WO2001040189A1 - Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis - Google Patents

Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis Download PDF

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Publication number
WO2001040189A1
WO2001040189A1 PCT/US2000/032621 US0032621W WO0140189A1 WO 2001040189 A1 WO2001040189 A1 WO 2001040189A1 US 0032621 W US0032621 W US 0032621W WO 0140189 A1 WO0140189 A1 WO 0140189A1
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Prior art keywords
group
pharmaceutically acceptable
compound
substituted
heterocycloalkyl
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PCT/US2000/032621
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French (fr)
Inventor
Peter S. Dragovich
Thomas J. Prins
Ru Zhou
Theodore O. Johnson, Jr.
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Agouron Pharmaceuticals, Inc.
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Priority to BR0016742-8A priority Critical patent/BR0016742A/en
Priority to PL00356062A priority patent/PL356062A1/en
Application filed by Agouron Pharmaceuticals, Inc. filed Critical Agouron Pharmaceuticals, Inc.
Priority to EP00980893A priority patent/EP1252145A1/en
Priority to EEP200200281A priority patent/EE200200281A/en
Priority to HU0204006A priority patent/HUP0204006A3/en
Priority to EA200200625A priority patent/EA200200625A1/en
Priority to MXPA02005124A priority patent/MXPA02005124A/en
Priority to JP2001541874A priority patent/JP2003515591A/en
Priority to IL14942700A priority patent/IL149427A0/en
Priority to NZ518934A priority patent/NZ518934A/en
Priority to KR1020027007099A priority patent/KR20020058076A/en
Priority to SK760-2002A priority patent/SK7602002A3/en
Priority to CA002392504A priority patent/CA2392504A1/en
Priority to AU18094/01A priority patent/AU777943B2/en
Priority to APAP/P/2002/002510A priority patent/AP2002002510A0/en
Publication of WO2001040189A1 publication Critical patent/WO2001040189A1/en
Priority to IS6378A priority patent/IS6378A/en
Priority to BG106754A priority patent/BG106754A/en
Priority to NO20022589A priority patent/NO20022589L/en
Priority to HK03104911.5A priority patent/HK1052933A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • 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
    • 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
    • C07D405/14—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 three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • 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 invention relates to pyridone-containing peptidomimetic compounds that advantageously inhibit the enzymatic activity of picornaviral 3C proteases, especially rhinovirus 3C proteases (RVPs), and that retard viral growth in cell culture.
  • the invention also relates to the use of these compounds in pharmaceutical compositions, methods of treatment of rhino viral infections using these compounds and compositions, and processes for the synthesis of these compounds and compounds useful in the syntheses thereof.
  • the picomaviruses are a family of tiny non-enveloped positive-stranded RNA-containing viruses that infect humans and other animals. These viruses include the human rhinoviruses, human polioviruses, human coxsackieviruses, human echoviruses, human and bovine enteroviruses, encephalomyocarditis viruses, meningitis virus, foot and mouth viruses, hepatitis A virus, and others.
  • the human rhinoviruses are a major cause of the common cold. To date, there are no effective therapies on the market that cure the common cold, only treatments that relieve the symptoms.
  • Picornaviral infections may be treated by inhibiting the proteolytic picornaviral 3C enzymes. These enzymes are required for the natural maturation of the picomaviruses. They are responsible for the autocatalytic cleavage of the genomic, large polyprotein into the essential viral proteins. Members of the 3C protease family are cysteine proteases, where the sulfhydryl group most often cleaves the glutamine-glycine . amide bond. Inhibition of 3C proteases is believed to block proteolytic cleavage of the viral polyprotein, which in turn can retard the maturation and replication of the viruses by interfering with viral particle production.
  • This invention relates to compounds useful for inhibiting the activity of picornaviral 3C proteases having the general formula:
  • R a is substituted or unsubstituted heterocycloalkyl or heterocycloalkylalkyl; R b is a substituent having the formula:
  • R f and R g are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5;
  • A is CH or N
  • a 2 is C(R h )(R), N(R), S, S(O), S(O) 2 , or O; where each R h , R' and R J is independently H or lower alkyl; each A 3 present is independently C(R h )(R'), N(R J ), S, S(O), S(O) 2 , or O; where each R h , R 1 and R J is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A 4 is N(R k ), C(R h )(R i ), or O; and when p is 0 (i.e., A 3 is not present), A 4 is N(R k )(R'), C(R h )(R i )(R j ), and O(R'), where each R h , R and R j is independently H or lower alkyl, each R is H, alkyl, aryl, or acyl, and each R 1 is H,
  • R c is H, halogen or a substituted or unsubstituted lower alkyl group
  • R d is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group
  • R e is H or a substituted or unsubstituted alkylgroup
  • Z and Z 1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R n , -CO 2 R n , -CN, -C(O)NR n R°, -C(O)NROR°, -C(S)R n , -C(S)OR n , -C(S)NR n R°, -NO 2 , -SOR°, -SO 2 R n , -SO 2 NR n R°, -SO 2 (NR n )(OR°), -SONR", -SO 3 R n , -PO(OR n ) 2 , -PO(OR n )(OR°) > -PO(NR n R°)(OR p ) ⁇ -PO(NR n R°)(NR p R q
  • R a is substituted or unsubstituted heterocycloalkylalkyl
  • the alkyl moiety thereof is a substituted or unsubstituted saturated alkyl moiety.
  • this invention relates to compounds useful for inhibiting the activity of picornaviral 3C proteases having the general Formula I:
  • R 1 is H, a substituted or unsubstituted lower alkyl group or a suitable nitrogen protecting group
  • R 2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties in the above groups may be substituted or unsubstituted, or a suitable nitrogen protecting group;
  • R 3 is H or a suitable substituent
  • R 1 together with R 2 form a suitable nitrogen protecting group
  • R 2 together with R 3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
  • R 4 is H or a suitable substituent; the dotted line represents an optional bond;
  • R 5 is H or a suitable substituent
  • R 6 is H or a substituted or unsubstituted alkyl group
  • R 5 together with R 6 form a heterocycloalkyl ring, which may be optionally substituted;
  • R 7 and R 10 are independently H, halogen or a substituted or unsubstituted lower alkyl group
  • R 8 is H or a substituted or unsubstituted lower alkyl group
  • R 11 is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
  • R 9 is a substituent having the formula:
  • R 12 and R 13 are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5; A, is CH or N;
  • a 2 is C(R 14 )(R 15 ), N(R 16 ), S, S(O), S(O) 2 , or O; where each R 14 , R 15 and R 16 is independently H or lower alkyl; each A 3 present is independently C(R 1 )(R 15 ), N(R 16 ), S, S(O), S(O) 2 , or O; where each R 14 , R 15 and R 16 is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A 4 is N(R 17 ), C(R 14 )(R 15 ), or O; and when p is 0 (i.e., A 3 is not present), A 4 is N(R 17 )(R 18 ), C(R 14 )(R 15 )(R 16 ), and O(R 18 ), where each R 14 , R 15 and R 16 is independently H or lower alkyl, each R 17 is H, alkyl, aryl, or acyl, and each R 18 is H, alkyl,
  • Z and Z 1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R 19 , -CO 2 R 19 , -CN, -C(O)NR 19 R 20 , -C(O)NR 19 OR 20 , -C(S)R 19 , -C(S)OR 19 , -C(S)NR 19 R 20 , -NO 2 , -SOR 20 , -SO 2 R 19 , -SO 2 NR ,9 R 20 , -SO 2 (NR 19 )(OR 20 ), -SONR 19 , -SO 3 R 19 , -PO(OR 19 ) 2 , -PO(OR 19 )(OR 20 ) ⁇ -PO(NR 19 R 20 )(OR 21 ) , -PO(NR 19 R 20 )(NR 21 R 22 ),
  • Z and Z' are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R 19 , -CO 2 R 19 , -CN, -C(O)NR' 9 R 20 , -C(O)NR ,9 OR 20 , -C(S)R 19 , -C(S)NR 19 R 20 , -NO 2 , -SOR 20 , -SO 2 R 19 , -SO 2 NR 19 R 20 , -SO 2 (NR 19 )(OR 20 ), -SONR 19 , -SO 3 R 19 , -PO(OR 19 ) 2 , -PO(OR 19 )(OR 20 ) ⁇ -PO(NR 19 R 20 )(OR 2, ) ⁇ -PO(NR 19 R 20 )(NR 21 R 22 ).
  • Z and Z 1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R 19 , -CO 2 R 19 , -CN, -C(O)NR ,9 R 20 , -C(O)NR l9 OR 20 , -C(S)R 19 , -C(S)NR 19 R 20 , -NO 2 , -SOR 20 , -SO 2 R 19 , -SO 2 NR 19 R 20 , -SO 2 (NR 19 )(OR 20 ), -SONR' 9 , -SO 3 R 19 , -PO(OR 19 ) 2 , -PO(OR 19 )(OR 20 ) ,
  • antipicomaviral agents of the invention include prodrugs, pharmaceutically active metabolites, and pharmaceutically acceptable salts and solvates of such compounds.
  • the compounds of this invention useful for inhibiting the activity of picornaviral 3C proteases have the Formula I-A:
  • R 1 , R 2 , R ⁇ R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R", Z and Z 1 , and the variables contained therein, are as defined above, or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
  • R 1 , R 7 , R 8 , R 10 and R 11 are independently H or a substituted or unsubstituted lower alkyl group
  • R 2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
  • R 3 is H or a suitable substituent
  • R 2 together with R 3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
  • R 4 is H or a suitable substituent
  • R 5 is H or a suitable substituent
  • R 6 is H or an unsubstituted alkyl group or a lower alkyl group optionally substituted with a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an alkoxyl group, an aryloxy group, an alkylthio group, an arylthio group, wherein each alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl moiety thereof may be optionally substituted; or
  • R 9 is an aminocarbonylalkyl group, an alkylcarbonylaminoalkyl group, an alkylaminocarbonylalkyl group or a substituent having the formula:
  • R 12 and R 13 are independently H or lower alkyl; m is 1; p is 1 or 2; A, is CH or N;
  • a 2 is C(R 14 )(R 15 ), N(R 16 ), S, S(O), S(O) 2 , or O; each A 3 present is independently C(R 14 )(R 15 ), N(R 16 ), S, S(O), S(O) 2 , or O;
  • Z and Z 1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R 19 ,
  • R 3 , R 4 and R 5 may be independently selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, amino, cyano, halogen, haloalkyl
  • alkyl represents a straight- or branched-chain saturated or unsaturated hydrocarbon, containing 1 to 10 carbon atoms which may be unsubstituted or substituted by one or more of the substituents described below.
  • alkyl substituents include, but are not limited to methyl (Me), ethyl (Et), propyl, isopropyl, butyl, isobutyl, t-butyl, ethenyl, propenyl, butenyl, pentenyl, ethynyl, butynyl, propynyl (propargyl, isopropynyl), pentynyl, hexynyl and the like.
  • the term "lower alkyl” refers to an alkyl group containing from 1 to 4 carbon atoms.
  • Cycloalkyl represents a group comprising a non-aromatic monocyclic, bicyclic, or tricyclic hydrocarbon containing from 3 to 14 carbon atoms which may be unsubstituted or substituted by one or more of the substituents described below and may be saturated or unsaturated.
  • exemplary cycloalkyls include monocyclic rings having from 3-7, preferably 3-6, carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like, that may be fully saturated or partially unsaturated.
  • Illustrative examples of cycloalkyl groups include the following:
  • Heterocycloalkyl represents a group comprising a non-aromatic, monovalent monocyclic, bicyclic, or tricyclic radical, which is saturated or partially unsaturated, containing 3 to 18 ring atoms, which includes 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, and which may be unsubstituted or substituted by one or more of the substituents described below.
  • heterocycloalkyl groups include, but are not limited to, azetidinyl, pyrrolidyl, piperidyl, piperazinyl, morpholinyl, tetrahydro-2H-l,4-thiazinyl, tetrahydrofuryl, dihydrofuryl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxolanyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, azabicylo[3.2.1]octyl, azabicylo[3.3.1]nonyl, azabicylo[4.3.0]nonyl, oxabicylo[2.2.1]heptyl, 1,5,9-triazacyclododecyl, and the like.
  • Aryl represents a group comprising an aromatic, monovalent monocyclic, bicyclic, or tricyclic radical containing from 6 to 18 carbon ring atoms, which may be unsubstituted or substituted by one or more of the substituents described below, and to which may be fused one or more cycloalkyl groups, heterocycloalkyl groups or heteroaryl groups, which themselves may be unsubstituted or substituted by one or more suitable substituents.
  • aryl groups include the following moieties:
  • Heteroaryl represents a group comprising an aromatic monovalent monocyclic, bicyclic, or tricyclic radical, containing 5 to 18 ring atoms, including 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, which may be unsubstituted or substituted by one or more of the substituents described below.
  • heteroaryl groups include, but are not limited to, thienyl, pyrrolyl, imidazolyl, pyrazolyl, furyl, isothiazolyl, furazanyl, isoxazolyl, thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, benzo[b]thienyl, naphtho[2,3-b]thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl, indolizinyl, isoindolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxyalinyl, quinzolinyl, benzothiazolyl, benzimidazolyl, te
  • suitable substituents that may be present on any of the above alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl groups are described herein and include alkyl (except for alkyl), aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy,
  • Preferred "suitable substituents" include alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, halogen, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heteroaryloxy, and carboxyl.
  • alkyl, alkylene, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl moieties of any of the above substituents may be optionally substituted by one or more of alkyl (except for alkyl), haloalkyl, aryl, nitro, amino, alkylamino, dialkylamino, halogen, hydroxyl, alkoxy, haloalkoxy, aryloxy, mercapto, alkylthio or arylthio groups.
  • the substituent may be protected with a suitable protecting group that is stable to the reaction conditions used in these methods.
  • the protecting group may be removed at a suitable point in the reaction sequence of the method to provide a desired intermediate or target compound.
  • suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3 rd ed.), John Wiley & Sons, NY (1999), which is incorporated herein by reference in its entirety.
  • a substituent may be specifically selected to be reactive under the reaction conditions used in the methods of this invention. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful in an intermediate compound in the methods of this invention or is a desired substituent in a target compound.
  • R 1 and R 2 may be a suitable nitrogen protecting group.
  • nitrogen protecting groups are well known in the art and any nitrogen protecting group that is useful in the methods of preparing the compounds of this invention or maybe useful in the antipicomaviral compounds of this invention may be used.
  • Exemplary nitrogen protecting groups include alkyl, substituted alkyl, carbamate, urea, amide, imide, enamine, sulfenyl, sulfonyl, nitro, nitroso, oxide, phosphinyl, phosphoryl, silyl, organometallic, borinic acid and boronic acid groups.
  • R 1 and/or R 2 are independently suitable nitrogen protecting groups
  • suitable R 1 and R 2 substituents include, but are not limited to, carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc: t-butyloxycarbonyl) and aryloxycarbonyl (e.g., Cbz: benzyloxycarbonyl, or FMOC: fiuorene-9-methyloxycarbonyl), alkyloxycarbonyls (e.g., methyloxycarbonyl), alkyl or arylcarbonyl, substituted alkyl, especially arylalkyl (e.g., trityl (triphenylmethyl), benzyl and substituted benzyl), and the like.
  • carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc: t-butyloxycarbonyl) and aryloxycarbonyl (e.g., Cbz: benzyloxycarbonyl, or FMOC:
  • R 1 and R 2 taken together are a suitable nitrogen protecting group
  • suitable R'/R 2 substituents include phthalimido and a stabase (1,2-bis (dialkylsilyl))ethylene).
  • halogen and “halo” represent chloro, fluoro, bromo or iodo substituents.
  • Heterocycle is intended to mean a heteroaryl or heterocycloalkyl group.
  • Acyl is intended to mean a -C(O)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group.
  • Acyloxy is intended to mean an -OC(O)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group.
  • Thioacyl is intended to mean a -C(S)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group.
  • Sulfonyl is intended to mean an -SO 2 - biradical.
  • Sulfenyl is intended to mean an -SO- biradical.
  • Sulfo is intended to mean an -SO 2 H radical.
  • Haldroxy is intended to mean the radical -OH.
  • Amin or “amino” is intended to mean the radical -NH 2 .
  • Alkylamino is intended to mean the radical -NHR_, where R_ is an alkyl group.
  • Dialkylamino is intended to mean the radical -NR_R b , where R_ and R,, are each independently an alkyl group, and is intended to include heterocycloalkyl groups, wherein R_ and R b , taken together, form a heterocyclic ring that includes the amine nitrogen.
  • Alkoxy is intended to mean the radical -OR-, where R_ is an alkyl group.
  • alkoxy groups include methoxy, ethoxy, propoxy, and the like.
  • “Lower alkoxy” groups have alkyl moieties having from 1 to 4 carbons.
  • “Alkoxycarbonyl” is intended to mean the radical -C(O)OR_, where R. is an alkyl group.
  • “Alkylsulfonyl” is intended to mean the radical -SO 2 R a , where R a is an alkyl group.
  • Alkylenedioxy is intended to mean the divalent radical -OR a O- which is bonded to adjacent atoms (e.g., adjacent atoms on a phenyl or naphthyl ring) , wherein R- is a lower alkyl group.
  • Alkylaminocarbonyl is intended to mean the radical -C(O)NHR a , where R a is an alkyl group.
  • Dialkylaminocarbonyl is intended to mean the radical -C(O)NR a R b , where R a and R b are each independently an alkyl group.
  • Mercapto is intended to mean the radical -SH.
  • Alkylthio is intended to mean the radical -SR-, where R a is an alkyl group.
  • Carboxy is intended to mean the radical -C(O)OH.
  • Carbamoyl is intended to mean the radical -C(O)NH 2 .
  • Cycloalkylalkyl is intended to mean the radical -alkyl-cycloalkyl, wherein alkyl and cycloalkyl are defined as above, and is represented by the bonding arrangement present in the groups -CH 2 -cyclohexane or -CH 2 -cyclohexene.
  • Arylalkyl is intended to mean the radical -alkylaryl, wherein alkyl and aryl are defined as above, and is represented by the bonding arrangement present in a benzyl group.
  • Amocarbonylalkyl is intended to mean the radical -alkylC(O) NH 2 and is represented by the bonding arrangement present in the group -CH 2 CH 2 C(O)NH 2 .
  • Alkylaminocarbonylalkyl is intended to mean the radical -alkylC(O)NHR a , where R_ is an alkyl group and is represented by the bonding arrangement present in the group -CH 2 CH 2 C(O)NHCH 3 .
  • Alkylcarbonylaminoalkyl is intended to mean the radical -alkylNHC(O)-alkyl and is represented by the bonding arrangement present in the group -CH 2 NHC(O)CH 3 .
  • Dialkylaminocarbonylalkyl is intended to mean the radical -alkylC(O)NR a R b , where R_ and R b are each independently an alkyl group.
  • Aryloxy is intended to mean the radical -OR c , where R c is an aryl group.
  • Heteroaryloxy is intended to mean the radical -OR d , where R d is a heteroaryl group.
  • Arylthio is intended to mean the radical -SR C , where R c is an aryl group.
  • Heteroarylthio is intended to mean the radical -SR d , where R d is a heteroaryl group.
  • a desired salt may be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such as glucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citric acid or tartaric acid, amino acid, such as aspartic acid or glutamic acid, aromatic acid, such as benzoic acid or cinnamic acid, sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
  • an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid,
  • a desired salt may be prepared by any suitable method known to the art, including treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary); an alkali metal or alkaline earth metal hydroxide; or the like.
  • suitable salts include organic salts derived from amino acids such as glycine and arginine; ammonia; primary, secondary, and tertiary amines; and cyclic amines, such as piperidine, morpholine, and piperazine; as well as inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.
  • All compounds of this invention contain at least one chiral center and may exist as single stereoisomers (e.g., single enantiomers or single diastereomers), any mixture of stereosisomers (e.g., any mixture of enantiomers or diastereomers) or racemic mixtures thereof. All such single stereoisomers, mixtures and racemates are intended to be encompassed within the broad scope of the present invention.
  • Compounds identified herein as single stereoisomers are meant to describe compounds that are present in a form that contains at least 90% of a single stereoisomer of each chiral center present in the compounds.
  • the chemical structure is intended to encompass compounds containing either stereoisomer of each chiral center present in the compound.
  • the inventive compounds are used in optically pure, that is, stereoisomerically pure, form or substantially optically pure (substantially stereoisomerically pure) form.
  • stereoisomeric purity refers to the "enantiomeric” purity and/or “diastereomeric” purity of a compound.
  • Compounds that are substantially enatiomerically pure contain at least 90% of a single isomer and preferably contain at least 95% of a single isomer of each chiral center present in the enantiomer.
  • Compounds that are substantially diastereomerically pure contain at least 90% of a single isomer of each chiral center present in the diastereomer, and preferably contain at least 95% of a single isomer of each chiral center. More preferably, the substantially enantiomerically and diasteriomerically pure compounds in this invention contain at least 97.5% of a single isomer and most preferably contain at least 99% of a single isomer of each chiral center in the compound.
  • racemic or “racemic mixture” refers to a mixture of equal amounts of enantiomeric compounds, which encompasses mixtures of enantiomers and mixtures of enantiomeric diastereomers.
  • the compounds of this invention may be obtained in stereoisomerically pure (i.e., enantiomerically and/or diastereomerically pure) or substantially stereoisomerically pure (i.e., substantially enantiomerically and/or diastereomerically pure) form.
  • Such compounds may be obtained synthetically, according to the procedures described herein using optically pure or substantially optically pure materials. Alternatively, these compounds may be obtained by resolution/separation of a mixture of stereoisomers, including racemic mixtures, using conventional procedures.
  • Exemplary methods that may be useful for the resolution/separation of stereoisomeric mixtures include chromatography and crystallization/re-crystallization. Other useful methods may be found in "Enantiomers, Racemates, and Resolutions," J. Jacques et al., 1981, John Wiley and Sons, New York, NY, the disclosure of which is incorporated herein by reference. Preferred stereoisomers of the compounds of this invention are described herein.
  • Another embodiment of this invention comprises the compounds depicted by Formula I-a (as represented by Formula I, wherein the dotted line represents a bond):
  • R 2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
  • R 3 , R 4 , R 5 are independently H or a suitable substituent described above, and
  • R 2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
  • R x represents H or one or more suitable substituents
  • R 3 and R 4 are independently H or a suitable substituent described above;
  • R 1 , R 9 , R 10 , Z and Z 1 , and the variables contained therein, are as defined above.
  • a further embodiment of this invention comprises the compounds depicted by
  • R , R and R are independently H or a substituted or unsubstituted lower alkyl group
  • R ⁇ represents H or one or more suitable substituents
  • R 4 and R 5 are independently H or a suitable substituent described above; and R 1 , R 6 , R' ⁇ Z and Z 1 , and the variables contained therein, are as defined above.
  • Another embodiment of this invention comprises the compounds depicted by Formula I-d:
  • R 7 , R 8 and R 10 are independently H or a substituted or unsubstituted lower alkyl group
  • R ⁇ represents H or one or more suitable substituents
  • R 4 and R 5 are independently H or a suitable substituent described above;
  • R 1 , R 6 , R 9 , Z and Z 1 are as defined above.
  • Yet another embodiment of this invention comprises the compounds depicted by Formula I-e (as represented by Formula I, wherein the dotted line does not represent a bond):
  • R , R and R are independently H or a substituted or unsubstituted lower alkyl group
  • R ⁇ represents H or one or more suitable substituents
  • R 4 and R 5 are independently H or a suitable substituent described above;
  • R 1 , R 6 , R 9 , Z and Z 1 , and the variables contained therein, are as defined above.
  • R 2 is selected from a substituted or unsubstituted alkyloxycarbonyl group, alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocycloalkylcarbonyl or heteroarylcarbonyl group.
  • R 2 is an unsubstituted or substituted arylalkyloxycarbonyl group.
  • R 2 groups include, but are not limited to benzyloxycarbonyl, methylcarbonyl, t-butylcarbonyl, trifluoromethylcarbonyl, cyclopentylcarbonyl, tetrahydrofuran-2-carbonyl, l,3-dithiolane-2-carbonyl and the like.
  • R 2 is a substituted or unsubstituted benzyloxycarbonyl, arylcarbonyl, or heteroarylcarbonyl group.
  • R 2 is an unsubstituted or substituted benzyloxycarbonyl or heteroarylcarbonyl group, wherein the heteroaryl moiety is a five-membered heterocycle having from one to three heteroatoms selected from O, N, and S, more preferably a five-membered heterocycle having at least one nitrogen heteroatom and at least one oxygen heteroatom (e.g., unsubstituted or substituted 1,2-oxazolyl (i.e., isoxazolyl), 1,3-oxazolyl (i.e., oxazolyl), or oxadiazolyl (1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, or 1,2,5-oxadiazolyl).
  • 1,2-oxazolyl i.e., isoxazolyl
  • 1,3-oxazolyl i.e., oxazolyl
  • oxadiazolyl 1,2,3-oxadiazolyl
  • heteroaryl moiety When the heteroaryl moiety is oxadiazolyl, unsubstituted and monomethyl-substituted 1,2,4-oxadiazolyl are preferred.
  • the heteroaryl moiety is 3-isoxazolyl or 5-isoxazolyl, either unsubstituted or substituted with one or two methyl groups and/or halogens (F, Cl, Br or I), with chlorine and fluorine being preferred.
  • the heteroarylcarbonylgroup in the especially preferred embodiments is an unsubstituted or substituted 3-carbonyl- 1,2-oxazolyl group (i.e., 3-carbonyl-isoxazolyl) or a 5-carbonyl- 1,2-oxazolyl group (i.e., 5-carbonyl-isoxazolyl).
  • the substituent variables R 3 , R 4 and R 5 as present in the compounds of Formulas I-a, I-b, I-c, I-d and I-e, are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto, alkoxy, aryloxy, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, or aryloxy.
  • the substituent variables R_ in the compounds of Formula I-b are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoy
  • the substituent variable R y in the compounds of formula Formulas I-c, I-d and I-e are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxy, heterocycloalkyloxy, heterocycl
  • R 6 is H or an unsubstituted alkyl group or an optionally substituted lower alkyl group, wherein these groups are comprised of a straight- or branched-chain saturated hydrocarbon group, a straight- or branched-chain substituted saturated hydrocarbon group, or group comprised of a straight- or branched-chain saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety.
  • R 6 is a substituted alkyl group, the point of attachment of R 6 is via a saturated hydrocarbon moiety.
  • R 6 is a substituted saturated hydrocarbon group
  • the saturated hydrocarbon group may be optionally substituted with a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an alkoxy group, an aryloxy group, an alkylthio group, an arylthio group, wherein each alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl moiety thereof may be optionally substituted.
  • R 6 is comprised of a saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety
  • the saturated hydrocarbon moiety may be bound to an unsaturated hydrocarbon moiety containing one or more double-bonds or triple-bonds, the terminal positions of which may be substituted by the substituents described above, or may contain additional straight- or branched-chain saturated hydrocarbon moieties.
  • the unsaturated hydrocarbon moiety contains one double-bond or one triple-bond, the terminal position(s) of which may optionally contain a straight- or branched-chain saturated hydrocarbon moiety.
  • both terminal positions of the double bond contain a straight- or branched-chain saturated hydrocarbon moiety.
  • R 6 is H or a substituted or unsubstituted lower alkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl group, or a group comprised of a straight-chain saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety.
  • R 6 is H, methyl, substituted methyl, ethyl, n-propyl, n-butyl, sec-butyl, 2-propyn-l-yl, 3-methyl-3-buten-l-yl, -methylcyclohexyl, substituted or unsubstituted -methylthienyl or substituted or unsubstituted benzyl, wherein the phenyl moiety of the substituted benzyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino or halogen and the thienyl moiety of the substituted -methylthienyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino or halogen.
  • R 6 When R 6 is substituted methyl, the methyl (methylene) moiety may be substituted with an alkoxy group, an aryloxy group, an alkylthio group or an arylthio group. Most preferably, R 6 is H, ethyl, 2-propyn-l-yl, -methylcyclohexyl, or substituted or unsubstituted benzyl, wherein the phenyl moiety of the substituted benzyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy and halogen.
  • R b and R 9 are defined as above and m is 1 and p is 1 or 2 (i.e., both A 2 and A 3 are present) or when p is 0, m is 0 (i.e, both A 2 and A 3 are absent). More preferably, in R b and R 9 , when m is 1 and p is 1 or 2, A 2 and A 3 are both C(R h )(R') or C(R 14 )(R 15 ), respectively. More preferably, when m is 1, p is 1.
  • R 9 is selected from -CH 2 CH 2 C(O)NH 2 ; -CH 2 CH 2 C(O)NH-alkyl; -CH 2 NHC(O)CH 3 ; and
  • n 1 or 2. More preferably, R 9 is
  • Z and Z 1 are independently H, substituted or unsubstituted alkyl, -CO 2 R n or -CO 2 R 19 , as appropriate, wherein R n and R 19 are as defined above, or Z and Z 1 , taken together with the atom to which they are attached, form a heterocycloalkyl group, as defined above.
  • Z and/or Z 1 may be -C(S)OR n or -C(S)OR 19 , wherein R n and R 19 are as defined above.
  • Such compounds may be prepared using procedures described in K.
  • Z and Z 1 are independently H, substituted or unsubstituted alkyl, -CO 2 R 19 , or taken together with the atom to which they are attached, form a heterocycloalkyl group, which may be optionally substituted.
  • Z and Z 1 are independently selected from H, -CO 2 H, substituted or unsubstituted lower alkyl, -CO 2 -alkyl, -CO 2 -cycloalkyl, -CO 2 -alkylaryl (e.g., -CO 2 -benzyl), -CO 2 -alkylheteroaryl (e.g., -CO 2 -(CH 2 ) n pyridyl) or taken together with the atom to which they are attached form a heterocycloalkyl group , which may be optionally substituted.
  • the heterocycloalkyl group may optionally contain O, N, S and/or P and may be substituted by one or more of oxo (keto) or thioketo.
  • Z and Z 1 are not both H.
  • Z 1 is H or lower alkyl and Z is -CO 2 H, substituted or unsubstituted-CO 2 -alkyl, -CO 2 -alkylaryl, -CO 2 -alkylheteroaryl, -CO 2 -cycloalkyl, or or taken together with the atom to which they are attached form a heterocycloalkyl group, which may be optionally substituted.
