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WO2008141385A1 - Inhibiteurs de polymérases virales - Google Patents

Inhibiteurs de polymérases virales Download PDF

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Publication number
WO2008141385A1
WO2008141385A1 PCT/AU2008/000714 AU2008000714W WO2008141385A1 WO 2008141385 A1 WO2008141385 A1 WO 2008141385A1 AU 2008000714 W AU2008000714 W AU 2008000714W WO 2008141385 A1 WO2008141385 A1 WO 2008141385A1
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optionally substituted
alkyl
group
compound
formula
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PCT/AU2008/000714
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English (en)
Inventor
Jeffrey Peter Mitchell
Michael John Lilly
John Nicholas Lambert
Alistair George Draffan
Silas Bond
Richard Hufton
Saba Jahangiri
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Biota Scientific Management Pty Ltd
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Priority claimed from AU2007902702A external-priority patent/AU2007902702A0/en
Application filed by Biota Scientific Management Pty Ltd filed Critical Biota Scientific Management Pty Ltd
Publication of WO2008141385A1 publication Critical patent/WO2008141385A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the present invention relates to viral polymerase inhibitors, in particular inhibitors of viral polymerases within the Flaviviridae family such as hepatitis C virus (HCV) , processes for their preparation and their use in the treatment of infections.
  • HCV hepatitis C virus
  • the Flaviviridae is a group of positive single- stranded RNA viruses with a genome size from 9-15 kb.
  • the Flaviviridae consists of various genera including: 1. Hepaciviruses : This genus contains only one species, the Hepatitis C virus (HCV) , which is composed of many genotypes and subtypes .
  • HCV Hepatitis C virus
  • Flaviviruses This genus includes the Dengue virus, Japanese Tick-Borne and the Yellow Fever virus.
  • Flaviviruses Apart from these major groups, there are some additional Flaviviruses that are unclassified.
  • Pestiviruses This genus includes three serotypes of bovine viral diarrhoea virus, but no known human pathogens .
  • HCV Hepatitis C virus
  • the present invention relates to compounds which are viral polymerase inhibitors , in particular inhibitors of viral polymerases within the Flaviviridae family such as HCV .
  • Y is independently selected from the group consisting of N, NO and CR in which R is selected from the group consisting of H, halogen, SH, S-optionally substituted Ci- 6 alkyl , OH, optionally substituted d- 6 alkoxy , NH 2 , NH- optionally substituted Ci -6 alkyl, N-di-optionally substituted Ci -6 alkyl and NHCOR 1 in which R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl, optionally substituted C 2 - 6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3- 8 cycloalkyl, optionally substituted aryl and optionally substituted heterocyclyl;
  • Z is selected from the group consisting of 0, S and NR 1 ' in which R 1 ' is selected from the group consisting of H, OH, optionally substituted Ci -6 alkyl, optionally substituted C 2 - 6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl and optionally substituted heterocyclyl; R 1 is selected from the group consisting of optionally substituted C 2-e alkyl, d -6 alkylene optionally substituted C 3-8 cycloalkyl, Ci -6 alkylene optionally substituted aryl, C 1-6 alkylene optionally substituted heterocyclyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl and optionally substituted heterocyclyl; R 2 is -L-R"' in which L is absent or a linker and R"' is selected from the group consisting of optionally substituted C 3-8 cycloalkyl, optional
  • R 3 is selected from the group consisting of C 2 - 6 alkyl; substituted Ci -6 alkyl; halo,- CN; optionally substituted aryl; optionally substituted heterocyclyl; NO 2 ; NR 12 R 13 in which R 12 and R 13 are independently selected from the group consisting of H provided that not both are H, optionally substituted Ci- 6 alkyl, S(O) P R 14 in which p is 0 to 2 and R 14 is selected from the group consisting of H, optionally substituted Ci -6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl optionally substituted heterocyclyl and NR 15 R 16 in which R 15 and R 16 are independently selected from the group consisting of H, optionally substituted Ci_ 6 alkyl, optionally substituted C 3-6 cycloalkyl, optionally substituted aryl and optionally substituted heterocycl
  • R 4 is selected from the group consisting of amino acid; optionally substituted C 3 - 8 cycloalkyl; optionally substituted aryl; optionally substituted heterocyclyl; OR 7 in which R 7 is selected from the group consisting of H, optionally substituted Ci- 6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl and optionally substituted heterocyclyl; and NR 5 R 6 ; in which R 5 and R 6 are independently selected from the group consisting of H; optionally substituted Ci -6 alkyl; optionally substituted C 2-6 alkenyl; optionally substituted C 2-6 alkynyl; optionally substituted (CH 2 ) m optionally substituted aryl; optionally substituted (CH 2 ) m optionally substituted heterocyclyl in which m is 0 to 4; S(O) P R 8 in which p is as defined above
  • the compounds of formula I are inhibitors of hepatitis C.
  • the compounds of formula I inhibit RNA synthesis by the RNA dependent RNA polymerase of HCV (the NS5B protein encoded by HCV) .
  • NS5B inhibitors have been clinically validated as potential antiviral agents for the treatment of HCV infection.
  • a HCV polymerase inhibitor such as a NS5B inhibitor comprising the compound of formula I defined above.
  • an antiviral agent comprising the compound formula I defined above.
  • the compound of formula I may be administered in the form of a pharmaceutical composition together with a pharmaceutically acceptable carrier.
  • a pharmaceutical composition comprising the compound of formula I defined above and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition additionally comprises a therapeutically effective amount of one or more antiviral agents such as at least one other anti-HCV agent.
  • a method for the treatment of a Flaviviridae viral infection such as a HCV infection which comprises administering an effective amount of the compound of formula I defined above to a subject in need thereof.
  • a method of inhibiting the RNA-dependent RNA polymerase activity of the enzyme NS5B, encoded by HCV comprising exposing the enzyme NS5B to an effective amount of the compound of formula I defined above under conditions where the RNA-dependent RNA polymerase activity of the enzyme NS5B is inhibited.
  • a method of inhibiting HCV replication comprising exposing a cell infected with HCV to an effective amount of the compound of formula I defined above under conditions where replication of HCV is inhibited.
  • the present invention relates to compounds of formula I which inhibit viral polymerases and are useful in the treatment of Flaviviridae viral infections such as hepatitis C.
  • the present invention relates to compounds of formula I, salts, solvates, derivatives, prodrugs, tautomers and/or isomers thereof as defined above.
  • each Y is CH or one or two of Y is N and the others are CH and n is 1 to 4, more preferably n is 1.
  • compounds of formula I have the formula Ia
  • R 1 , R 2 and R 3 are as defined above; Y 1 is CH; Y 2 is CH; and Y 3 and Y 4 are independently selected from the group consisting of N and CH.
  • R 1 is preferably optionally substituted C 3-8 cycloalkyl such as cyclohexyl, optionally substituted aryl such as phenyl or optionally substituted heterocyclyl .
  • L is absent optionally substituted Ci- 6 alkylene or optionally substituted Ci -6 acyl in R 2 .
  • R 3 is preferably located at any one or more of positions 4, 5 or 6, preferably the 5 position.
  • R 3 is selected from the group consisting of C 2-6 alkyl; substituted Ci -6 alkyl; CN; optionally substituted aryl; optionally substituted heterocyclyl; NR 12 R 13 in which R 12 and R 13 are independently selected from the group consisting of H provided that not both are H, optionally substituted Ci -6 alkyl, S(O) P R 14 in which p is 0 to 2 and R 14 is selected from the group consisting of H, C 2- 6 alkyl, substituted Ci- S alkyl, optionally substituted C 2- 6 alkenyl, optionally substituted C 2 _ 6 a-lkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl optionally substituted heterocyclyl and NR 15 R 16 in which R 15 and R 16 are independently selected from the group consisting of H, optionally substituted Ci -6 alkyl, optionally substituted C 3 - 6 cycoalkyl, optionally substituted aryl and optionally
  • R 4 is selected from the group consisting of amino acid; optionally substituted C 3 - 8 cycloalkyl; optionally substituted aryl; optionally substituted heterocyclyl; OR 7 in which R 7 is selected from the group consisting of C 2 - 6 alkyl, substituted Ci -6 alkyl, optionally substituted C 2 - 6 alkenyl, optionally substituted C 2 .
  • R 5 and R 6 are independently selected from the group consisting of H; optionally substituted Ci_ 6 alkyl; optionally substituted C 2-6 alkenyl; optionally substituted C 2 - 6 alkynyl; optionally substituted (CH 2 ) ra optionally substituted aryl; optionally substituted (CH 2 ) m optionally substituted heterocyclyl in which m is 0 to 4; S(O) P R 8 in which p is as defined above and R 8 is selected from the group consisting of H, optionally substituted C !
  • R 15 and R 16 are as defined above;
  • CR 17 R 18 COR 19 in which R 17 and R 18 are independently selected from the group consisting of H and optionally substituted C 1-6 alkyl or R 17 and R 18 together with the C to which they are attached form an optionally substituted 3 to 6-membered ring and R 19 is selected from the group consisting of 0R 15 in which R 15 is as defined above and NR 20 R 21 in which R 20 and R 21 are selected from the group consisting of H, optionally substituted d- 6 alkyl, optionally substituted aryl and optionally substituted heterocyclyl; and CR 22 (CO 2 R 23 ) CH 2 R 24 in which R 22 and R 23 are independently selected from the group consist
  • R 5 and R 6 together with the N to which they are attached form an optionally substituted 5 or 6-membered ring.
