gradient of EtOAc/hexanes (40 to 70% EtOAc) afforded 30 a.

Step 2The procedure of example 1, step 4, with POCl₃Conversion of 20a to 30b

Step 3To a vial was added 30b (0.025g,0.064mmol), uracil (0.022g,0.19mmol), Cs₂CO₃(0.042g,0.128mmol) and DMSO (0.5mL), sealed and heated at 100 ℃ for 2.5 hours. The reaction mixture was cooled, diluted with EtOAc and successively with H₂O and brine, dried (MgSO)₄) Filtering and vacuum concentrating. Passing the crude product through SiO₂Purification was performed eluting with a gradient of EtOAc/hexanes (30 to 50% EtOAc/hexanes) to give 14mg of pure I-2 as a white solid.

Example 3

N- {4- [ 2-tert-butyl-4- (2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (I-3) was prepared by aminating 24 with uracil according to the method of example 2, step 3.

Example 4

N- {4- [ 2-tert-butyl-4- (6-hydroxy-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (I-4)

N- {4- [ 2-tert-butyl-4- (2, 6-dimethoxy-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (32) was prepared by palladium catalyzed coupling of 24 with 2, 6-dimethoxy-pyridin-3-ylboronic acid according to the procedure disclosed in example 1, step 4.

Step 1To a sealable tube was added 32(115mg,2330.223mmol), HOAc (1mL) and HBr (56.7mg,0.700 mmol). The tube was sealed and heated at 60 ℃ for 2 hours. After stirring overnight at room temperature, an aliquot of HBr was added and the solution was stirred at 70 ℃ for an additional 3 hours. No starting material was detected by TLC. The reaction mixture was cooled and washed with H₂O dilution with saturated NaHCO₃The aqueous solution was neutralized, resulting in an orange precipitate. The solid was washed out of the tube with EtOAc and the filtrate was evaporated. Collecting the yellow precipitate with H₂O, DCM and Et₂O was washed thoroughly and dried in a vacuum oven at 70 deg.C for 1 hour to give 75.5mg (69.6%) of I-4.

Example 5

N- {4- [ 2-tert-butyl-4- (2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl) -pyrido [3,2-d ] pyrimidin-7-yl ] -phenyl } -methanesulfonamide (I-5)

Step 1To a solution of 34a (1.05g,4.86mmol, CASRN669066-89-1), TEA (639mg,881 μ L,6.32mmol) in DCM (15mL) and THF (5mL) was added pivaloyl chloride (586mg,598 μ L,4.86mmol) dropwise and the resulting solution was stirred at room temperature. Another equivalent of TEA and 0.5eq pivaloyl chloride was added and the solution was stirred over the weekend. The mixture was washed with DCM and H₂Partitioning between O, and neutralizing with 1N HCl. The organic extracts were washed with brine and dried (MgSO)₄) Filtration and concentration gave 1.43g (98%) of 34b, which was used without further purification.

Step 2A solution of 34b (4.73g,15.8mmol) and NaOH (3.15mL,31.5mmol,1MEtOH solution) and EtOH (31.5mL) was heated to reflux for 2.5 hours, resulting in a homogeneous solution. The reaction was cooled, diluted with EtOAc and successively with H₂O and brine wash. The organic extract was dried (MgSO)₄) Filtering and vacuum concentrating. The resulting orange solid was treated with Et₂Trituration with O gave 2.63g (59.2%) of 36a as a white solid which was used without additional purification.

Step 3-adding 36a (463mg,1.64mmol), 4- (methylsulfonylamino) phenylboronic acid (370mg,1.72mmol) and Na to a microwave tube₂CO₃(521mg,4.92mmol), MeOH (3mL), toluene (1.00mL), and H₂O (500. mu.L). Argon was bubbled into the mixture for 10 minutes, and Pd (PPh) was added₃)₄(94.8mg, 82.0. mu. mol). Argon was continued to be introduced for another 5 minutes. The vial was sealed and the reaction was heated at 120 ℃ for 8 hours. The mixture formed a non-stirrable yellow opaque mixture. Subjecting the mixture to hydrogenation with H₂Dilution with O and stirring with aqueous acid yielded an insoluble white solid which was filtered and dried in vacuo to yield 0.606g of 36 b. The NMR corresponds to the expected product.