  • Exemplary Z groups include, but are not limited to substituted and unsubstituted -CO 2 -alkyl groups, which include straight- and branched-chain alkyl groups such as ethoxycarbonyl, t-butoxycarbonyl, isopropoxycarbonyl, (2,2-dimethylpropyl)- oxycarbonyl, and the like, and which include straight and branched-chain arylalkyl and heteroarylalkyl groups, such as benzyloxycarbonyl, pyridylmethyleneoxycarbonyl, and the like, substituted and unsubstituted -CO 2 -cycloalkyl groups such as cyclobutyloxycarbonyl, cyclopentyloxycarbonyl,, cyclohexyloxycarbonyl, cycloheptyloxycarbonyl, and the like, or taken together with Z 1 and the atom to which they are
  • R 1 , R 2 , R 3 , R 4 , R , R 6 , R 9 , Z and Z 1 are as previously defined;
  • R , R , R , R , R x , Z and Z are as previously defined;
  • R 4 , R 5 , R 6 , R 9 , R y , Z and Z 1 are as previously defined;
  • R , R , R 6 , R , R_,, Z and Z are as previously defined;
  • R 4 , R 5 , R 6 , R 9 , R y , Z and Z 1 are as previously defined.
  • R 2 is unsubstituted or substituted benzyloxycarbonyl, arylcarbonyl, or heteroarylcarbonyl, more preferably heteroarylcarbonyl, where the heteroaryl moiety is a five-membered heterocycle having from one to three heteroatoms selected from O, N, and S.
  • R 2 is heteroarylcarbonyl wherein the heteroaryl moiety is a five-membered heterocycle having at least one nitrogen heteroatom and at least one oxygen heteroatom (e.g., unsubstituted or substituted 1,2-oxazolyl (i.e., isoxazolyl), 1,3-oxazolyl (i.e., oxazolyl), or oxadiazolyl (1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, or 1,2,5-oxadiazolyl); preferred oxadiazolyls are unsubstituted and monomethyl-substituted 1,2,4-oxadiazolyl.
  • 1,2-oxazolyl i.e., isoxazolyl
  • 1,3-oxazolyl i.e., oxazolyl
  • oxadiazolyl 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl,
  • the heteroaryl moiety is 3-isoxazolyl or 5-isoxazolyl, either unsubstituted or substituted with one or two substituents selected from methyl and halogen, with chloro and fluoro being preferred halogen substituents.
  • R 6 is selected from H or:
  • R ' may be H or alkyl and R" may be H or independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino, and halogen.
  • a particularly preferred embodiment of this invention comprises a compound having the formula:
  • One preferred stereoisomer of this compound may be represented by the formula:
  • Another preferred stereoisomer of this compound may be represented by the formula:
  • An especially preferred stereoisomer of this compound may be represented by the formula:
  • Preferred specific compounds include those of the Examples below, especially:
  • the invention is also directed to the intermediates of Formula ⁇ , which are useful in the synthesis of certain compounds of Formula I:
  • R 1 , R 2 , R 3 , R 4 and R x are as defined above, and B is H, -OR 24
  • R 1 and/or R 2 are independently suitable nitrogen protecting groups, any suitable nitrogen-protecting group known in the art may be used (see, e.g., Greene & Wuts, supra).
  • Suitable R 1 and R 2 substituents include, but are not limited to, carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc) and aryloxycarbonyl (e.g., Cbz or FMOC), alkyloxycarbonyls (e.g., methyloxycarbonyl), alkyl or arylcarbonyl, substituted alkyl, especially arylalkyl (e.g., trityl (triphenylmethyl), benzyl and substituted benzyl) and the like.
  • carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc) and aryloxycarbonyl (e.g., Cbz or FMOC), alkyloxycarbonyls (e.g., methyloxycarbonyl), alkyl or arylcarbonyl, substituted alkyl, especially arylalkyl (e.g., trityl (triphenylmethyl), benzyl and substitute
  • R 1 and R 2 are independently suitable nitrogen protecting groups
  • suitable R 1 and R 2 substituents include, but are not limited to, Boc, Cbz, FMOC, methyloxycarbonyl and trityl.
  • suitable R'/R 2 substituents include phthalimido and a stabase (1,2-bis (dialkylsilyl))ethylene).
  • R 24 may be H or a suitable protecting group for a carboxyl moiety.
  • Suitable carboxyl protecting groups are also well known in the art, examples of which may be found in Greene and Wuts, supra, and include, but are not limited to, protecting groups where R 24 is alkyl, substituted or unsubstituted aryl, alkyl and/or aryl substituted silyl (e.g., t-butyldimethylsilyl (TBS)), and the like.
  • R 24 is alkyl, substituted or unsubstituted aryl, alkyl and/or aryl substituted silyl (e.g., t-butyldimethylsilyl (TBS)), and the like.
  • TBS t-butyldimethylsilyl
  • the invention is also directed to pharmaceutically acceptable salts of the compounds ol Formula II.
  • Preferred examples of the Formula II useful as intermediates include the following:
  • the antipicomaviral compounds of this invention include prodrugs, the pharmaceutically active metabolites, and the pharmaceutically acceptable salts and solvates thereof.
  • the compounds of Formula I, prodrugs, pharmaceutically acceptable salts, and pharmaceutically active metabolites and solvates thereof have an antipicomaviral activity, more preferably antirhinoviral activity, corresponding to an EC 50 less than or equal to 100 ⁇ M in the Hl-HeLa cell culture assay.
  • a "prodrug” is intended to mean a compound that is converted under physiological conditions or by solvolysis or metabolically to a specified compound that is pharmaceutically active.
  • a prodrug may be a derivative of one of the compounds of this invention that contains a moiety, such as for example -CO 2 R, or -PO(OR) 2 , that may be cleaved under physiological conditions or by solvolysis. Any suitable R substituent may be used that provides a pharmaceutically acceptable solvolysis or cleavage product.
  • a prodrug containing such a moiety may be prepared according to conventional procedures by treatment of a compound of this invention containing, for example, an amido, carboxylic acid, or hydroxyl moiety with a suitable reagent.
  • a "pharmaceutically active metabolite" is intended to mean a pharmacologically active compound produced through metabolism in the body of a specified compound.
  • a "pharmaceutically acceptable salt” is intended to mean a salt that retains the biological effectiveness of the free acids and bases of a specified compound and that is not biologically or otherwise undesirable.
  • pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-l,4-dioates, hexyne-l,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates
  • a “solvate” is intended to mean a pharmaceutically acceptable solvate form of a specified compound that retains the biological effectiveness of such compound.
  • solvates include compounds of the invention in combination with water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine.
  • inventive compounds, salts, and solvates may exist in different crystal forms, all of which are intended to be within the scope of the present invention and specified formulas.
  • the present invention is also directed to a method of inhibiting picornaviral 3C protease activity, comprising contacting the protease with an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, prodrug, pharmaceutically active metabolite, or solvate thereof.
  • picornaviral 3C protease activity may be inhibited in mammalian tissue by administering a compound of Formula I or a pharmaceutically acceptable salt, prodrug, pharmaceutically active metabolite, or solvate thereof.
  • the present method is directed at inhibiting rhinviral protease activity.
  • Treating is intended to mean at least the mitigation of a disease condition in a mammal, such as a human, that is alleviated by the inhibition of the activity of one or more picornaviral 3C proteases, including, but not limited to human rhinoviruses, human poliovirus, human coxsackieviruses, encephalomyocarditis viruses, meningitis virus, and hepatitis A virus.
  • the methods of treatment for mitigation of a disease condition include the use of the compounds in this invention in any conventionally acceptable manner, for example, as a prophylactic.
  • the activity of the inventive compounds as inhibitors of picornaviral 3C protease activity may be measured by any of the suitable methods known to those skilled in the art, including in vivo and in vitro assays.
  • An example of a suitable assay for activity measurements is the antiviral Hl-HeLa cell culture assay described herein.
  • Administration of the compounds of the Formula I and their pharmaceutically acceptable prodrugs, salts, active metabolites, and solvates may be performed according to any of the generally accepted modes of administration available to those skilled in the art.
  • suitable modes of administration include oral, nasal, parenteral, topical, transdermal, and rectal.
  • An inventive compound of Formula I or a pharmaceutically acceptable salt, prodrug, active metabolite, or solvate thereof may be administered as a pharmaceutical composition in any pharmaceutical form recognizable to the skilled artisan as being suitable.
  • Suitable pharmaceutical forms include solid, semisolid, liquid, or lyophilized formulations, such as tablets, powders, capsules, suppositories, suspensions, liposomes, and aerosols.
  • compositions of the invention may also include suitable excipients, diluents, vehicles, and carriers, as well as other pharmaceutically active agents, depending upon the intended use or mode of administration.
  • inventive pharmaceutical compositions are delivered orally, or intranasally in the form of suspensions.
  • Acceptable methods of preparing suitable pharmaceutical forms of the pharmaceutical compositions may be routinely determined by those skilled in the art.
  • pharmaceutical preparations may be prepared following conventional techniques of the pharmaceutical chemist involving steps such as mixing, granulating, and compressing when necessary for tablet forms, or mixing, filling, and dissolving the ingredients as appropriate, to give the desired products for oral, parenteral, topical, intravaginal, intranasal, intrabronchial, intraocular, intraaural, and/or rectal administration.
  • Solid or liquid pharmaceutically acceptable carriers, diluents, vehicles, or excipients may be employed in the pharmaceutical compositions.
  • Illustrative solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, pectin, acacia, magnesium stearate, and stearic acid.
  • Illustrative liquid carriers include syrup, peanut oil, olive oil, saline solution, and water.
  • the carrier or diluent may include a suitable prolonged-release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax.
  • the preparation may be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid (e.g., solution), or a nonaqueous or aqueous liquid suspension.
  • a dose of the pharmaceutical composition contains at least a therapeutically effective amount of the active compound (i.e., a compound of Formula I or a pharmaceutically acceptable salt, prodrug, active metabolite, or solvate thereof), and preferably is made up of one or more pharmaceutical dosage units.
  • the selected dose may be administered to a mammal, for example, a human patient, in need of treatment mediated by inhibition of picornaviral 3C protease activity, by any known or suitable method of administering the dose, including: topically, for example, as an ointment or cream; orally; rectally, for example, as a suppository; parenterally by injection; or continuously by intravaginal, intranasal, intrabronchial, intraaural, or intraocular infusion.
  • a “therapeutically effective amount” is intended to mean the amount of an inventive agent that, when administered to a mammal in need thereof, is sufficient to effect treatment for disease conditions alleviated by the inhibition of the activity of one or more picornaviral 3C proteases, such as human rhinoviruses, human poliovirus, human coxsackieviruses, encephalomyocarditis viruses, menigovirus, and hepatitis A virus.
  • the amount of a given compound of the invention that will be therapeutically effective will vary depending upon factors such as the particular compound, the disease condition and the severity thereof, the identity of the mammal in need thereof, which amount may be routinely determined by artisans.
  • inventive compounds of Formulas I and II are prepared by the methods of the present invention, including the General Methods shown below.
  • stereochemistry is not specified in chemical structures, either stereocenter may be utilized.
  • 2-hydroxypyridines it is understood that such entities may also exist as the corresponding 2-pyridone tautomers.
  • Boc tert-butoxycarbonyl
  • Ac acetyl
  • Cbz benzyloxycarbonyl
  • DMB 2,4-dimethoxybenzyl
  • TBS tert-butyldimethylsilyl
  • TBDPS tert-butyldiphenylsilyl
  • Ms methanesulfonate
  • Ts toluenesulfonate
  • Bn benzyl
  • Tr triphenylmethyl
  • the alcohol functionality present in D is then converted to an appropriate leaving group (e.g., mesylate, tosylate) E and is coupled with 2-hydroxypyridine F (which incorporates Ri, R2, R3, R4, and R5) to give intermediate G.
  • an appropriate leaving group e.g., mesylate, tosylate
  • 2-hydroxypyridine F which incorporates Ri, R2, R3, R4, and R5
  • the Ri and R2 moieties present in F may be an appropriate protecting groups for the amine functionality.
  • the P3 protecting group is subsequently removed from G and the resulting alcohol (H) is oxidized to the corresponding aldehyde (not shown) and subjected to an olefin-forming reaction to afford intermediate I (which incorporates Ri 1, Z, and Zi).
  • the P2 protecting group present in I is then removed to give product J.
  • Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates G, H, or I or product J and replaced with a different Ri and/or R2 substituent to afford alternate intermediates G, H, or I or products J.
  • amino alcohol K (prepared by methods described in the chemical literature), which incorporates R9 and in which Rio is H, Pi is an appropriate protecting group for amine functionality (e.g, Cbz, Boc, or Ac), and P2 is an appropriate protecting group for the amide nitrogen (e.g, DMB), is converted to compound L where P3 is an appropriate protecting group for the alcohol functionality (e.g., TBDPS).
  • P3 is an appropriate protecting group for the alcohol functionality (e.g., TBDPS).
  • the Pi protecting group present in L is then removed and the resulting amine or salt thereof (not shown) is subjected to an amide bond forming reaction with an appropriate ⁇ -hydroxycarboxylic acid (which incorporates R6 and in which R7 is H; also not shown) to provide intermediate M.
  • the alcohol functionality present in M is then converted to an appropriate leaving group (e.g., mesylate, tosylate) N and is coupled with 2-hydroxypyridine F (which incorporates Ri, R2, R3, R 4 , and R5) to give intermediate O.
  • an appropriate leaving group e.g., mesylate, tosylate
  • 2-hydroxypyridine F which incorporates Ri, R2, R3, R 4 , and R5
  • the Ri and R2 moieties present in F may be an appropriate protecting group for the amine functionality.
  • the P3 protecting group is subsequently removed from O and the resulting alcohol (P) is oxidized to the corresponding aldehyde (not shown) and subjected to an olefin-forming reaction to afford intermediate Q (which incorporates Ri 1 , Z, and Z ⁇ ).
  • the P2 protecting group present in Q is then removed to give product R.
  • Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates O, P, or Q or product R and replaced with a different Ri and/or R2 substituent to afford alternate intermediates O, P, or Q or products R.
  • the P4 protecting group is subsequently removed from V to afford carboxylic acid W.
  • intermediates X and Z prepared by methods described in the chemical literature) which incorporate Ri 1, Z, and Zi and in which Pi is an appropriate protecting group for the amine functionality (e.g., Cbz, Boc, or Ac) and P2 is an appropriate protecting group for the amide nitrogen (e.g., Tr), are converted to their corresponding amines Y and AA (or salts thereof), respectively.
  • Amines AA and Y are then independently coupled with carboxylic acid W to afford intermediate I and product R, respectively.
  • the P2 protecting group present in intermediate I is subsequently removed to afford product J.
  • the lactam nitrogen present in intermediate X may be protected with a suitable protecting group (e.g., DMB). If such a moiety is present in X, it may be removed after coupling of W with Y to afford product R.
  • a suitable protecting group e.g., DMB
  • an appropriate pyridine g (which incorporates Ri, R2, R3, and R4, and which may be optionally substituted with R x as shown) (an example of which may prepared as described in Specific Method 7 below) in which Pi 1 and P12 are protecting groups for the alcohol functionality (e.g., silyl ether, methyl) is subjected to an intramolecular cychzation/deprotection protocol in which both Pi 1 and P 12 are removed to give pyridone h.
  • the Ri and R2 moieties present in g may be an appropriate protecting group for the amine functionality (e.g., Cbz).
  • the alcohol moiety present in h is then oxidized to the corresponding carboxylic acid i.
  • This oxidation may be accomplished via an aldehyde intermediate (not shown).
  • Carboxylic acid i is then independently coupled with amines AA and Y (or salts thereof) (prepared as described in General Method 3 above) to afford intermediate j and product 1, respectively.
  • the P2 protecting group present in intermediate j is subsequently removed to afford product k.
  • the lactam nitrogen present in intermediate Y may be protected with a suitable protecting group (e.g., DMB). If such a moiety is present in Y, it may be removed after coupling of i with Y to afford product 1.
  • a suitable protecting group e.g., DMB
  • Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates g, h, i, or j or products k or 1 and replaced with a different Ri and/or R2 substituent to afford alternate intermediates g, h, i, or j or products k or 1.
  • the hydroxyl group present in o is converted to the corresponding trifluoromethane sulfonate (OTf) p, and this intermediate is further transformed to the amine r via imine q.
  • the amine present in r is derivatized with an appropriate moiety to afford intermediate s (which contains the R2 functional group and in which Ri is H).
  • Intermediate s is subjected to a desulfurization reaction to give intermediate t and this entity is deprotected to give carboxylic acid i (in which R l , R3, R 4 , and R x are H).
  • intermediate s may be deprotected to give carboxylic acid u (in which Ri, R3, and R x are H) which may be utilized in place of i in General Method 4 above.
  • intermediate z (which contains the R2 functional group and in which Ri is H).
  • Intermediate z is reduced to give intermediate aa and this entity is transformed to intermediate bb in which X is a halogen.
  • Intermediate bb is subsequently deprotected to give carboxylic acid cc.
  • intermediate aa may also be transformed into intermediate t (General Method 5 above).
  • intermediate aa is transformed to intermediate dd which is subsequently deprotected to give carboxylic acid ee.
  • intermediate aa is transformed to intermediate ff in which R is alkyl, acyl, sulfonyl, or acyloxy.
  • Intermediate ff is subsequently deprotected to give carboxylic acid gg.
  • Carboxylic acids cc, ee, and gg may each be utilized in place of i in General Method 4 above.
  • hydroxyl group present in hh is converted to the corresponding trifluoromethane sulfonate (OTf) ii, and this intermediate is further transformed to the amine kk via imine jj.
  • the amine present in kk is derivatized with an appropriate moiety to afford intermediate 11 (which contains the R2 functional group and in which Ri is H).
  • Intermediate 11 is subsequently deprotected to give carboxylic acid mm which may be utilized in place of i in General Method 4 above.
  • Z2 is alkoxy or benzyloxy it may be replaced with a hydroxyl functionality.
  • the resulting carboxylic acid may subsequently be rearranged to the corresponding amine using established methods and the amine-containing intermediates may be utilized as depicted in General Method 6.
  • intermediate 11 when Z2 is OH
  • intermediate aa General Method 6
  • R x is/are H and P13 is a suitable protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl) to give intermediate ss.
  • Intermediate ss is subsequently oxidized (by a variety of literature methods) to afford intermediate z which may be utilized as described above in General Method 6.
  • General Method 10 depicts the preparation of the bicyclic pyrrole vv, starting with aldehyde tt, prepared according to the procedure described in Smith, K. M., J. Chem. Soc, Perkin Trans. I, 1973, p. 516. This compound is subjected to reductive amination conditions with an amine to give compound uu. The carboxylic acid protecting group Pi6 is removed, and the resulting acid is condensed mtramolecularly with the secondary amine to give bicycle w.
  • This bicyclic compound may be coupled to amine Y or AA according to the method described in General Method 3.
  • Specific Method 1 describes the preparation of specific compound Jl (compound 5).
  • commercially available amino acid Al was reduced to alcohol BI which, in turn, was transformed into compound Cl .
  • the Cbz moiety present in Cl was removed by hydrogenation and the resulting amine (not shown) was coupled with D-3-phenyllactic acid (commercially available) to afford intermediate Dl .
  • This latter entity was subsequently transformed to the corresponding methanesulfonate (mesylate) (El) and was coupled with the sodium salt of 2-hydroxypyridine FI to provide intermediate Gl.
  • the 2-hydroxypyridine FI was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent Boc protection of the resulting amine.
  • Specific Method 3 describes the preparation of specific compound R2 (compound 23).
  • Boc-D-(4-F)Phe-OH was deprotected under acidic conditions and the resulting amine salt was subjected to a diazotization/displacement protocol to provide (2R)-3-(4'-fluorophenyl)-2-hydroxypropionic acid (SI).
  • This material was subsequently transformed into the corresponding trifluoromethane sulfonate (triflate) UI via the methyl ester Tl and was coupled with the sodium salt of 2-hydroxypyridine F3 to provide intermediate VI.
  • the 2-hydroxypyridine F3 was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent derivatization of the resulting amine with 5-chloroisoxazole-3-carbonyl chloride (prepared as described in the Experimental Section of this work).
  • the methyl ester present in VI was subsequently hydrolyzed under basic conditions and the resulting carboxylic acid (Wl) was coupled with amine Yl (or salt thereof) to complete the preparation of specific compound R2 (compound 23).
  • Amine Yl (or salt thereof) was prepared by deprotection of intermediate XI (prepared in a manner analogous to that described in Baldwin et al., J. Org. Chem. 1971, 36, 1441).
  • Specific Method 4 describes the preparation of specific compound J2 (compound 24).
  • (2R)-3-(4'-fluorophenyl)-2-hydroxypropionic acid (SI, prepared as described above in Specific Method 3) was transformed into the corresponding trifluoromethane sulfonate (triflate) U2 via the benzyl ester T2 and was coupled with the sodium salt of 2-hydroxypyridine F4 to provide intermediate V2.
  • the 2-hydroxypyridine F4 was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent derivatization of the resulting amine with trifluoroacetic anhydride.
  • Specific Method 5 describes the preparation of specific compound R3 (compound 26).
  • R3 commercially available tert-butyl (R)-2-hydroxybutyrate (T3) was transformed into the corresponding trifluoromethane sulfonate (triflate) U3 and was coupled with the sodium salt of 2-hydroxypyridine F2 (prepared as described in Specific Method 2 above) to provide intermediate V3.
  • the tert-butyl ester present in V3 was subsequently hydro lyzed under acidic conditions and the resulting carboxylic acid (W3) was coupled with amine Y2 (or salt thereof) to complete the preparation of specific compound R3 (compound 26).
  • Amine Y2 (or salt thereof) was prepared by deprotection of intermediate X2, prepared according to the method disclosed in the co-pending application, U.S. Provisional Patent Application No. 60/150,358, filed August 24, 1999, the disclosure of which is inco ⁇ orated herein by reference.
  • R4 Y3 Specific Method 6 describes the preparation of specific compound R4 (compound 29).
  • trifluoromethane sulfonate (triflate) U3 prepared as described in Specific method 5 above
  • 2-hydroxypyridine F3 prepared as described in Specific Method 3 above
  • the tert-butyl ester present in V4 was subsequently hydrolyzed under acidic conditions and the resulting carboxylic acid (W4) was coupled with amine Y3 (or salt thereof) to complete the preparation of specific compound R4 (compound 29).
  • Amine Y3 (or salt thereof) was synthesized from alcohol Kl (prepared as described in Dragovich, et al., J. Med Chem.
  • Alcohol Kl was oxidized to the corresponding aldehyde (not shown) and subjected to an olefin- forming reaction to give intermediate X3.
  • the DMB moiety contained in X3 was then deprotected to provide intermediate X4, and this entity was deprotected under acidic conditions to afford amine Y3 (or salt thereof).
  • the hydroxyl moiety contained in dl was selectively methylated and the resulting methyl ether (el) was subjected to an asymmetric dihydroxylation reaction to give diol fl .
  • This asymmetric dihydroxylation reaction can be effected utilizing a variety of commercial and non-commercial chiral additives.
  • the primary hydroxyl moiety contained in fl was selectively protected as the corresponding tert-butyldimethylsilyl ether (gl).
  • This intermediate was treated with trifluoromethanesulfonic anhydride in the presence of 2,6-lutidine at low temperature followed by exposure to tetrabutylammonium fluoride to effect (i) intramolecular cyclization and (ii) silyl ether deprotection and afford alcohol hi.
  • Alcohol hi was oxidized to the corresponding aldehyde (not shown), and this intermediate was further oxidized to the corresponding carboxylic acid il.
  • Acid il was coupled with amine AAl (or salt thereof) to give intermediate j 1.
  • the trityl protecting group present in j 1 was then removed under acidic conditions to complete the preparation of specific compound kl (compound 36).
  • Amine AAl (or salt thereof) was prepared by deprotection of intermediate Zl (prepared as described in Dragovich, et al. J. Med. Chem. 1998, 41, 2806).
  • Specific Method 9 describes the synthesis of a bicyclic pyrrole.
  • Alcohol ml was oxidized with 2-iodoxybenzoic acid to give aldehyde nl, then reductively aminated with phenylalanine methyl ester and sodium cyanoborohydride to give amine ol.
  • the t-butyl- protecting group was selectively removed, and the resulting amino acid was cyclized with DCC-HOBT to give pi .
  • the methyl ester was cleaved with lithium hydroxide to give acid ql .
  • Boc-protected 4S-amino-5-(2-oxo-pyrrolidin-3S-yl)-pent-2(trans)-enoic acid ethyl ester X2 was deprotected with HC1, then coupled to acid ql, using HATU, to complete the preparation of compound 43.
  • Flash column chromatography was performed using Silica gel 60 (Merck Art 9385). Analytical thin layer chromatography (TLC) was performed using precoated sheets of Silica 60 F254 (Merck Art 5719). Melting points were determined on a Mel-Temp apparatus and are uncorrected. All reactions were performed in septum-sealed flasks under a slight positive pressure of argon unless otherwise noted. All commercial reagents were used as received from their respective suppliers with the following exceptions. Tetrahydrofuran (THF) was distilled from sodium-benzophenone ketyl prior to use. Dichloromethane (CH2CI2) was distilled from calcium hydride prior to use. Et2 ⁇ refers to diethyl ether.
  • THF Tetrahydrofuran
  • CH2CI2 Dichloromethane
  • Et2 ⁇ refers to diethyl ether.
  • DMF refers to N,N-dimethylformamide.
  • DMSO refers to dimethylsulfoxide.
  • MTBE refers to tert-butyl methyl ether.
  • Other abbreviations include: CH3OH (methanol), EtOH (ethanol), EtOAc (ethyl acetate), DME (ethylene glycol dimethyl ether) Ac (acetyl), Me (methyl), Ph (phenyl), Tr (triphenylmethyl), Cbz (benzyloxycarbonyl), Boc (tert-butoxycarbonyl), TFA (trifluoroacetic acid), DIEA (N,N-diisopropylethylamine), TMEDA (N,N,N',N'-tetramethylethylenediamine), AcOH (acetic acid), AC2O (acetic anhydride), NMM (4-methylmorpholine), HOBt (1-hydroxybenzotriazole hydrate), HATU [O-(7-azabenzotriazol- 1
  • Methanesulfonyl chloride (0.286 mL, 3.70 mmol, 1.2 equiv) was added dropwise, slowly, with vigorous stirring. After 30 min, the reaction mixture was diluted with CH2CI2 (200 mL), washed with brine (50 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
  • the title compound was prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate [l,3]dithiolane-2-carbonyl chloride (prepared as described in Helv. Chim.
  • Boc-D-3,4-Difluorophenylalanine (3.05 g, 11.0 mmol, 1 equiv) was dissolved in 1,4- dioxane (10 mL), and a solution of ⁇ C1 in 1,4-dioxane (4.0 M, 10 mL) was added. The reaction was stirred at 23 °C for 5 h, then the solvent was removed under reduced pressure. The residue was dissolved in 1 M ⁇ 2 SO 4 (22 mL), cooled to 0 °C and 2 N NaNO 2 (22 mL) was added via addition funnel under argon. The reaction mixture was stirred at 0 °C for 3 h and warmed to room temperature overnight.
  • Boc-D-Cyclohexylalanine-OH (3.00 g, 11.1 mmol, 1 equiv) was dissolved in 1,4- dioxane (10 mL), and a solution of HC1 in 1,4-dioxane (4.0 M, 10 mL) was added. The reaction was stirred at 23 °C for 5 h, then the solvent was removed under reduced pressure. The residue was dissolved in 1 M H 2 SO 4 (22 mL), cooled to 0 °C and 2 N NaNO 2 (22 mL) was added via addition funnel under argon. The reaction mixture was stirred at 0 °C for 3 h and warmed to room temperature overnight.
  • reaction mixture was allowed to cool, diluted with CH2CI2 (250 mL), and washed sequentially with a mixture of brine and 1 N HCl (2:1, 80 mL), a mixture of brine and NaHCO3 (2:1, 80 mL) and brine (80 mL).
  • the organic phase was dried over MgSO 4 and evaporated.
  • T3 (0.575 g, 3.59 mmol, 1 equiv) was dissolved in CH2CI2 (25 mL) and cooled in an ice bath. 2,6-Lutidine (0.836 mL, 7.18 mmol, 2 equiv) and trifluoromethanesulfonic anhydride (1.15 mL, 6.84 mmol, 1.9 equiv) were added and the reaction mixture was stirred 30 min. It was then diluted with MTBE (400 mL), washed with a mixture of brine and 1 N HCl (2:1, 100 mL) and brine (100 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
  • reaction mixture was diluted with CH2CI2 (600 mL) and washed with a mixture of brine and 10% KHSO 4 (1 :1, 150 mL) and a mixture of brine and NaHCO3 (2 x 200 mL). The organic phase was dried over MgSO 4 and evaporated.
  • Hydrogen peroxide (30 wt. % solution in water, 45 mL) was added to a solution of al (12.13 g, 70.2 mmol) in a mixture of EtOH (150 mL) and 10% aqueous NaOH (280 mL) at 23 °C.
  • the reaction mixture was heated to 50 °C for 18 h, then was cooled to 23 °C and the volatiles were removed under reduced pressure.
  • Triethylamine (13.9 mL, 99.7 mmol, 2.0 equiv) and diphenylphosphoryl azide (16.1 mL, 74.7 mmol, 1.5 equiv) were added sequentially to a suspension of cl (9.63 g, 49.8 mmol, 1 equiv) in 1,4-dioxane (450 mL) at 23 °C.
  • the resulting solution was heated to reflux for 7.5 h, then benzyl alcohol (10.3 mL, 99.5 mmol, 2.0 equiv) was added and reflux was continued
  • Trimethyloxonium tetrafluoroborate 2.0 g, 13.5 mmol, 1.2 equiv
  • 2,6-di-tert- butylpyridine (1.52 mL, 6.76 mmol, 0.6 equiv) were added to a solution of dl (3.36 g, 11.26 mmol, 1 equiv) in CH2CI2 (80 mL) at 23 °C.
  • the reaction mixture was stirred at that temperature for 65 h, then was partitioned between water (2 x 50 mL) and CH2CI2 (2 x 200 mL) and combined organic layers were dried over Na2SO 4 and concentrated.
  • Triethylamine (1.55 mL, 1 1.1 mmol, 2.5 equiv)
  • tert-butyldimethylsilyl chloride (1.07 g, 7.10 mmol, 1.6 equiv)
  • 4-dimethylaminopyridine 0.025 g, 0.20 mmol, 0.045 equiv
  • reaction mixture was stirred for 19 h at 23 °C, then was partitioned between 0.5 M HCl (150 mL) and a 1 :1 mixture of EtOAc and hexanes (2 x 150 mL). The combined organic layers were dried over Na2SO 4 and were concentrated.
  • the reaction mixture was maintained at -78 °C for 1.5 h, then acetic acid (15.57 mmol, 0.891 mL, 5.5 equiv) was added.
  • the reaction mixture was warmed to 0 °C for 5 min, then was washed with water (50 mL), saturated NaHCO3 (50 mL) and brine (50 mL).
  • the organic layer was dried over Na2SO 4 and concentrated to afford the crude aldehyde product as off-white foam. This material was utilized without further purification.