  • R 3 is optionally substituted heterocyclyl, it is preferably a 5 or 6 membered N-containing heterocyclyl such as tetrazolyl .
  • R 3 is optionally substituted Ci- 6 alkyl, halo, CN, NO 2 or optionally substituted heterocyclyl.
  • R 2 may be in which
  • A is selected from the group consisting of C 3 - 8 cycloalkyl, aryl and heterocyclyl; and R 8 is selected from the group consisting of hydrogen, halo, cyano, nitro, amino, optionally substituted Ci -6 alkanoylamino, optionally substituted Ci -6 alkylsulfonyl, optionally substituted Ci -6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl, COOR 9 in which R 9 is hydrogen or optionally substituted Ci -6 alkyl, CONH- (CH 2 ) n -R 10 in which R 10 is optionally substituted C 1-6 alkyl, Ci -6 alkoxycarbonyl or Ci -6 alkanoylamino and n is 1 to 6 and OR 11 in which R 11 is hydrogen, optionally substituted Ci -6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl optionally substituted C 3
  • A is preferably cyclohexyl, phenyl or a 5 or 6 membered N-containing heterocyclyl such as pyridyl .
  • Z is 0.
  • R 1 is preferably optionally substituted C 3- scycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl or C 2-6 alkyl substituted with optionally substituted C 3 - 8 cylcoalkyl, optionally substituted aryl or optionally substituted heterocyclyl.
  • R 8 is OR 11 in which R 11 is C 1- 6 alkyl or benzyl .
  • Ic has the formula Ig
  • R 1 , R 2 and R 4 are as defined above.
  • R 1 is C 5 . 7 cycloalkyl such as cyclohexyl; phenyl; or a 5 or 6 membered N-containing heterocyclyl such as pyridyl and R 2 is C 5-7 cycloalkyl such as cyclohexyl; optionally substituted phenyl; or heterocyclyl containing an N, 0 or S such as pyridyl, furyl or thienyl .
  • Subclasses of compounds of this type are as follows: (i) 7 in which R 1 , R 2 and R 7 are as defined above.
  • R 1 is cyclohexyl or phenyl
  • R 2 is phenyl optionally substituted with Ci- S alkoxy or benzyloxy
  • R 7 is H, Ci -6 alkyl such as Ci -4 alkyl for example methyl; optionally substituted aryl such as phenyl optionally substituted with a d- 6 alkyl which is substituted with amino, acyl and/or Ci -6 alkoxy; or N-containing heterocyclyl such as indolyl optionally substituted with a Ci -6 alkyl which is substituted with amino, acyl and/or Ci -6 alkoxy.
  • Representative examples are as follows: Table 1
  • R 1 , R 2 , R 5 and R 6 are as defined above.
  • R 1 is cyclohexyl or phenyl
  • R 2 is phenyl
  • one of R 5 and R 6 is H and the other is Ci -6 alkyl substituted with optionally substituted phenyl or optionally substituted heterocyclyl, both R 5 and R 6 are H or together with the N to which they are attached form an optionally substituted 5 or 6-membered heterocyclyl.
  • the optional substituents on the phenyl are preferably Ci -6 alkyl, halo such as Cl or F, d- S alkoxy, hydroxy, C 2 - 6 alkenyleneacylCi- 6 alkoxy or acylCi_ 6 alkoxy .
  • the heterocyclyl is preferably an unsaturated 5 or 6- membered heteromonocyclic group containing 1 or 2 N atoms such as imidazolyl or pyridinyl; unsaturated condensed heterocyclyl containing 1 to 3 N atoms and optionally S and 0 atoms such as benzothiazol-2-yl, 5- (3-oxo-3 , 4- dihydro-2H-benzo [1, 4] oxazin-6-yl) , benzoxazolyl and benzotriazolyl; unsaturated 5 or 6 -membered heteromonocyclic group containing N and S such as thiazolyl; or unsaturated condensed heterocyclic group containing 0 atoms such as 2-oxo-2H-chromen-7-yl .
  • the optional substituents on the heterocyclyl are preferably Ci -6 alkyl, Ci- ⁇ alkyleneacylCi-ealkoxy, acylCi- 6 alkoxy, CF 3 , Ci -6 alkyleneCi -6 alkoxy, carboxy or C 1 - 6 alkenyenecarboxy.
  • the optionally substituted 5 or 6-membered heterocyclyl is preferably a saturated N-containing 5 or 6-membered heterocyclyl such as pyrrolidinyl .
  • R 1 , R 2 , R 6 , R 17 , R 18 and R 19 are as defined above.
  • R 1 is cyclohexyl, phenyl or pyridyl
  • R 2 is cyclohexyl or phenyl
  • R 6 is H
  • R 17 and R 18 are both H or Ci. 6 alkyl, one is H and the other is Ci- S alkyl or Ci -6 alkyl substituted with optionally substituted phenyl or optionally substituted heterocyclyl or together with the C to which they are attached form a 4-membered ring
  • R 19 is OR 15 in which R 15 is as defined above or NR 20 R 21 in which one of R 20 and R 21 is H and the other is optionally substituted phenyl or optionally substituted heterocyclyl.
  • the optionally substituent on the phenyl or heterocyclyl is preferably hydroxy, acylCi -6 alkoxy, C 2- 6 alkenyleneacyl Ci -6 alkoxy, d- 6 alkylenecarboxy, C 1 - 6 alkylenecarboxy, carboxy or Ci- ⁇ alkylacylCi-ealkoxy.
  • the heterocyclyl is preferably an unsaturated condensed heterocyclyl containing 1 to 3 N atoms such as indolyl or quinolinyl.
  • R 1 , R 2 , R 6 , p and R 8 are as defined above.
  • R 1 is cyclohexyl or phenyl;
  • R 2 is optionally substituted phenyl;
  • R 6 is H;
  • p is 2 and
  • R 8 is optionally substituted C 3 - 8 cycloalkyl, optionally substituted phenyl, optionally substituted heterocyclyl, Ci -6 alkyl, d- 6 alkyl substituted with optionally substituted phenyl, C 2-6 alkenyl substituted with optionally substituted phenyl or NR 15 R 16 in which R 15 and R 16 are as defined above.
  • the optional substituents on the phenyl or heterocyclyl are preferably Ci- 6 alkoxy, halo such as Cl, Br or F, acylCi -6 alkoxy, nitro, carboxy, amidoC 1-6 alkyl or CF 3 .
  • the C 3-8 cycloalkyl is preferably C 3 cycloalkyl.
  • the heterocyclyl is preferably an unsaturated 5- membered heteromonocyclic group containing 1 to 2 S atoms such as thiophenyl ; unsaturated condensed heterocyclyl containing 1 to 3 N atoms and optionally S atoms such as benzothiazol-2-yl,benzo [1,2,5] oxodiazol-4-yl and benzothiazol-2-yl or an unsaturated condensed heterocyclyl containing S atoms such as benzothiophenyl .
  • Representative examples are as follows : Table 4
  • the compounds are of formula Ih
  • both R 1 and R 2 are phenyl and R 9 and R 10 are both H or one is H and the other is SO P R 14 in which p and R 14 are as defined above.
  • Subclasses of compound of this type are as follows: (i) in which R 1 , R 2 , p and R 14 are as defined above.
  • R 1 and R 2 are phenyl; p is 2; and R 14 is optionally substituted phenyl.
  • the substituents on the phenyl are preferably nitro, acylCi -6 alkyl, Ci -6 alkyl or halo such as Cl.
  • R 1 , R 2 and R 25 are as defined above.
  • R 1 is heterocyclyl such as pyridyl
  • R 2 is cyclohexyl
  • R 25 is Ci- 6 alkylene substituted with carboxylic acid or phenyl substituted with C 1-6 alkyl, carboxylic acid or methoxy carboxylic acid.
  • the compounds are of the formula Ii
  • R 1 is phenyl or cyclohexyl
  • R 2 is optionally substituted phenyl
  • R 3' is C 1-6 alkylene substituted with CO 2 CF 3 , Br, CN, NO 2 or tetrazolyl .
  • Representative examples are as follows:
  • Ci -6 alkyl refers to straight chain or branched chain hydrocarbon groups having from 1 to 6 carbon atoms. Examples include ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert -butyl, pentyl, neopentyl and hexyl .
  • C 2-6 alkenyl refers to straight chain or branched chain hydrocarbon groups having at least one double bond of either E or Z stereochemistry where applicable and 2 to 6 carbon atoms. Examples include vinyl, 1-propenyl, 1- and 2-butenyl and 2-methyl-2- propenyl .
  • C 2-6 alkynyl refers to straight chain or branched chain hydrocarbon groups having at least one triple bond and 2 to 6 carbon atoms. Examples include ethynyl, 1- or 2-propynyl, 1-, 2- or 3- butynyl and methyl-2-propynyl.
  • C 3-8 cycloalkyl refers to non-aromatic cyclic hydrocarbon groups having from 3 to 8 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloctyl, preferably cyclopentyl or cyclohexyl.