Step 4To a suspension containing 36b (0.4g,1.07mmol), DIPEA (236mg, 316. mu.l, 1.83mmol) in benzene (3mL) was added POCl₃(98.8mg, 60.1. mu.L, 644. mu. mol) and the mixture was heated under reflux for 2 hours. Adding additional POCl₃(20uL) and refluxing was continued for another 3 hours. The reaction was slow. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was diluted with EtOAc, stirred with ice, and acidified to help dissolve the resulting brown solid. Extracting the mixture, and sequentially extracting the organic extract with H₂O, saturated NaHCO₃Aqueous solution, H₂O and brine. The resulting solution was dried (MgSO)₄) Filtered, evaporated and dried under high vacuum. Passing the crude product through SiO₂Chromatography, eluting with a gradient of EtOAc/hexanes (35 to 60% EtOAc), afforded 0.151g of 36 c.

Step 5-addition of 36c (62.5mg, 160. mu. mol), uracil (53.8mg, 480. mu. mol) and Cs to a microwave vial₂CO₃(104mg, 320. mu. mol) and DMSO (1.14mL) to give a light brown suspension. The mixture was heated in a closed vial at 120 ℃ for 4 hours. All the raw materials are consumed. The crude product is prepared into SiO₂Purification was performed on TLC plates with 7% MeOH/DCM to afford 7.3 mgI-5.

Example 6

N- {4- [ 2-tert-butyl-4- (6-hydroxy-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide (I-6)1

Step (ii) of1-to vial 36c (37mg, 94.7. mu. mol, Eq), 2-methoxy-6-methylpyridin-3-ylboronic acid (19.0mg, 114. mu. mol) and Cs were added₂CO₃(92.5mg, 284. mu. mol), bisAlkane (2mL) and H₂O (500. mu.L). Argon is introduced into the suspension, PdCl is added₂(dppf) (3.46mg, 4.73. mu. mol), the vial was sealed and heated at 120 ℃ for 1 hour. The mixture was dissolved in EtOAc and H₂Partitioning between O, and neutralizing with 1N HCl. The organic extracts were washed with brine and dried (MgSO)₄) Filtering, and concentrating. The crude product is prepared into SiO₂Purification on TLC plate using 30% EtOAc/hexanes gave 24.3mg (53.8%) of N- (4- (2-tert-butyl-4-chloropyrido [3, 2-d)]Pyrimidin-7-yl) phenyl) methanesulfonamide (38).

Step 2Add 38(24.3mg, 50.9. mu. mol), HBr (25.7mg, 17.3. mu.L, 153. mu. mol) and HOAc (0.5mL) to a sealable vial, fill with argon, seal, and heat at 60 ℃ for 3 hours. The mixture was cooled with EtOAc and H₂O dilution with saturated NaHCO₃And (4) drying the aqueous solution. The organic layer was separated, washed with brine and dried (MgSO)₄) Filtering and vacuum concentrating. The crude product is prepared into SiO₂Purification on TLC plate with 50% EtOAc in hexanes afforded 20.4 mgI-6.

Example 7

HCV NS5BRNA polymerase activity

The enzymatic activity of HCV polymerase (NS5B570n-Con1) was determined by incorporation of radiolabeled monophosphate nucleotides into acid-insoluble RNA products. Unincorporated radiolabeled substrate is removed by filtration and a scintillant is added to the washed and dried filter plate containing radiolabeled RNA product. The amount of RNA product produced by NS5B570n-Con1 at the end of the reaction was directly proportional to the amount of light emitted by the scintillator.

The 1B genotype N-terminal histidine-tagged HCV polymerase (NS5B570N-Con1) derived from HCVCon1 strain had a 21 amino acid deletion at the C-terminus relative to the full-length HCV polymerase, which was purified from E.coli strain BL21(DE) pLysS. A construct containing the coding sequence of hcv ns5BCon1(GenBank accession No. AJ242654) was inserted into the plasmid construct pET17b (pET17b is downstream of the T7 promoter expression cassette) and transformed into e. Individual colonies were grown overnight at 37 ℃ for use as starter cultures and later for inoculation of 10LLB medium supplemented with 100. mu.g/mL ampicillin. Protein expression was induced by the addition of 0.25mM isopropyl-beta-D-thiogalactopyranoside (IPTG) when the optical density of the culture was between 0.6 and 0.8 at 600nM, and cells were harvested after 16 to 18 hours at 30 ℃. NS5B570n-Con1 was purified to homogeneity using a three-step protocol comprising column chromatography on Ni-NTA, SP-Sepharose HP and Superdex75 resin in that order.