  • This material (15 mg, 0.05 mmol) was coupled to 4S-amino-5-(2-oxo-pyrrolidin-3S-yl)-pent- 2(trans)-enoic acid ethyl ester (11 mg, 0.05 mmol) in DMF (1 mL) by treatment with diisopropylethyl amine (0.02 mL, 0.01 mmol) and HATU (19 mg, 0.05 mmol) at room temperature for 2 h. The resulting solution was washed with brine (10 mL), and extracted with EtOAc (30 mL).
  • the title compounds were prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid pyridin-2-ylmethyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate.
  • Rhinovirus 3C Protease Stock solutions (50 mM, in DMSO) of various compounds were prepared; dilutions were in the same solvent.
  • Recombinant rhinovirus 3C proteases (see Birch et al., "Purification of recombinant human rhinovims 14 3C protease expressed in Escherichia coli," Protein Expr. Pur. (1995), vol. 6(5), 609-618) from serotypes 14, 16, and 2 were prepared by the following standard chromatographic procedures: (1) ion exchange using Q Sepharose Fast Flow from Pharmacia; (2) affinity chromatography using Affi-Gel Blue from Biorad; and (3) sizing using Sephadex G-100 from Pharmacia.
  • Each assay sample contained 2% DMSO, 50 mM tris pH 7.6, 1 mM EDTA, a test compound at the indicated concentration, approximately 1 ⁇ M substrate, and 50-100 nM protease.
  • the k 0 bs/I values were obtained from reactions initiated by addition of enzyme rather than substrate.
  • RVP activity was measured in the fluorescence resonance energy transfer assay.
  • the substrate was (N-terminal) DABCYL-(Gly-Arg-Ala-Val- Phe-Gln-Gly-Pro-Val-Gly)-EDANS. In the uncleaved peptide, the EDANS fluorescence was quenched by the proximal DABCYL moiety.
  • Hl-HeLa cells were infected with HRV-14 at a multiplicity of infection (m.o.i.) of 0.13 (vims particles/cell) or mock-infected with medium only. Infected or mock-infected cells were resuspended at 8 x 10 ⁇ cells per mL, and incubated with appropriate concentrations of the compounds to be tested.
  • the EC50 value was calculated as the concentration of compound that increased the percentage of formazan production in compound-treated, virus- infected cells to 50% of that produced by compound- free, mock-infected cells.
  • the 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased the percentage of formazan produced in compound-treated, mock-infected cells to 50% of that produced by compound-free, mock-infected cells.
  • the therapeutic index (Tl) was calculated by dividing the CC50 value by the EC50 value. All strains of human rhinovirus (HRV) for use in this assay were purchased from
  • HRV serotype-14 produced from the infectious cDNA clone constructed by Dr. Robert Rueckert, Institute for Molecular Virology, University of Wisconsin, Madison, Wisconsin. HRV stocks were propagated and viral assays were performed in Hl-HeLa cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum, available from Life Technologies (Gaithersburg, MD). Test results for the HRV assay are shown in the table below.
  • Anticoxsackieviral Cell Culture Assay is shown in the table below.
  • Coxsackievims types A-21 (CAV-21) and B3 (CVB3) were purchased from American Type Culture Collection (ATCC, Rockville, MD). Vims stocks were propagated and antiviral assays were performed in Hl-HeLa cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum (Life Technologies, Gaithersburg, MD). The ability of the compounds of this invention to protect cells against either CAV-21 or CVB3 infection was measured by the XTT dye reduction method. This method is described in Weislow et al., 7 Natl. Cancer Ins t. (1989), vol. 81, 577-586.
  • Hl-HeLa cells were infected with CAV-21 or CVB3 at a multiplicity of infection (m.o.i.) of 0.025 or 0.075, respectively, or mock-infected with medium only. Hl-HeLa cells were plated at 4 x 10 ⁇ cells per well in a 96- well plate and incubated with appropriate concentrations of the test compound. One day (CVB3) or two days (CAV-21) later, XTT /PMS was added to test plates and the amount of formazan produced was quantified spectrophotometrically at 450/650 nm.
  • CVB3 multiplicity of infection
  • CAV-21 CAV-21
  • the EC50 was calculated as the concentration of compound that increased the formazan production in compound-treated, vims-infected cells to 50% of that produced by compound- free, uninfected cells.
  • the 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased formazan production in compound-treated, uninfected cells to 50% of that produced in compound-free, uninfected cells.
  • the therapeutic index (Tl) was calculated by dividing the CC50 by the EC50-
  • Echovirus type 11 was purchased from ATCC (Rockville, MD). Vims stocks were propagated and antiviral assays were performed in MRC-5 cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum (Life Technologies, Gaithersburg, MD). The ability of the compounds of this invention to protect cells against ECHO 11 infection was measured by the XTT dye reduction method (Weislow et al., 7. Natl. Cancer Inst. (1989), vol. 81, 577-586). MRC-5 cells were infected with ECHO 11 at an m.o.i. of 0.003 or 0.004, respectively, or mock-infected with medium only.
  • Infected or uninfected cells were added at 1 x 10 ⁇ cells per well and incubated with appropriate concentrations of compound. Four days later, XTT/PMS was added to test plates, and the amount of formazan produced was quantified spectrophotometrically at 450/650 nm.
  • the EC50 was calculated as the concentration of compound that increased the formazan production in compound-treated, vims-infected cells to 50% of that produced by compound- free, uninfected cells.
  • the 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased formazan production in compound-treated, uninfected cells to 50% of that produced in compound- free, uninfected cells.
  • the therapeutic index (Tl) was calculated by dividing the CC50 by the ECso- Activity of the compounds against enterovims type 70 (EV 70) may be measured by the same assay as described above in this section.
  • Enterovims type 70 (EV 70) may be obtained from the American Type Culture Collection ATCC (Rockville, MD).
  • Results obtained for the compounds of the invention may be compared to results obtained in the same manner for control compounds WEN 51711, WEN 52084, and WIN 54954 (obtained from Sterling- Winthrop Pharmaceuticals), Pirodavir (obtained from Janssen Pharmaceuticals), and Pleconaril (prepared according to the method described in Diana et al., 7. Med. Chem. (1995), vol. 38, 1355).
  • Antiviral data obtained for the test compounds are shown in the table below. The designation "ND” indicates that a value was not determined for that compound, and the designation "NA” means not applicable.
  • ⁇ HRV human rhinovirus of designated serotype.
  • C CAV-21 coxsackievirus A21.
  • ⁇ ECHO-11 echovirus 11.
  • e CVB3 coxsackievirus B3.
  • JENT-70 enterovirus 70.
  • SCVB2 coxsackievirus B2.
  • ⁇ CVB5 coxsackievirus B5.
  • ⁇ CHO-6 echovirus 6.
  • ECHO-9 echovirus 9.

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Abstract

Compounds of formula (I), where the formula variables are as defined herein, are disclosed that advantageously inhibit or block the biological activity of the picornaviral 3C protease. Also disclosed are compounds of formula (II) where the formula variables are as defined herein that advantageously inhibit or block the biological activity of the picornaviral 3C protease. These compounds, as well as pharmaceutical compositions containing these compounds, are useful for treating patients or hosts infected with one or more picornaviruses, such as rhinovirus 3C proteases. Intermediates and synthetic methods for preparing such compounds are also described.

Description

TITLE
ANTIPICORNAVIRAL COMPOUNDS AND COMPOSITIONS, THEIR PHARMACEUTICAL USES, AND MATERIALS FOR THEIR SYNTHESIS
BACKGROUND OF THE INVENTION
Field of the Invention The invention relates to pyridone-containing peptidomimetic compounds that advantageously inhibit the enzymatic activity of picornaviral 3C proteases, especially rhinovirus 3C proteases (RVPs), and that retard viral growth in cell culture. The invention also relates to the use of these compounds in pharmaceutical compositions, methods of treatment of rhino viral infections using these compounds and compositions, and processes for the synthesis of these compounds and compounds useful in the syntheses thereof. Related Background Art
The picomaviruses are a family of tiny non-enveloped positive-stranded RNA-containing viruses that infect humans and other animals. These viruses include the human rhinoviruses, human polioviruses, human coxsackieviruses, human echoviruses, human and bovine enteroviruses, encephalomyocarditis viruses, meningitis virus, foot and mouth viruses, hepatitis A virus, and others. The human rhinoviruses are a major cause of the common cold. To date, there are no effective therapies on the market that cure the common cold, only treatments that relieve the symptoms.
Picornaviral infections may be treated by inhibiting the proteolytic picornaviral 3C enzymes. These enzymes are required for the natural maturation of the picomaviruses. They are responsible for the autocatalytic cleavage of the genomic, large polyprotein into the essential viral proteins. Members of the 3C protease family are cysteine proteases, where the sulfhydryl group most often cleaves the glutamine-glycine . amide bond. Inhibition of 3C proteases is believed to block proteolytic cleavage of the viral polyprotein, which in turn can retard the maturation and replication of the viruses by interfering with viral particle production. Therefore, inhibiting the processing of this cysteine protease with selective small molecules that are specifically recognized should represent an important and useful approach to treat and cure viral infections of this nature and, in particular, the common cold. Some small-molecule inhibitors of the enzymatic activity of picornaviral 3C proteases (i.e., antipicomaviral compounds) have been recently discovered. See, for example: U.S. Patent No. 5,856,530; U.S Patent No. 5,962,487; U.S. Patent Application No. 08/991,282, filed December 16, 1997, by Dragovich et al.; and U.S. Patent Application No. 09/301,977, filed April 29, 1999, by Dragovich et al. See also: Dragovich et al., "Structure-Based Design, Synthesis, and Biological Evaluation of
Irreversible Human Rhinovirus 3C Protease Inhibitors . . . ," J Med. Chem. (1999), Vol. 42, No. 7, 1203-1212, 1213-1224; and Dragovich et al., "Solid-phase Synthesis of Irreversible Human Rhinovirus 3C Protease Inhibitors . . . ," Bioorg. & Med. Chem. (1999), Vol. 7, 589-598. There is still a desire, however, to discover small-molecule compounds that are especially potent antipicomaviral agents. Inhibitors of other related cysteine proteases such as cathepsins have been described in, e.g., U.S. Patent No. 5,374,623; U.S. Patent No. 5,498,616; and WIPO International Publication Nos. WO 94/04172, WO 95/15749, WO 97/19231, and WO 97/49668. There yet remains a need for inhibitors targeting the picornaviral 3C cysteine protease with desirable pharmaceutical properties, such as high specificity.
SUMMARY OF THE INVENTION
This invention relates to compounds useful for inhibiting the activity of picornaviral 3C proteases having the general formula:
Figure imgf000004_0001
wherein:
Rais substituted or unsubstituted heterocycloalkyl or heterocycloalkylalkyl; Rb is a substituent having the formula:
Figure imgf000004_0002
wherein:
Rf and Rg are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5;
A, is CH or N;
A2 is C(Rh)(R), N(R), S, S(O), S(O)2, or O; where each Rh, R' and RJ is independently H or lower alkyl; each A3 present is independently C(Rh)(R'), N(RJ), S, S(O), S(O)2, or O; where each Rh, R1 and RJ is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A4 is N(Rk), C(Rh)(Ri), or O; and when p is 0 (i.e., A3 is not present), A4 is N(Rk)(R'), C(Rh)(Ri)(Rj), and O(R'), where each Rh, R and Rj is independently H or lower alkyl, each R is H, alkyl, aryl, or acyl, and each R1 is H, alkyl, or aryl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond when A2 is present (i.e., m - 1) and a hydrogen atom when A2 is absent (i.e., m = 0);
Rc is H, halogen or a substituted or unsubstituted lower alkyl group; Rd is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
Re is H or a substituted or unsubstituted alkylgroup; and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)Rn, -CO2Rn, -CN, -C(O)NRnR°, -C(O)NROR°, -C(S)Rn, -C(S)ORn, -C(S)NRnR°, -NO2, -SOR°, -SO2Rn, -SO2NRnR°, -SO2(NRn)(OR°), -SONR", -SO3Rn, -PO(ORn)2, -PO(ORn)(OR°)> -PO(NRnR°)(ORp)ι -PO(NRnR°)(NRpRq), -C(O)NRnNR°Rp, or -C(S)NRnNR°Rp, wherein Rn, R°, R and Rq are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the Rn, R°, Rp and Rq, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Rd, together with the atoms to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Rd are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Z1 are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group.
Preferably, when Ra is substituted or unsubstituted heterocycloalkylalkyl, the alkyl moiety thereof is a substituted or unsubstituted saturated alkyl moiety. Specifically, this invention relates to compounds useful for inhibiting the activity of picornaviral 3C proteases having the general Formula I:
Figure imgf000006_0001
I wherein:
R1 is H, a substituted or unsubstituted lower alkyl group or a suitable nitrogen protecting group;
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties in the above groups may be substituted or unsubstituted, or a suitable nitrogen protecting group;
R3 is H or a suitable substituent; or
R1 together with R2 form a suitable nitrogen protecting group; or
R2 together with R3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
R4 is H or a suitable substituent; the dotted line represents an optional bond;
R5 is H or a suitable substituent;
R6 is H or a substituted or unsubstituted alkyl group; or
R5 together with R6 form a heterocycloalkyl ring, which may be optionally substituted;
R7 and R10 are independently H, halogen or a substituted or unsubstituted lower alkyl group;
R8is H or a substituted or unsubstituted lower alkyl group;
R11 is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
R9 is a substituent having the formula:
Figure imgf000007_0001
wherein: R12 and R13 are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5; A, is CH or N;
A2 is C(R14)(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; each A3 present is independently C(R1 )(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A4 is N(R17), C(R14)(R15), or O; and when p is 0 (i.e., A3 is not present), A4 is N(R17)(R18), C(R14)(R15)(R16), and O(R18), where each R14, R15 and R16 is independently H or lower alkyl, each R17 is H, alkyl, aryl, or acyl, and each R18 is H, alkyl, or aryl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond when A2 is present (i.e., m = 1) and a hydrogen atom when A2 is absent (i.e., m = 0); and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)OR19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR,9R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20)ι -PO(NR19R20)(OR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and R", together with the atoms to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and R1 ' are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Z1 are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group.
In another embodiment of the compounds of the above formulae, Z and Z' are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR'9R20, -C(O)NR,9OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20)ι -PO(NR19R20)(OR2,)ι -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR,9NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
In yet another embodiment of the compounds of the above formulae, Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR,9R20, -C(O)NRl9OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR'9, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20),
-PO(NR19R20)(OR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR,9NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted. In addition to compounds of the above formulae, antipicomaviral agents of the invention include prodrugs, pharmaceutically active metabolites, and pharmaceutically acceptable salts and solvates of such compounds.
DETAILED DESCRIPTION OF INVENTION AND PREFERRED EMBODIMENTS
In one embodiment, the compounds of this invention useful for inhibiting the activity of picornaviral 3C proteases have the Formula I-A:
Figure imgf000009_0001
I-A wherein R1, R2, R\ R4, R5, R6, R7, R8, R9, R10, R", Z and Z1 , and the variables contained therein, are as defined above, or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
Preferably, in the compounds of Formula I-A:
R1, R7, R8, R10 and R11 are independently H or a substituted or unsubstituted lower alkyl group;
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
R3 is H or a suitable substituent; or
R2 together with R3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
R4 is H or a suitable substituent; R5 is H or a suitable substituent;
R6 is H or an unsubstituted alkyl group or a lower alkyl group optionally substituted with a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an alkoxyl group, an aryloxy group, an alkylthio group, an arylthio group, wherein each alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl moiety thereof may be optionally substituted; or
R5 together with R6 form a substituted or unsubstituted five- or six-membered heterocycloalkyl ring; wherein when R3, R4 and R5 are suitable substituents, said suitable substituents may be independently selected from alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl (tri luoromethyl), hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto (oxo), thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more suitable substituents, preferably selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, or aryloxy;
R9 is an aminocarbonylalkyl group, an alkylcarbonylaminoalkyl group, an alkylaminocarbonylalkyl group or a substituent having the formula:
Figure imgf000011_0001
wherein:
R12 and R13 are independently H or lower alkyl; m is 1; p is 1 or 2; A, is CH or N;
A2 is C(R14)(R15), N(R16), S, S(O), S(O)2, or O; each A3 present is independently C(R14)(R15), N(R16), S, S(O), S(O)2, or O;
A4 is N(R17), C(R1 )(R15), or O ; wherein each R14, R15 and R16 is independently H or lower alkyl, and each R17 is H, alkyl, aryl, or acyl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond; and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19,
-CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R'9, -C(S)NR,9R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20χ -PO(NR19R20)(OR21)ι -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R'9, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Z1 are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group.
Preferably, R3, R4 and R5 may be independently selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, amino, cyano, halogen, haloalkyl
(trifluoromethyl), hydroxyl, keto, alkoxy, aryloxy, cycloalkoxy, heterocycloalkoxy, alkyloxycarbonyl, aryloxycarbonyl, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroaryloxycarbonyl, heteroaryl carbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, alkylamino, arylamino, dialkylamino, alkylaminocarbonyl, alkylsulfonyl, or arylsulfonyl, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl moieties of any of the above substituents may be optionally substituted by one or more of haloalkyl, nitro, amino, cyano, halogen, hydroxyl, haloalkoxy, mercapto, keto or unsubstituted alkyl (except for alkyl), cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, aryloxy, alkylamino, dialkylamino, alkylthio or arylthio groups.
In accordance with a convention used in the art,
Figure imgf000012_0001
is used in structural
formulas herein to depict the bond that is the point of attachment of the moiety or substituent to the core or backbone structure.
As used herein, the term "alkyl" represents a straight- or branched-chain saturated or unsaturated hydrocarbon, containing 1 to 10 carbon atoms which may be unsubstituted or substituted by one or more of the substituents described below.
Exemplary alkyl substituents include, but are not limited to methyl (Me), ethyl (Et), propyl, isopropyl, butyl, isobutyl, t-butyl, ethenyl, propenyl, butenyl, pentenyl, ethynyl, butynyl, propynyl (propargyl, isopropynyl), pentynyl, hexynyl and the like. The term "lower alkyl" refers to an alkyl group containing from 1 to 4 carbon atoms. "Cycloalkyl" represents a group comprising a non-aromatic monocyclic, bicyclic, or tricyclic hydrocarbon containing from 3 to 14 carbon atoms which may be unsubstituted or substituted by one or more of the substituents described below and may be saturated or unsaturated. Exemplary cycloalkyls include monocyclic rings having from 3-7, preferably 3-6, carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like, that may be fully saturated or partially unsaturated. Illustrative examples of cycloalkyl groups include the following:
Figure imgf000013_0001
"Heterocycloalkyl" represents a group comprising a non-aromatic, monovalent monocyclic, bicyclic, or tricyclic radical, which is saturated or partially unsaturated, containing 3 to 18 ring atoms, which includes 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, and which may be unsubstituted or substituted by one or more of the substituents described below. Illustrative examples of heterocycloalkyl groups include, but are not limited to, azetidinyl, pyrrolidyl, piperidyl, piperazinyl, morpholinyl, tetrahydro-2H-l,4-thiazinyl, tetrahydrofuryl, dihydrofuryl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxolanyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, azabicylo[3.2.1]octyl, azabicylo[3.3.1]nonyl, azabicylo[4.3.0]nonyl, oxabicylo[2.2.1]heptyl, 1,5,9-triazacyclododecyl, and the like. Illustrative examples of heterocycloalkyl groups include the following moieties:
Figure imgf000013_0002
Figure imgf000014_0001
"Aryl" represents a group comprising an aromatic, monovalent monocyclic, bicyclic, or tricyclic radical containing from 6 to 18 carbon ring atoms, which may be unsubstituted or substituted by one or more of the substituents described below, and to which may be fused one or more cycloalkyl groups, heterocycloalkyl groups or heteroaryl groups, which themselves may be unsubstituted or substituted by one or more suitable substituents. Illustrative examples of aryl groups include the following moieties:
Figure imgf000014_0002
"Heteroaryl" represents a group comprising an aromatic monovalent monocyclic, bicyclic, or tricyclic radical, containing 5 to 18 ring atoms, including 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, which may be unsubstituted or substituted by one or more of the substituents described below. Illustrative examples of heteroaryl groups include, but are not limited to, thienyl, pyrrolyl, imidazolyl, pyrazolyl, furyl, isothiazolyl, furazanyl, isoxazolyl, thiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, benzo[b]thienyl, naphtho[2,3-b]thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl, indolizinyl, isoindolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxyalinyl, quinzolinyl, benzothiazolyl, benzimidazolyl, tetrahydroquinolinyl, cinnolinyl, pteridinyl, carbazolyl, beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, and phenoxazinyl. Further examples of heteroaryl groups include the following moieties:
Figure imgf000015_0001
Figure imgf000015_0002
Figure imgf000015_0004
Figure imgf000015_0003
and
Figure imgf000016_0001
Exemplary "suitable substituents" that may be present on any of the above alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl groups are described herein and include alkyl (except for alkyl), aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyoxycarbonyl, carboxyl, carbamoyl, formyl, keto (oxo), thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio, heteroarylthio, wherein any of the alkyl, alkylene, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted. Preferred "suitable substituents" include alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, halogen, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heteroaryloxy, and carboxyl. The alkyl, alkylene, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl moieties of any of the above substituents may be optionally substituted by one or more of alkyl (except for alkyl), haloalkyl, aryl, nitro, amino, alkylamino, dialkylamino, halogen, hydroxyl, alkoxy, haloalkoxy, aryloxy, mercapto, alkylthio or arylthio groups.
If the substituents themselves are not compatible with the synthetic methods of this invention, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions used in these methods. The protecting group may be removed at a suitable point in the reaction sequence of the method to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999), which is incorporated herein by reference in its entirety. In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used in the methods of this invention. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful in an intermediate compound in the methods of this invention or is a desired substituent in a target compound.
In the compounds of this invention, R1 and R2, independently or taken together, may be a suitable nitrogen protecting group. As indicated above, nitrogen protecting groups are well known in the art and any nitrogen protecting group that is useful in the methods of preparing the compounds of this invention or maybe useful in the antipicomaviral compounds of this invention may be used. Exemplary nitrogen protecting groups include alkyl, substituted alkyl, carbamate, urea, amide, imide, enamine, sulfenyl, sulfonyl, nitro, nitroso, oxide, phosphinyl, phosphoryl, silyl, organometallic, borinic acid and boronic acid groups. Examples of each of these groups, methods for protecting nitrogen moieties using these groups and methods for removing these groups from nitrogen moieties are disclosed in T. Greene and P. Wuts, supra. Preferably, when R1 and/or R2 are independently suitable nitrogen protecting groups, suitable R1 and R2 substituents include, but are not limited to, carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc: t-butyloxycarbonyl) and aryloxycarbonyl (e.g., Cbz: benzyloxycarbonyl, or FMOC: fiuorene-9-methyloxycarbonyl), alkyloxycarbonyls (e.g., methyloxycarbonyl), alkyl or arylcarbonyl, substituted alkyl, especially arylalkyl (e.g., trityl (triphenylmethyl), benzyl and substituted benzyl), and the like. When R1 and R2 taken together are a suitable nitrogen protecting group, suitable R'/R2 substituents include phthalimido and a stabase (1,2-bis (dialkylsilyl))ethylene). The terms "halogen" and "halo" represent chloro, fluoro, bromo or iodo substituents. "Heterocycle" is intended to mean a heteroaryl or heterocycloalkyl group. "Acyl" is intended to mean a -C(O)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group. "Acyloxy" is intended to mean an -OC(O)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group. "Thioacyl" is intended to mean a -C(S)-R radical, where R is a substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl group. "Sulfonyl" is intended to mean an -SO2- biradical. "Sulfenyl" is intended to mean an -SO- biradical. "Sulfo" is intended to mean an -SO2H radical. "Hydroxy" is intended to mean the radical -OH. "Amine" or "amino" is intended to mean the radical -NH2. "Alkylamino" is intended to mean the radical -NHR_, where R_ is an alkyl group. "Dialkylamino" is intended to mean the radical -NR_Rb, where R_ and R,, are each independently an alkyl group, and is intended to include heterocycloalkyl groups, wherein R_ and Rb, taken together, form a heterocyclic ring that includes the amine nitrogen. "Alkoxy" is intended to mean the radical -OR-, where R_ is an alkyl group. Exemplary alkoxy groups include methoxy, ethoxy, propoxy, and the like. "Lower alkoxy" groups have alkyl moieties having from 1 to 4 carbons. "Alkoxycarbonyl" is intended to mean the radical -C(O)OR_, where R. is an alkyl group. "Alkylsulfonyl" is intended to mean the radical -SO2Ra, where Ra is an alkyl group. "Alkylenedioxy" is intended to mean the divalent radical -ORaO- which is bonded to adjacent atoms (e.g., adjacent atoms on a phenyl or naphthyl ring) , wherein R- is a lower alkyl group. "Alkylaminocarbonyl" is intended to mean the radical -C(O)NHRa, where Ra is an alkyl group. "Dialkylaminocarbonyl" is intended to mean the radical -C(O)NRaRb, where Ra and Rb are each independently an alkyl group. "Mercapto" is intended to mean the radical -SH. "Alkylthio" is intended to mean the radical -SR-, where Ra is an alkyl group. "Carboxy" is intended to mean the radical -C(O)OH. "Keto" or "oxo" is intended to mean the diradical =O. "Thioketo" is intended to mean the diradical =S. "Carbamoyl" is intended to mean the radical -C(O)NH2. "Cycloalkylalkyl" is intended to mean the radical -alkyl-cycloalkyl, wherein alkyl and cycloalkyl are defined as above, and is represented by the bonding arrangement present in the groups -CH2-cyclohexane or -CH2-cyclohexene. "Arylalkyl" is intended to mean the radical -alkylaryl, wherein alkyl and aryl are defined as above, and is represented by the bonding arrangement present in a benzyl group. "Aminocarbonylalkyl" is intended to mean the radical -alkylC(O) NH2 and is represented by the bonding arrangement present in the group -CH2CH2C(O)NH2. "Alkylaminocarbonylalkyl" is intended to mean the radical -alkylC(O)NHRa, where R_ is an alkyl group and is represented by the bonding arrangement present in the group -CH2CH2C(O)NHCH3. "Alkylcarbonylaminoalkyl is intended to mean the radical -alkylNHC(O)-alkyl and is represented by the bonding arrangement present in the group -CH2NHC(O)CH3. "Dialkylaminocarbonylalkyl" is intended to mean the radical -alkylC(O)NRaRb, where R_ and Rb are each independently an alkyl group. "Aryloxy" is intended to mean the radical -ORc, where Rc is an aryl group. "Heteroaryloxy" is intended to mean the radical -ORd, where Rd is a heteroaryl group. "Arylthio" is intended to mean the radical -SRC, where Rc is an aryl group. "Heteroarylthio" is intended to mean the radical -SRd, where Rd is a heteroaryl group.
If an inventive compound is a base, a desired salt may be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such as glucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citric acid or tartaric acid, amino acid, such as aspartic acid or glutamic acid, aromatic acid, such as benzoic acid or cinnamic acid, sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
If an inventive compound is an acid, a desired salt may be prepared by any suitable method known to the art, including treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary); an alkali metal or alkaline earth metal hydroxide; or the like. Illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine; ammonia; primary, secondary, and tertiary amines; and cyclic amines, such as piperidine, morpholine, and piperazine; as well as inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.
All compounds of this invention contain at least one chiral center and may exist as single stereoisomers (e.g., single enantiomers or single diastereomers), any mixture of stereosisomers (e.g., any mixture of enantiomers or diastereomers) or racemic mixtures thereof. All such single stereoisomers, mixtures and racemates are intended to be encompassed within the broad scope of the present invention. Compounds identified herein as single stereoisomers are meant to describe compounds that are present in a form that contains at least 90% of a single stereoisomer of each chiral center present in the compounds. Where the stereochemistry of the chiral carbons present in the chemical structures illustrated herein is not specified, the chemical structure is intended to encompass compounds containing either stereoisomer of each chiral center present in the compound. Preferably, however, the inventive compounds are used in optically pure, that is, stereoisomerically pure, form or substantially optically pure (substantially stereoisomerically pure) form. As used herein, the term "stereoisomeric" purity (or "optical" purity) refers to the "enantiomeric" purity and/or "diastereomeric" purity of a compound. Compounds that are substantially enatiomerically pure contain at least 90% of a single isomer and preferably contain at least 95% of a single isomer of each chiral center present in the enantiomer. Compounds that are substantially diastereomerically pure contain at least 90% of a single isomer of each chiral center present in the diastereomer, and preferably contain at least 95% of a single isomer of each chiral center. More preferably, the substantially enantiomerically and diasteriomerically pure compounds in this invention contain at least 97.5% of a single isomer and most preferably contain at least 99% of a single isomer of each chiral center in the compound. The term "racemic" or "racemic mixture" refers to a mixture of equal amounts of enantiomeric compounds, which encompasses mixtures of enantiomers and mixtures of enantiomeric diastereomers. The compounds of this invention may be obtained in stereoisomerically pure (i.e., enantiomerically and/or diastereomerically pure) or substantially stereoisomerically pure (i.e., substantially enantiomerically and/or diastereomerically pure) form. Such compounds may be obtained synthetically, according to the procedures described herein using optically pure or substantially optically pure materials. Alternatively, these compounds may be obtained by resolution/separation of a mixture of stereoisomers, including racemic mixtures, using conventional procedures. Exemplary methods that may be useful for the resolution/separation of stereoisomeric mixtures include chromatography and crystallization/re-crystallization. Other useful methods may be found in "Enantiomers, Racemates, and Resolutions," J. Jacques et al., 1981, John Wiley and Sons, New York, NY, the disclosure of which is incorporated herein by reference. Preferred stereoisomers of the compounds of this invention are described herein.
Another embodiment of this invention comprises the compounds depicted by Formula I-a (as represented by Formula I, wherein the dotted line represents a bond):
Figure imgf000021_0001
I-a wherein:
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
R3, R4, R5 are independently H or a suitable substituent described above, and
R\ R6, R7, R8, R9, R10, Z and Z1 , and the variables contained therein, are as defined above. Yet another embodiment of this invention comprises the compounds depicted by Formula I-b:
Figure imgf000022_0001
I-b wherein:
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties of the above groups may be substituted or unsubstituted;
Rx represents H or one or more suitable substituents;
R3 and R4 are independently H or a suitable substituent described above; and
R1, R9, R10, Z and Z1 , and the variables contained therein, are as defined above. A further embodiment of this invention comprises the compounds depicted by
Formula I-c:
Figure imgf000022_0002
I-c wherein:
R , R and R "are independently H or a substituted or unsubstituted lower alkyl group;
Rγ represents H or one or more suitable substituents;
R4 and R5 are independently H or a suitable substituent described above; and R1, R6, R'^Z and Z1 , and the variables contained therein, are as defined above. Another embodiment of this invention comprises the compounds depicted by Formula I-d:
Figure imgf000023_0001
I-d wherein:
R7, R8 and R10 are independently H or a substituted or unsubstituted lower alkyl group;
Rγ represents H or one or more suitable substituents;
R4 and R5 are independently H or a suitable substituent described above; and
R1, R6, R9, Z and Z1 , and the variables contained therein, are as defined above.