  • Ci -6 alkoxy refers to an oxy-containing radical having Ci -6 alkyl as defined above. Examples include methoxy, ethoxy, propoxy, butoxy, tert-butoxy and pentoxy.
  • aryl refers to single, polynuclear, conjugated or fused residues of aromatic hydrocarbons. Examples include phenyl, biphenyl, terphenyl, quaterphenyl , naphthyl, tetrahydronaphthyl , anthracenyl, dihydroanthracenyl , benzanthracenyl, dibenxanthracenyl and phenanthrenyl . A preferred aryl is phenyl.
  • heterocyclyl refers to saturated or unsaturated, monocyclic or polycyclic hydrocarbon groups containing at least one heteroatom atom selected from the group consisting of nitrogen, sulphur and oxygen.
  • Suitable heterocyclyls include N-containing heterocyclic groups, such as, unsaturated 3 to 6-membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl; saturated 3 to 6-membered heteromonocyclic groups containing 1 to 4 nitrogen atoms, such as, pyrrolidinyl, imidazolidinyl, piperidino or piperazinyl; unsaturated condensed heterocyclic groups containing 1 to 5 nitrogen atoms, such as indolyl, isoindo
  • Preferred heterocyclyls are pyridinyl, indolyl, benzoxazolyl, benzotriazolyl, thiazolyl, pyrrolidinyl, quinolinyl, thiophenyl, benzothiophenyl, tetrazolyl, furyl, morpholinyl, piperidinyl and 5-oxo-pyrrolidinyl .
  • linker group refers to a linear or branched chain of atoms connecting two distinct molecular subunits.
  • Suitable linker groups include optionally substituted Ci -6 acyl, optionally substituted Ci -6 alkylene, optionally substituted C 2-6 alkenylene, optionally substituted C 2 - 6 alkynylene, optionally substituted C 3-8 cycloalkylene, optionally substituted C 3-8 cycloalkenylene, optionally substituted divalent aryl, optionally substituted divalent heterocyclyl, optionally substituted Ci -6 SuIfonamido, optionally substituted Ci -6 amido and optionally substituted Ci -6 acylamino.
  • Preferred linker groups for R 2 are optionally substituted Ci -6 alkylene or optionally substituted Ci -6 acyl.
  • Preferred linker groups for R 3 are optionally substituted C x-6 alkylene, optionally substituted C 2-6 alkenylene, optionally substituted divalent aryl, optionally substituted divalent heterocyclyl, optionally substituted Ci -6 acyl or optionally substituted C 3 _ 8 cycloalkyl.
  • amino acid refers to any of the twenty (20) common amino acids generally accepted in the peptide art. Examples include alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glycine, histidine, 5-hydroxyIysine, 4-hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, ⁇ — aminoadipic acid, ⁇ —amino-n-butyric acid, 3,4- dihydroxyphenylalanine , homoserine, ⁇ —methylserine, ornithine, pipecolic acid and thyroxine.
  • halo refers to fluorine, chlorine, bromine or iodine .
  • Ci -6 alkanoyl refers to a radical provided by the residue after removal of hydroxyl from an organic acid. Examples include formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl and trifluoroacetyl .
  • optionally substituted refers to a group that may or may not be further substituted with one or more groups selected from Ci -6 alkyl, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, aryl, CF 3 , heterocyclyl, halo, haloCi_ 6 alkyl, haloC 3-6 cycloalkyl, haloC 2-6 alkenyl, - S l -
  • Preferred optional substituents are selected from the group consisting of Ci -6 alkyl, CF 3 , C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, aryl, heterocyclyl, halo, haloaryl, haloheterocycylyl, hydroxy, Ci -4 alkoxy, aryloxy, carboxy, amino, acyl, arylacyl, heterocyclylacyl, acylamino, acyloxy, acylCi -s alkoxy, nitro, arylsulphonyl, cyano and the like.
  • salts of the compound of formula I are preferably pharmaceutically acceptable, but it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the present invention, since these are useful as intermediates in the preparation of pharmaceutically acceptable salts.
  • pharmaceutically acceptable salts include salts of pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium; acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric , sulphuric, phosphoric, nitric, carbonic, boric, sulfamic and hydrobromic acids; or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, trihalomethanesulphonic , toluenesulphonic ,
  • Salts of amine groups may also comprise quaternary ammonium salts in which the amino nitrogen atom carries a suitable organic group such as an alkyl , alkenyl , alkynyl or aralkyl moiety.
  • the salts may be formed by conventional means, such as by reacting the free base form of the compound with one or more equivalents of the appropriate acid in a solvent or medium in which the salt is insoluble, or in a solvent such as water which is removed in vacuo or by freeze drying or by exchanging the anions of an existing salt for another anion on a suitable ion exchange resin.
  • solvates may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.
  • the derivatives of the compound of formula I are preferably pharmaceutically acceptable.
  • pharmaceutically acceptable derivative refers to any pharmaceutically acceptable salt, hydrate, ester, amide, active metabolite, analogue, residue or any other compound which is not biologically or otherwise undesirable and induces the desired pharmacological and/or physiological effect.
  • pro-drug refers to functional derivatives of the compound of formula I which are readily convertible in vivo into the required compound of formula I. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs” ed. H. Bundgaard, Elsevier, 1985.
  • a prodrug may be a pharmacologically inactive derivative of the active compound that requires transformation within the body in order to release the active compound, and that has improved delivery properties over the active compound.
  • the transformation in vivo may be, for example, as the result of some metabolic process, such as chemical or enzymatic hydrolysis of a carboxylic, phosphoric or sulphate ester, or reduction or oxidation of a susceptible functionality.
  • tautomer refers to compounds of formula I which are capable of existing in a state of equilibrium between two isomeric forms. Such compounds may differ in the bond connecting two atoms or groups and the position of these atoms or groups in the compound.
  • the formylation step (a) may be carried out using any suitable known process such as a Vilsmeyer formylation or modified version thereof.
  • the cycloaddition step (b) may involve a diene and a dienophile or functionally equivalent groups thereof.
  • R 3 can be introduced using any suitable known technique such as aromatic substitution chemistry or by appropriate choice of reagents in the cycloaddition reaction.
  • RNA dependent RNA polymerase of HCV RNA dependent RNA polymerase of HCV
  • the compounds of formula I act by specific inhibition of NS5B
  • the compounds may be tested for the lack of inhibitory activity in an assay measuring the activity of an RNA-dependent RNA polymerase other than HCV polymerase or in a DNA dependent RNA polymerase assay.
  • the administration of the compound of the present invention may be any suitable means that results in a concentration of the compound that is effective to yield the desired therapeutic or prophylactic response.
  • the compound may be contained in any appropriate amount in any suitable carrier and is generally present in an amount of 1-95% by weight of the total weight of the composition.
  • the carrier must be "pharmaceutically acceptable" in the sense of being compatible with other ingredients of the composition and not injurious to the subject.
  • the pharmaceutical composition may additionally comprise a therapeutically effective amount of one or more antiviral agents such as ribavirin, an antiviral nucleoside, polymerase inhibitor, protease inhibitor and/or inhibitor of viral entry, assembly or egress.
  • the composition may also additionally comprise at least one immunomodulatory agent for example an interferon or interferon derivative and/or an inhibitor of inosine-5 ' -monophosphate dehydrogenase (IMPDH) .
  • IMPDH inosine-5 ' -monophosphate dehydrogenase
  • the composition may be provided in a dosage form that is suitable for oral, parenteral (including intravenous, intramuscular, subcutaneous and intradermal), rectal, vaginal, nasal, inhalation, topical or ocular administration routes.
  • the composition may be in form of tablets, capsules, pills, powders, granulates, suspensions, emulsions, liquids, gels including hydrogels, pastes, ointments, creams, plasters, drenches, delivery devices, suppositories, enemas, injectables, implants, sprays or aerosols.
  • the pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see , e.g., Remington: The Science and Practice of Pharmacy, (19 th ed.). A. R. Gennaro, 1995, Mack Publishing Company, Easton, PA. and Encyclopedia of Pharmaceutical Technology, eds . J.Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York) .
  • compositions may be formulated to release the active compound substantially immediately upon administration or at any predetermined time or time period after administration.
  • controlled release formulations include (i) formulations that create a substantially constant concentration of the active compound within the body over an extended period of time; (ii) formulations that after a predetermined lay time create a substantially constant concentration of the active compound within the body over an extended period of time; (iii) formulations that sustain active compound action during a predetermined time period by maintaining a relatively, constant, effective active compound level in the body with concomitant minimization of undesirable side effects associated with fluctuations in the plasma level of the active compound (sawtooth kinetic pattern) ; (iv) formulations that localise active compound action by, e.g., special placement of a controlled release composition adjacent to or in the diseased tissue or organ; and (v) formulations that target active compound action by using carriers or chemical derivatives to deliver the active compound to a particular target cell type.
  • Administration of compounds in the form of a controlled release formulation is especially preferred in cases in which the compound has (i) a narrow therapeutic index (i.e., the difference between the plasma concentration leading to harmful side effects or toxic reactions and the plasma concentration leading to a therapeutic effect is small; in general, the therapeutic index, TI, is defined as the ratio of median lethal dose (LD 50 ) to median effective dose (ED 50 ) ) ; (ii) a narrow absorption window in the gastro-intestinal tract; or (iii) a very short biological half-like so that frequent dosing during a day is required in order to sustain the plasma level at a therapeutic level.