Each 50. mu.L of enzyme reaction contained 20nM of RNA template derived from the complementary sequence of the internal ribosome entry site (cIRES), 20nMN 5B570n-Con1 enzyme, 0.5. mu. Ci tritium-labeled UTP (PerkinElmer) catalog number TRK-412; specific activity: 30 to 60 Ci/mmol; stock concentration 7.5 × 10^-5M to 20.6 × 10^-6M), 1. mu.M each of ATP, CTP and GTP, 40mM Tris-HCl (pH8.0), 40mM NaCl, 4mM DTT (dithiothreitol), 4mM MgCl₂And 5 μ l serial dilutions of the compound in DMSO. The reaction mixtures were pooled in 96-well filter plates (catalog number MADVN0B, Millipore Co.) and incubated for 2h at 30 ℃. The reaction was stopped by the addition of 10% final (v/v) trichloroacetic acid and incubated at 4 ℃ for 40 min. The reaction was filtered, washed with 8 reaction volumes of 10% (v/v) trichloroacetic acid, 4 reaction volumes of 70% (v/v) ethanol, air dried, and 25. mu.L of a scintillator (Microscint20, Perkin Elmer) was added to each reaction well.

In thatThe amount of light emitted from the scintillator was converted to Counts Per Minute (CPM) on a plate reader (Perkin Elmer, energy Range: Low, Performance mode: Normal, count time: 1min, background subtract: No, reduced cross-talk: OFF).

In thatAndthe data is analyzed. Reactions in the absence of enzymesThe background signal is determined and subtracted from the enzyme reaction. A positive control reaction was performed in the absence of compound, from which the background corrected activity was set to 100% polymerase activity. All data are expressed as a percentage of the positive control. Substituting the data into equation (i), the concentration of compound (IC) that reduces the rate of enzyme-catalyzed RNA synthesis by 50% was calculated₅₀)：

Where "Y" corresponds to relative enzyme activity (%), "% Min" is the residual relative activity at saturating compound concentration, "% Max" is the relative maximum enzyme activity, "X" corresponds to compound concentration, and "S" is the Hill coefficient (or slope).

Example 8

HCV replicon assay

The present assay measures the ability of compounds of formula I to inhibit HCV rna replication, thereby measuring their potential use for the treatment of HCV infection. The assay uses a reporter gene as a simple readout of intracellular HCV replicon RNA levels. Immediately after the Internal Ribosome Entry Site (IRES) sequence, the Renilllaluciferase gene was introduced into the first open reading frame of the 1b genotype replicon construct NK5.1 (N.Krieger et al, J.Virol.200175(10):4614) via fusion of the self-cleaving peptide 2A from foot-and-mouth disease virus with the Neomycin Phosphotransferase (NPTII) gene (M.D.Ryan & J.Drew, EMBO199413(4): 928-933). Following in vitro transcription, RNA was electroporated into human hepatoma Huh7 cells, and G418-tolerant colonies were isolated and expanded. Stably selected cell line 2209-23 contains replicative HCV subgenomic RNA, the renilla luciferase activity expressed by the replicon reflecting its intracellular RNA levels. The assay was performed in duplicate plates, one opaque white and one clear, in order to measure the antiviral activity and cytotoxicity of the compounds in parallel, ensuring that the observed activity was not due to reduced cell proliferation or due to cell death.

HCV replicon cells (2209-23) expressing the Renillaluciferase reporter gene were cultured in Dulbecco's MEM (Invitrogen Cat. No. 10569-010) containing 5% fetal calf serum (FCS, Invitrogen Cat. No. 10082- & 147), plated at 5000 cells/well in 96-well plates, incubated overnight. after 24 hours, different dilutions of the compound in growth medium were added to the cells, followed by further incubation at 37 ℃ for three days.at the end of the incubation period, the cells were harvested from the white plates, luciferase activity was measured using the R.luciferase assay system (Promega Cat. No. E2820). all reagents described in the following paragraphs were included in the manufacturer's kit, prepared as per the manufacturer's instructions.the reagents were washed once with 100. mu.l/well of phosphate buffered saline (pH7.0) (PBS), lysed with 25. mu.l 1 × R.luciferase lysis buffer, then inserted into the incubation plate at 20 minutes. the luciferase assay concentration was measured by the luminescence signal reduction procedure described in Bertec. mu.2. mu.S.S.RTM.S.S.S.2. the luminescence signal was measured using the luminescence-signal-luminescence-2-luminescence-assay procedure₅₀I.e., the concentration of drug required to reduce replicon levels by 50% relative to control values for untreated cells.