Yet another embodiment of this invention comprises the compounds depicted by Formula I-e (as represented by Formula I, wherein the dotted line does not represent a bond):
Figure imgf000023_0002
I-e wherein:
R , R and R are independently H or a substituted or unsubstituted lower alkyl group;
Rγ represents H or one or more suitable substituents;
R4 and R5 are independently H or a suitable substituent described above; and
R1, R6, R9, Z and Z1 , and the variables contained therein, are as defined above. In preferred embodiments of Formulas I-a and I-b, R2 is selected from a substituted or unsubstituted alkyloxycarbonyl group, alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocycloalkylcarbonyl or heteroarylcarbonyl group. Preferably, when R2 is a substituted alkyloxycarbonyl group, R2 is an unsubstituted or substituted arylalkyloxycarbonyl group. Exemplary R2 groups include, but are not limited to benzyloxycarbonyl, methylcarbonyl, t-butylcarbonyl, trifluoromethylcarbonyl, cyclopentylcarbonyl, tetrahydrofuran-2-carbonyl, l,3-dithiolane-2-carbonyl and the like. Preferably, R2 is a substituted or unsubstituted benzyloxycarbonyl, arylcarbonyl, or heteroarylcarbonyl group. Even more preferably, R2 is an unsubstituted or substituted benzyloxycarbonyl or heteroarylcarbonyl group, wherein the heteroaryl moiety is a five-membered heterocycle having from one to three heteroatoms selected from O, N, and S, more preferably a five-membered heterocycle having at least one nitrogen heteroatom and at least one oxygen heteroatom (e.g., unsubstituted or substituted 1,2-oxazolyl (i.e., isoxazolyl), 1,3-oxazolyl (i.e., oxazolyl), or oxadiazolyl (1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, or 1,2,5-oxadiazolyl). When the heteroaryl moiety is oxadiazolyl, unsubstituted and monomethyl-substituted 1,2,4-oxadiazolyl are preferred. In especially preferred embodiments, the heteroaryl moiety is 3-isoxazolyl or 5-isoxazolyl, either unsubstituted or substituted with one or two methyl groups and/or halogens (F, Cl, Br or I), with chlorine and fluorine being preferred. Accordingly, the heteroarylcarbonylgroup in the especially preferred embodiments is an unsubstituted or substituted 3-carbonyl- 1,2-oxazolyl group (i.e., 3-carbonyl-isoxazolyl) or a 5-carbonyl- 1,2-oxazolyl group (i.e., 5-carbonyl-isoxazolyl).
In preferred embodiments of the compounds of this invention, the substituent variables R3, R4 and R5, as present in the compounds of Formulas I-a, I-b, I-c, I-d and I-e, are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto, alkoxy, aryloxy, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, or aryloxy. In preferred embodiments of the compounds of this invention, the substituent variables R_ in the compounds of Formula I-b are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto, thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, or aryloxy. In preferred embodiments of the compounds of this invention, the substituent variable Ry in the compounds of formula Formulas I-c, I-d and I-e, are selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto, thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, or aryloxy.
In especially preferred embodiments of Formulas I-a, I-c, I-d and I-e, R6 is H or an unsubstituted alkyl group or an optionally substituted lower alkyl group, wherein these groups are comprised of a straight- or branched-chain saturated hydrocarbon group, a straight- or branched-chain substituted saturated hydrocarbon group, or group comprised of a straight- or branched-chain saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety. When R6 is a substituted alkyl group, the point of attachment of R6 is via a saturated hydrocarbon moiety. When R6 is a substituted saturated hydrocarbon group, the saturated hydrocarbon group may be optionally substituted with a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an alkoxy group, an aryloxy group, an alkylthio group, an arylthio group, wherein each alkyl, cycloalkyl, aryl, heterocycloalkyl or heteroaryl moiety thereof may be optionally substituted. When R6 is comprised of a saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety, the saturated hydrocarbon moiety may be bound to an unsaturated hydrocarbon moiety containing one or more double-bonds or triple-bonds, the terminal positions of which may be substituted by the substituents described above, or may contain additional straight- or branched-chain saturated hydrocarbon moieties. Preferably, the unsaturated hydrocarbon moiety contains one double-bond or one triple-bond, the terminal position(s) of which may optionally contain a straight- or branched-chain saturated hydrocarbon moiety. Preferably, if the unsaturated hydrocarbon moiety contains a double-bond, both terminal positions of the double bond contain a straight- or branched-chain saturated hydrocarbon moiety. In especially preferred embodiments, R6 is H or a substituted or unsubstituted lower alkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl group, or a group comprised of a straight-chain saturated hydrocarbon moiety and an unsaturated hydrocarbon moiety. Preferably, R6 is H, methyl, substituted methyl, ethyl, n-propyl, n-butyl, sec-butyl, 2-propyn-l-yl, 3-methyl-3-buten-l-yl, -methylcyclohexyl, substituted or unsubstituted -methylthienyl or substituted or unsubstituted benzyl, wherein the phenyl moiety of the substituted benzyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino or halogen and the thienyl moiety of the substituted -methylthienyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino or halogen. When R6 is substituted methyl, the methyl (methylene) moiety may be substituted with an alkoxy group, an aryloxy group, an alkylthio group or an arylthio group. Most preferably, R6 is H, ethyl, 2-propyn-l-yl, -methylcyclohexyl, or substituted or unsubstituted benzyl, wherein the phenyl moiety of the substituted benzyl is substituted by one or more substituents independently selected from lower alkyl, lower alkoxy and halogen.
In the preferred embodiments of the compounds, prodrugs, pharmaceutically acceptable salts, pharmaceutically active metabolites, or pharmaceutically acceptable solvates of this invention Rb and R9 are defined as above and m is 1 and p is 1 or 2 (i.e., both A2 and A3 are present) or when p is 0, m is 0 (i.e, both A2 and A3 are absent). More preferably, in Rb and R9, when m is 1 and p is 1 or 2, A2 and A3 are both C(Rh)(R') or C(R14)(R15), respectively. More preferably, when m is 1, p is 1. In especially preferred embodiments of Formulas I-a, I-b, I-c, I-d and I-e, R9 is selected from -CH2CH2C(O)NH2; -CH2CH2C(O)NH-alkyl; -CH2NHC(O)CH3; and
Figure imgf000028_0001
, where n is 1 or 2. More preferably, R9 is
In the preferred embodiments of the compounds prodrugs, pharmaceutically acceptable salts, pharmaceutically active metabolites, or pharmaceutically acceptable solvates of this invention, Z and Z1 are independently H, substituted or unsubstituted alkyl, -CO2Rn or -CO2R19, as appropriate, wherein Rn and R19 are as defined above, or Z and Z1, taken together with the atom to which they are attached, form a heterocycloalkyl group, as defined above. In another useful embodiment of the compounds of this invention, Z and/or Z1 may be -C(S)ORn or -C(S)OR19, wherein Rn and R19 are as defined above. Such compounds may be prepared using procedures described in K. Hartke, et al., Leibigs Ann. Chem., 321-330 (1989) and K. Hartke, et al., Synthesis, 960- 961 (1985). Preferably, in the compounds of Formulas I-a, I-b, I-c, I-d and I-e, Z and Z1 are independently H, substituted or unsubstituted alkyl, -CO2R19, or taken together with the atom to which they are attached, form a heterocycloalkyl group, which may be optionally substituted. More preferably, Z and Z1 are independently selected from H, -CO2H, substituted or unsubstituted lower alkyl, -CO2-alkyl, -CO2-cycloalkyl, -CO2-alkylaryl (e.g., -CO2-benzyl), -CO2-alkylheteroaryl (e.g., -CO2-(CH2)npyridyl) or taken together with the atom to which they are attached form a heterocycloalkyl group , which may be optionally substituted. The heterocycloalkyl group may optionally contain O, N, S and/or P and may be substituted by one or more of oxo (keto) or thioketo. In preferred embodiments of this invention, Z and Z1 are not both H. Most preferably, Z1 is H or lower alkyl and Z is -CO2H, substituted or unsubstituted-CO2-alkyl, -CO2-alkylaryl, -CO2-alkylheteroaryl, -CO2-cycloalkyl, or or taken together with the atom to which they are attached form a heterocycloalkyl group, which may be optionally substituted. Exemplary Z groups include, but are not limited to substituted and unsubstituted -CO2-alkyl groups, which include straight- and branched-chain alkyl groups such as ethoxycarbonyl, t-butoxycarbonyl, isopropoxycarbonyl, (2,2-dimethylpropyl)- oxycarbonyl, and the like, and which include straight and branched-chain arylalkyl and heteroarylalkyl groups, such as benzyloxycarbonyl, pyridylmethyleneoxycarbonyl, and the like, substituted and unsubstituted -CO2-cycloalkyl groups such as cyclobutyloxycarbonyl, cyclopentyloxycarbonyl,, cyclohexyloxycarbonyl, cycloheptyloxycarbonyl, and the like, or taken together with Z1 and the atom to which they are
attached form
Figure imgf000029_0001
A preferred embodiment of this invention comprises stereoisomers of the subject compounds having the formula:
Figure imgf000029_0002
Another preferred embodiment of this invention comprises stereoisomers of the subject compounds having the formula:
Figure imgf000029_0003
Especially preferred embodiments of this invention comprise stereoisomers of the subject compounds having the formula:
Figure imgf000030_0001
Specific especially preferred embodiments of this invention comprise compounds of Formulas I-a', I-b', I-c', I-d' and I-e' as follows:
Figure imgf000030_0002
I-a'
wherein R1, R2, R3, R4, R , R6, R9, Z and Z1 are as previously defined;
Figure imgf000030_0003
I-b'
wherein R , R , R , R , R , Rx, Z and Z are as previously defined;
Figure imgf000031_0001
I-c'
wherein R4, R5, R6, R9, Ry, Z and Z1 are as previously defined;
Figure imgf000031_0002
I-d'
wherein R , R , R6, R , R_,, Z and Z are as previously defined; and
Figure imgf000031_0003
I-e'
wherein R4, R5, R6, R9, Ry, Z and Z1 are as previously defined.
In especially preferred embodiments of Formulas I-a' and I-b', R2 is unsubstituted or substituted benzyloxycarbonyl, arylcarbonyl, or heteroarylcarbonyl, more preferably heteroarylcarbonyl, where the heteroaryl moiety is a five-membered heterocycle having from one to three heteroatoms selected from O, N, and S. More preferably R2 is heteroarylcarbonyl wherein the heteroaryl moiety is a five-membered heterocycle having at least one nitrogen heteroatom and at least one oxygen heteroatom (e.g., unsubstituted or substituted 1,2-oxazolyl (i.e., isoxazolyl), 1,3-oxazolyl (i.e., oxazolyl), or oxadiazolyl (1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, or 1,2,5-oxadiazolyl); preferred oxadiazolyls are unsubstituted and monomethyl-substituted 1,2,4-oxadiazolyl. In especially preferred embodiments, the heteroaryl moiety is 3-isoxazolyl or 5-isoxazolyl, either unsubstituted or substituted with one or two substituents selected from methyl and halogen, with chloro and fluoro being preferred halogen substituents.
In the especially preferred embodiments of Formulas I-a', I-b', I-c', I-d' and I-e', R6 is selected from H or:
Figure imgf000032_0001
Figure imgf000032_0002
Figure imgf000032_0003
wherein R' may be H or alkyl and R" may be H or independently selected from lower alkyl, lower alkoxy, hydroxy, amino, alkylamino or dialkylamino, and halogen.
Preferred groups are
Figure imgf000032_0004
Figure imgf000032_0005
A particularly preferred embodiment of this invention comprises a compound having the formula:
Figure imgf000033_0001
and prodrugs, pharmaceutically active metabolites, and pharmaceutically acceptable salts and solvates thereof. Other useful embodiments of this invention comprise any stereoisomer or mixture of stereoisomers of the above-noted compound.
One preferred stereoisomer of this compound may be represented by the formula:
Figure imgf000033_0002
Another preferred stereoisomer of this compound may be represented by the formula:
Figure imgf000033_0003
An especially preferred stereoisomer of this compound may be represented by the formula:
Figure imgf000034_0001
Preferred specific compounds include those of the Examples below, especially:
Figure imgf000034_0002
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
and
Figure imgf000040_0002
The invention is also directed to the intermediates of Formula π, which are useful in the synthesis of certain compounds of Formula I:
Figure imgf000041_0001
wherein the variables R1, R2, R3, R4 and Rx are as defined above, and B is H, -OR24 When R1 and/or R2 are independently suitable nitrogen protecting groups, any suitable nitrogen-protecting group known in the art may be used (see, e.g., Greene & Wuts, supra). Suitable R1 and R2 substituents include, but are not limited to, carbamate protecting groups such as alkyloxycarbonyl (e.g., Boc) and aryloxycarbonyl (e.g., Cbz or FMOC), alkyloxycarbonyls (e.g., methyloxycarbonyl), alkyl or arylcarbonyl, substituted alkyl, especially arylalkyl (e.g., trityl (triphenylmethyl), benzyl and substituted benzyl) and the like. Preferably, when R1 and R2 are independently suitable nitrogen protecting groups, suitable R1 and R2 substituents include, but are not limited to, Boc, Cbz, FMOC, methyloxycarbonyl and trityl. When R1 and R2 taken together are a suitable nitrogen protecting group, suitable R'/R2 substituents include phthalimido and a stabase (1,2-bis (dialkylsilyl))ethylene). R24 may be H or a suitable protecting group for a carboxyl moiety. Suitable carboxyl protecting groups are also well known in the art, examples of which may be found in Greene and Wuts, supra, and include, but are not limited to, protecting groups where R24 is alkyl, substituted or unsubstituted aryl, alkyl and/or aryl substituted silyl (e.g., t-butyldimethylsilyl (TBS)), and the like. c_. ■ 41
The invention is also directed to pharmaceutically acceptable salts of the compounds ol Formula II. Preferred examples of the Formula II useful as intermediates include the following:
Figure imgf000042_0001
and pharmaceutically acceptable salts thereof.
The antipicomaviral compounds of this invention include prodrugs, the pharmaceutically active metabolites, and the pharmaceutically acceptable salts and solvates thereof. In preferred embodiments, the compounds of Formula I, prodrugs, pharmaceutically acceptable salts, and pharmaceutically active metabolites and solvates thereof have an antipicomaviral activity, more preferably antirhinoviral activity, corresponding to an EC50 less than or equal to 100 μM in the Hl-HeLa cell culture assay. A "prodrug" is intended to mean a compound that is converted under physiological conditions or by solvolysis or metabolically to a specified compound that is pharmaceutically active. A prodrug may be a derivative of one of the compounds of this invention that contains a moiety, such as for example -CO2R, or -PO(OR)2, that may be cleaved under physiological conditions or by solvolysis. Any suitable R substituent may be used that provides a pharmaceutically acceptable solvolysis or cleavage product. A prodrug containing such a moiety may be prepared according to conventional procedures by treatment of a compound of this invention containing, for example, an amido, carboxylic acid, or hydroxyl moiety with a suitable reagent. A "pharmaceutically active metabolite" is intended to mean a pharmacologically active compound produced through metabolism in the body of a specified compound. A "pharmaceutically acceptable salt" is intended to mean a salt that retains the biological effectiveness of the free acids and bases of a specified compound and that is not biologically or otherwise undesirable. Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-l,4-dioates, hexyne-l,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycollates, tartrates, methane-sulfonates, propanesulfonates, naphthalene- 1 -sulfonates, naphthalene-2-sulfonates, and mandelates. A "solvate" is intended to mean a pharmaceutically acceptable solvate form of a specified compound that retains the biological effectiveness of such compound. Examples of solvates include compounds of the invention in combination with water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine. In the case of compounds, salts, or solvates that are solids, it is understood by those skilled in the art that the inventive compounds, salts, and solvates may exist in different crystal forms, all of which are intended to be within the scope of the present invention and specified formulas.
The present invention is also directed to a method of inhibiting picornaviral 3C protease activity, comprising contacting the protease with an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, prodrug, pharmaceutically active metabolite, or solvate thereof. For example, picornaviral 3C protease activity may be inhibited in mammalian tissue by administering a compound of Formula I or a pharmaceutically acceptable salt, prodrug, pharmaceutically active metabolite, or solvate thereof. More preferably, the present method is directed at inhibiting rhinviral protease activity. "Treating" or "treatment" is intended to mean at least the mitigation of a disease condition in a mammal, such as a human, that is alleviated by the inhibition of the activity of one or more picornaviral 3C proteases, including, but not limited to human rhinoviruses, human poliovirus, human coxsackieviruses, encephalomyocarditis viruses, meningitis virus, and hepatitis A virus. The methods of treatment for mitigation of a disease condition include the use of the compounds in this invention in any conventionally acceptable manner, for example, as a prophylactic. The activity of the inventive compounds as inhibitors of picornaviral 3C protease activity may be measured by any of the suitable methods known to those skilled in the art, including in vivo and in vitro assays. An example of a suitable assay for activity measurements is the antiviral Hl-HeLa cell culture assay described herein. Administration of the compounds of the Formula I and their pharmaceutically acceptable prodrugs, salts, active metabolites, and solvates may be performed according to any of the generally accepted modes of administration available to those skilled in the art. Illustrative examples of suitable modes of administration include oral, nasal, parenteral, topical, transdermal, and rectal.
An inventive compound of Formula I or a pharmaceutically acceptable salt, prodrug, active metabolite, or solvate thereof may be administered as a pharmaceutical composition in any pharmaceutical form recognizable to the skilled artisan as being suitable. Suitable pharmaceutical forms include solid, semisolid, liquid, or lyophilized formulations, such as tablets, powders, capsules, suppositories, suspensions, liposomes, and aerosols.
Pharmaceutical compositions of the invention may also include suitable excipients, diluents, vehicles, and carriers, as well as other pharmaceutically active agents, depending upon the intended use or mode of administration. In preferred embodiments, the inventive pharmaceutical compositions are delivered orally, or intranasally in the form of suspensions. Acceptable methods of preparing suitable pharmaceutical forms of the pharmaceutical compositions may be routinely determined by those skilled in the art. For example, pharmaceutical preparations may be prepared following conventional techniques of the pharmaceutical chemist involving steps such as mixing, granulating, and compressing when necessary for tablet forms, or mixing, filling, and dissolving the ingredients as appropriate, to give the desired products for oral, parenteral, topical, intravaginal, intranasal, intrabronchial, intraocular, intraaural, and/or rectal administration.
Solid or liquid pharmaceutically acceptable carriers, diluents, vehicles, or excipients may be employed in the pharmaceutical compositions. Illustrative solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, pectin, acacia, magnesium stearate, and stearic acid. Illustrative liquid carriers include syrup, peanut oil, olive oil, saline solution, and water. The carrier or diluent may include a suitable prolonged-release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. When a liquid carrier is used, the preparation may be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid (e.g., solution), or a nonaqueous or aqueous liquid suspension. A dose of the pharmaceutical composition contains at least a therapeutically effective amount of the active compound (i.e., a compound of Formula I or a pharmaceutically acceptable salt, prodrug, active metabolite, or solvate thereof), and preferably is made up of one or more pharmaceutical dosage units. The selected dose may be administered to a mammal, for example, a human patient, in need of treatment mediated by inhibition of picornaviral 3C protease activity, by any known or suitable method of administering the dose, including: topically, for example, as an ointment or cream; orally; rectally, for example, as a suppository; parenterally by injection; or continuously by intravaginal, intranasal, intrabronchial, intraaural, or intraocular infusion. A "therapeutically effective amount" is intended to mean the amount of an inventive agent that, when administered to a mammal in need thereof, is sufficient to effect treatment for disease conditions alleviated by the inhibition of the activity of one or more picornaviral 3C proteases, such as human rhinoviruses, human poliovirus, human coxsackieviruses, encephalomyocarditis viruses, menigovirus, and hepatitis A virus. The amount of a given compound of the invention that will be therapeutically effective will vary depending upon factors such as the particular compound, the disease condition and the severity thereof, the identity of the mammal in need thereof, which amount may be routinely determined by artisans.
GENERAL SYNTHETIC METHODS
Preferably, the inventive compounds of Formulas I and II are prepared by the methods of the present invention, including the General Methods shown below. When stereochemistry is not specified in chemical structures, either stereocenter may be utilized. Although several intermediates are described and depicted as 2-hydroxypyridines, it is understood that such entities may also exist as the corresponding 2-pyridone tautomers. The following abbreviations also apply: Boc (tert-butoxycarbonyl), Ac (acetyl), Cbz (benzyloxycarbonyl), DMB (2,4-dimethoxybenzyl), TBS (tert-butyldimethylsilyl), TBDPS (tert-butyldiphenylsilyl), Ms (methanesulfonate), Ts (toluenesulfonate), Bn (benzyl), and Tr (triphenylmethyl). General Method 1
Figure imgf000046_0001
B
Figure imgf000046_0002
Figure imgf000046_0003
In General Method 1, an amino acid A (commercially available or prepared by methods described in the chemical literature) where ?\ is an appropriate protecting group for the amine functionality (e.g, Cbz, Boc, or Ac), and P2 is an appropriate protecting group for the amide nitrogen (e.g, Tr), is reductively transformed into alcohol B. Compound B is subsequently converted to compound C where P3 is an appropriate protecting group for the alcohol functionality (e.g., TBS). At this point, the Pi protecting group present in C is removed and the resulting amine or salt thereof (not shown) is subjected to an amide bond forming reaction with an appropriate cc-hydroxycarboxylic acid (which incorporates R and in which R7 is H; also not shown) to provide imtermediate D. The alcohol functionality present in D is then converted to an appropriate leaving group (e.g., mesylate, tosylate) E and is coupled with 2-hydroxypyridine F (which incorporates Ri, R2, R3, R4, and R5) to give intermediate G. Note that the Ri and R2 moieties present in F may be an appropriate protecting groups for the amine functionality. The P3 protecting group is subsequently removed from G and the resulting alcohol (H) is oxidized to the corresponding aldehyde (not shown) and subjected to an olefin-forming reaction to afford intermediate I (which incorporates Ri 1, Z, and Zi). The P2 protecting group present in I is then removed to give product J. If Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates G, H, or I or product J and replaced with a different Ri and/or R2 substituent to afford alternate intermediates G, H, or I or products J.
General Method 2
Figure imgf000048_0001
K M
Figure imgf000048_0002
O N
Figure imgf000048_0003
Figure imgf000048_0004
In General Method 2, amino alcohol K (prepared by methods described in the chemical literature), which incorporates R9 and in which Rio is H, Pi is an appropriate protecting group for amine functionality (e.g, Cbz, Boc, or Ac), and P2 is an appropriate protecting group for the amide nitrogen (e.g, DMB), is converted to compound L where P3 is an appropriate protecting group for the alcohol functionality (e.g., TBDPS). The Pi protecting group present in L is then removed and the resulting amine or salt thereof (not shown) is subjected to an amide bond forming reaction with an appropriate α-hydroxycarboxylic acid (which incorporates R6 and in which R7 is H; also not shown) to provide intermediate M. The alcohol functionality present in M is then converted to an appropriate leaving group (e.g., mesylate, tosylate) N and is coupled with 2-hydroxypyridine F (which incorporates Ri, R2, R3, R4, and R5) to give intermediate O. Note that the Ri and R2 moieties present in F may be an appropriate protecting group for the amine functionality. The P3 protecting group is subsequently removed from O and the resulting alcohol (P) is oxidized to the corresponding aldehyde (not shown) and subjected to an olefin-forming reaction to afford intermediate Q (which incorporates Ri 1, Z, and Z\). The P2 protecting group present in Q is then removed to give product R. If Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates O, P, or Q or product R and replaced with a different Ri and/or R2 substituent to afford alternate intermediates O, P, or Q or products R.
General Method 3
Figure imgf000050_0001
w
Figure imgf000050_0002
Figure imgf000050_0003
Figure imgf000050_0004
An alternate method for preparing either product J or product R is illustrated in
General Method 3. An α-hydroxycarboxylic acid S (either commercially available or prepared by methods described in the chemical literature) which incorporates R and in which Rγ is H is converted to α-hydroxyester T where P4 is an appropriate protecting group for the carboxylic acid functional group (e.g, methyl, benzyl, or tert-butyl). The alcohol functionality present in T is then converted to an appropriate leaving group (e.g., mesylate, tosylate, triflate) U and is coupled with 2-hydroxypyridine F (which incorporates R\, R2, R3, R4, and R5) to give intermediate V. Note that the Ri and R2 moieties present in F may be an appropriate protecting group for the amine functionality. The P4 protecting group is subsequently removed from V to afford carboxylic acid W. Independently, intermediates X and Z (prepared by methods described in the chemical literature) which incorporate Ri 1, Z, and Zi and in which Pi is an appropriate protecting group for the amine functionality (e.g., Cbz, Boc, or Ac) and P2 is an appropriate protecting group for the amide nitrogen (e.g., Tr), are converted to their corresponding amines Y and AA (or salts thereof), respectively. Amines AA and Y are then independently coupled with carboxylic acid W to afford intermediate I and product R, respectively. The P2 protecting group present in intermediate I is subsequently removed to afford product J. Note that, although not depicted in General Method 3, the lactam nitrogen present in intermediate X may be protected with a suitable protecting group (e.g., DMB). If such a moiety is present in X, it may be removed after coupling of W with Y to afford product R.
General Method 4
Figure imgf000052_0001
Figure imgf000052_0002
Figure imgf000052_0003
A method for preparing bicyclic products k and 1 is illustrated in General Method 4.
Thus, an appropriate pyridine g (which incorporates Ri, R2, R3, and R4, and which may be optionally substituted with Rx as shown) (an example of which may prepared as described in Specific Method 7 below) in which Pi 1 and P12 are protecting groups for the alcohol functionality (e.g., silyl ether, methyl) is subjected to an intramolecular cychzation/deprotection protocol in which both Pi 1 and P 12 are removed to give pyridone h. Note that the Ri and R2 moieties present in g may be an appropriate protecting group for the amine functionality (e.g., Cbz). The alcohol moiety present in h is then oxidized to the corresponding carboxylic acid i. This oxidation may be accomplished via an aldehyde intermediate (not shown). Carboxylic acid i is then independently coupled with amines AA and Y (or salts thereof) (prepared as described in General Method 3 above) to afford intermediate j and product 1, respectively. The P2 protecting group present in intermediate j is subsequently removed to afford product k. Note that, although not depicted in General Method 4, the lactam nitrogen present in intermediate Y may be protected with a suitable protecting group (e.g., DMB). If such a moiety is present in Y, it may be removed after coupling of i with Y to afford product 1. In addition, if Ri and/or R2 is/are initially a protecting group for the amine functionality, it/they may be removed from intermediates g, h, i, or j or products k or 1 and replaced with a different Ri and/or R2 substituent to afford alternate intermediates g, h, i, or j or products k or 1.
General Method 5
Figure imgf000054_0001
m
Figure imgf000054_0002
Figure imgf000054_0003
Figure imgf000054_0004
An alternate method for preparing a particular type of intermediate is illustrated in
General Method 5. Thus, the sulfone m (which may be prepared by the method described in Org. Lett. 1999, 1, 83) in which Rx is/are H and P13 is an appropriate protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl ester) is transformed into diazo compound n. This intermediate is subjected to a rhodium-catalyzed cyclization reaction involving phenylvinyl sulfone to give intermediate o (in which R3 is H). The hydroxyl group present in o is converted to the corresponding trifluoromethane sulfonate (OTf) p, and this intermediate is further transformed to the amine r via imine q. The amine present in r is derivatized with an appropriate moiety to afford intermediate s (which contains the R2 functional group and in which Ri is H). Intermediate s is subjected to a desulfurization reaction to give intermediate t and this entity is deprotected to give carboxylic acid i (in which Rl, R3, R4, and Rx are H). Alternatively, intermediate s may be deprotected to give carboxylic acid u (in which Ri, R3, and Rx are H) which may be utilized in place of i in General Method 4 above.
General Method 6
Figure imgf000056_0001
w
Figure imgf000056_0002
Figure imgf000056_0003
aa bb cc
Figure imgf000056_0004
gg An additional method for preparing intermediates related to i is illustrated in General Method 6. Thus, the diazo compound n (prepared in General Method 5 above) in which Rx is/are H and in which P13 is an appropriate protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl ester) is subjected to a rhodium-catalyzed cyclization reaction involving nitroethylene to give intermediate v (in which R3 is H). The hydroxyl group present in v is converted to the corresponding trifluoromethane sulfonate (OTf) w, and this intermediate is further transformed to the amine y via imine x. The amine present in y is derivatized with an appropriate moiety to afford intermediate z (which contains the R2 functional group and in which Ri is H). Intermediate z is reduced to give intermediate aa and this entity is transformed to intermediate bb in which X is a halogen. Intermediate bb is subsequently deprotected to give carboxylic acid cc. If desired, intermediate aa may also be transformed into intermediate t (General Method 5 above). Alternatively, intermediate aa is transformed to intermediate dd which is subsequently deprotected to give carboxylic acid ee. Alternatively, intermediate aa is transformed to intermediate ff in which R is alkyl, acyl, sulfonyl, or acyloxy. Intermediate ff is subsequently deprotected to give carboxylic acid gg. Carboxylic acids cc, ee, and gg may each be utilized in place of i in General Method 4 above.
General Method 7
Figure imgf000058_0001
Figure imgf000058_0002
kk
Figure imgf000058_0003
mm An additional method for preparing intermediates related to i is illustrated in General
Method 7. Thus, the diazo compound n (prepared in General Method 5 above) in which Rx is/are H and in which Pi 3 is an appropriate protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl ester) is subjected to a rhodium-catalyzed cyclization reaction involving acrylate esters or vinyl ketones to give intermediate hh (in which R3 is H and Z2 is alkyl, aryl, alkoxy, and benzyloxy). The hydroxyl group present in hh is converted to the corresponding trifluoromethane sulfonate (OTf) ii, and this intermediate is further transformed to the amine kk via imine jj. The amine present in kk is derivatized with an appropriate moiety to afford intermediate 11 (which contains the R2 functional group and in which Ri is H). Intermediate 11 is subsequently deprotected to give carboxylic acid mm which may be utilized in place of i in General Method 4 above. At any point in the above sequence, if Z2 is alkoxy or benzyloxy it may be replaced with a hydroxyl functionality. The resulting carboxylic acid may subsequently be rearranged to the corresponding amine using established methods and the amine-containing intermediates may be utilized as depicted in General Method 6. For example, intermediate 11 (when Z2 is OH) may be rearranged to intermediate aa (General Method 6).