  • a narrow therapeutic index i.e., the difference between the plasma concentration leading to harmful side effects or toxic reactions and the plasma concentration leading to a therapeutic effect is small
  • the therapeutic index, TI is defined as the ratio of median lethal dose (LD 50 ) to median effective dose (ED 50 ) ) ; (ii) a narrow absorption window in the gastro-intestinal tract; or (i
  • controlled release is obtained by appropriate selection of various formulation parameters and ingredients, including, e.g., various types of controlled release compositions and coatings.
  • the active compound is formulated with appropriate excipients into a pharmaceutical composition, that, upon administration to the subject, releases the active compound in a controlled manner. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches and liposomes.
  • Formulations for oral use include tablets containing the active compound in a mixture with non-toxic pharmaceutically acceptable excipients.
  • excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, mirocrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate) ; granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates or alginic acid); binding agents (e. g.
  • Other pharmaceutically acceptable excipients can be colourants, flavouring agents, plasticisers, humectants, buffering agents and the like.
  • the tablets may be uncoated or they may be coated by known techniques, optionally to delay disintegration and absorption in the gastrointestinal tract and thereby providing a sustained action over a longer period.
  • the coating may be adapted to release the active compound in a predetermined pattern (e.g., in order to achieve a controlled release formulation) or it may be adapted not to release the active compound until after passage of the stomach (enteric coating) .
  • the coating may be a sugar coating, a film coating (e.g., based on hydroxypropyl methylcellulose, methylcellulose, methyl hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers, polyethylene glycols and/or polyvinylpyrrolidone) , or an enteric coating (e.g., based on methacrylic acid copolymer, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, shellac and/or ethylcellulose) .
  • a film coating e.g., based on hydroxypropyl methylcellulose, methylcellulose, methyl hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers, polyethylene glycols and/or polyvinylpyrrolidone
  • enteric coating e.g
  • a time delay material such as, glyceryl monostearate or glyceryl distearate may be employed.
  • the solid tablet compositions may include a coating adapted to protect the composition from unwanted chemical changes, (e.g., chemical degradation prior to the release of the active compound) .
  • the coating may be applied on the solid dosage form in a similar manner as that described in Encyclopedia of Pharmaceutical Technology, supra.
  • Formulations for oral use may also be presented as chewing tablets or as hard gelatin capsules wherein the active compound is mixed with an inert solid diluent ⁇ e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active compound is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.
  • an inert solid diluent e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin
  • water or an oil medium for example, peanut oil, liquid paraffin or olive oil.
  • Powders and granulates may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g, a mixer, a fluid bed apparatus or a spray drying equipment.
  • Powders, dispersible powders, or granules suitable for preparation of an aqueous suspension by addition of water are convenient dosage forms for oral administration.
  • Formulation as a suspension provides the active compound in a mixture with a dispersing or wetting agent, suspending agent, and one or more preservatives.
  • Suitable dispersing or wetting agents are, for example, naturally- occurring phosphatides ⁇ e.g., lecithin or condensation products of ethylene oxide with a fatty acid, a long chain aliphatic alcohol or a partial ester derived from fatty acids) and a hexitol or a hexitol anhydride ⁇ e.g., polyoxyethylene stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan monooleate and the like) .
  • Suitable suspending agents are, for example, sodium carboxymethylcellulose, methylcellulose, sodium alginate and the like.
  • the compound may be administered parenterally by injection, infusion or implantation (intravenous, intramuscular, subcutaneous or the like) in dosage forms, formulations or via suitable delivery devices or implants containing conventional, non-toxic pharmaceutically acceptable carriers.
  • injection, infusion or implantation intravenous, intramuscular, subcutaneous or the like
  • suitable delivery devices or implants containing conventional, non-toxic pharmaceutically acceptable carriers.
  • the formulation and preparation of such compositions is well known to those skilled in the art of pharmaceutical formulation. Specific formulations can be found in Remington: The Science and Practice of Pharmacy, supra .
  • compositions for parenteral use may be presented in unit dosage forms (e.g., in single-dose ampoules) or in vials containing several doses and in which a suitable preservative may be added (see below) .
  • the composition may be in form of a solution, a suspension, an emulsion, an infusion device or a delivery device for implantation or it may be presented as a dry powder to be reconstituted with water or another suitable vehicle before use.
  • the composition may include suitable parenterally acceptable carriers.
  • the active compound may be incorporated into microspheres, microcapsules, nanoparticles, liposomes or the like for controlled release.
  • the composition may include suspending, solubilizing, stabilizing, pH-adjusting agents and/or dispersing agents.
  • the pharmaceutical compositions may be in the form suitable for sterile injection.
  • a parenterally acceptable liquid vehicle that may be employed are water, water adjusted to a suitable pH by addition of an appropriate amount of hydrochloric acid, sodium hydroxide or a suitable buffer, 1, 3-butanediol, Ringer's solution and isotonic sodium chloride solution.
  • the aqueous formulation may also contain one or more preservatives (e.g., methyl, ethyl or n-propyl p-hydroxybenzoate) .
  • a dissolution enhancing or solubilising agent can be added or the solvent may include 10- 60% w/w of propylene glycol or the like.
  • suitable dosage forms for a composition include suppositories (emulsion or suspension type) and rectal gelatin capsules (solutions or suspensions) .
  • the active compound is combined with an appropriate pharmaceutically acceptable suppository base such as cocoa butter, esterified fatty acids, glycerinated gelatin and various water-soluble or dispersible bases like polyethylene glycols and polyoxyethylene sorbitan fatty acid esters.
  • an appropriate pharmaceutically acceptable suppository base such as cocoa butter, esterified fatty acids, glycerinated gelatin and various water-soluble or dispersible bases like polyethylene glycols and polyoxyethylene sorbitan fatty acid esters.
  • Various additives, enhancers or surfactants may be incorporated.
  • compositions suitable for vaginal administration may be presented as 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 active compound may be administered by any of the methods and formulations employed in the art for administration to the respiratory tract.
  • the active compound may be administered in the form of a solution or a suspension or as a dry powder.
  • Solutions and suspensions will generally be aqueous, for example prepared from water alone (for example sterile or pyrogen-free water) or water and a physiologically acceptable co-solvent (for example ethanol, propylene glycol or polyethylene glycols such as PEG 400) .
  • a physiologically acceptable co-solvent for example ethanol, propylene glycol or polyethylene glycols such as PEG 400
  • Such solutions or suspensions may additionally contain other excipients for example preservatives (such as benzalkonium chloride) , solubilising agents/surfactants such as polysorbates (eg. Tween 80, Span 80, benzalkonium chloride) , buffering agents, isotonicity-adjusting agents (for example sodium chloride) , absorption enhancers and viscosity enhancers.
  • Suspensions may additionally contain suspending agents (for example microcrystalline cellulose and carboxymethyl cellulose sodium) .
  • Solutions or suspensions are applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or spray.
  • the formulations may be provided in single or multidose form. In the latter case a means of dose metering is desirably provided.
  • a dropper or pipette this may be achieved by the subject administering an appropriate, predetermined volume of the solution or suspension.
  • a spray this may be achieved for example by means of a metering atomising spray pump.
  • Administration to the respiratory tract may also be achieved by means of an aerosol formulation in which the compound is provided in a pressurised pack with a suitable propellant, such as a chlorofluorocarbon (CFC) , for example dichlorodifluoromethane , trichlorofluoromethane or dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant such as a chlorofluorocarbon (CFC) , for example dichlorodifluoromethane , trichlorofluoromethane or dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • CFC chlorofluorocarbon
  • the aerosol may conveniently also contain a surfactant such as lecithin.
  • the dose of active compound may be controlled by provision of a metered valve.
  • the active compound may be provided in the form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidine (PVP) .
  • a powder base such as lactose, starch, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidine (PVP) .
  • PVP polyvinylpyrrolidine
  • the powder composition may be presented in unit dose form, for example in capsules or cartridges of eg. gelatin, or blister packs from which the powder may be administered by means of an inhaler.
  • the active compound In formulations intended for administration to the respiratory tract, including intranasal formulations, the active compound will generally have a small particle size, for example of the order of 5 microns or less. Such a particle size may be obtained by means known in the art, for example by micronisation. When desired, formulations adapted to give sustained release of the active compound may be employed.
  • the active compound may be administered by oral inhalation as a free-flow powder via a "Diskhaler” (trade mark of Glaxo Wellcome pic or a meter dose aerosol inhaler.
  • a "Diskhaler” trade mark of Glaxo Wellcome pic or a meter dose aerosol inhaler.
  • compositions may also be administered topically on the skin for percutaneous absorption in dosage forms or formulations containing conventionally non-toxic pharmaceutical acceptable carriers and excipients including microspheres and liposomes.
  • the formulations include creams, ointments, lotions, liniments, gels, hydrogels, solutions, suspensions, sticks, sprays, pastes, plasters and other kinds of transdermal drug delivery systems.
  • the pharmaceutically acceptable carriers may include emulsifying agents, antioxidants, buffering agents, preservatives, humectants, penetration enhancers, chelating agents, gel forming agents, ointment bases, perfumes and skin protective agents.