For the cytotoxicity assay, use is made ofWST-1 reagent from Roche diagnostics (catalog No. 1644807). Add 10. mu.l of WST-1 reagent to each well of a transparent plate comprising wells containing only medium as blank. The cells were then incubated at 37 ℃ for 2 hours and the OD at 450nm was measured using an MRXRevaluation microplate reader (LabSystem) (reference filter at 650 nm). Also, CC can be calculated from the% reduction in WST-1 values vs. drug concentration plot as described above₅₀I.e., the concentration of drug required to reduce cell proliferation by 50% relative to a control value for untreated cells.

TABLE II

Example 9

Pharmaceutical compositions of the subject compounds administered via several routes were prepared as described in this example.

Composition for oral administration (A))

Mixing the above materials, and packaging into capsule containing about 100mg per capsule; one capsule will approximate the total daily dose.

Composition for oral administration (B)

The ingredients are combined and granulated with a solvent such as methanol. The formulation is then dried and tableted with a suitable tabletting machine (containing about 20mg of active compound).

Composition for oral administration(C)

Mixing the above components, and making into suspension for oral administration.

Parenteral preparation (D)

The active ingredient is dissolved in a portion of the water for injection. Sodium chloride is then added with stirring in an amount sufficient to make the solution isotonic. The remaining water for injection was added to the solution to the desired weight, filtered through a 0.2 micron filter and packaged under sterile conditions.

The features disclosed in the foregoing description, or the following claims, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for attaining the disclosed result, as appropriate, may, separately, or in any combination of such features, be utilised for realising the invention in diverse forms thereof.

The foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding. It will be apparent to those skilled in the art that changes and modifications may be made within the scope of the appended claims. Accordingly, it is to be understood that the above description is intended to be illustrative, and not restrictive. The scope of the invention should, therefore, be determined not with reference to the above description, but instead should be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled.

The patents, patent applications, and scientific literature cited herein establish the knowledge of those skilled in the art and are hereby incorporated by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. Any divergence between any reference cited herein and the specific teachings of this specification should be addressed in favor of the latter. Likewise, any divergence between the definition of a word or phrase as understood in the art and the definition of that word or phrase as specifically taught in this specification should also be resolved in favor of the latter.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

n- {4- [ 2-tert-butyl-4- (6-methyl-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide;

n- {4- [ 2-tert-butyl-4- (2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide;

n- { (S) -1- [ 2-tert-butyl-4- (2, 4-dioxo-3, 4-dihydro-2H-pyrimidin-1-yl) -quinazolin-7-yl ] -pyrrolidin-3-ylmethyl } -methanesulfonamide; and

n- {4- [ 2-tert-butyl-4- (6-hydroxy-2-oxo-1, 2-dihydro-pyridin-3-yl) -quinazolin-7-yl ] -phenyl } -methanesulfonamide.

2. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prevention of Hepatitis C Virus (HCV).

3. The use of claim 2, further comprising co-administration of at least one immune system modulator and/or at least one antiviral agent that inhibits replication of HCV.

4. Use according to claim 3, wherein the immune system modulator is an interferon, interleukin, tumor necrosis factor or colony stimulating factor.

5. Use according to claim 3, wherein the immune system modulator is an interferon or a chemically derivatized interferon.

6. The use according to claim 3, wherein the antiviral agent is selected from the group consisting of HCV protease inhibitors, other HCV polymerase inhibitors, HCV helicase inhibitors, HCV primase inhibitors and HCV fusion inhibitors.

7. A non-therapeutic method of inhibiting HCV replication in a cell by delivering a compound of claim 1, or a pharmaceutically acceptable salt thereof.

8. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, in admixture with at least one pharmaceutically acceptable carrier.

9. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, in admixture with at least one pharmaceutically acceptable diluent.

10. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, in admixture with at least one pharmaceutically acceptable excipient.