General Method 8
Figure imgf000059_0001
Figure imgf000059_0002
II
Yet another method for preparing intermediate 11 is depicted in General Method 8. Thus the sodium salt of α-dehydroalanine derivative nn (which incorporates R2 and in which Rl is H) (which is either commercially available or may be prepared from serine by a variety of literature techniques) is condensed with intermediate oo (which may be prepared as described in Tetrahedron Lett. 1989, 30, 3621) in which Rx is/are H and P13 is an appropriate protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl ester) and in which Z2 is alkyl, aryl, alkoxy, or benzyloxy to give intermediate pp. Intermediate pp is subsequently oxidized (by a variety of literature methods) to afford intermediate 11 which may be utilized as described above in General Method 7. General Method 9
Figure imgf000060_0001
qq rr SS
Figure imgf000060_0002
An alternate method for preparing intermediate z (from General Method 6 above) is depicted in General Method 9. Thus the nitro compound qq (prepared by analogy with the method described in: J. Chem. Soc, Perkin Trans. 1 1998, 1113) which incorporates R2 and in which Ri is H and P14 is a suitable protecting group for the carboxylic acid functionality (e.g., methyl or ethyl) is condensed with intermediate rr (prepared according to: J. Heterocyclic Chem. 1992, 29, 1285) in which Rx is/are H and P13 is a suitable protecting group for the carboxylic acid functionality (e.g., methyl, ethyl, benzyl, or tert-butyl) to give intermediate ss. Intermediate ss is subsequently oxidized (by a variety of literature methods) to afford intermediate z which may be utilized as described above in General Method 6.
General Method 10
Figure imgf000061_0001
tt uu vv
General Method 10 depicts the preparation of the bicyclic pyrrole vv, starting with aldehyde tt, prepared according to the procedure described in Smith, K. M., J. Chem. Soc, Perkin Trans. I, 1973, p. 516. This compound is subjected to reductive amination conditions with an amine to give compound uu. The carboxylic acid protecting group Pi6 is removed, and the resulting acid is condensed mtramolecularly with the secondary amine to give bicycle w. This bicyclic compound may be coupled to amine Y or AA according to the method described in General Method 3.
SPECIFIC METHODS
The following Specific Methods may also be utilized to prepare some of the compounds described in this invention.
Specific Method 1
Figure imgf000062_0001
H1 G1
Figure imgf000062_0002
Specific Method 1 describes the preparation of specific compound Jl (compound 5). Thus, commercially available amino acid Al was reduced to alcohol BI which, in turn, was transformed into compound Cl . The Cbz moiety present in Cl was removed by hydrogenation and the resulting amine (not shown) was coupled with D-3-phenyllactic acid (commercially available) to afford intermediate Dl . This latter entity was subsequently transformed to the corresponding methanesulfonate (mesylate) (El) and was coupled with the sodium salt of 2-hydroxypyridine FI to provide intermediate Gl. The 2-hydroxypyridine FI was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent Boc protection of the resulting amine. The silyl protecting group present in Gl was removed and the alcohol thus obtained (HI) was oxidized to the corresponding aldehyde (not shown) and subjected to an olefm-forming reaction to give intermediate II . The Boc moiety contained in II was then thermally deprotected and the resulting amine (not shown) was derivatized with commercially available 5-methylisoxazole-3-carbonyl chloride to give intermediate 12. The trityl protecting group present in 12 was subsequently removed under acidic conditions to complete the preparation of specific compound Jl (compound 5).
Specific Method 2
Figure imgf000064_0001
M1
Figure imgf000064_0002
N1
Figure imgf000064_0003
P1 01
Figure imgf000064_0004
Specific Method 2 describes the preparation of specific compound Rl (compound 20).
Thus, alcohol Kl (prepared as described in Dragovich, et al., J. Med Chem. 1999, 42, 1213) was protected to give intermediate LI . The Boc protecting group present in LI was removed under acidic conditions and the resulting amine salt (not shown) was coupled with (2R)-3-(4'-fluorophenyl)-2-hydroxypropionic acid (SI, prepared as described in Specific Method 3 below) to afford intermediate Ml. This latter entity was subsequently transformed to the corresponding methanesulfonate (mesylate) (Nl) and was coupled with the sodium salt of 2-hydroxypyridine F2 to provide intermediate 01. The 2-hydroxypyridine F2 was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent derivatization of the resulting amine with commercially available
5-methylisoxazole-3-carbonyl chloride. The silyl protecting group present in 01 was removed and the alcohol thus obtained (PI) was oxidized to the corresponding aldehyde (not shown) and subjected to an olefin-forming reaction to give intermediate Ql . The DMB moiety contained in Ql was then deprotected to complete the preparation of specific compound Rl (compound 20).
Figure imgf000066_0001
S1 T1
Figure imgf000066_0002
Specific Method 3 describes the preparation of specific compound R2 (compound 23). Thus, commercially available Boc-D-(4-F)Phe-OH was deprotected under acidic conditions and the resulting amine salt was subjected to a diazotization/displacement protocol to provide (2R)-3-(4'-fluorophenyl)-2-hydroxypropionic acid (SI). This material was subsequently transformed into the corresponding trifluoromethane sulfonate (triflate) UI via the methyl ester Tl and was coupled with the sodium salt of 2-hydroxypyridine F3 to provide intermediate VI. The 2-hydroxypyridine F3 was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent derivatization of the resulting amine with 5-chloroisoxazole-3-carbonyl chloride (prepared as described in the Experimental Section of this work). The methyl ester present in VI was subsequently hydrolyzed under basic conditions and the resulting carboxylic acid (Wl) was coupled with amine Yl (or salt thereof) to complete the preparation of specific compound R2 (compound 23). Amine Yl (or salt thereof) was prepared by deprotection of intermediate XI (prepared in a manner analogous to that described in Baldwin et al., J. Org. Chem. 1971, 36, 1441).
Specific Method 4
Figure imgf000068_0001
S1 T2
Figure imgf000068_0002
U2
Figure imgf000068_0003
Specific Method 4 describes the preparation of specific compound J2 (compound 24). Thus, (2R)-3-(4'-fluorophenyl)-2-hydroxypropionic acid (SI, prepared as described above in Specific Method 3) was transformed into the corresponding trifluoromethane sulfonate (triflate) U2 via the benzyl ester T2 and was coupled with the sodium salt of 2-hydroxypyridine F4 to provide intermediate V2. The 2-hydroxypyridine F4 was prepared from commercially available 2-hydroxy-3-nitropyridine by reduction and subsequent derivatization of the resulting amine with trifluoroacetic anhydride. The benzyl ester present in V2 was subsequently removed by hydrogenation and the resulting carboxylic acid (W2) was coupled with amine AA1 (or salt thereof) to give intermediate 13. The trityl protecting group present in 13 was then removed under acidic conditions to complete the preparation of specific compound J2 (compound 24). Amine AA1 (or salt thereof) was prepared by deprotection of intermediate Zl (prepared as described in Dragovich, et al. J. Med. Chem. 1998, 41, 2806).
:ific Method 5
Figure imgf000070_0001
T3 U3 F2
Figure imgf000070_0002
W3 V3
Figure imgf000070_0003
Specific Method 5 describes the preparation of specific compound R3 (compound 26). Thus, commercially available tert-butyl (R)-2-hydroxybutyrate (T3) was transformed into the corresponding trifluoromethane sulfonate (triflate) U3 and was coupled with the sodium salt of 2-hydroxypyridine F2 (prepared as described in Specific Method 2 above) to provide intermediate V3. The tert-butyl ester present in V3 was subsequently hydro lyzed under acidic conditions and the resulting carboxylic acid (W3) was coupled with amine Y2 (or salt thereof) to complete the preparation of specific compound R3 (compound 26). Amine Y2 (or salt thereof) was prepared by deprotection of intermediate X2, prepared according to the method disclosed in the co-pending application, U.S. Provisional Patent Application No. 60/150,358, filed August 24, 1999, the disclosure of which is incoφorated herein by reference.
Specific Method 6
Figure imgf000071_0001
T3 U3 F3
Figure imgf000071_0002
W4 V4
Figure imgf000071_0003
Figure imgf000071_0004
R4 Y3 Specific Method 6 describes the preparation of specific compound R4 (compound 29). Thus, trifluoromethane sulfonate (triflate) U3 (prepared as described in Specific method 5 above) was coupled with the sodium salt of 2-hydroxypyridine F3 (prepared as described in Specific Method 3 above) to provide intermediate V4. The tert-butyl ester present in V4 was subsequently hydrolyzed under acidic conditions and the resulting carboxylic acid (W4) was coupled with amine Y3 (or salt thereof) to complete the preparation of specific compound R4 (compound 29). Amine Y3 (or salt thereof) was synthesized from alcohol Kl (prepared as described in Dragovich, et al., J. Med Chem. 1999, 42, 1213) by the following method. Alcohol Kl was oxidized to the corresponding aldehyde (not shown) and subjected to an olefin- forming reaction to give intermediate X3. The DMB moiety contained in X3 was then deprotected to provide intermediate X4, and this entity was deprotected under acidic conditions to afford amine Y3 (or salt thereof).
Specific Method 7
Figure imgf000073_0001
gi hi i1
Figure imgf000073_0002
k1
Specific Method 7 describes the preparation of specific compound kl (compound 36).
Thus, the dianion of commercially available 2-hydroxy-6-methylnicotinonitrile was converted to intermediate al by methods related to those described in the literature (DeJohn, D.; Domagala, J. M.; Kaltenbroitn, J. S.; Krolls, U. J. Heterocyclic Chem. 1983, 20, 1295). The nitrile functionality present in this intermediate was then converted to the corresponding carboxylic acid cl via the primary amide bl. Intermediate cl was subjected to a Curtius rearrangement and the resulting isocyanate was trapped with benzyl alcohol to provide carbamate dl. The hydroxyl moiety contained in dl was selectively methylated and the resulting methyl ether (el) was subjected to an asymmetric dihydroxylation reaction to give diol fl . This asymmetric dihydroxylation reaction can be effected utilizing a variety of commercial and non-commercial chiral additives. The primary hydroxyl moiety contained in fl was selectively protected as the corresponding tert-butyldimethylsilyl ether (gl). This intermediate was treated with trifluoromethanesulfonic anhydride in the presence of 2,6-lutidine at low temperature followed by exposure to tetrabutylammonium fluoride to effect (i) intramolecular cyclization and (ii) silyl ether deprotection and afford alcohol hi. Alcohol hi was oxidized to the corresponding aldehyde (not shown), and this intermediate was further oxidized to the corresponding carboxylic acid il. Acid il was coupled with amine AAl (or salt thereof) to give intermediate j 1. The trityl protecting group present in j 1 was then removed under acidic conditions to complete the preparation of specific compound kl (compound 36). Amine AAl (or salt thereof) was prepared by deprotection of intermediate Zl (prepared as described in Dragovich, et al. J. Med. Chem. 1998, 41, 2806).
Specific Method 8
Figure imgf000075_0001
gi g2
Ϊ2 i3
Figure imgf000075_0003
5 Specific Method 8 describes the preparation of specific compounds 11 and 12
(compounds 37 and 39, respectively). Thus, the Cbz moiety present in intermediate gl (prepared as described in Specific Method 7 above) was removed and the resulting amine (not shown) was derivatized with commercially available 5-methylisoxazole-3-carbonyl chloride to provide intermediate g2. This intermediate was converted to specific compound 11 (compound 10 37) by a process analogous to that described in Specific Method 7 for the conversion of intermediate gl to intermediate j 1 utilizing amine Y2 (Specific Method 5) where appropriate. During this process, a small amount of intermediate i3 was also serendipitously generated. This intermediate was transformed to specific compound 12 (compound 39) by a process analogous to that described in Specific Method 7 for the conversion of intermediate gl to intermediate j 1 utilizing amine Y2 (Specific Method 5) where appropriate.
Specific Method 9
Figure imgf000076_0001
ml
Figure imgf000076_0002
Figure imgf000076_0003
Specific Method 9 describes the synthesis of a bicyclic pyrrole. Alcohol ml was oxidized with 2-iodoxybenzoic acid to give aldehyde nl, then reductively aminated with phenylalanine methyl ester and sodium cyanoborohydride to give amine ol. The t-butyl- protecting group was selectively removed, and the resulting amino acid was cyclized with DCC-HOBT to give pi . The methyl ester was cleaved with lithium hydroxide to give acid ql . Boc-protected 4S-amino-5-(2-oxo-pyrrolidin-3S-yl)-pent-2(trans)-enoic acid ethyl ester X2 was deprotected with HC1, then coupled to acid ql, using HATU, to complete the preparation of compound 43.
EXAMPLES Examples of the processes used to make several of the compounds of Formulas I and II are set forth below. The structures of the compounds of the following examples were confirmed by one or more of the following: proton magnetic resonance spectroscopy, infrared spectroscopy, elemental microanalysis and melting point. Proton magnetic resonance (JH NMR) spectra were determined using either a Varian IJNYIYplus 300 or a General Electric QE-300 spectrometer operating at a field strength of 300 megahertz (MHz). Chemical shifts are reported in parts per million (ppm, δ) downfield from an internal tetramethylsilane standard. Alternatively, *H NMR spectra were referenced to residual pro tic solvent signals as follows: CHCI3 = 7.26 ppm; DMSO = 2.49 ppm, C6HD5 = 7.15 ppm. Peak multiplicities are designated as follows: s, singlet; d, doublet; dd, doublet of doublets; t, triplet; q, quartet; br, broad resonance; m, multiplet. Coupling constants are given in Hertz. Infrared absorption (IR) spectra were obtained using a Perkin-Elmer 1600 series FTIR spectrometer. Elemental microanalyses were performed by Atlantic Microlab Inc., Norcross, GA and gave results for the elements stated within ±0.4% of the theoretical values. Flash column chromatography was performed using Silica gel 60 (Merck Art 9385). Analytical thin layer chromatography (TLC) was performed using precoated sheets of Silica 60 F254 (Merck Art 5719). Melting points were determined on a Mel-Temp apparatus and are uncorrected. All reactions were performed in septum-sealed flasks under a slight positive pressure of argon unless otherwise noted. All commercial reagents were used as received from their respective suppliers with the following exceptions. Tetrahydrofuran (THF) was distilled from sodium-benzophenone ketyl prior to use. Dichloromethane (CH2CI2) was distilled from calcium hydride prior to use. Et2θ refers to diethyl ether. DMF refers to N,N-dimethylformamide. DMSO refers to dimethylsulfoxide. MTBE refers to tert-butyl methyl ether. Other abbreviations include: CH3OH (methanol), EtOH (ethanol), EtOAc (ethyl acetate), DME (ethylene glycol dimethyl ether) Ac (acetyl), Me (methyl), Ph (phenyl), Tr (triphenylmethyl), Cbz (benzyloxycarbonyl), Boc (tert-butoxycarbonyl), TFA (trifluoroacetic acid), DIEA (N,N-diisopropylethylamine), TMEDA (N,N,N',N'-tetramethylethylenediamine), AcOH (acetic acid), AC2O (acetic anhydride), NMM (4-methylmorpholine), HOBt (1-hydroxybenzotriazole hydrate), HATU [O-(7-azabenzotriazol- 1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate], EDC [ 1 -(3-dimethylaminopropyl)-3-ethylcarbarbodiimide hydrochloride], DCC (dicyclohexyl-carbodiimide), DDQ (2,3-dichloro-5,6-dicyano-l,4-benzoquinone), DMAP (4-dimethylaminopyridine), Gin (glutamine), Leu (leucine), Phe (phenylalanine), Phe(4-F) (4-fluorophenylalanine), Val (valine), amino-Ala (2,3-diaminopropionic acid), and (S)-Pyrrol-Ala [(2S,3'S)-2-amino-3-(2'-oxopyrrolidin-3'-yl)-propionic acid]. Additionally, "L" represents the configuration of naturally occurring amino acids.
EXAMPLE 1
Preparation of Compound 1: trans-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino- 2"-oxo-2"H-pyridin-l"-yl)-3'-phenylpropionylamino]-6- carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000078_0001
Preparation of Intermediate (2-Hydroxypyridin-3-yl)carbamic Acid Benzyl Ester
A suspension of 10% palladium on carbon (0.20 g) and 2-hydroxy-3-nitropyridine
(3.00 g, 21.4 mmol, 1 equiv) in EtOH (100 mL) was subjected to one atmosphere of hydrogen for 4 hours. After purging the reaction vessel with argon, the mixture was filtered and the filtrate evaporated to give 2-hydroxy-3-aminopyridine which was used without further purification. This crude material was stirred in THF (70 mL) at 23 °C. Benzyl chloroformate (3.37 mL, 23.6 mmol, 1.1 equiv) and Na2CO3 (5.00 g, 47.2 mmol, 2.2 equiv) were added and the reaction mixture was stirred for 60 h then diluted with EtOAc (250 mL) and washed sequentially with saturated NaHCO3 and brine (50 mL each). The organic phase was dried over MgSO4, filtered, and concentrated. The residue was purified by flash column chromatography (gradient elution, 3→5% CH3OH in CH2CI2) to provide the title intermediate (3.69 g, 71%) as a white solid: *H NMR (CDCI3) δ 5.21 (s, 2H), 6.33 (t, 1H, J= 6.8), 7.00 (dd, 1H, J= 6.8, 1.6), 7.31-7.43 (m, 5H), 7.80 (s, 1H), 8.12-8.17 (m, 1H), 12.97 (s, 1H). Preparation of Intermediate 5-(3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-l-yl)-6-phenyl- 4-[2-(tritylcarbamoyl)ethyl]hex-2-enoic Acid Ethyl Ester
(2-Ηydroxypyridin-3-yl)carbamic acid benzyl ester was converted to the title intermediate via condensation with El (see Example 5) by a process analogous to that described in Example 5 for the conversion of Fl to intermediate II : *H NMR (CDCI3) δ 1.28 (t, 3H, J= 7.2), 1.55-1.69 (m, IH), 1.86-2.00 (m, IH), 2.06-2.23 (m, 2H), 3.12 (dd, 1H, J= 13.6, 7.8), 3.46 (dd, IH, J= 13.6, 8.2), 4.17 (q, 2H, J= 7.2), 4.43-4.54 (m, IH), 5.14 (d, IH, J = 12.1), 5.18 (d, IH, J= 12.1), 5.49-5.57 (m, IH), 5.62 (dd, IH, J= 15.7, 1.7), 6.14 (t, IH, J = 7.3), 6.59-6.75 (m, 3H), 7.08-7.41 (m, 26H), 7.73 (s, IH), 7.93-7.98 (m, IH).
Preparation of Compound 1
The preceding intermediate was converted to compound 1 by a process analogous to that described in Example 5 for the conversion of 12 to product l: IR (cm-1) 3298, 1713, 1655, 1590, 1508, 1 196; !H NMR (CDCI3) δ 1.30 (t, 3H, J= 7.1), 1.65-1.96 (m, 2H), 2.02- 2.19 (m, 2H), 3.15 (dd, IH, J= 13.8, 7.6), 3.49 (dd, IH, J= 13.8, 8.3), 4.18 (q, 2H, J= 7.1), 4.42-4.53 (m, IH), 5.17 (s, 2H), 5.62-5.81 (m, 4H), 6.28 (t, IH, J= 7.2), 6.66 (dd, IH, J = 15.6, 5.4), 7.12-7.40 (m, 12H), 7.81 (s, IH), 7.97-8.04 (m, IH); Anal. C31H34N4O7O.25H2O: C, H, N.
EXAMPLE 2 Preparation of a 1:1 mixture of Compound 1 (above) and Compound 2: trα«5-(2'R,4S)-4-[2'- (3"-Benzyloxycarbonylamino-2"-oxo-2"H-pyridin-l"-yl)-3'-phenylpropionylamino]-6- carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000080_0001
The title products were prepared from commercially available (2R)-2-hydroxy-3- phenylpropionic acid by a process analogous to that described in Example 22 for the conversion of SI to product R2 utilizing intermediates (2-hydroxypyridin-3-yl)carbamic acid benzyl ester (Example 1) and AAl (Example 23) where appropriate: *H NMR (CDCI3) (mixture of isomers) δ 1.20 (t, J= 7.0), 1.29 (t, J= 7.2), 1.64-2.25 (m), 3.08 (dd, J= 13.6, 6.4), 3.14 (dd, J= 13.6, 7.8), 3.41-3.52 (m), 4.09 (q, J= 7.2), 4.18 (q, J= 7.0), 4.39-4.50 (m),' 5.16 (s), 5.18 (s), 5.59-5.82 (m), 6.12-6.22 (m), 6.25-6.34 (m), 6.65 (dd, J= 15.6, 5.6), 6.70 (dd, J= 15.6, 5.3), 7.10-7.41 (m), 7.47-7.58 (m), 7.79-7.86 (m), 7.97-8.07 (m).
EXAMPLE 3 Preparation of Compound 3: tran^-(2'S,3'"S,4S)-4-[2'-(7"-Oxo-l",7"-dihydropyrrolo[2",3"-c- pyridin-6"-yl)-3'-phenylpropionylamino]-5-(2'"-oxopyrrolidin-3'"-yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000080_0002
Preparation of Intermediate (2S)-2-(2',2'-Diethyloxyethylamino)-3-phenylpropionic Acid Ethyl Ester
A solution of L-phenylalanine ethyl ester (8.37 g, 46.8 mmol, 1 equiv), bromoacetaldehyde diethyl acetal (10.6 mL, 70.1 mmol, 1.5 equiv), and DIEA (16.2 mL, 93.4 mmol, 2.0 equiv) in DMF (100 mL) was heated at 80 °C for 2 d. The resulting solution was cooled to 23 °C, taken up in chloroform (200 mL), washed with 10% aqueous potassium carbonate (3 x 50 mL), dried over potassium carbonate, and concentrated. Purification of the residue by silica gel chromatography yield 7.4 g (54%) of product. !H NMR (CDCI3) δ 1.13- 1.23 (m, 9H), 2.61 (dd, IH, J= 11.8, 5.9), 2.78 (dd, IH, J= 11.8, 5.9), 2.96-3.03 (m, 2H), 3.46-3.53 (m, IH), 3.55-3.70 (m, 4H), 4.18 (q, 2H, J= 7.1), 4.57 (t, IH, J= 5.5), 7.19-7.33 (m, 5H).
Preparation of Intermediate (2S)-2-[(2,2-Diethoxyethyl)-(lH-pyrrole-2-carbonyl)-amino]-3- phenylpropionic Acid Ethyl Ester
Pyrrole-2-carboxylic acid (0.82 g, 7.41 mmol, 1.42 equiv) in CH2CI2 (30 mL) was treated with oxalyl chloride (1.0 mL, 11.1 mmol, 2.13 equiv), followed by DMF (1 drop). The reaction mixture was held at 23 °C overnight, then was concentrated under reduced pressure. The resulting solid was taken up in CH2CI2 (30 mL), and was treated with the preceding intermediate (1.60 g, 5.2 mmol, 1 equiv), and collidine (1.96 mL, 14.8 mmol, 2.85 equiv). The resulting solution was held at 23 °C overnight, then was taken up in EtOAc (100 mL), and washed sequentially with saturated aqueous citric acid (20 mL), saturated aqueous sodium bicarbonate (20 mL), and brine (2 x 20 mL). The organic layer was dried over MgSO4, then was concentrated to yield 2.18 g of crude product. The product was purified by silica gel chromatography to give 1.42 g (68%) of product. iH NMR (CDCI3) δ 1.15-1.30 (m, 9H), 3.24 (d, 2H, J= 5.8), 3.75-4.43 (m, 2H), 4.15-4.28 (m, 6H), 4.50-4.57 (m, IH), 5.05 (dd, IH, J= 13.6, 5.8), 6.22-6.24 (m, IH), 6.56-6.59 (m, IH), 6.94-6.96 (m, IH), 7.19-7.33 (m, 5H), 9.76 (s, IH). Preparation of Intermediate (2S)-(7-Oxo-l,7-dihydropyrrolo[2,3-c]pyridin-6-yl)-3- phenylpropionic Acid Ethyl Ester
A solution of the preceding intermediate (1.04 g, 2.59 mmol) in toluene (30 mL) was treated with PTSA (5 mg), then heated to reflux for 1 h. The solution was taken up in EtOAc (100 mL), washed with saturated aqueous sodium bicarbonate (30 mL), then brine (30 mL), then concentrated. Purification of the residue by silica gel chromatography gave 0.14 g (18%) of product. iH NMR (CDC13) δ 1.11 (t, 3H, J = 7.1), 3.25 (dd, 1H, J= 14.2, 9.8), 3.49 (dd, IH, J= 14.2, 5.7), 4.11 (q, 2H, J= 7.1), 5.57 (dd, IH, J= 9.6, 5.7), 6.26 (t, IH, J= 2.3), 6.42 (d, IH, J= 7.2), 6.70 (d, IH, J= 7.2), 7.00-7.20 (m, 6H), 10.33 (s, IH),.
Preparation of Compound 3
A solution of the preceding intermediate (0.14 g, 0.45 mmol, 1 equiv) in 1 :1 1,4- dioxane-water (5 mL) was treated with LiOH-hydrate (28 mg, 0.68 mmol, 1.42 equiv), and heated to reflux for 30 min. The resulting solution was taken up in ethyl acetate (50 mL), washed with saturated aqueous citric acid (10 mL), followed by brine (2 x 15 mL), dried over MgSO4, then evaporated to yield 61 mg of the corresponding acid. This was taken up in DMF (5 mL), treated with intermediate Y2 (Example 25, 49 mg, 0.22 mmol, 0.48 equiv), DIEA (0.07 mL, 0.43 mmol, 0.95 equiv), and HATU (82 mg, 0.22 mmol, 0.49 equiv), then held at 23 °C overnight. The solution was taken up in ethyl acetate (30 mL), washed with brine (10 mL), then evaporated. Purification of the residue by silica gel chromatography yielded 0.12 g (56%) of product as a 2:1 mixture of 2S : 2R diastereomers (Compounds 3 and 4, respectively). The diastereomers were separated by preparative reverse phase HPLC (acetonitrile-H2θ gradient). Compound 3: Η NMR (CDCI3) δ 1.28 (t, 3H, J= 7.1), 1.42-1.65 (m, 2H), 1.90-2.10 (m, 2H), 2.20-2.35 (m, IH), 2.82-2.95 (m, IH), 3.05-3.25 (m, 2H), 3.53 (dd, 1H, J= 13.8, 7.8), 4.17 (q, 2H, J= 7.1), 4.45-4.58 (m, IH), 5.76-5.90 (m, 2H), 6.32 (t, IH, J= 2.1), 6.63 (d, IH, J= 7.3), 6.69-6.78 (m, 2H), 7.08-7.35 (m, 7H), 8.11 (d, 1H, J= 7.1), 11.22 (s, IH). EXAMPLE 4
Preparation of Compound 4: tra«5-(2'R,3'"S,4S)-4-[2'-(7"-Oxo-l",7"-dihydropyrrolo[2",3"-c- pyridin-6"-yl)-3'-phenylpropionylamino]-5-(2'"-oxopyrrolidin-3'"-yl)pent-2-enoic Acid Ethyl
Ester
Figure imgf000083_0001
See Preceding Example for preparation: iH NMR (CDCI3) δ 1.18 (t, 3H, J= 7.1), 1.42-1.52 (m, IH), 1.58-1.70 (m, IH), 1.90-2.28 (m, 3H), 3.12 (dd, 1H, J= 13.3, 7.1), 3.20- 3.28 (m, 2H), 3.57 (dd, IH, J= 13.3, 9.0), 4.07 (q, 2H, J= 7.0), 4.45-4.58 (m, IH), 5.80 (d, IH, J= 15.7), 6.04 (t, IH, J= 1.1), 6.37 (t, IH, J= 2.3), 6.48 (s, br, IH), 6.60-6.80 (m, 2H), 7.00-7.40 (m, 6H), 8.11 (d, 1H, J= 7.8), 10.61 (s, IH).
EXAMPLE 5 Preparation of Compound 5: trα«5-(2'S,4S)-6-Carbamoyl-4-(2'-{3"-[(5'"-methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin- 1 "-yl} -3'-phenylpropionylamino)hex-2-enoic Acid Ethyl Ester (Jl)
Figure imgf000083_0002
Preparation of Intermediate (lS)-[l-Hydroxymethyl-3-(tritylcarbamoyl)propyl]carbamic Acid Benzyl Ester (BI)
4-Methylmorpholine (1.89 mL, 17.2 mmol, 1 equiv) and ethyl chloroformate (1.65 mL, 17.3 mmol, 1 equiv) were added to a mixture of commercially available Cbz-Gln(Trt)-OH (9.00 g, 17.2 mmol, 1 equiv) in THF (23 mL) at -10 °C. After stirring 20 min, the reaction mixture was filtered and the filtrate was added dropwise to a suspension of NaBH4 (1.47 g, 38.9 mmol, 2.25 equiv) in H2O (10 mL) at 0 °C. The resulting mixture was allowed to warm to 23 °C and stirred for 5 h. It was then cooled again to 0 °C, quenched by the careful addition of 1 N HC1 (30 mL) and then partitioned between MTBE (500 mL) and brine (2 x 100 mL). The organic phase was dried over Na2SO4 and evaporated to provide the title intermediate 5 which was used without further purification.
Preparation of Intermediate (lS)-[l-(tert-Butyldimethylsilanyloxymethyl)- 3-(tritylcarbamoyl)propyl]carbamic Acid Benzyl Ester (Cl)
Intermediate BI, prepared above, was dissolved in DMF (10 mL). Imidazole (2.69 g,
10 39.5 mmol, 2.3 equiv) and tert-butyldimethylsilyl chloride (2.86 g, 19.0 mmol, 1.1 equiv) were added. The reaction mixture was stirred overnight, then diluted with MTBE (500 mL) and washed sequentially with 2.5% KHSO4, H2O, NaHCO3, H2O, and brine (100 mL each). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 25→40% EtOAc in hexanes) to provide the title
15 intermediate (7.6 g, 71%) as a white amorphous solid: IR (cm"1) 3307, 1708, 1660, 1496, 1249; *H NMR (CDCI3) δ -0.01-0.05 (m, 6H), 0.89 (s, 9H), 1.76-1.93 (m. 2H), 2.29-2.40 (m, 2H), 3.56-3.77 (m, 3H), 5.03-5.16 (m, 3H), 7.00 (s, IH), 7.18-7.39 (m, 20 H); Anal. C38H46N2O4Si: C, H, N.