  • emulsifying agents are naturally occurring gums (e.g., gum acacia or gum tragacanth) and naturally occurring phosphatides (e.g., soybean lecithin and sorbitan monooleate derivatives) .
  • antioxidants are butylated hydroxy anisole (BHA) , ascorbic acid and derivatives thereof, tocopherol and derivatives thereof, butylated hydroxy anisole and cysteine.
  • preservatives are parabens, such as methyl or propyl p- hydroxybenzoate and benzalonium chloride .
  • humectants are glycerin, propylene glycol, sorbitol and urea.
  • Examples of penetration enhancers are propylene glycol, DMSO, triethanolamine, N,N-dimethylacetamide, N, N- dimethylformamide, 2-pyrrolidone and derivatives thereof, tetrahydrofurfuryl alcohol and Azone.RTM.
  • Examples of chelating agents are sodium EDTA, citric acid and phosphoric acid.
  • Examples of gel forming agents are Carbopol, cellulose derivatives, bentonite, alginates, gelatin and polyvinylpyrrolidone.
  • ointment bases are beeswax, paraffin, cetyl palmitate, vegetable oils, sorbitan esters of fatty acids (Span) , polyethylene glycols and condensation products between sorbitan esters of fatty acids and ethylene oxide (e.g., polyoxyethylene sorbitan monooleate (Tween) ) .
  • compositions described above for topical administration on the skin may also be used in connection with topical administration onto or close to the part of the body that is to be treated.
  • the compositions may be adapted for direct application or for introduction into relevant orifice (s) of the body (e.g., rectal, urethral, vaginal or oral orifices) .
  • the composition may be applied by means of special delivery devices such as dressings or alternatively plasters, pads, sponges, strips or other forms of suitable flexible material .
  • the active compound may be in the form of a solution or suspension in a suitable sterile aqueous or non-aqueous vehicle.
  • Additives for instance buffers, preservatives including bactericidal and fungicidal agents, such as phenyl mercuric acetate or nitrate, benzalkonium chloride, or chlorohexidine and thickening agents such as hypromellose may also be included.
  • the active compounds may also be presented for use in the form of veterinary compositions, which may be prepared, for example, by methods that are conventional in the art.
  • veterinary compositions include those adapted for: (a) oral administration, external application, for example drenches (e.g. aqueous or non-aqueous solutions or suspensions) ; tablets or boluses; powders, granules or pellets for admixture with feed stuffs; pastes for application to the tongue;
  • parenteral administration for example by- subcutaneous, intramuscular or intravenous injection, e.g. as a sterile solution or suspension; or (when appropriate) by intramammary injection where a suspension or solution is introduced in the udder via the teat;
  • topical applications e.g. as a cream, ointment or spray applied to the skin;
  • the compounds of formula I may be used in the treatment of a Flaviviridae viral infection such as a HCV infection.
  • treatment means affecting a subject, tissue or cell to obtain a desired pharmacological and/or physiological effect and include: (a) preventing the infection from occurring in a subject that may be predisposed to the infection, but has not yet been diagnosed as having it; (b) inhibiting the infection, i.e., arresting its development; or (c) relieving or ameliorating the effects of the infection, i.e., cause regression of the effects of the infection.
  • the prevention of hepatitis C means, for example, administration of a pharmaceutical agent to a subject found to carry a HCV by a test and the like but without a symptom of infection, or to a subject who shows an improved disease state of hepatitis after a treatment of hepatitis C, but who still carries a HCV and is associated with a risk of recurrence of hepatitis.
  • subject refers to any animal having a disease or condition which requires treatment with a pharmaceutically-active agent.
  • the subject may be a mammal, preferably a human, or may be a non-human primate or non-primates such as used in animal model testing. While it is particularly contemplated that the compounds are suitable for use in medical treatment of humans, it is also applicable to veterinary treatment, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, ponies, donkeys, mules, llama, alpaca, pigs, cattle and sheep, or zoo animals such as primates, felids, canids, bovids and ungulates .
  • viral infection refers to the introduction of a virus into cells or tissues, e.g., hepatitis C virus (HCV) .
  • HCV hepatitis C virus
  • the introduction of a virus is also associated with replication.
  • Viral infection may be determined by measuring virus antibody titer in samples of a biological fluid, such as blood, using, e.g., enzyme immunoassay.
  • Other suitable diagnostic methods include molecular based techniques, such as RT-PCR, direct hybrid capture assay, nucleic acid sequence based amplification, and the like.
  • a virus may infect an organ, e.g., liver, and cause disease, e.g., hepatitis, cirrhosis, chronic liver disease and hepatocellular carcinoma.
  • Flaviviridae virus refers to a virus of the family Flaviviridae, which family includes the
  • Hepacivirus Flavivirus and Pestivirus or hepatitis C- like virus genera A representative species of the genus of hepatitis C-like viruses is hepatitis C virus.
  • Flavivirus include yellow fever virus, tick-borne encephalitis virus, Rio Bravo virus, Japanese encephalitis virus, Tyuleniy virus, Ntaya virus, Kenya S virus, Dengue virus and Modoc virus.
  • Representative species of the genus Pestivirus include bovine diarrhea virus, border disease virus and hog cholera virus. Unassigned viruses in the family
  • Flaviviridae are included in the meaning of Flaviviridae virus . Dosages
  • terapéuticaally effective amount means an amount of the compound of formula I effective to yield a desired therapeutic response.
  • Dosage levels of the compound of formula I are of the order of about 0.5 mg to about 20 mg per kilogram body weight, with a preferred dosage range between about 0.5 mg to about 10 mg per kilogram body weight per day (from about 0.5 gms to about 3 gms per patient per day) .
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage will vary depending upon the subject treated and the particular mode of administration.
  • a formulation intended for oral administration to humans may contain about 5 mg to Ig of an active compound with an appropriate and convenient amount of carrier material which may vary from about 5 to 95 percent of the total composition.
  • Dosage unit forms will generally contain between from about 5 mg to 500 mg of active ingredient.
  • the compounds are administered in a divided dose schedule, such that there are at least two administrations in total in the schedule. Administrations are given preferably at least every two hours for up to four hours or longer; for example the compound may be administered every hour or every half hour.
  • the divided-dose regimen comprises a second administration of the compound after an interval from the first administration sufficiently long that the level of active compound in the blood has decreased to approximately from 5-30% of the maximum plasma level reached after the first administration, so as to maintain an effective content of active compound in the blood.
  • one or more subsequent administrations may be given at a corresponding interval from each preceding administration, preferably when the plasma level has decreased to approximately from 10-50% of the immediately- preceding maximum.
  • the specific dose level for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, active compound combination and the severity of the particular infection undergoing therapy.
  • BINAP 2,2' -bis (diphenylphosphino) -1,1' -binaphthyl
  • DIPEA diisopropylethylamine
  • HATU O- (7-azabenzotriazol-l-yl) -N,N,N' ,N'- tetramethyluronium hexafluorophosphate
  • HBTU O-benzotriazol-l-yl-N, N, N' , N' -tetramethyluronium hexafluorophosphate
  • HMBC heteronuclear multiple bond correlation
  • TFFH fluoro-N, N,N' , N' -tetramethylformamidinium hexafluorophosphate
  • the resulting hydrazone is reduced by treatment with approximately 1.2 equivalents of sodium cyanoborohydride .
  • the crude N 1 N' -disubstituted hydrazine is then condensed with at least one equivalent of ethyl acetoacetate by heating under dehydrating conditions to yield a pyrazolone derivative.
  • the pyrazolone is formylated by treatment with POCl 3 and DMF.
  • a solution of the resulting aldehyde and approximately 1.1 equivalents of LDA is then treated with an excess of methyl acrylate or other suitable Diels-Alder substrate.
  • the product thus obtained is then dehydrated with pyridinium tosylate and aromatised using DDQ.
  • the ester may be hydrolysed by treatment with an aqueous base .
  • Phenylhydrazine (4.0 mL, 40.7 mmol) was treated with cyclohexanone (4.2 mL, 40.7 mmol, 1 eq) then dissolved in ethanol (20 mL) and stirred for 10 min at ambient temperature to give hydrazone 1-1.
  • Sodium cyanoborohydride (3.1 g, 40.8mmol, 1.2 eq) was added and the mixture was stirred. After 30 minutes LCMS showed complete reduction to the hydrazine 1-2.
  • Ethyl acetoacetate (5.2 mL, 40.7mmol, 1 eq) was added and the stirred mixture was heated to reflux under Dean-Stark conditions.
  • More ethyl acetoacetate (5.2 mL, 40.7 mmol, 1 eq) was added and heating was continued overnight. The next morning, more ethyl acetoacetate (2 mL, 16.3 mmol, 0.4 eq) was added, along with molecular sieves (-1OmL) . After heating for a further 2 h, more ethyl acetoacetate (3 mL, 24.4 mmol, 0.6 eq) was added, along with more molecular sieves (-1OmL) .
  • the Diels-Alder adduct was treated with pyridinium tosylate (530 mg, 2.11mmol, 1.2 eq) in toluene (30 mL) at 90 0 C for 30 min. The mixture was diluted with water, extracted with CH2C12, dried (MgSO4), filtered and concentrated to give dehydrated adduct 1-6.