0 Preparation of Intermediate (2'R,4S)-5-(tert-Butyldimethylsilanyloxy)-4-(2'-hydroxy-3'- phenylpropionylamino)pentanoic Acid Tritylamide (Dl)
Intermediate Cl from above (7.6 g, 12 mmol, 1 equiv) and 10% palladium on carbon (0.45 g) were suspended in EtOH (140 mL) and hydrogenated at 50 psi overnight. The reaction mixture was filtered through Whatman #3 paper, the paper was washed with EtOH 5 (120 mL) and the combined filtrates were evaporated. The residue was combined with D-3- phenyllactic acid (1.42 g, 12.2 mmol, 1 equiv), iP^NEt (4.25 mL, 24.4 mmol, 2 equiv) and HATU (4.64 g, 12.2 mmol, 1 equiv) in DMF (35 mL) at 0 °C. After stirring 1 h, the reaction mixture was allowed to warm to 23 °C and stirred 20 min more. Then 5% KHSO4 (80 mL) and MTBE (600 mL) were added and the phases were separated. The organic phase was 0 washed with H2O (80 mL) and brine (70 mL), dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (50% EtOAc in hexanes) to provide the title intermediate as a white foam (4.85 g, 62%): IR (cm"1) 3394, 3295, 1666, 1649, 1519, 1255, 1114, 1085; lU NMR (CDCI3) δ -0.03-0.02 (m, 6H), 0.85 (s, 9H), 1.70-1.89 (m, 2H), 2.18-2.42 (m, 3H), 2.83 (dd, IH, J= 13.8, 8.1), 3.15 (dd, IH, J= 13.8, 4.0), 3.40 (dd, IH, J= 10.0, 4.7), 3.51 (dd, IH, J= 10.0, 3.1), 3.83-3.94 (m, IH), 4.17-4.23 (m, IH), 6.79 (d, IH, J = 8.7), 7.09 (s, IH), 7.17-7.32 (m, 20H); Anal. C39H48N2θ4Si'0.30H2O: C, H, N.
Preparation of Intermediate (l'S,2R)-Methanesulfonic Acid l-[l'-(tert- butyldimethylsilanyloxymethyl)-3 '-(tritylcarbamoyl)propylcarbamoyl] -2-phenylethyl Ester (El) Intermediate Dl from above (1.96 g, 3.08 mmol, 1 equiv) and iP^NEt (0.752 mL, 4.32 mmol, 1.4 equiv) were dissolved in CH2CI2 (30 mL) and cooled to -10 °C. Methanesulfonyl chloride (0.286 mL, 3.70 mmol, 1.2 equiv) was added dropwise, slowly, with vigorous stirring. After 30 min, the reaction mixture was diluted with CH2CI2 (200 mL), washed with brine (50 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate (2-Hydroxypyridin-3-yl)carbamic acid tert-Butyl Ester (Fl)
A suspension of 10% palladium on carbon (0.35 g) and 2-hydroxy-3-nitropyridine (5.00 g, 35.7 mmol, 1 equiv) in EtOH (170 mL) was subjected to one atmosphere of hydrogen for 16 hours. After purging the reaction vessel with argon, the mixture was filtered through Whatman #3 paper and the filtrate was evaporated to give 2-hydroxy-3-aminopyridine which was used without further purification. This crude material was stirred in THF (100 mL) at 23 °C. Di-tert-butyl dicarbonate (7.79 g, 35.7 mmol, 1 equiv) was added and the reaction mixture was heated to reflux for 4 h. More di-tert-butyl dicarbonate (6.0 g, 27 mmol, 0.8 equiv) was added and the reaction mixture was heated to reflux overnight. The solvent was evaporated and the residue was purified by flash column chromatography (gradient elution, 50→60% EtOAc in hexanes) to provide the title intermediate as a white solid (6.48 g, 83%): IR (cm-1) 3225, 1725, 1649, 1514; *H NMR (CDCI3) δ 1.52 (s, 9H), 6.33 (dd, IH, J= 7.4, 6.6), 7.01 (dd, IH, J= 6.6, 1.8), 7.56 (s, IH), 8.11 (d, IH, J= 7.1), 12.61 (s, IH); Anal. C10H14N2O3: C, H, N. Preparation of Intermediate (1 "S,2'S)-(1 - { 1 '-[ 1 "-(tert-Butyldimethylsilanyloxymethyl)-3"- (tritylcarbamoyl)propylcarbamoyl]-2'-phenylethyl}-2-oxo-l,2-dihydropyridin-3-yl)-carbamic Acid tert-Butyl Ester (Gl)
Intermediate Fl from above (0.838 g, 3.99 mmol, 1.3 equiv) was stirred in THF (20 mL). Sodium hydride (60% dispersion in mineral oil, 0.148 g, 3.70 mmol, 1.2 equiv) was added. After stirring 20 min, a solution of intermediate El from above (1 equiv based on Dl) in THF (15 mL) was added. The resulting mixture was heated to reflux for 40 h, but thin layer chromatography showed the reaction to be only 50% complete. In a separate flask, more sodium hydride (60% dispersion in mineral oil, 0.111 g, 2.78 mmol, 0.9 equiv) was added to a suspension of Fl (0.647 g, 3.08 mmol, 1 equiv) in THF (10 mL). After stirring 20 min, this mixture was added to original reaction vessel and the resulting mixture was heated to reflux for 21 h. After stirring at 23 °C over the weekend, the crude reaction mixture was diluted with MTBE (600 mL) and washed with a mixture of brine and 10% KHSO4 (3:1, 80 mL) and brine (80 mL), then dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 30→35% EtOAc in hexanes) to provide the desired intermediate Gl as a white foam (1.98 g, 77%): IR (cm"1) 3389, 3307, 1725, 1678, 1649, 1590, 1502; *H NMR (CDCI3) δ -0.02-0.04 (m, 6H), 0.86 (s, 9H), 1.52 (s, 9H), 1.55-1.88 (m, 2H), 2.08-2.14 (m, 2H), 3.19 (dd, IH, J= 13.7, 8.1), 3.39-3.51 (m, 2H), 3.53 (dd, IH, J= 14.2, 7.8), 3.82-3.93 (m, IH), 5.60-5.67 (m, IH), 6.17 (t, IH, J= 7.3), 6.44 (d, IH, J= 8.3), 7.04 (s, IH), 7.12-7.36 (m, 21H), 7.59 (s, IH), 7.94 (d, 1H, J= 7.1); Anal. C49H6oN4O6Si: C, H, N.
Preparation of Intermediate (1"S,2'S)-(1 - { 1 '-[ 1 "-Hydroxymethyl-3"-
(tritylcarbamoyl)propylcarbamoyl]-2'-phenylethyl}-2-oxo-l,2-dihydropyridin-3-yl)carbamic Acid tert-Butyl Ester (HI) Intermediate Gl from above (1.92 g, 2.32 mmol, 1 equiv) was dissolved in a mixture of CH3CN (30 mL) and H2O (3 mL) in a plastic tube. Triethylamine trihydro fluoride (21 drops) was added and the reaction solution was stirred overnight. It was then diluted with EtOAc (750 mL), washed with brine (3 x 80 mL), dried over MgSO4 and evaporated to provide the title intermediate which was used without further purification. Preparation of Intermediate tr «5-(2'S,4S)-4-[2'-(3"-tert-Butoxycarbonylamino-2"-oxo-2"H- pyridin-l"-yl)-3'-phenylpropionylamino]-6-(tritylcarbamoyl)-hex-2-enoic Acid Ethyl Ester (II) Intermediate Ηl from above (1 equiv based on Gl) was dissolved in CΗ2CI2 (20 mL) . Commercially available Dess-Martin periodinane (1.09 g, 2.55 mmol, 1.1 equiv) was added. After stirring 2 h, the solvent was evaporated and the residue was suspended in toluene and again evaporated (70 mL, then 2 x 20 mL) to give a yellow foam. One half of this material (1.16 mmol based on Gl) was dissolved in THF (17 mL). (Carbethoxymethylene) triphenylphosphorane (0.563 g, 1.62 mmol, 1.4 equiv) was added and the reaction mixture was heated to reflux for 1 h and then stirred at 23 °C overnight. After evaporating the solvent, the residue was purified by flash column chromatography (gradient elution, 40→50% EtOAc in hexanes) to provide the title intermediate (0.710 g, 77%): IR (cm-1) 3378, 3284, 1719, 1649, 1596, 1508, 1267; Η NMR (CDCI3) δ 1.28 (t, 3H, J= 7.1), 1.47 (s, 9H), 1.54-1.69 (m, IH), 1.87-2.02 (m, IH), 2.09-2.22 (m, 2H), 3.12 (dd, IH, J= 13.7, 7.7), 3.47 (dd, IH, J= 13.7, 8.1), 4.17 (q, 2H, J= 7.1), 4.43-4.54 (m, IH), 5.51-5.58 (m, IH), 5.64 (dd, IH, J= 15.7, 1.6), 6.12 (t, 1H, J= 7.2), 6.60-6.68 (m, 3H), 7.08-7.31 (m, 21H), 7.51 (s, IH), 7.90 (d, 1H, J= 7.1); Anal. C47H50N4O7O.5OH2O: C, H, N.
Preparation of Intermediate tran5-(2'S,4S)-4-(2'- {3"-[(5'"-Methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin- 1 "-yl} -3'-phenylpropionylamino)-6-(tritylcarbamoyl)hex- 2-enoic Acid Ethyl Ester (12)
Intermediate II from above (0.088 g, 0.11 mmol, 1 equiv) was heated (neat) to between 190 and 200 °C for 65 minutes, then allowed to cool providing the crude amine as a dark residue which was dissolved in CΗ3CN (2 mL) and cooled to 0 °C. 5-Methylisoxazole-3- carbonyl chloride (0.033 g, 0.23 mmol, 2 equiv) and 4-methylmorpholine (0.025 mL, 0.23 mmol, 2 equiv) were added and the reaction mixture was allowed to warm to 23 °C. After stirring 40 min, a mixture of 10% KHSO4 and brine (1 :1, 15 mL) and EtOAc (70 mL) were added. The phases were separated and the organic phase was washed with brine, dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (50% EtOAc in hexanes) to provide the title intermediate (0.049 g, 55%): IR (cm"1) 3331, 1678 (br), 1590, 1525; Η NMR CDC ) δ 1.28 (t, 3H, J= 7.1), 1.58-1.72 (m, IH), 1.87-2.03 (m, IH), 2.10-2.26 (m, 2H), 2.48 (s, 3H), 3.15 (dd, IH, J= 13.7, 7.8), 3.47 (dd, IH, J= 13.7, 8.1), 4.17 (q, 2H, J= 7.1), 4.43-4.55 (m, IH), 5.54-5.61 (m, IH), 5.65 (dd, IH, J= 15.8, 1.5), 6.17 (t, IH, J= 7.3), 6.45 (s, IH), 6.65 (dd, IH, J= 15.8, 5.4), 6.72 (s, IH), 6.84 (d, IH, J= 8.0), 7.08- 7.32 (m., 22 H), 8.35 (dd, IH, J= 7.3, 1.5), 9.49 (s, IH); Anal. C47H46N5O7*0.25H2O: C, H, N.
Preparation of Product Jl (Compound 5)
Intermediate 12 from above (0.047 g, 0.059 mmol, 1 equiv) was dissolved in CH2CI2 (3 mL). Triisopropylsilane (0.036 mL, 0.176 mmol, 3 equiv) and TFA (2 mL) were added. The bright yellow solution was stirred 25 min, then diluted with CCI4 (3 mL) and all the volatiles were evaporated. The residue was purified by flash column chromatography (3% CH3OH in CH C1 ) to give the desired product Jl (0.028 g, 85%): IR (cm"1) 3342, 1666 (br), 1590, 1531, 1455; !H NMR (CDCI3) δ 1.30 (t, 3H, J= 7.1), 1.70-1.84 (m, IH), 1.85-1.99 (m, IH), 2.17-2.24 (m, 2H), 2.48 (s, 3H), 3.18 (dd, IH, J= 13.7, 7.8), 3.50 (dd, IH, J= 13.7, 8.1), 4.19 (q, 2H, j = 7.1), 4.43-4.54 (m, IH), 5.68 (dd, IH, J = 15.7, 1.3), 5.74-5.82 (m, IH), 6.00 (s, IH), 6.19 (s, IH), 6.32 (t, 1H, J= 7.3), 6.46 (s, IH), 6.69 (dd, IH, J= 15.7, 5.5), 7.13-7.30 (m, 5H), 7.48 (dd, IH, J= 7.3, 1.6), 7.62 (d, IH, J= 7.6), 8.39 (dd, IH, J= 7.3, 1.6), 9.46 (s, IH); Anal. C28H3iN5O7*0.50H2O: C, H, N.
EXAMPLE 6 Preparation of Compound 6: trαns-(2'S,4S)-6-Carbamoyl-4-{2'-[3"- cyclopentanecarbonylamino-2"-oxo-2"H-pyridin-l"-yl]-3'-phenylpropionylamino}hex-2-enoic Acid Ethyl Ester
Figure imgf000088_0001
The title compound was prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate cyclopentanecarbonyl chloride where appropriate: IR (cm-1) 3319, 1713, 1666, 1590, 1514; ]H NMR (CDCI3) δ 1.31 (t, 3H, J= 7.1), 1.55-2.02 (m, 10H), 2.04-2.22 (m, 2H)2.68-2.80 (m, 1H), 3.16 (dd, IH, J= 13.7, 7.7), 3.51 (dd, J= 13.7, 8.1), 4.19 (q, 2H, J= 7.1), 4.45-4.56 (m, IH), 5.57-5.74 (m, 4H), 6.29 (t, IH, J= 7.4), 6.68 (dd, IH, J= 15.8, 5.5), 7.10-7.32 (m, 7H), 8.30 (s, IH), 8.35 (dd, IH, J= 7.4, 1.7); Anal. C29H36N O6*0.50H2O: C, H, N.
EXAMPLE 7
Preparation of Compound 7: tra«_'-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo-2"H- pyridin-l"-yl)-3'-(4'"-fluorophenyl)propionylamino]-6-carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000089_0001
Preparation of Intermediate tran5-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo-2"H- pyridin- 1 "-yl)-3'-(4",-fluorophenyl)propionylamino]-6-(tritylcarbamoyl)hex-2-enoic Acid Ethyl Ester
Intermediate Cl (Example 5) was converted to the title intermediate by a process analogous to that described in Example 5 for the conversion of Cl to intermediate II utilizing SI (Example 22) where appropriate: IR (cm-1) 3283, 1722, 1651, 1604, 1513; *Η NMR (CDCl3) δ 1.32 (t, 3Η, J= 6.9), 1.62-1.71 (m, IH), 1.94-2.01 (m, IH), 2.17-2.24 (m, 2H), 3.06-3.13 (m, IH), 3.43-3.50 (m, IH), 4.21 (q, 2H, J= 6.9), 4.50-4.57 (m, IH), 5.16-5.25 (m, 2H), 5.52 (t, IH, J= 8.1), 5.63 (dd, IH, J= 15.6, 1.5), 6.18 (t, IH, J= 7.2), 6.65 (s, br. IH), 6.69 (dd, IH, J= 15.6, 5.4), 6.84 (d, IH, J= 8.4), 6.96 (t, 2H, J= 8.4), 7.09-7.18 (m, 7H), 7.21-7.33 (m, 12H), 7.38-7.42 (m, 4H), 7.76 (s, br. IH), 7.99 (d, IH, J= 7.5); Anal. C50H47N4O7O.5OH2O: C, H, N.
Preparation of Compound 7
The preceding intermediate was converted to compound 7 by a process analogous to that described in Example 5 for the conversion of 12 to product Jl : mp = 99-101 °C; IR (cm-1) 3308, 1714, 1650, 1511, 1199; !H NMR (CDC13) δ 1.33 (t, 3H, J= 6.9), 1.72 -1.82 (m, IH), 1.86-1.96 (m, IH), 2.10-2.18 (m, 2H), 3.10-3.17 (m, 2H), 3.10-3.17 (m, 2H), 3.46-3.53 (m, 1H), 3.63-3.74 (m, 2H), 4.22 (q, 2H, J= 6.9), 4.48-4.57 (m, IH), 5.21 (s, 2H), 5.58-5.65 (m, 2H), 5.91-5.97 (m, IH), 6.19-6.22 (m, IH), 6.35 (t, IH, J= 7.2), 6.69 (dd, IH, J= 15.6, 5.1), 6.98 (t, 2H, J= 8.7), 7.13-7.18 (m, 2H), 7.36-7.40 (m, 5H), 7.85 (s, br. IH), 8.06-8.09 (m, IH); Anal. C33H33N4θ7«1.25H2O: C, H, N.
EXAMPLE 8 Preparation of Compound 8: tra«5-(2,S,4S)-4-[2'-(3"-Acetylamino-2"-oxo-2"H-pyridin-l"-yl)- 3'-phenypropionylamino]-6-carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000090_0001
The title compound was prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate acetyl chloride where appropriate: IR (cm"1) 3307, 1708, 1666, 1643, 1590, 1519; iH NMR (CDCI3) δ 1.30 (t, 3H, J= 7.1), 1.70-1.86 (m, 2H), 2.05-2.24 (m, 5H), 3.15 (dd, 1H, J= 13.7, 8.1), 3.50 (dd, IH, J= 13.7, 7.8), 4.19 (q, 2H, J= 7.1), 4.45-4.56 (m, IH), 5.66-5.77 (m, 2H), 5.82 (s,
IH), 5.94 (s, IH), 6.28 (t, IH, J= 7.2), 6.69 (dd, IH, J= 15.7, 5.6), 7.10-7.29 (m, 5H), 7.32-
7.45 (m, 2H), 8.28-8.36 (m, 2H); Anal. C25H3oN4O6'0.50H2O: C, H, N.
EXAMPLE 9 Preparation of Compound 9: traws-(2'S,4S)-6-Carbamoyl-4-{2,-[3"-(2,",2"'- dimethylpropionylamino)-2"-oxo-2"H-pyridin-l"-yl]-3'-phenylpropionylamino}hex-2-enoic Acid Ethyl Ester
Figure imgf000091_0001
The title compound was prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate 2,2- dimethylpropionyl chloride where appropriate: JR (cm-1) 3378, 3307, 3213, 1713, 1666, 1643, 10 1590, 1514, 1273; ]H NMR (CDC13) δ 1.18-1.37 (m, 12H), 1.67-1.98 (m, 2H), 2.05-2.20 (m, 2H), 3.17 (dd, IH, J= 13.6, 7.7), 3.50 (dd, IH, J= 13.6, 8.2), 4.19 (q, 2H, J= 7.1), 4.43-4.54 (m, IH), 5.62-5.72 (m, 2H), 5.81-5.92 (m, 2H), 6.29 (t, IH, J= 7.2), 6.66 (dd, IH, J= 15.8, 5.7), 7.13-7.39 (m, 7H), 8.33-8.38 (m, IH), 8.59 (s, IH); Anal. C28H36N4O6*0.50H2O: C, H, N.
15
EXAMPLE 10 Preparation of Compound 10: tra«5-(2'S,4S)-6-Carbamoyl-4-(2'-{3"-[([l'",3'"]dithiolane-2'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}-3'-phenylpropionylamino)hex-2-enoic Acid Ethyl Ester
Figure imgf000091_0002
The title compound was prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate [l,3]dithiolane-2-carbonyl chloride (prepared as described in Helv. Chim. Ada 1975, 58, 2509) where appropriate: IR (cπr1) 3295, 1672 (br), 1590, 1519 (br), 1273; lR NMR (CDC13) δ 1.30 (t, 3H, J= 7.1), 1.66-2.22 (m, 4H), 3.17 (dd, 1H, J= 13.7, 7.8), 3.28-3.45 (m, 4H), 3.52 (dd, IH, J= 13.7, 8.1), 4.19 (q, 2H, J= 7.1), 4.43-4.55 (m, IH), 5.01 (s, IH), 5.66 (dd, 1H, J= 15.8, 1.5), 5.67 (s, IH), 5.86 (s, 2H), 6.29 (t, IH, J= 7.3), 6.67 (dd, IH, J= 15.8, 5.5), 7.12- 7.40 (m, 7H), 8.31 (dd, IH, J= 7.3, 1.6), 9.57 (s, IH); Anal. C27H32N4θ6S4O.50H2O: C, H, N.
EXAMPLE 11 Preparation of Compound 11: trα«5-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo-2"H- pyridin-r'-yl)-3'-(3'",4"l-difluorophenyl)propionylamino]-6-carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000092_0001
Preparation of Intermediate (2R)-3-(3',4'-Difluorophenyl)-2-hydroxypropionic Acid
Boc-D-3,4-Difluorophenylalanine (3.05 g, 11.0 mmol, 1 equiv) was dissolved in 1,4- dioxane (10 mL), and a solution of ΗC1 in 1,4-dioxane (4.0 M, 10 mL) was added. The reaction was stirred at 23 °C for 5 h, then the solvent was removed under reduced pressure. The residue was dissolved in 1 M Η2SO4 (22 mL), cooled to 0 °C and 2 N NaNO2 (22 mL) was added via addition funnel under argon. The reaction mixture was stirred at 0 °C for 3 h and warmed to room temperature overnight. The resulting mixture was extracted with MTBE (3 x 40 mL), dried over Na2SO4 and concentrated. The residue was recrystallized from ether/petroleum ether to afford the title intermediate (1.29 g, 58%) as off-white solid. lR NMR (DMSO-./6) δ 0.82-0.86 (m, 2H), 1.10-1.27 (m, 4H), 1.40-1.48 (m, 2H), 1.63-1.78 (m, 5H), 3.95-4.00 (m, IH). Preparation of Compound 11
Intermediate Cl (Example 5) was converted to compound 11 by a process analogous to that described in Example 7 for the conversion of Cl to compound 7 utilizing (2R)-3-(3',4'- difluorophenyl)-2-hydroxypropionic acid (Example 11) where appropriate: mp = 175-178 °C; IR (cm-1) 3298, 1661, 1516, 1266; !H NMR (CDC13) δ 1.32 (t, 3H, J= 7.2), 1.74-1.95 (m, 2H), 2.12-2.20 (m, 2H), 3.05-3.12 (m, IH), 3.42-3.50 (m, IH), 4.21 (q, 2H, J= 7.2), 4.45-4.56 (m, IH), 5.21 (s, 2H), 5.58-5.71 (m, 3H), 6.07-6.10 (m, IH), 6.32-6.34 (m, IH), 6.71 (dd, IH, J= 15.6, 5.1), 6.88-6.91 (m, IH), 6.99-7.11 (m, 2H), 7.30-7.33 (m, IH), 7.36-7.39 (m, 5H), 7.53 (s, br. IH), 7.86 (s, br. IH), 8.04 (s, br. IH); Anal. C3ιH32N4θ7'0.50H2θ: C, H, N.
EXAMPLE 12 Preparation of Compound 12: tra«5-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino-4"-methyl-2"- oxo-2"H-pyridin-l"-yl)-3'-phenylpropionylamino]-6-carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000093_0001
Preparation of Intermediate (2-Hydroxy-6-methylphenyl)carbamic Acid Benzyl Ester
A suspension of 10% palladium on carbon (0.045 g) and 2-hydroxy-4-methyl-3- nitropyridine (0.600 g, 3.89 mmol, 1 equiv) in EtOH (20 mL) was subjected to one atmosphere of hydrogen for 16 hours. After purging the reaction vessel with argon, the mixture was filtered through Whatman #3 paper and the filtrate was evaporated to give 2-hydroxy-4- methyl-3-aminoρyridine which was used without further purification. This crude material was stirred in THF (18 mL) at 23 °C. Benzyl chloroformate (0.611 mL, 4.28 mmol, 1.1 equiv) and Na2CO3 (0.907 g, 8.56 mmol, 2.2 equiv) were added and the reaction mixture was stirred for 60 h, but TLC showed the reaction to be only 25% complete. The reaction mixture was heated to reflux for 24 h. More benzyl chloroformate (0.611 mL, 4.28 mmol, 1.1 equiv) and Na2CO3 (0.454 g, 4.28 mmol, 1.1 equiv) were added. The reaction mixture was heated to reflux for 24 h more, allowed to cool, diluted with EtOAc (200 mL), washed with half-saturated brine (2 x 40 mL), dried over MgSO4, filtered and evaporated. The residue was purified by flash column chromatography (gradient elution, 3→5% CH3OH in CH2CI2) to provide the title intermediate (0.363 g, 35%) as a white solid: lH NMR (OMSO-d6) δ 2.02 (s, 3H), 3.31-3.35 (m, 2H), 5.06 (s, 2H), 6.83 (d, IH, J= 6.7), 7.17 (d, IH, J= 6.7), 7.28-7.39 (m, 5H); Anal. C14H14N2O3: C, H, N.
Preparation of Compound 12
The preceding intermediate was converted to compound 12 by a process analogous to that described in Example 7 for the conversion of Cl to compound 7 utilizing commercially available (2R)-2-hydroxy-3-phenylpropionic acid and Cl (Example 5) where appropriate: IR (cm-1) 3284, 1684 (br), 1596, 1327, (br), 1308, 1267, 1237; JH NMR (CDCI3) δ 1.29 (t, 3H, J = 7.1), 1.55-1.68 (m, IH), 1.82-2.07 (m, 4H), 2.14 (s, 3H), 3.08 (dd, 1H, J= 13.6, 7.1), 3.47 (dd, J= 13.6, 8.8), 4.17 (q, 2H, J= 7.1), 4.41-4.52 (m, IH), 5.06 (d, IH, J= 12.3), 5.12 (d, IH, J= 12.3), 5.59 (dd, IH, J= 15.7, 1.6), 5.67-5.74 (m, IH), 5.90 (s, IH), 6.02 (s, IH), 6.17 (d, IH, J= 7.3), 6.68 (dd, IH, J= 15.7, 5.1), 7.01 (s, IH), 7.13-7.38 (m, 10H), 7.48 (d, IH, J = 7.3); Anal. C32H36N4O7O.75H2O: C, H, N.
EXAMPLE 1 Preparation of a 1 :1 mixture of Compound 13: trαn5-(2S,2"S,3S)-2-(2'-Naphthalen-l'-yl-7'- oxo-l',7'-dihydropyrrolo[2',3'-c]pyridin-6'-yl)-N-[2"-(2'"-oxodihydrofuran-3'"-ylidene)-l"-(2""- oxopyrrolidin-3""-ylmethyl)ethyl]-3-phenylpropionamide and Compound 14: tran^-
(2R,2"S,3S)-2-(2•-Νa hthalen-l•-yl-7*-oxo-^,7•-dihydropyrrolo[2•,3•-c]pyridin-6•-yl)-N-[2"-(2",- oxodihydrofuran-3"'-ylidene)-r'-(2'"'-oxopyrrolidin-3'"'-ylmethyl)ethyl]-3-phenylpropionamide
Figure imgf000094_0001
Preparation of Intermediate Pyrrole-2-carboxylic Acid Methyl Ester
Pyrrole-2-carboxylic acid (10.0 g, 90.0 mmol) in diethyl ether (200 mL) was treated with diazomethane (270 mmol, generated from N-nitroso-N-methyl urea), then back titrated with acetic acid until the yellow color dissipated. The solution was washed with saturated aqueous sodium bicarbonate (3 x 20 mL), followed by brine (3 x 20 mL), then was evaporated to obtain 10 g (88%) of product. *H NMR (CDC13) δ 3.86 (s, 3H), 6.26-6.29 (m, IH), 6.91- 6.94 (m, IH), 6.65-6.98 (m, IH), 9.14 (s, IH).
Preparation of Intermediate 5-Bromo-lH-pyrrole-2-carboxylic Acid Methyl Ester A solution of the preceding intermediate (10.0 g, 79.9 mmol, 1 equiv) in carbon tetrachloride (300 mL) was heated to 70 °C, then treated dropwise with a solution of bromine (126.0 mL, 99.9 mmol, 1.25 equiv) in carbon tetrachloride (200 mL). The reaction was initiated by the addition of iodine (40 mg). After the addition was complete, the reaction was held at 70 °C for 10 min, then cooled to 23 °C using an ice bath. The mixture was washed with 10% aqueous sodium carbonate (100 mL), followed by water (100 mL). The volatiles were evaporated, and the residue was purified by silica gel chromatography to obtain 4.5 g (27%) of product: JH NMR (CDCI3) δ 3.88 (s, 3H), 6.23 (dd, IH, J= 3.8, 2.6), 6.80 (dd, IH, J = 3.9, 2.7), 9.29 (s, IH).
Preparation of Intermediate 5 -Naphthalene- l-yl-lH-pyrrole-2-carboxylic Acid Methyl Ester Argon gas was bubbled for 15 min through a solution of the preceding intermediate (2.04 g, 10.0 mmol, 1 equiv) in 2 M aqueous sodium carbonate (20 mL), and DMF (150 mL). The mixture was then treated with tris(dibenzylidienacetone)dipalladium (0) (0.46 g, 0.50 mmol, 0.05 equiv), and triphenylarsine (0.61 g, 2.0 mmol, 0.20 equiv), then heated to reflux, under argon, for 12 h. The mixture was taken up in ethyl acetate (500 mL) and water (150 mL), then filtered through celite. The organic layer was washed with brine (3 x 50 mL), then evaporated. Purification of the residue by silica gel chromatography yielded 2.05 g (81%) of product. iH NMR (CDCI3) δ 3.88 (s, 3H), 6.22 (dd, IH, J= 3.8. 2.6), 7.50-7.59 (m, 4H), 7.88 (dd, IH, J= 3.9, 2.7), 7.89-8.16 (m, 2H), 8.17-8.22 (m, IH), 9.37 (s, IH). This material was taken up in 1:1 1 ,4-dioxane-water (30 mL), and treated with lithium hydroxide hydrate (1.02 g, 24.4 mmol, 2.4 equiv), then heated to reflux for 15 min. The solution was acidified with 20% aqueous citric acid (30 mL), then extracted with ethyl acetate (75 mL). The organic layer was washed with brine (2 x 20 mL), then evaporated. The residue was taken up in CH2CI2, (30 mL), and treated with oxalyl chloride (2.10 mL, 24.0 mmol, 2.4 equiv), and DMF (one drop), then heated to reflux for 30 min. Evaporation yielded 5-naphthalen-l-yl-lH-pyrrole-2- carboxylic acid chloride which was used in the subsequent procedure without additional purification.
Preparation of Products 13 and 14
The preceding intermediate was converted to an inseparable 1:1 mixture of the title compounds by a process that was analogous to that described in Example 3 for the conversion of pyrrole-2-carbonyl chloride to compounds 3 and 4 and utilizing intermediate Yl (Example 22) in lieu of Y2 where appropriate. JH NMR (CDCI3) δ 1.55-2.00 (m, 6H), 2.50-3.50 (m, 7H), 4.00-4.50 (m, 3H), 4.42-5.60 (m, IH), 5.90-8.40 (m, 15H), 10.49 (s, 0.5H), 11.54 (s, 0.5H).
EXAMPLE 14 Preparation of Compound 15: tra«5-(2'S,4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo-2"H- pyridin- 1 "-yl)-3'-cyclohexylpropionylamino]-6-carbamoyl-hex-2-enoicAcid Ethyl Ester
Figure imgf000096_0001
Preparation of Intermediate (2R)-3-Cyclohexyl-2-hydroxypropionic Acid
Boc-D-Cyclohexylalanine-OH (3.00 g, 11.1 mmol, 1 equiv) was dissolved in 1,4- dioxane (10 mL), and a solution of HC1 in 1,4-dioxane (4.0 M, 10 mL) was added. The reaction was stirred at 23 °C for 5 h, then the solvent was removed under reduced pressure. The residue was dissolved in 1 M H2SO4 (22 mL), cooled to 0 °C and 2 N NaNO2 (22 mL) was added via addition funnel under argon. The reaction mixture was stirred at 0 °C for 3 h and warmed to room temperature overnight. The resulting mixture was extracted with MTBE (3 x 40 mL), dried over Na2SO and concentrated. The residue was triturated with petroleum ether, filtered and dried in the air to afford the title intermediate (0.6 g, 31%) as off-white solid. !H NMR (OMSO-d6) δ 3.09-3.24 (m, 2H), 4.20-4.24 (m, IH), 7.13-7.18 (m, IH), 7.38- 7.47 (m, 2H), 8.49 (s, br. IH). Preparation of Compound 15
Intermediate Cl (Example 5) was converted to product 15 by a process analogous to that described in Example 7 for the conversion of Cl to compound 7 utilizing (2R)-3- cyclohexyl-2-hydroxypropionic acid (Example 14) where appropriate: mp = 64-66 °C; IR (cm- 1) 3302, 2925, 1721, 1651, 1197; H NMR (CDC13) δ 0.93-1.05 (m, 2H), 1.18-1.22 (m, 4H), 1.32 (t, 3H, J= 7.2), 1.66-1.75 (m, 4H), 1.82-1.91 (m, 2H), 1.94-2.06 (m, 2H), 2.10-2.17 (m, 2H), 2.90-2.92 (m, 2H), 4.23 (q, 2H, J= 7.2), 4.52-4.60 (m, IH), 5.23 (s, 2H), 5.65 (t, IH, J = 8.1), 5.82-5.86 (m, IH), 5.62 (dd, IH, J= 15.9, 1.8), 6.01-6.05 (m, IH), 6.37 (t, IH, J= 7.2), 6.85 (dd, 1H, J= 15.9, 5.7), 7.25-7.29 (m, IH), 7.36-7.43 (m, 5H), 7.88 (s, br. IH), 8.09 (d, 1H, J= 6.9); Anal. C31H40N4OVI.OH2O: C, H, N.
EXAMPLE 15 Preparation of Compound 16: trα«s-(2'S,2"'S,3"'"S)-Cyclopentanecarboxylic acid(l-{2'-(4"- fluorophenyl)-l'-[2'"-(2""-oxo-dihydrofuran-3""-ylidene)-l'"-(2 -oxopyrrolidin-3'""- ylmethyl)ethylcarbamoyl]ethyl}-2-oxo-l,2-dihydropyridin-3-yl)amide
Figure imgf000097_0001
Preparation of Intermediate Cyclopentanecarboxylic Acid (2-Hydroxypyridin-3-yl)amide A sample of 10% Pd on C (0.40 g) was added to a solution of 2-hydroxy-3- nitropyridine (3.52 g, 25.0 mmol, 1 equiv) in EtOH. The reaction mixture was stirred at room temperature under H2 atmosphere (balloon) overnight and then was filtered through Celite. The filtrate was concentrated under reduced pressure. The residue was dissolved in CH3CN (100 mL), cooled to 0 °C, and cyclopentanecarbonyl chloride (25 mmol, 3.04 mL, 1 equiv) and NMM (2.75 mL, 25 mmol, 1 equiv) were added sequentially. The resulting mixture was stirred at 0 °C for 20 min, then was partitioned between water (400 mL) and 10% CH3OH in - CH2CI2 (2 x 400 mL). The organic layers were dried over Na2SO4, concentrated, and the resulting residue recrystallized from C^C hexanes to afford the title intermediate as a off- white solid (3.36 g, 65%); mp = 242-243 °C; IR (cπr1) 3263, 1644, 1605, 1521, 1199; JH NMR (DMSO-flfc) δ 1.51-1.57 (m, 2H), 1.60-1.72 (m, 4H), 1.79-1.88 (m, 2H), 2.98-3.08 (m, IH), 6.20 (t, IH, J= 6.9), 7.08 (d, IH, J= 6.6), 8.23 (d, IH, J= 7.5), 9.02 (s, br. IH), 11.95 (s, br. IH); Anal. C11H14N2O2: C, H, N.
Preparation of Compound 16
The preceding intermediate was converted to compound 16 by a process that was analogous to that described in Example 19 for the conversion of F2 to Rl : mp = 135-137 °C; IR (cm-1) 3288, 2954, 1754, 1682, 1511, 1219; lR NMR (CDCI3) δ 0.84-0.93 (m, 2H), 1.55- 1.66 (m, 4H), 1.82-2.02 (m, 5H), 2.27-3.22 (m, 2H), 2.70-2.89 (m, 3H), 3.09-3.17 (m, IH), 3.20-3.37 (m, 2H), 3.42-3.39 (m, IH), 5.63 (t, IH, J= 7.2), 6.03 (s, br. IH), 6.25-6.35 (m, 2H), 6.94 (t, 2H, J= 8.7), 7.08-7.12 (m, 2H), 7.25 (d, IH, J- 9.0), 8.30-8.35 (m, 2H), 8.44 (d, 1H, J= 6.0); Anal. C3iH35FN4O6*1.0H2O: C, H, N.
EXAMPLE 16
Preparation of Compound 17: trα«s-(4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo-2"H- pyridin-l"-yl)acetylamino]-6-carbamoylhex-2-enoic Acid Ethyl Ester
Figure imgf000098_0001
Preparation of intermediate (3-benzyloxycarbonylamino-2-oxo-2 j-pyridin-l-yl)acetic acid tert-butyl ester
Sodium hydride (0.070 g of a 60% suspension in mineral oil, 1.75 mmol, 1.0 equiv) was added to a solution of (2-hydroxypyridin-3-yl)carbamic acid benzyl ester (prepared as described in Example 1) (0.415 g, 1.70 mmol, 1 equiv) in THF (20 mL) at 0 °C. The reaction mixture was stirred for 20 min at 0 °C, then tert-butyl bromoacetate (0.275 mL, 1.86 mmol, 1.1 equiv) was added. The reaction mixture was warmed to 23 °C for 45 min, then was partitioned between 0.5 M HC1 (150 mL) and EtOAc (2 x 100 mL). The organic layers were dried over Na2SO4 and were concentrated. Purification of the residue by flash column chromatography (30% EtOAc in hexanes) provided the title compound (0.485 g, 85%) as an off-white solid: mp = 88-90 °C; IR (cm"1) 3380, 1739, 1652, 1604; JH NMR (CDC13) δ 1.48 (s, 9H), 4.57 (s, 2H), 5.20 (s, 2H), 6.25 (t, IH, J= 7.1), 6.87 (dd, IH, J= 6.9, 1.7), 7.27-7.41 (m, 5H), 7.86 (s, br, IH), 8.05 (d, br, 1H J= 6.8); Anal. C19H22N2O5: C, H, N. 5
Preparation of Intermediate (3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-l-yl)acetic Acid (3-Benzyloxycarbonylamino-2-oxo-2H-pyridin-l-yl)acetic acid tert-butyl ester (0.485 g, 1.35 mmol) was stirred in a 1 :1 mixture of trifluoroacetic acid and CΗ2CI2 at 23°C for 1 h. The volatiles were then removed under reduced pressure and the residue was triturated with 10 Et2θ (40 mL). The resulting solid was filtered through a medium frit, washed with Et2θ (20 mL) and air-dried to give the title intermediate (0.315 g, 77%): mp = 171-173 °C; ]H NMR (DMSO-έ β) δ 4.67 (s, 2H), 5.15 (s, 2H), 6.28 (t, IH, J= 7.1), 7.29-7.43 (m, 7H), 7.85 (dd, IH, .7= 7.4, 1.7), 8.47 (s, IH).
15 Preparation of Compound 17
The preceding intermediate was converted to compound 17 by a process analogous to that described in Example 23 for the conversion of W2 to J2: mp = 169-174 °C; IR (cm-1) 3273, 1719, 1649; JH NMR (OMSO-d6) δ 1.19 (t, 3H, J= 7.1), 1.64-1.85 (m, 2H), 2.10 (t, 2H, J= 7.6), 4.11 (q, 2H, J= 7.1), 4.38-4.41 (m, IH), 4.60 (d, IH, J= 15.5), 4.67 (d, IH, J =
20 15.5), 5.15 (s, 2H), 5.92 (dd, IH, J= 15.8, 1.3), 6.26 (t, 1H, J= 7.1), 6.76-6.83 (m, 2H), 7.09- 7.42 (m, 7H), 7.84 (d, IH, J= 7.3), 8.41-8.44 (m, 2H); Anal. C24H28N4O7O.5OH2O: C, H, N.
EXAMPLE 17 Preparation of Compound 18: trαn._-(2'S,4S)-6-Carbamoy_-4-(2'-{4"-methyl-3"-[(5'"- methylisoxazole-3'"-carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}-3'- phenylpropionylamino)hex-2-enoic Acid Ethyl Ester
Figure imgf000100_0001
Preparation of Intermediate 5-Methylisoxazole-3-carboxylic Acid (2'-hydroxy-4'- methylpyridin-3 '-yl)amide
A sample of 10% Pd on C (0.35g) was added to a solution of 2-hydroxy-3- nitropyridine (2.03 g, 14.5 mmol, 1 equiv) in EtOH. The reaction mixture was stirred at room temperature under H2 atmosphere (balloon) overnight and then was filtered through Celite. The filtrate was concentrated under reduced pressure. The residue was dissolved in CH3CN (100 mL), cooled to 0 °C, and 5-methylisoxazole-3-carbonyl chloride (2.1 lg, 14.5 mmol, 1 equiv) and NMM (1.51 mL, 14.5 mmol, 1 equiv) were added sequentially. The resulting mixture was stirred at 0 °C for 20 min, and then was partitioned between water (400 mL) and 10% CH3OH in CH2CI2 (2 x 400 mL). The organic layers were dried over Na2SO4, concentrated and the resulting residue was recrystallized from CH2Cl2/hexanes to afford the title intermediate as an off-white solid (2.42 g, 76%): IR (cm"1) 3330, 1650, 1536; JH NMR (OMSO-dβ) δ 3.34 (s, 3H), 6.31 (t, IH, J= 6.6), 6.73 (s, IH), 7.21 (d, 1H,J= 7.2), 8.29 (s, IH, 7= 7.2), 9.46 (s, br. IH), 12.23 (s, br. IH).
Preparation of Compound 18
The preceding intermediate was converted to Compound 18 by a process analogous to that described in Example 7 for the conversion of Cl to Compound 7 utilizing (2R)-2- hydroxy-3-phenylpropionic acid where appropriate: mp = 138-141 °C; IR (cm*1) 3289, 1663, 1542, 1203; !H NMR (CDCI3) δ 1.22 (t, 3H, J= 7.2), 1.62-1.89 (m, 2H), 1.99 (s, 3H), 2.02- 2.07 (m, IH), 2.47 (s, 3H), 3.03-3.39 (m, 5H), 4.12 (q, 2H, J= 7.2), 4.32-4.41 (m, IH), 5.76 (dd, IH, J= 15.6, 1.5), 5.80-5.83 (m, IH), 6.13 (d, IH, J= 7.5), 6.60 (s, br. IH), 6.75 (dd, IH, J= 15.6, 5.4), 7.15-7.24 (m, 5H), 7.76 (d, IH, J= 7.2), 8.65 (d, IH, J= 7.8), 9.59 (s, br. IH); Anal. C29H33N5OVI.5TFA: C, H, N.
EXAMPLE 18 Preparation of Compound 19: tra«5-(2'S,3'"S,4S)-4-[2'-(3"-Benzyloxy-carbonylamino-2"-oxo- 2"H-pyridin-l"-yl)-3'-phenylpropionylamino]-5-(2'"-oxopyrrolidin-3'"-yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000101_0001
The title compound was prepared from Kl (Example 19) by a process analogous to that described in Example 19 for the conversion of Kl to Rl utilizing commercially available (2R)-
2-hydroxy-3-phenylpropionic acid, (2-hydroxypyridin-3-yl)carbamic acid benzyl ester
(Example 2), and commercially available (carbethoxymethylene)-triphenylphosphorane where appropriate: IR (cm-1) 3272, 1684 (br), 1590, 1514, 1273, 1196; Η NMR CDCb) δ 1.30 (t, 3Η, J= 7.1), 1.42-1.55 (m, IH), 1.59-1.75 (m, IH), 2.00-2.27 (m, 3H), 3.07-3.28 (m, 3H), 3.43 (dd, IH, J= 13.7, 7.3), 4.19 (q, 2H, J= 7.1), 4.36-4.47 (m, IH), 5.12-5.21 (m, 2H), 5.75 (dd, 1H, J= 15.6, 1.2), 5.85-5.94 (m, IH), 6.26 (t, 1H, J= 7.2), 6.58 (s, IH), 6.70 (dd, 1H, J = 15.6, 5.7), 7.10-7.41 (m, 10H), 7.44-7.50 (m, IH), 7.71 (s, IH), 7.97 (d, IH J= 6.2), 8.28 (d, IH, J= 6.8); Anal. C33H36N O7'0.25H2O: C, H, N.
EXAMPLE 19
Preparation of Compound 20: trans-(2"S,2""S,3 'S)-5-Methylisoxazole-3-carboxylic Acid (1'-
{2"-(4'"-fluorophenyl)-l"-[2""-(2'""-oxodihydrofuran-3 -ylidene)-l""-(2 -oxopyrrolidin-3 '- ylmethyl)ethylcarbamoyl] ethyl} -2'-oxo- 1 ',2'-dihydropyridin-3'-yl)amide (Rl )
Figure imgf000102_0001
Preparation of Intermediate (1S,3'S)- {2-(tert-Butyldiρhenylsilanyloxy)-l -[ 1 '-(2",4"- dimethoxybenzyl)-2'-oxopyιτolidin-3'-ylmethyl]ethyl}carbamicAcid tert-Butyl Ester (LI)
Intermediate Kl (4.41 g, 10.8 mmol, 1 equiv) was stirred in CH2CI2 (50 mL) and the mixture was cooled to 0 °C. Triethylamine (7.52 mL, 54.0 mmol, 5 equiv), tert- butylchlorodiphenylsilane (5.53 mL, 21.6 mmol, 2 equiv) and 4-(dimethylamino)pyridine (0.330 g, 2.70 mmol, 0.25 equiv) were added successively. The mixture was allowed to warm to 23 °C and was stirred for 2 h. It was then diluted with MTBE (400 mL), washed with brine (2 x 100 mL), dried over MgSO4 and evaporated. The residue was purified by flash column chromatography to provide the title intermediate (3.51 g, 50%) as a white foam: JR (cm-1) 3319, 1678, 1508; Η NMR CDC ) δ 1.05 (s, 9H), 1.42 (s, 9H), 1.44-1.65 (m, 2H), 2.05- 2.17 (m, IH), 2.23-2.35 (m, IH), 2.44-2.56 (m, IH), 3.14-3.21 (m, 2H), 3.55-3.68 (m, 2H), 3.69-3.81 (m, IH), 3.79 (s, 3H), 3.79 (s, 3H), 4.42 (s, 2H), 4.77 (d, IH, J= 9.3), 6.41-6.46 (m, 2H), 7.09-7.13 (m, IH), 7.34-7.46 (m, 6H), 7.61-7.67 (m, 4H); Anal. C37H50N2θ6Si: C, H, N.
Preparation of Intermediate ( 1 'S,2R,3"S)-N- {2'-(tert-Butyldiphenylsilanyloxy)- 1 '-[ 1 M-(2"',4'"- dimethoxybenzyl)-2"-oxopyrrolidin-3"-ylmethyl]ethyl}-2-hydroxy-3-(4""- fiuorophenyl)propionamide (Ml)
Intermediate LI from above (3.4 g, 5.26 mmol, 1 equiv) was dissolved in 1,4-dioxane (20 mL) at 23 °C. A solution of HCl in the same solvent (4.0 M, 20 mL) was added. After stirring 75 min, the volatiles were evaporated to provide a residue which was dissolved in CH3CN (25 mL) and cooled to 0 °C. (2R)-3-(4'-Fluorophenyl)-2-hydroxypropionic acid (intermediate SI, see below, 0.968 g, 5.26 mmol, 1 equiv), 4-methylmorpholine (1.91 mL, 17.4 mmol, 3.3 equiv) and HATU (2.20 g, 5.79 mmol, 1.1 equiv) were added successively and the reaction mixture was allowed to warm to 23 °C and stirred 2.5 h. It was then diluted with EtOAc (500 mL) and washed with a mixture of brine and 10% KHSO4 (3:1, 100 mL) and a mixture of brine and NaHCO3 (1 :1, 100 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (first in 5% CH3OH in CH2CI2, then in 3% CH3OH in CH2CI2) to provide the title intermediate (0.90 g, 51%) as a white foam: IR (cm-1) 3389, 3319, 1660, 1508; *H NMR (CDCI3) δ 1.04 (s, 9H), 1.52-1.66 (m, 2H), 2.02-2.13 (m, 2H), 2.20-2.39 (m, 2H), 2.77-2.86 (m, IH), 3.07-3.23 (m, 3H), 3.47 (dd, IH, J= 9.9, 5.9), 3.59 (dd, IH, J= 9.9, 3.6), 3.77 (s, 3H), 3.78 (s, 3H), 3.99-4.11 (m, IH), 4.22-4.29 (m, IH), 4.31-4.41 (m, 2H), 6.36-6.45 (m, 2H), 6.81-6.89 (m, 2H), 7.05 (d, 1H, J= 8.0), 7.13-7.22 (m, 3H), 7.34-7.46 (m, 6H), 7.59-7.67 (m, 4H); Anal. C4iH49FN2O6Si'0.25H2O: C, H, N.
Preparation of Intermediate (l'S,2R,3"S)-Methanesulfonic Acid l-{2'-(tert- butyldiphenylsilanyloxy)-r-[l"-(2"',4"'-dimethoxybenzyl)-2"-oxopyιτolidin-3"- ylmethyl]ethylcarbamoyl}-2-(4""-fluorophenyl)ethyl Ester (Nl)
Intermediate Ml from above (2.13 g, 2.99 mmol, 1 equiv) and iP^NEt (0.729 mL,
4.19 mmol, 1.4 equiv) were dissolved in CH2C2 (35 mL) and cooled to -10 °C. Methanesulfonyl chloride (0.277 mL, 3.58 mmol, 1.2 equiv) was added dropwise, slowly, with vigorous stirring. After 30 min, the reaction mixture was diluted with MTBE (500 mL), washed with a mixture of brine and 10% KHSO4 (2:1, 100 mL) and brine (100 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate 5-Methylisoxazole-3-carboxylic Acid(2'-hydroxypyridin-3'- yl)amide (F2)
A suspension of 10% palladium on carbon (0.45 g) and 2-hydroxy-3-nitropyridine
(7.00 g, 50.0 mmol, 1 equiv) in EtOH (210 mL) was subjected to one atmosphere of hydrogen for 16 hours. After purging the reaction vessel with argon, the mixture was filtered through
Whatman #3 paper and the filtrate was evaporated to give 2-hydroxy-3-aminopyridine which was used without further purification. This crude material was suspended in CH3CN (170 mL) and cooled to 0 °C. 5-Methylisoxazole-3-carbonyl chloride (8.00 g, 55.0 mmol, 1 equiv) was added in one portion. After 25 min at 0 °C, the reaction mixture was allowed to warm to 23 °C and stirred for an additional 75 min. The thick mixture was then poured into dilute HCl (0.02 M, 150 mL) and mixed thoroughly. The undissolved solid was collected by filtration and washed with H2O (2 x 20 mL) then dried under vacuum overnight to provide the title intermediate (7.1 g, 65%): !H NMR (DMSO-_/6) δ 2.48 (s, 3H), 6.29 (dd, IH, J= 7.2, 6.6), 6.69 (s, IH), 7.19 (dd, IH, J= 6.6, 1.8), 8.26 (dd, IH, 7= 7.2, 1.8), 9.43 (s, IH), 12.20 (s, IH).
Preparation of Intermediate (l'"S,2"S,3""S)-5-Methylisoxazole-3-carboxylic Acid{l'-[l"-{2"'-
(tert-butyldiphenylsilanyloxy)-l'"-[l""-(2 ,4 -dimethoxybenzyl)-2""-oxopyrrolidin-3""- ylmethyl]ethylcarbamoyl}-2"-(4' -fluorophenyl)ethyl]-2'-oxo-l',2'-dihydroρyridin-3'-yl}amide
(01)
Intermediate F2 from above (1.11 g, 5.06 mmol, 1.7 equiv) was stirred in THF (14 mL). Sodium hydride (60% dispersion in mineral oil, 0.173 g, 4.32 mmol, 1.45 equiv) was added. After stirring 30 min, a solution of intermediate Nl from above (1 equiv based on Ml) in THF (11 mL) was added. The resulting mixture was heated to reflux overnight, then diluted with EtOAc (500 mL) and washed with a mixture of brine and 10% KHSO4 (3:1, 100 mL) and brine and NaHCO3 (3:1, 100 mL). The organic phase was dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 2→3% CH3OH in CH2CI2) to provide the desired intermediate OI as a white foam (2.58 g, 95%): IR (cm-1) 3331, 3284, 1666, 1596, 1531, 1455; !H NMR (CDCI3) δ 1.06 (s, 9H), 1.45-1.71 (m, 2H), 1.94-2.32 (m, 3H), 2.52 (s, 3H), 3.07-3.19 (m, 3H), 3.42-3.52 (m, 2H), 3.67-3.74 (m, IH), 3.79 (s, 3H), 3.81 (s, 3H), 3.99-4.10 (m, IH), 4.30 (d, IH, J= 14.6), 4.39 (d, IH, J= 14.6), 5.54-5.63 (m, IH), 6.25 (t, IH, J= 7.3), 6.42-6.50 (m, 3H), 6.81-6.88 (m, 2H), 7.04-7.13 (m, 3H), 7.20-7.26 (m, IH), 7.36-7.48 (m, 7H), 7.60-7.68 (m, 4H), 8.40 (dd, IH, J= 7.3, 1.6), 9.60 (s, IH); Anal. C5iH56FN5O8Si: C, H, N.
Preparation of Intermediate (2"S,2'"S,3""S)-5-Methylisoxazole-3-carboxylic Acid {1'-[1"-{1'"- [l""-(2 ,4 -dimethoxybenzyl)-2""-oxopyrrolidin-3""-ylmethyl]-2'"-hydroxyethylcarbamoyl}-
2"-(4 -fluorophenyl)ethyl]-2,-oxo-l',2'-dihydropyridin-3'-yl}amide (Pl) Intermediate 01 from above (2.51 g, 2.75 mmol, 1 equiv) was dissolved in a mixture of CH3CN (30 mL) and H2O (1 mL) in a plastic tube. Hydrofluoric acid (48%, 5 mL) was added dropwise. After 1 h and 1.5 h, more hydrofluoric acid (2.5 mL and 2 mL respectively) was added. After 3.5 h total, the reaction mixture was poured into saturated aqueous NaHCO3 (250 mL), extracted with CH2CI2 (3 x 400 mL), dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (5% CH3OH in CH2CI2) to provide the title intermediate (1.84 g, 99%) as a white foam: IR (cm"1) 3401 (br), 3331, 1655, 1590, 1531, 1508, 1455, 1208; Η NMR CCDCb) δ 1.47-1.63 (m, 2H), 1.87-1.99 (m, IH), 2.07-2.18 (m, IH), 2.33-2.44 (m, IH), 2.49 (s, 3H), 3.10-3.24 (m, 3H), 3.46-3.56 (m, 3H), 3.76 (s, 3H), 3.78 (s, 3H), 3.84-3.89 (m, IH), 3.92-4.03 (m, IH), 4.19 (d, IH, J= 14.5), 4.39 (d, IH, J = 14.5), 5.54-5.61 (m, IH), 6.22 (t, IH, J= 7.2), 6.38-6.46 (m, 3H), 6.85-6.93 (m, 2H), 6.99-7.04 (m, IH), 7.06-7.13 (m, 2H), 7.24 (dd, 1H, J= 7.2, 1.7), 8.33 (d, 1H, J= 7.3), 8.38 (dd, IH, .7= 7.2, 1.7), 9.54 (s, IH); Anal. C35H38FN5O8*0.50H2O: C, H, N.
Preparation of Intermediate tra«5-(2"S,2'"S,3""S)-5-Methylisoxazole-3-carboxylic Acid { 1 '-[ 1 "- [ 1 "'-[ 1 ""-(2""',4""'-dimethoxybenzyl)-2""-oxopyrrolidin-3""-ylmethyl]-2"'-(2- oxocyclopentylidene)ethylcarbamoyl]-2"-(4"""-fluorophenyl)ethyl]-2'-oxo-r,2'-dihydropyridin- 3'-yl} amide (Ql)
Intermediate PI from above (1.76 g, 2.60 mmol, 1 equiv) and commercially available Dess-Martin periodinane (1.11 g, 2.60 mmol, 1 equiv) were combined in CH2CI2 (20 mL) at 23 °C and stirred 2 h. The volatiles were evaporated. The residue was suspended in toluene and concentrated to dryness (2 x 25 mL). The resulting residue was dissolved in a mixture of ethylene glycol dimethyl ether and DMF (5:1, 24 mL). 3-(triphenyl-l5-phosphanylidene)- dihydrofuran-2-one (prepared in a manner analogous to that described in Baldwin et al., J. Org. Chem. 1971, vol. 36, 1441) (0.902 g, 2.60 mmol, 1 equiv) was added and the reaction mixture was heated to 100 °C for 1.5 h. It was then allowed to cool, diluted with EtOAc (500 mL), washed with brine (200 mL, 100 mL), dried over MgSO and evaporated. The residue was chromatographed (gradient elution, 2→3% CH3OH in CH2CI2) then rechromatographed (50% EtOAc in CH2CI2) to give the title intermediate (1.46 g) contaminated with triphenylphosphine oxide (approximately 35% by weight): !H NMR (CDCI3) δ 1.47-1.62 (m, 2H), 1.87-1.98 (m, IH), 2.21-2.31 (m, IH), 2.38-2.52 (m, IH), 2.50 (s, 3H), 2.81-2.93 (m, 1H), 3.10-3.33 (m, 4H), 3.50 (dd, IH, 7= 14.1, 7.0), 3.76 (s, 3H), 3.77 (s, 3H), 4.19 (d, IH, J = 14.3), 4.29-4.52 (m, 3H), 4.38 (d, IH, J= 14.3), 5.42-5.49 (m, IH), 6.23 (t, IH, J= 7.2), 6.27-6.33 (m, IH), 6.40-6.48 (m, 3H), 6.90-6.97 (m, 2H), 7.00-7.12 (m, 3H), 7.15 (dd, 1H, J= 7.2, 1.6), 8.39 (dd, 1H, J= 7.2, 1.6), 8.83 (d, 1H, J= 5.7), 9.56 (s, IH).
Preparation of Product Rl (Compound 20)
Intermediate Ql, prepared above, was dissolved in a mixture of CHCI3 (30 mL) and H2O (3 mL). DDQ (0.390 g, 1.72 mmol, 1.4 equiv) was added and the reaction mixture was warmed in a 60 °C oil bath for 1.5 h. More DDQ (0.390 g, 1.72 mmol, 1.4 equiv) was added and the reaction vessel was held in the 60 °C oil bath for 1.5 h more. DDQ (0.390 g, 1.72 mmol, 1.4 equiv) was again added. After stirring 2 h more, the reaction mixture was allowed to cool, diluted with CH2CI2 (250 mL), and washed sequentially with a mixture of brine and 1 N HCl (2:1, 80 mL), a mixture of brine and NaHCO3 (2:1, 80 mL) and brine (80 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (2.5% CH3OH in CH2CI2) to provide the desired product (0.530 g, 73%) as an off-white amorphous solid: IR (cm'1) 3336, 1750, 1682, 1530; JH NMR (CDCI3) δ 1.39-1.49 (m, IH), 1.63-1.80 (m, IH), 2.09-2.37 (m, 3H), 2.49 (s, 3H), 2.74-2.86 (m, IH), 3.11 (dd, IH, J= 13.8, 8.5), 3.14-3.38 (m, 3H), 3.39 (dd, IH, J= 13.8, 7.5), 4.30-4.46 (m, 3H), 5.72-5.79 (m, IH), 6.29 (t, IH, J= 7.2), 6.30-6.36 (m, IH), 6.44-6.47 (m, IH), 6.77 (s, IH), 6.88-6.97 (m, 2H), 7.04-7.11 (m, 2H), 7.44-7.50 (m, IH), 8.37 (dd, 1H, J= 7.2, 1.6), 8.66 (d, IH, J= 6.2), 9.47 (s, IH); Anal. C30H30FN5O7O.25H2O: C, H, N.
EXAMPLE 20 Preparation of Compound 21: trα/25-(2'S,3'"R,4S)-4-[2'-(3"-Benzyloxycarbonylamino-2"-oxo- 2"H-pyridin-l"-yl)-3,-phenylpropionylamino]-5-(2"'-oxopyrrolidin-3"'-yl)pent-2-enoic Acid
Ethyl Ester
Figure imgf000106_0001
The title compound was prepared from (2S,3'R)-{l-[l,-(2",4"-dimethoxybenzyl)-2'- oxopyrrolidin-3'-ylmethyl]-2-hydroxyethyl}carbamic acid tert-butyl ester (prepared as described in Dragovich, et al. J. Med. Chem. 1999, 42, 1213) by a process analogous to that described in Example 19 for the conversion of Kl to Rl utilizing commercially available (2R)- 2-hydroxy-3-phenylpropionic acid, (2-hydroxypyridin-3-yl)carbamic acid benzyl ester
(Example 2), and commercially available (triphenyl-l5-phosphanylidene)acetic acid ethyl ester where appropriate: IR (cnr1) 3483, 3272, 1684 (br), 1514, 1267, 1196; Η NMR (CDCI3) δ 1.31 (t, 3H, 7= 7.1), 1.48-1.79 (m, 2H), 2.02-2.24 (m, 2H), 2.27-2.39 (m, IH), 3.12 (dd, 1H, J = 13.7, 8.2), 3.19-3.34 (m, 2H), 3.48 (dd, IH, J= 13.7, 7.8), 4.19 (q, 2H, 7 = 7.1), 4.43-4.53 (m, IH), 5.17 (s, 2H), 5.73 (dd, 1H, J= 15.6, 1.3), 5.90-5.98 (m, IH), 6.27 (t, 1H, J= 7.1), 6.63 (dd, 1H, J= 15.6, 6.0), 6.65-6.71 (m, IH), 7.13-7.27 (m, 6H), 7.31-7.40 (m, 4H), 7.50 (dd, IH, J= 7.1, 1.6), 7.75 (s, IH), 7.97 (d, IH, J= 6.6), 8.69 (d, IH, 7= 7.0); Anal. C33H36N4O7O.5OH2O: C, H, N.