  • the dehydrated adduct was treated with DDQ (480 mg, 2.11mmol, 1.2 eq) in dioxane (50 mL) at reflux (-110 0 C) for 3 h. The mixture was cooled and allowed to stand overnight. It was then diluted with CH2C12 and concentrated to dryness. The residue was diluted with CH2C12 and purified by flash chromatography to yield the target indazolinone 103 as an off-white solid (106 mg, 0.302mmol, 17% for three steps).
  • Step 1 HATU (1.25 eq) was added to 2-amino-5- bromobenzoic acid (5 g) , 4-benzyloxyaniline hydrochloride (2 eq) and DIPEA (about 10 eq) in CH 2 Cl 2 (about 200 mL) and the solution allowed to stand for 16 h. The mixture was concentrated and chromatographed on silica to yield amide 2-2.
  • Step 2 Pd 2 (dba) 3 (30 mg) and BINAP (30 mg) were added to toluene (10 mL) and the mixture stirred for 10 min. Bromobenzene (200 ⁇ L) , KO 12 Bu (60 mg) and amine 2-2 (100 mg) were added and the mixture heated to reflux for 2 h. The mixture was filtered through Celite, concentrated and chromatographed on silica to yield compound 2-3 as a yellow powder (40 mg, 32%)
  • Step 3 PIFA (60 mg) was added to a solution of amide 2- 3 (30 mg) in TFA (5 drops) and CH 2 Cl 2 (5 mL) . Progress was monitored by TLC and when the reaction was complete the mixture was concentrated and chromatographed on silica to yield indazolinone 809 as a yellow gum (18 mg, 60%) .
  • Example 2 may be used to prepare other compounds of Formula I by replacing bromobenzene with an appropriate reagent of the formula R 3 ⁇ -Br, or by replacing 4-benzyloxyaniline with an appropriate reagent of the formula R 2 -NH 2 , where R 1 and R 2 are defined herein.
  • Step 1 2-Chloro-5-nitrobenzoic acid (1.5 g) was dissolved in cyclohexylamine (4 mL) with stirring and warming. The bright yellow solution was heated in a microwave reactor at 120 0 C for 6 minutes. The resulting gel was diluted with CH 2 Cl 2 (-100 mL) and extracted with 0.2M NaOH (4 x -100 mL) . The aqueous extracts were acidified with 5M HCl, causing precipitation of a white solid. The solid was recovered by filtration and dried. The solid was recrystallised from methanol/water to yield light yellow crystals of amine 3-1 (0.6 g, 31%) .
  • Step 2 2-Cyclohexylamino-5-nitrobenzoic acid (3-1) (0.6 g) was dissolved in CH 2 Cl 2 (12 mL) and TEA (2 mL) and treated with BOP (1 g) . After stirring for 2 minutes at room temperature, 2-aminopyridine (0.5 g) was added. The mixture was stirred for 64 h, then another portion of BOP (1 g) was added. After an additional 30 minutes, LCMS indicated almost complete reaction.
  • Step 3 Compound 3-2(270 mg) was dissolved in acetonitrile and treated with PIFA (341 mg, 1 eq) at room temperature. After 2h, TFA (1 mL) and additional PIFA (1 eq) were added. After 3h, more PIFA (leq) was added and the mixture was stirred at room temperature for 64 h.
  • Example 3 may be used to prepare other compounds of Formula I by replacing cyclohexylamine with an appropriate reagent of the formula R 3 ⁇ -NH 2 , or by replacing 2-aminopyridine with an appropriate reagent of the formula R 2 -NH 2 , where R 1 and R 2 are defined herein.
  • Step 1 Amine 2-2 was prepared as described in Example 2.
  • Step 2 Amine 2-2 (9.3 g) was treated with an excess of cyclohexanone in acetic acid at reflux for 1 h. The mixture was diluted with EtOAc, washed with aqueous NaOH, dried and concentrated to yield imine 4-1 (10.1 g) .
  • Step 3 Imine 4-1 (10.1 g) was treated with sodium borohydride ( ⁇ 5 eq) in EtOH and the reaction heated at reflux for 3 h. Concentration and silica chromatography yielded the target amine 4-2 (8.2 g) .
  • Step 4 Amide 4-2 was converted to indazolinone 811 using the analogous procedure to that described in Step 3 of Example 2.
  • Step 1 To a stirred solution of 2-amino-4-bromobenzoic acid (1 g) in acetic acid (20 mL) was added cyclohexanone (1.2 eq) and the mixture was stirred at room temperature for Ih. Sodium borohydride (3 eq) was added in portions over 2 h at 40 0 C and the reaction mixture was stirred for an additional 2 h. Concentration gave an oil, which was diluted with water. The precipitated solids were isolated by filtration and purified by chromatography to yield the target 5-1 (1.2 g, 86%) .
  • Step 2 Compound 5-2 was prepared from acid 5-1 by the procedure in Step 1 of Example 2, using aniline in place of 4-benzyloxyaniline hydrochloride.
  • Step 3 Amide 5-2 was converted to indazolinone 817 using the analogous procedure to that described in Step 3 of Example 2.
  • Step 1 2, 5-Dichloropyridine was added to a THF solution of n-BuLi and PMDTA and stirred for 2 hours at -78 0 C before quenching with dry ice. The product was isolated by acid-base work-up in 47% yield.
  • Step 2 To a methanolic solution of 2, 5-dichloro- isonicotinic acid was added H 2 SO 4 (cone) (0.5 mL) and the solution was heated to reflux for 12 h. The mixture was cooled to room temperature, neutralized with aqueous NaHCO 3 , extracted into EtOAc and evaporated to afford the methyl ester (6-2) in 89% yield.
  • Step 3 To a THF solution of methyl 2, 5-dichloro- isonicotinate was added Fe(acac) 3 (0.05 eq) and NMP (-10 vol) . After cooling to O 0 C, CH 3 MgBr (1.2 eq) was added and the mixture was stirred at 0 0 C for 1 h.
  • Step 4 Methyl 5-chloro-2-methylisonicotinate was suspended in a solution of THF/MeOH/water (1:1:1) and to that LiOH (3 eq) was added. The mixture was stirred for 1 h and the resultant carboxylic acid was isolated by aqueous workup .
  • Step 5 To a CH 2 Cl 2 solution of acid 6-4 (0.8 g) was added EDC (1.3 eq) and aniline (2 eq) at room temperature. After 10 minutes, the reaction was diluted with CH 2 Cl 2 and washed with dilute HCl. The organic layer was dried and concentrated to yield amide 6-5 (0.9 g, 80%) .
  • Step 6 Amide 6-5 was taken up in DME and to this was added NaO'Bu (1.4 eq) , Pd(OAc) 2 (0.05 eq) , CyPF( 11 Bu) (0.05 eq) and aniline (1.2 eq) . The mixture was heated at 100 0 C in a sealed tube for 12 hours and the product was purified by column chromatography.
  • Step 7 Amide 6-6 was converted to indazolinone 819 using the analogous procedure to that described in Step 3 of Example 2 but using MeCN as the solvent instead of CH 2 Cl 2 .
  • Step 1 Compound 7-1 was prepared from 2-amino-5- methylbenzoic acid using the analogous procedure set out in Step 1 of Example 2, using p-anisidine in place of 4- benzyloxyaniline hydrochloride.
  • Step 2 Imine 7-2 was prepared from amine 7-1 by the analogous procedure set out in Step 2 of Example 4.
  • Step 3 Imine 7-2 was reduced to amine 7-3 by the analogous procedure set out in Step 3 of Example 4.
  • Step 4 Amide 7-3 was converted to an indazolinone using the analogous procedure to that described in Step 3 of Example 2 but in the current example, the substrate was observed to have simultaneously undergone both ring- closure and partial benzylic oxidation to yield trifluoroacetoxy derivative 816.
  • Step 1 2-Hydroxynicotinic acid (100 g) was converted to 5-bromo-2-hydroxynicotinic acid (8-1) in 79% yield using the procedure reported in Letavic, M.A. et al., Bioorganic and Medicinal Chemistry Letters, 17, 2007, 2566-2569.
  • Step 2 A stirred suspension of 5-bromo-2-hydroxy- nicotinic acid (8-1) (67 g) in POCl 3 (1 vol) was heated to 100 0 C for 14 h. The reaction mixture was cooled to room temperature and quenched by stirring with ice-water for 10 min. The product was extracted into CH 2 Cl 2 and treated with aniline for 1 h. After washing with water, concentration of the organic layer gave crude amide 8-2.
  • Step 3 A solution of 8-2 (10 g) in cyclohexylamine (10 eq) was heated to 100 0 C for 14 h, after which time TLC indicated complete conversion of the starting material. The mixture was diluted with dilute HCl solution and the product extracted into CH 2 Cl 2 . The organic extracts were concentrated and recrystallised from diethyl ether to give amine 8-3 (6.5 g, 54%).
  • Step 4 Amide 8-3 was converted to indazolinone 807 using the analogous procedure to that described in Step 3 of Example 2.
  • ESI-MS m/z calc . for [M+H] + : 374 (Br isotope); found: 374.0
  • Bromide 811 was converted to nitrile 812 using the procedure described for Example 9, except that DMF was used as the solvent instead of NMP and the mixture was heated only to 140 0 C.
  • Bromide 817 was converted to nitrile 818 using the procedure described for Example 9.
  • Bromide 804 was converted to nitrile 810 using the procedure described for Example 9, except that DMF was used as the solvent instead of NMP and the mixture was heated only to 140 0 C.
  • Nitro-indazolinone 814 (90 mg) was dissolved in ethanol (30 mL) , treated with 10%Pd/C (5 mg) and placed under an atmosphere of hydrogen. The mixture was stirred vigourously for 16 h before filtering through celite. The filtrate was concentrated to dryness and the residue purified by silica chromatography (eluting with EtOAc/hexanes) to yield amine 815 (70 mg)
  • Nitrile 818 (2.6 g) was added to a solution of sulfuric acid and water (1:1) and the reaction mixture was heated to 100 0 C for 1 h. The mixture was diluted with water and the product extracted into chloroform. The organic extracts were concentrated and purified by silica chromatography to yield acid 111 (l.Og, 35%) .
  • Nitrile 820 was converted to acid 112 using the procedure described for Example 15.
  • Nitrile 812 (130 mg) was treated with acetic acid (0.5 mL and concentrated HCl (3 mL) and stirred at reflux for 50 h. After cooling, the mixture was diluted with water and the product extracted into CH 2 Cl 2 . The organic layer was concentrated and the residue purified by silica chromatography to yield acid 110 (48 mg, 54%) .
  • Nitrile 810 was converted to the title tetrazole using trimethylaluminium and trimethylsilylazide according to the analogous procedure reported in B. E. Huff and M. A. Staszak, Tetrahedron Letters, 34, 8011 (1993).
  • Indazolinone 816 (40 mg) and NaOH (100 mg) were dissolved in acetone/water (3:1, 4 mL) and heated to reflux. Portions of KMnO 4 were added whenever the purple colouration subsided. After 3 h, TLC indicated that the benzylic oxidation was complete. The mixture was filtered through celite, rinsing the filter with 5M NaOH and acetone . The acetone was removed in vacuo and the resulting aqueous solution was washed with CH 2 Cl 2 . The aqueous layer was then acidified with cone. HCl and extracted with CH 2 Cl 2 , which was dried and concentrated to give acid 107 as a pale tan powder (18 mg, 53%) . ESI-MS m/z calculated for [M+H] + : 367; found: 367.1
  • a solution of an appropriate indazolinone- carboxylic acid in an inert solvent is treated with the desired sulfonamide (at least 1 equivalent) , an activating agent such as EDC (about 1 to 2 eq) and, if required, DMAP ( ⁇ 2 eq) , such as in the manner reported by Stansfield et al. in Bioorg Med Chem Lett 17(18), 2007, 5143.
  • an activating agent such as EDC (about 1 to 2 eq)
  • DMAP ⁇ 2 eq
  • DIPEA non-nucleophilic base
  • sulfonamides may be purchased or prepared from their corresponding sulfonyl chlorides using techniques that are well known to those skilled in the art. It will also be apparent to one skilled in the art that the general procedure of Examples 20 and 21 may be used to prepare other compounds of Formula I by replacing 2-chlorobenzenesulfonamide (Example 21) or 4 -methoxybenzenesulfonamide (Example 20) with alternative nucleophiles of the formula R 6 -NHS(O) P R 8 , where R 6 , R 8 and p are defined herein.
  • certain amides of formula I may be prepared from an appropriate carboxylic acid as set out in the scheme above.
  • the indazolinone-carboxylic acid and approximately 1.2 equivalents of an activating agent such as HBTU are dissolved in an appropriate solvent.
  • a suitable non-nucleophilic base is added together with the desired amine and the solution is allowed to stand at room temperature until condensation has occurred.
  • the products may be isolated by chromatography.
  • carboxylic acid 104 (40.5 mg, 0.12 mmol) and HBTU (55 mg, 0.14 mmol, 1.2 eq) were dissolved in DMF (400 ⁇ L) .
  • DIPEA 105 ⁇ L, 0.6 mmol, 5 eq
  • L-tryptophan methyl ester hydrochloride 37 mg, 0.14 mmol, 1.2 eq
  • the solution was allowed to stand at room temperature. After 20 min the reaction mixture was diluted with CH2C12 (2 mL) and washed with water (3 x 1 mL) . The organic layer was filtered through cotton wool and concentrated.
  • the crude residue was purified by flash chromatography using ethyl acetate :hexanes (1:1) as eluent to yield the target amide 303 as a white solid (60 mg, 0.11 mmol, 93%) .
  • a suitable carboxylic acid substrate is dissolved in H 2 SO 4 , treated with sodium azide and heated to approximately 70° C until the reaction is complete. The reaction mixture is then poured onto an aqueous solution of NaOH and extracted with a suitable organic solvent.
  • the amine substrate is dissolved in a suitable solvent such as THF, CH 2 Cl 2 or pyridine and treated with an excess of a suitable electrophile, such as a carboxylic acid halide, an anhydride, a sulfonyl halide or an isocyanate.
  • a suitable electrophile such as a carboxylic acid halide, an anhydride, a sulfonyl halide or an isocyanate.
  • a non-nucleophilic base such as TEA may be added and the mixture may be heated.
  • the volatile components are evaporated and the residue may be purified by chromatography.
  • a stirred solution of the indazolinone (5 mg) in pyridine (100 ⁇ l) was treated with benzenesulfonyl chloride (5 ⁇ l) .
  • reaction mixture was then heated at 60 0 C for 18 h, cooled to room temperature and the volatiles removed in vacuo. The residue was filtered through a plug of silica gel, eluting with EtOAc. The material was purified further by preparative HPLC to yield the title compound (708) .
  • Example 28 (Method T) l-Cyclohexyl-3-oxo-2-phenyl-2 , 3-dihydro-lH-indazole-5- carboxylic acid (Compound 104, Table 1)
  • Indazolinone 103 (68 mg, 0.194mmol) was dissolved in a mixture of methanol (5 mL) and 1 M aqueous NaOH (2 mL) and warmed to 60 0 C for 1 h. The mixture was evaporated to dryness and the residue partitioned between 1 M HCl (aq) and CH2C12. The organic layer was filtered through cotton wool and concentrated to yield carboxylic acid 104 as a light brown solid (64 mg, 0.190mmol, 98%).
  • the phenol 110 was dissolved in acetone and treated with benzyl bromide (2 eq) and K 2 CO 3 (excess) . After 1 h at room temperature, LCMS indicated complete conversion to the desired ether. The mixture was concentrated, resuspended in dichloromethane and washed with water. The organic phase was dried (MgSO 4 ) and concentrated to yield the target phenol, 114.
  • Indazolinone 804 (50 mg) was dissolved in 1-butanol (0.5 tnL) , treated with DIPEA (3 eq) , Pd(OAc) 2 (0.2 eq) and Xantphos (0.2 eq) and placed under an atmosphere of carbon monoxide. The mixture was stirred and heated to 100 0 C for 3 days. After liquid-liquid work-up, the major product was purified by flash chromatography to yield the title ester (108) as a colourless, waxy solid.
  • HCV polymerase reactions were carried out using a modified method of Howe et al . , Antimicrobial Agents and Chemotherapy 2004 48(12): 4813-4821. Reactions contained 50 nM NS5Bb ⁇ 21, 20 mM Tris-HCl pH 7.5 , 5 mM MgC12, 3 mM DTT, 25 mM KCl, 0.05% BSA, 0.5% DMSO, 0.4U/ ⁇ L RNasin, and 20 nM Poly r(C) (Amersham) . GTP was present at Km concentration including 0.04 ⁇ Ci/ ⁇ L 33P-GTP, with a total reaction volume of 50 ⁇ L.
  • Ava5 cells Human cells containing a genotype Ib subgenomic HCV replicon, Blight et al . , Science 2000 290: 1972-1974 were used in these studies. Cultures were maintained in a sub-confluent state in DMEM with glutamine, non-essential amino acids, 10% heat-inactivated foetal bovine serum and G418.
  • HCV RNA analysis was seeded at a concentration of 2000 cells per well into 96 well tissue culture trays and grown overnight in the presence of G418. The following day the cell monolayers were exposed to dilutions of compound. Stock solutions (5OmM) of each compound were made in DMSO and diluted in culture media lacking G418 on the day of addition. Compounds were tested in a final concentration range of 20-100 uM downwards. Cells were treated for three consecutive days with the test compounds. Medium was replaced daily with fresh test compounds. Analysis of HCV RNA was performed 24 h following the last addition of test compound.
  • HCV replicon RNA was assayed by real-time reverse transcription (RT-PCR) using primers located in the HCV NS5B gene and quantified on the basis of HCV RNA standards and relative amounts of cellular actin mRNA.
  • Example 36 Cytotoxicity of compounds against Ava5 cells was assessed using the MTT assay (Watanabe et al . , Journal of Virological Methods 1994 48:257-265). Plates were prepared as described for the HCV Replicon assay, and cytotoxicity analysis was performed 24 h following the last addition of test compound. Culture media in each test well was replaced with lmg/mL MTT solution, and the cells incubated at 37°C for several hours. The MTT solution was then replaced with isopropanol, and cytotoxicity assessed by reading the plates at 540/690 nm and determining colour development in each test well.
  • HCV polymerase reactions were carried out using a modified method of Howe et al., Antimicrobial Agents and Chemotherapy 2004 48(12): 4813-4821. Reactions contained 25 nM NS5b ⁇ 21, 20 mM Tris-HCl pH 7.5 , 5 mM MgCl 2 , 3 mM DTT, 0.05% BSA, 0.4U/ ⁇ L RNasin, and 5 nM RNA (500 nt template) . All NTPs used were at Km concentrations, and 0.04 ⁇ Ci/ ⁇ L 33 P-UTP, with a total reaction volume of 50 ⁇ L.
  • the in vivo efficacy of the compounds may be evaluated using known methods such as those described in: Lanford, R. E. "The chimpanzee model of hepatitis C virus. Framing the Knowledge of Viral Hepatitis", 197-218 IHL Press, 2006,- Lanford, R. E., Bigger, C, Bassett, S., Klimpel, G. , "The chimpanzee model of hepatitis C virus infections", Institute for Laboratory Animal Research
  • a model that employs a surrogate virus — in particular GB virus B — may be used according to known methods such as described in Haqshenas, G. et al., "A chimeric GB virus B encoding the hepatitis C virus hypervariable region 1 is infectious in vivo” , Journal of General Virology 88, 2007, 895 and Bright, H. et al . , "Development of a GB Virus B Marmoset Model and Its Validation with a Novel Series of Hepatitis C Virus NS3 Protease Inhibitors", Journal of Virology, 78(4), 2004, 2062.

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Abstract

L'invention porte sur des inhibiteurs des polymérases virales, et en particulier sur de telles polymérases de la famille des Flaviviridae dont le virus de l'hépatite C (VHC), sur leur préparation, et sur leur utilisation dans le traitement d'infections.
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WO2013014038A1 (fr) 2011-07-26 2013-01-31 Nerviano Medical Sciences S.R.L. Dérivés de 3-oxo-2,3-dihydro-1h-indazole-4-carboxamide utilisés comme inhibiteurs de la parp-1
WO2013036994A1 (fr) * 2011-09-16 2013-03-21 Biota Scientific Management Pty Ltd Composés pour le traitement des infections par vhc
WO2013152391A1 (fr) * 2012-04-11 2013-10-17 Biota Scientific Management Pty Ltd Inhibiteurs de polymérase virale
CN103483274A (zh) * 2013-09-25 2014-01-01 江苏快达农化股份有限公司 一种制备苄嘧磺隆的方法
JP2014528412A (ja) * 2011-09-30 2014-10-27 キネタ・インコーポレイテツド 抗ウイルス化合物
US9782408B2 (en) 2014-10-06 2017-10-10 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US9856253B2 (en) 2015-04-17 2018-01-02 Abbvie, Inc. Tricyclic modulators of TNF signaling
US9879016B2 (en) 2015-04-17 2018-01-30 Abbvie Inc. Indazolones as modulators of TNF signaling
US10160748B2 (en) 2015-04-17 2018-12-25 Abbvie Inc. Indazolones as modulators of tnf signaling
US10570115B2 (en) 2016-09-30 2020-02-25 Vertex Pharmaceuticals Incorporated Modulator of cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US10654829B2 (en) 2017-10-19 2020-05-19 Vertex Pharmaceuticals Incorporated Crystalline forms and compositions of CFTR modulators
US10738030B2 (en) 2016-03-31 2020-08-11 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US10793547B2 (en) 2016-12-09 2020-10-06 Vertex Pharmaceuticals Incorporated Modulator of the cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11179367B2 (en) 2018-02-05 2021-11-23 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions for treating cystic fibrosis
US11253509B2 (en) 2017-06-08 2022-02-22 Vertex Pharmaceuticals Incorporated Methods of treatment for cystic fibrosis
US11414439B2 (en) 2018-04-13 2022-08-16 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11434201B2 (en) 2017-08-02 2022-09-06 Vertex Pharmaceuticals Incorporated Processes for preparing pyrrolidine compounds
US11465985B2 (en) 2017-12-08 2022-10-11 Vertex Pharmaceuticals Incorporated Processes for making modulators of cystic fibrosis transmembrane conductance regulator
US11517564B2 (en) 2017-07-17 2022-12-06 Vertex Pharmaceuticals Incorporated Methods of treatment for cystic fibrosis
US11786528B2 (en) 2018-06-04 2023-10-17 Exscientia Ltd. Pyrazolopyrimidine compounds as adenosine receptor antagonists
EP4345102A1 (fr) * 2022-09-30 2024-04-03 Medizinische Universität Innsbruck Synthèse améliorée d'otviciclib

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WO2013014038A1 (fr) 2011-07-26 2013-01-31 Nerviano Medical Sciences S.R.L. Dérivés de 3-oxo-2,3-dihydro-1h-indazole-4-carboxamide utilisés comme inhibiteurs de la parp-1
JP2014521620A (ja) * 2011-07-26 2014-08-28 ネルビアーノ・メデイカル・サイエンシーズ・エツセ・エルレ・エルレ Ｐａｒｐ−１阻害剤としての３−オキソ−２，３−ジヒドロ−１ｈ−インダゾール−４−カルボキサミド誘導体
US9073893B2 (en) 2011-07-26 2015-07-07 Nerviano Medical Sciences S.R.L. 3-Oxo-2,3-dihydro-1H-indazole-4-carboxamide derivatives as PARP-1 inhibitors
WO2013036994A1 (fr) * 2011-09-16 2013-03-21 Biota Scientific Management Pty Ltd Composés pour le traitement des infections par vhc
JP2014528412A (ja) * 2011-09-30 2014-10-27 キネタ・インコーポレイテツド 抗ウイルス化合物
WO2013152391A1 (fr) * 2012-04-11 2013-10-17 Biota Scientific Management Pty Ltd Inhibiteurs de polymérase virale
US9447080B2 (en) 2012-04-11 2016-09-20 Biota Scientific Management Pty Ltd. Viral polymerase inhibitors
CN103483274A (zh) * 2013-09-25 2014-01-01 江苏快达农化股份有限公司 一种制备苄嘧磺隆的方法
US10258624B2 (en) 2014-10-06 2019-04-16 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US10758534B2 (en) 2014-10-06 2020-09-01 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US11426407B2 (en) 2014-10-06 2022-08-30 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US12168009B2 (en) 2014-10-06 2024-12-17 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US9782408B2 (en) 2014-10-06 2017-10-10 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US9879016B2 (en) 2015-04-17 2018-01-30 Abbvie Inc. Indazolones as modulators of TNF signaling
US10160748B2 (en) 2015-04-17 2018-12-25 Abbvie Inc. Indazolones as modulators of tnf signaling
US10266532B2 (en) 2015-04-17 2019-04-23 Abbvie Inc. Tricyclic modulators of TNF signaling
US10273238B2 (en) 2015-04-17 2019-04-30 Abbvie Inc. Indazolones as modulators of TNF signaling
US9856253B2 (en) 2015-04-17 2018-01-02 Abbvie, Inc. Tricyclic modulators of TNF signaling
US10738030B2 (en) 2016-03-31 2020-08-11 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator
US10570115B2 (en) 2016-09-30 2020-02-25 Vertex Pharmaceuticals Incorporated Modulator of cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11186566B2 (en) 2016-09-30 2021-11-30 Vertex Pharmaceuticals Incorporated Modulator of cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US10793547B2 (en) 2016-12-09 2020-10-06 Vertex Pharmaceuticals Incorporated Modulator of the cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11453655B2 (en) 2016-12-09 2022-09-27 Vertex Pharmaceuticals Incorporated Modulator of the cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11253509B2 (en) 2017-06-08 2022-02-22 Vertex Pharmaceuticals Incorporated Methods of treatment for cystic fibrosis
US11517564B2 (en) 2017-07-17 2022-12-06 Vertex Pharmaceuticals Incorporated Methods of treatment for cystic fibrosis
US11434201B2 (en) 2017-08-02 2022-09-06 Vertex Pharmaceuticals Incorporated Processes for preparing pyrrolidine compounds
US10654829B2 (en) 2017-10-19 2020-05-19 Vertex Pharmaceuticals Incorporated Crystalline forms and compositions of CFTR modulators
US11155533B2 (en) 2017-10-19 2021-10-26 Vertex Pharmaceuticals Incorporated Crystalline forms and compositions of CFTR modulators
US11465985B2 (en) 2017-12-08 2022-10-11 Vertex Pharmaceuticals Incorporated Processes for making modulators of cystic fibrosis transmembrane conductance regulator
US11179367B2 (en) 2018-02-05 2021-11-23 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions for treating cystic fibrosis
US11414439B2 (en) 2018-04-13 2022-08-16 Vertex Pharmaceuticals Incorporated Modulators of cystic fibrosis transmembrane conductance regulator, pharmaceutical compositions, methods of treatment, and process for making the modulator
US11786528B2 (en) 2018-06-04 2023-10-17 Exscientia Ltd. Pyrazolopyrimidine compounds as adenosine receptor antagonists
EP4345102A1 (fr) * 2022-09-30 2024-04-03 Medizinische Universität Innsbruck Synthèse améliorée d'otviciclib
WO2024068853A1 (fr) * 2022-09-30 2024-04-04 Medizinische Universität Innsbruck Synthèse améliorée d'otviciclib

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