EXAMPLE 21 Preparation of Compound 22: tra«5-(4S,3""S)-4-(2'-{3"-[(5'"-Methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin- 1 "-yl} acetylamino)-5-(2""-oxopyrrilidin-3""-yl)pent-2- enoic Acid Ethyl Ester
Figure imgf000107_0001
Preparation of Intermediate {3-[(5'-Methylisoxazole-3'-carbonyl)amino]-2-oxo-2H-pyridin-l- yl} acetic Acid tert-Butyl Ester
To a solution of 5-methylisoxazole-3-carboxylic acid (2'-hydroxy-4'-methylpyridin-3'- yl)amide (F2, Example 19) (0.520 g, 2.37 mmol, 1 equiv) in TΗF (20 mL) at 0 °C was added NaΗ (0.095 g, 2.37 mmol, 1.0 equiv). The resulting mixture was stirred at 0 °C for 20 min, and then t-butyl bromoacetate (0.385 mL, 2.61 mmol, 1.1 equiv) was added. The reaction mixture was stirred and warmed to room temperature for 30 min, then was partitioned between 0.5 N ΗC1 (100 mL) and EtOAc (2 x 100 mL). The combined organic layers were dried over Na2SO and were concentrated. Purification of the residue by flash column chromatography (30% EtOAc in hexanes) provided the title intermediate (0.628 g, 79%) as a white solid: IR (cm-1) 3343, 1743, 1651, 1581, 1156; Η NMR (CDC13) δ 1.52 (s, 9H), 2.53 (s, 3H), 4.65 (s, 2H), 6.32 (t, 1H, 7= 7.2), 6.51 (s, IH), 7.01 (dd, 1H, 7= 6.9, 1.8), 8.50 (dd, 1H, 7= 7.5, 1.8), 9.63 (s, br. IH); Anal. C16H19N3O5: C, H, N.
Preparation of Compound 22
The preceding intermediate was transformed into Compound 22 by a process that was analogous to that described in Example 25 for the transformation of V3 to product R3: mp = 102-106 °C; IR (cm"1) 3336, 1684, 1534, 1457; JH NMR (CDCI3) δ 1.27 (t, 3H, 7= 7.2), 1.67-1.75 (m, IH), 1.98-2.09 (m, IH), 2.37-2.49 (m, IH), 2.53 (s, 3H), 2.55-2.61 (m, IH), 3.34-3.46 (m, 2H), 3.51-3.52 (m, IH), 4.17 (q, 2H, 7= 7.2), 4.61-4.78 (m, 3H), 5.98 (dd, IH, 7 = 15.6, 1.5), 6.20 (s, br. IH), 6.35 (t, 1H, 7= 7.8), 6.51 (s, IH), 6.85 (dd, IH, 7= 15.6, 5.1), 7.17 (d, IH, 7= 7.2), 8.33 (d, IH, 7= 7.2), 8.49 (d, IH, 7= 7.5), 9.57 (s, br. IH); Anal. C23H27N5O7: C, H, N.
EXAMPLE 22 Preparation of Compound 23: tran.s-(2"S,2""S,3"""S)-5-Chloroisoxazole-3-carboxylic Acid (1'-
{2"-(4'"-fluorophenyl)-l"-[2""-(2'""-oxodihydrofuran-3 -ylidene)-l""-(2 -oxopyrrolidin-3"""- ylmethyl)ethylcarbamoyl] ethyl }-2'-oxo-l',2'-dihydropyridin-3'-yl)amide (R2)
Figure imgf000108_0001
Preparation of Intermediate (2R)-3-(4'-Fluorophenyl)-2-hydroxypropionic Acid (SI)
This intermediate was prepared according to the method disclosed in the co-pending application, U.S. Provisional Patent Application No. 60/150,365, filed August 24, 1999. Preparation of Intermediate (2R)-3-(4'-Fluorophenyl)-2-hydroxypropionic Acid Methyl Ester (Tl)
This intermediate was prepared as described in co-pending U.S. Provisional Patent Application No. 60/150,365, filed August 24, 1999.
Preparation of Intermediate (2R)-3-(4-Fluorophenyl)-2-trifluoromethane-sulfonyloxypropionic Acid Methyl Ester (UI)
Intermediate Tl from above (0.198 g, 1.00 mmol, 1 equiv) was dissolved in CH2CI2 (6 mL) and cooled in an ice bath. 2,6-Lutidine (0.198 mL, 1.70 mmol, 1.7 equiv) and trifluoromethanesulfonic anhydride (0.269 mL, 1.60 mmol, 1.6 equiv) were added and the reaction mixture was stirred 30 min. It was then diluted with MTBE (200 mL), washed with a mixture of brine and 1 N HCl (2:1, 75 mL) and brine (75 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate 5-Chloroisoxazole-3-carboxylic Acid Ethyl Ester
To a solution of ethyl chlorooximidoacetate (11.0 g, 72.6 mmol, 1 equiv) in 1,1- dichloroethylene (350 mL) was added Et3N (25.3 mL, 181 mmol, 2.5 equiv) in 1,1- dichloroethylene (100 mL) via addition funnel over 30 min. The reaction mixture was stirred at room temperature for 23 h, then was partitioned between water (150 mL) and CH2CI2 (2 x 150 mL). The combined organic layers were dried over Na2SO4 and were concentrated. The residue was purified by flash column chromatography (10% EtOAc in hexanes) to give the title intermediate (4.49 g, 35%) as a pale yellow oil: IR (cm"1) 1735, 1436, 1253; ]H NMR (CDCI3) δ 1.45 (t, 3H, 7= 7.2), 4.49 (q, 2H, 7= 7.2), 7.29 (s, IH).
Preparation of Intermediate 5-Chloroisoxazole-3-carboxylic Acid
An aqueous solution of LiOH (2.0 M, 22.5 mL, 45.0 mmol, 2.0 equiv) was added to a solution of the preceding intermediate (3.95 g, 22.5 mmol, 1 equiv) in EtOH (80 mL) at 23 °C. The resulting mixture was stirred at room temperature for 45 min, then the volatiles were removed under reduced pressure. The residue thus obtained was partitioned between 1.0 N HCl (150 mL) and EtOAc (2 x 150 mL). The combined organic layers were dried over Na2SO4 and were concentrated to give the title intermediate (crude) (2.84 g, 86%) as a off- white solid which was used without further purification: !H NMR (DMSO-Jό) δ 7.15 (s, IH).
Preparation of Intermediate 5-Chloroisoxazole-3-carbonyl Chloride
A solution of 5-chloroisoxazole-3-carboxylic acid (0.750 g, 4.32 mmol, 1 equiv) and thionyl chloride (9.0 mL, 120 mmol, 28 equiv) in CHCI3 (21 mL) was heated to reflux for 28 h. The volatiles were evaporated to provide a residue which was dissolved in benzene (40 mL), which was subsequently evaporated to give the title intermediate as a yellow oil which was used without further purification.
Preparation of Intermediate 5-Chloroisoxazole-3-carboxylic Acid (2'-hydroxypyridin-3'- yl)amide (F3)
A suspension of 10% palladium on carbon (0.15 g) and 2-hydroxy-3-nitropyridine (1.70 g, 12.1 mmol, 2.8 equiv) in EtOH (60 mL) was subjected to one atmosphere of hydrogen for 16 hours. After purging the reaction vessel with argon, the mixture was filtered through Whatman #3 paper and the filtrate was evaporated to give 2-hydroxy-3-aminopyridine (1.33 g, 99%) which was used without further purification. A portion of this material (0.524 g, 4.76 mmol, 1.1 equiv) was combined with the 5-chloroisoxazole-3-carbonyl chloride prepared above (4.32 mmol, 1 equiv based on theoretical yield) in THF (50 mL) and stirred 60 min. The volatiles were evaporated and the residue was stirred in half saturated NaHCO3 (20 mL) for 30 min. The undissolved solid was collected by filtration, washed with H2O (3 x 5 mL) and dried under vacuum overnight to provide the title intermediate (0.900 g, 87%) as a tan solid which was used without further purification. ]H NMR (DMSO-.-6) δ 6.30 (t, IH, 7= 6.9), 7.22 (dd, IH, 7= 6.9, 1.7), 7.25 (s, IH), 8.24 (dd, IH, 7= 6.9, 1.7), 9.47 (s, IH), 12.22 (s, IH).
Preparation of Intermediate (2S)-2-{3'-[(5"-Chloroisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyridin- -yl}-3-(4'"-fluorophenyl)propionic Acid Methyl Ester (VI)
Intermediate F3 from above (0.280 g, 1.17 mmol, 1.17 equiv) was suspended in TΗF (7 mL). Sodium hydride (60% dispersion in mineral oil, 0.044 g, 1.1 mmol, 1.1 equiv) was added in one portion. After stirring 30 min, a solution of intermediate UI from above (1.00 mmol, 1 equiv, based on Tl) in THF (10 mL) was added dropwise. The resulting mixture was stirred 16 hours, then diluted with MTBE (250 mL) and washed with brine (2 x 80 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (33% EtOAc in hexanes) to provide the title intermediate (0.316 g, 75%) as a white foam: IR (cm"1) 3331, 1743, 1690, 1643, 1602, 1531, 1437; !H MR (CDC13) δ 3.33 (dd, IH, 7= 14.5, 10.3), 3.53 (dd, IH, 7= 14.5, 5.5), 3.78 (s, 3H), 5.36 (dd, IH, 7= 10.3, 5.5), 6.18 (t, IH, 7= 7.2), 6.67 (s, IH), 6.82 (dd, IH, 7= 7.2, 1.7), 6.89-6.97 (m, 2H), 7.01-7.07 (m, 2H), 8.38 (dd, 1H, 7= 7.2, 1.7), 9.52 (s, IH); Anal. C19H15CIFN3O5: C, H, N.
Preparation of Intermediate (2S)-2- {3'-[(5"-Chloroisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyridin- -yl}-3-(4'"-fluorophenyl)propionic Acid (Wl)
Intermediate VI from above (0.445 g, 1.06 mmol, 1 equiv) was dissolved in CΗ3OΗ (10 mL). An aqueous solution of NaOH (1.0 M, 4.2 mL, 4.2 mmol, 4 equiv) was added dropwise. The reaction mixture was stirred 2 h, then poured into a mixture of brine and 10% KHSO4 (1 :1, 50 mL) and extracted with CH2CI2 (3 x 100 mL). The combined organic phases were dried over MgSO4 and evaporated to provide the title intermediate as a yellow foam which was used without further purification.
Preparation of Intermediate trα«5-(2'S,3"S)-3-[2*-Amino-3'-(2"-oxodihydrofuran-3"- ylidene)propyl]pyrrolidin-2-one (Y 1 )
Intermediate XI (prepared in a manner analogous to that described in Baldwin et al., 7 Org. Chem. 1971, 36, 1441) (0.413 g, 1.27 mmol, 1.2 equiv) was stirred for 45 min in a mixture of TFA (5 mL) in CH2CI2 (10 mL). Then the volatiles were evaporated to provide the title intermediate which was used without purification.
Preparation of Product R2 (Compound 23)
Intermediates Wl (1.06 mmol, 1 equiv based on VI) and Yl (1.27 mmol, 1.2 equiv based on XI) were combined in CH3CN (14 mL) and cooled in an ice bath. 4- Methylmorpholine (0.583 mL, 5.30 mmol, 5 equiv) and HATU (0.403 g, 1.06 mmol, 1 equiv) were added and the reaction mixture was allowed to warm to 23 °C and stirred 1.5 h. It was - I l l -
then diluted with EtOAc (500 mL) and washed with a mixture of brine and 10% KHSO4 (3:1, 80 mL), a mixture of brine and NaHCO3 (3:1, 80 mL) and brine (80 mL). The organic phase was dried over Na2SO4 and evaporated. The residue was chromatographed (5% CH3OH in CH2O2) then rechromatographed (2.5% CH3OH in CH2CI2) to provide the desired product 5 R2 (0.418 g, 64%) contaminated with a diastereomer (approximately 30%, presumably the 2"R epimer): JH NMR (CDCI3) (mixture of isomers) δ 1.33-1.51 (m), 1.62-1.88 (m), 2.06-2.39 (m), 2.73-2.88 (m), 2.98-3.45 (m), 4.27-4.44 (m), 5.71-5.88 (m), 6.27-6.43 (m), 6.66 (s), 6.68 (s), 6.81 (s), 6.88-6.98 (m), 7.03-7.24 (m), 7.48 (d, 7= 7.3), 7.55-7.62 (m), 8.34-8.42 (m), 8.65 (d, 7= 6.2), 8.77 (d, 7= 7.0), 9.41-9.48 (m).
10
EXAMPLE 23
Preparation of Compound 24: trα«5-(2'S,4S)-6-Carbamoyl-4-{3'-(4"-fluorophenyl)-2'-[2'"-oxo-
3"'-(2"",2"",2""-trifluoroacetylamino)-2"'H-pyridin-l"'-yl]proρionylamino}hex-2-enoic Acid
Ethyl Ester (J2)
Figure imgf000112_0001
Preparation of Intermediate (2R)-3-(4'-Fluorophenyl)-2-hydroxyproρionic Acid Benzyl Ester (T2)
Intermediate SI (1.00 g, 5.43 mmol, 1 equiv) was dissolved in CΗ3CN (8 mL). 1,8-
20 Diazabicyclo[5.4.0]undec-7-ene (0.812 mL, 5.43 mmol, 1 equiv) and benzyl bromide (0.646 mL, 5.43 mmol, 1 equiv) were added successively. The resulting solution was stirred 40 h, then diluted with MTBE (250 mL) and washed with 5% KHSO4 and brine (75 mL each). The organic phase was dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (25% EtOAc in hexanes) to provide the title intermediate (1.25 g,
25 84%) as a colorless oil which solidified over several days: IR (cm"1) 3450, 1954, 1890, 1725, 1602; JH NMR (CDCI3) δ 2.76-2.81 (m, IH), 2.94 (dd, IH, 7= 14.1, 6.2), 3.08 (dd, IH, 7 = 14.1, 4.6), 4.42-4.49 (m, IH), 5.14 (d, 1H, 7= 12.1), 5.21 (d, 1H, 7= 12.1), 6.86-6.94 (m, 2H), 7.03-7.11 (m, 2H), 7.29-7.42 (m, 5H); Anal. C16H15FO3: C, H.
Preparation of Intermediate (2R)-3-(4-Fluorophenyl)-2-trifluoromethane-sulfonyloxypropionic Acid Benzyl Ester (U2)
Intermediate T2 from above (0.134 g, 0.489 mmol, 1 equiv) was dissolved in CH2CI2 (5 mL) and cooled in an ice bath. 2,6-Lutidine (0.114 mL, 0.979 mmol, 2 equiv) and trifluoromethanesulfonic anhydride (0.156 mL, 0.927 mmol, 1.9 equiv) were added and the reaction mixture was stirred 30 min. It was then diluted with MTBE (150 mL), washed with a mixture of brine and 1 N HCl (2:1, 75 mL) and brine (50 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate 2,2,2-Trifluoro-N-(2'-hydroxypyridin-3'-yl)acetamide (F4)
TFA (4 mL) was added to a solution of intermediate Fl (0.700 g, 3.33 mmol, 1 equiv) in CH2CI2 (6 mL). After stirring 30 min, the volatiles were evaporated. The residue was suspended in toluene and concentrated to dryness to provide crude 2-hydroxy-3-aminopyridine which was (without further purification) suspended in CH2CI2 (14 mL) and cooled in an ice bath. 4-Methylmorpholine (1.10 mL, 10.0 mmol, 3 equiv) and trifluoroacetic anhydride (0.471 mL, 3.33 mmol, 1 equiv) were added successively. After 30 min, more 4- methylmorpholine (0.550 mL, 5 mmol, 1.5 equiv) was added. The reaction mixture was stirred an additional 1.5 h, then diluted with EtOAc (250 mL) and washed with brine, H2O and brine (25 mL each). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (5% CH3OH in CH2CI2) to provide the title intermediate (0.173 g, 25%) as a white amorphous solid: ]H ΝMR (OMSO-dβ) δ 6.27 (t, IH, 7 = 6.9), 7.32 (dd, IH, 7= 6.9, 1.8), 7.94 (dd, IH, 7= 6.9, 1.8), 10.16 (s, IH), 12.20 (s, IH); Anal. C7H5F3Ν2O2: C, H, N.
Preparation of Intermediate (2S)-3-(4'-Fluorophenyl)-2-[2"-oxo-3"-(2'",2'",2'"- trifluoroacetylamino)-2"H-pyridin-l"-yl]propionic Acid Benzyl Ester (V2) Intermediate F4 from above (0.111 g, 0.539 mmol, 1.1 equiv) was suspended in TΗF
(3 mL). Sodium hydride (60% dispersion in mineral oil, 0.020 g, 0.50 mmol, 1 equiv) was added in one portion. After stirring 30 min, a solution of intermediate U2 (0.489 mmol, 1 equiv, based on T2) in THF (5 mL) was added dropwise. The resulting mixture was stirred 2 hours, then diluted with MTBE (150 mL) and washed with brine (2 x 50 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (25% EtOAc in hexanes) to provide the title intermediate (0.190 g, 84%) as a thick oil: IR (cm-1) 3343, 1743, 1727, 1655, 1602, 1296, 1214, 1161 ; *H NMR (CDC13) δ 3.31 (dd, 1H, 7= 14.4, 10.0), 3.52 (dd, 1H, 7= 14.4, 5.4), 5.21 (s, 2H), 5.39 (dd, 1H, 7= 10.0, 5.4), 6.18 (t, IH, 7= 7.3), 6.85 (dd, IH, 7= 7.3, 1.7), 6.87-7.04 (m, 4H), 7.25-7.38 (m, 5H), 8.31 (dd, IH, 7= 7.3, 1.7), 9.03 (s, IH); Anal. C23Hι8F4N O4: C, H, N.
Preparation of Intermediate (2S)-3-(4'-Fluorophenyl)-2-[2"-oxo-3"-(2"',2"*,2'"- trifluoroacetylamino)-2"H-pyridin-l"-yl]propionic Acid (W2)
A suspension of 10% palladium on carbon (0.030 g) and intermediate V2 from above (0.151 g, 0.327 mmol, 1 equiv) in EtOΗ (5 mL) was subjected to one atmosphere of hydrogen for 16 hours. After purging the reaction vessel with argon, the mixture was filtered through Whatman #3 paper and the filtrate was evaporated to give the title intermediate which was used without further purification.
Preparation of Intermediate tr_m_ (4S)-4-Amino-6-(tritylcarbamoyl)hex-2-enoic Acid Ethyl Ester (AAl)
Intermediate Zl (prepared according to Dragovich, et al., 7 Med. Chem. 1998, 41,
2806) (0.177 g, 0.326 mmol, 1 equiv) was stirred for 2 h in a solution of ΗC1 in 1,4-dioxane
(2.0 M, 8 mL). Then the volatiles were evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate trans-(7S,4S)-4- {3'-(4"-Fluorophenyl)-2'-[2,"-oxo-3'"-(2"",2"",2""- trifluoroacetylamino)-2'"H-pyridin- 1 '"-yljpropionylamino} -6-(tritylcarbamoyl)hex-2-enoic
Acid Ethyl Ester (13)
Intermediates W2 and AAl (as prepared above) were combined in CΗ2CI2 (5 mL) and cooled in an ice bath. HOBt (0.049 g, 0.36 mmol, 1.1 equiv), iP^NEt (0.171 mL, 0.982 mmol, 3 equiv) and EDC (0.063 g, 0.33 mmol, 1 equiv) were added sequentially. The reaction mixture was allowed to warm to 23 °C overnight, then diluted with EtOAc (200 mL) and washed with 5% KHSO4 and brine (50 mL each). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (44% EtOAc in hexanes) to provide the title intermediate (0.138 g, 53%) as a white amorphous solid: !H NMR (CDCI3) δ 1.30 (t, 3H, 7= 7.1), 1.70-1.84 (m, IH), 1.85-1.99 (m, IH), 2.26-2.34 (m, 2H), 3.01-3.10 (m, IH), 3.31-3.40 (m, IH), 4.19 (q, 2H, 7= 7.1), 4.46-4.56 (m, IH), 5.42-5.49 (m, IH), 5.63 (dd, 1H, 7= 15.7, 1.6), 6.18 (t, 1H, 7= 7.3), 6.62-6.71 (m, 2H), 6.88-6.96 (m, 2H), 7.00-7.06 (m, 2H), 7.10-7.34 (m, 17H), 8.25 (dd, 1H, 7= 7.3, 1.6), 9.02 (s, IH); Anal. C44H40F4N4O6: C, H, N.
Preparation of Product J2 (Compound 24)
Intermediate 13 from above (0.112 g, 0.141 mmol, 1 equiv) and triisopropylsilane (0.086 mL, 0.420 mmol, 3 equiv) were combined in CH2CI2 (4 mL). TFA (2 mL) was added. After stirring 30 min, CCI4 (4 mL) was added and the volatiles were evaporated. The residue was purified by flash column chromatography (5% CH3OH in CH2CI2) to provide the desired product (0.078 g, 100%) as a tan amorphous powder: JH NMR (CDCI3) δ 1.31 (t, 3H, 7 = 7.1), 1.75-1.96 (m, 2H), 2.13-2.30 (m, 2H), 3.12 (dd, IH, 7= 13.7, 7.5), 3.46 (dd, IH, J = 13.7, 8.5), 4.20 (d, 2H, 7= 7.1), 4.45-4.55 (m, IH), 5.63 (dd, IH, J = 15.7, 1.6), 5.66-5.74 (m, IH), 5.92 (s, IH), 6.28 (s, IH), 6.37 (t, IH, 7= 7.3), 6.67 (dd, IH, 7= 15.7, 5.7), 6.92-7.00 (m, 2H), 7.09-7.17 (m, 2H), 7.54 (dd, IH, 7= 7.3, 1.6), 7.60 (d, IH, 7= 7.5), 8.34 (dd, IH, 7= 7.3, 1.6), 9.04 (s, IH); Anal. C25H26F4N4O6*1.25H2O: C, H, N.
EXAMPLE 24 Preparation of Compound 25: trans-(2'S,3""'S,4S)-4-(3,-(4"-Fluorophenyl)-2'-{3"'-[(5""- methylisoxazole-3""-carbonyl)amino]-2'"-oxo-2'"H-pyridin- "-yl}propionylamino)-5-(2" oxopyrrolidin-3'""-yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000116_0001
The title compound was prepared from F2 (Example 19) in a manner analogous to that described for the conversion of U2 to 13 in Example 23 utilizing intermediate Y2 (Example 25) where appropriate: IR (cm-1) 3331, 1690, 1590, 1531, 1455; !H NMR (CDCI3) δ 1.30 (t, 3H, 7= 7.0), 1.45-1.55 (m, IH), 1.64-1.75 (m, IH), 2.03-2.31 (m, 3H), 2.49 (s, 3H), 3.10 (dd, IH, 7= 13.7, 7.9), 3.20-3.46 (m, 3H), 4.20 (q, 2H, 7= 7.0), 4.36-4.47 (m, IH), 5.67 (dd, IH, 7 = 15.7, 1.4), 5.85-5.92 (m, IH), 6.29 (t, 1H, 7= 7.2), 6.45 (s, IH), 6.70 (dd, IH, 7= 15.7, 5.7), 6.86 (s, IH), 6.90-6.97 (m, 2H), 7.10-7.16 (m, 2H), 7.60 (dd, IH, 7= 7.2, 1.6), 8.37 (dd, IH, 7 = 7.2, 1.6), 8.51 (d, IH, 7= 6.6), 9.47 (s, IH).
EXAMPLE 25 Preparation of Compound 26: tr_.«5-(2'S,3""S,4S)-4-(2'-{3"-[(5"'-Methyl- isoxazole-3'"-carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}butyrylamino)- 5-(2""-oxopyrrolidin-3""-yl)pent-2-enoic Acid Ethyl Ester (R3)
Figure imgf000117_0001
Preparation of Intermediate (2R)-2-Trifluoromethanesulfonyl-oxybutyric acid tert-butyl ester (U3)
Commercially available T3 (0.575 g, 3.59 mmol, 1 equiv) was dissolved in CH2CI2 (25 mL) and cooled in an ice bath. 2,6-Lutidine (0.836 mL, 7.18 mmol, 2 equiv) and trifluoromethanesulfonic anhydride (1.15 mL, 6.84 mmol, 1.9 equiv) were added and the reaction mixture was stirred 30 min. It was then diluted with MTBE (400 mL), washed with a mixture of brine and 1 N HCl (2:1, 100 mL) and brine (100 mL), dried over Na2SO4 and evaporated to provide the title intermediate which was used without further purification.
Preparation of Intermediate (2S)-2- {3'-[(5"-Methylisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyri din- l'-yl} butyric Acid tert-Butyl Ester (V3)
Intermediate F2 from above (0.200 g, 0.912 mmol, 1.1 equiv) was suspended in TΗF (6 mL). Sodium hydride (60% dispersion in mineral oil, 0.0332 g, 0.830 mmol, 1 equiv) was added in one portion. After stirring 30 min, a solution of intermediate U3 (0.830 mmol, 1 equiv, based on T3) in TΗF (7 mL) was added dropwise. The resulting mixture was stirred 2 hours, then diluted with EtOAc (200 mL) and washed with brine (2 x 50 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (25% EtOAc in hexanes) to provide the title intermediate (0.178 g, 59%) as an oil: R/= 0.30 (25% EtOAc in hexanes); IR (cm"1) 3331, 1731, 1690, 1649, 1602, 1531 ; *Η NMR (CDCI3) δ 0.93 (t, 3H, 7= 7.3), 1.45 (s, 9H), 1.83-2.01 (m, IH), 2.17-2.31 (m, IH), 2.50 (s, 3H), 5.44-5.51 (m, IH), 6.32 (t, IH, 7= 7.2), 6.48 (s, IH), 7.10 (dd, IH, 7= 7.2, 1.8), 8.45 (dd, 1H, 7= 7.2, 1.8), 9.64 (s, IH); Anal. C18H23N3O5: C, H, N. Preparation of Intermediate (2S)-2- {3'-[(5"-Methylisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyridin-l'-yl}butyric Acid (W3)
Intermediate V3 from above (0.143 g, 0.397 mmol, 1 equiv) was stirred for 1 h in a solution of TFA (2 mL) in CΗ2CI2 (3 mL). The volatiles were evaporated. The residue was suspended in toluene (10 mL) and concentrated to dryness, providing the title intermediate which was used without further purification.
Preparation of Intermediate trα«5-(3'S,4S)-4-Amino-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic Acid Ethyl Ester (Y2) Intermediate X2, prepared according to the method disclosed in the co-pending application, U.S. Provisional Patent Application No. 60/150,358, filed August 24, 1999(0.130 g, 0.398 mmol, 1 equiv), was stirred for 30 min in a solution of TFA (2 mL) in CH2CI2 (3 mL). The volatiles were evaporated. The residue was suspended in toluene (10 mL) and concentrated to dryness, providing the title intermediate which was used without further purification.
Preparation of Product R3 (Compound 26)
Intermediates W3 and Y2 (as prepared above) were combined in CH2CI2 (7 mL) and cooled in an ice bath. HOBt (0.064 g, 0.47 mmol, 1.2 equiv), iP^NEt (0.484 mL, 2.78 mmol, 7 equiv) and EDC (0.084 g, 0.44 mmol, 1.1 equiv) were added sequentially. The reaction mixture was allowed to warm to 23 °C overnight, then diluted with EtOAc (500 mL) and washed with 5% KHSO4 , half saturated NaHCO3, and brine (100 mL each). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 2→3% CH3OH in CH2CI2) to provide the title intermediate (0.119 g, 58%) as a white foam: IR (cm"1) 3331, 1684, 1649, 1590, 1531; JH NMR (CDCI3) δ 0.92 (t, 3H, J = 7.3), 1.29 (t, 3H, J = 7.1), 1.47-1.58 (m, IH), 1.62-1.77 (m, IH), 1.85-2.00 (m, IH), 2.08-2.33 (m, 4H), 2.49 (s, 3H), 3.25-3.42 (m, 2H), 4.19 (q, 2H, J = 7.1), 4.39-4.50 (m, IH), 5.73 (dd, IH, J = 8.8, 6.8), 5.97 (dd, IH, J = 15.7, 1.4), 6.34 (t, IH, J = 7.2), 6.46 (s, IH), 6.86 (dd, IH, J = 15.7, 5.9), 7.18 (s, IH), 7.59 (dd, IH, J = 7.2, 1.8), 8.42 (dd, IH, J = 7.2, 1.8), 8.58-8.62 (m, IH), 9.56 (s, 1); Anal. C25H31N5O7O.5OH2O: C, H, N. EXAMPLE 26 Preparation of a 1:1 Mixture of Compound 27: trø>w-(2'S,2"'S,4S)-6-Carbamoyl-4-(2'-{2"-oxo- 3"-[(tetrahydrofuran-2'"-carbonyl)amino]-2"H-pyridin-l"-yl}-3'-phenylpropionylamino)hex-2- enoic Acid Ethyl Ester and Compound 28: tra«s-(2'S,2"'R,4S)-6-Carbamoyl-4-(2,-{2"-oxo-3"- [(tetrahydrofuran-2'"-carbonyl)amino]-2"H-pyridin-l"-yl}-3'-phenylpropionylamino)hex-2- enoic Acid Ethyl Ester
Figure imgf000119_0001
The title compounds were prepared from II (Example 5) by a process analogous to that described in Example 5 for the conversion of II to product Jl utilizing intermediate tetrahydrofuran-2-carbonyl chloride (racemic, Aldrich) where appropriate: IR (cm-1) 3344, 1646, 1519, 1178; >Η NMR (CDC13) δ 1.34 (t, 3H, 7= 7.2), 1.72-1.82 (m, IH), 1.95-2.04 (m, 2H), 2.16-2.23 (m, 2H), 2.32-2.43 (m, IH), 3.18-3.27 (m, IH), 3.51-3.60 (m, 5H), 3.93-4.00 (m, IH), 4.05-4.12 (m, IH), 4.22 (q, 2H, 7= 7.20, 4.46-4.55 (m, 2H), 5.54-5.69 (m, 2H), 6.34- 6.41 (m, 2H), 6.68 (dd, 1H, 7= 15.6, 5.4), 6.86-6.93 (m, IH), 7.17-7.41 (m, 5H), 8.42-8.45 (m, IH), 9.37 (d, 1H, 7= 10.2); Anal. C28H34N4O7'1.5TFA: C, H, N.
EXAMPLE 27 Preparation of Compound 29: trα«5-(2'S,3""S,4S)-4-(2'-{3"-[(5'"-Chloroisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}butyrylamino)-5-(2""-oxopyrroldin-3""-yl)pent-2- enoic Acid Cyclopentyl Ester (R4)
Figure imgf000120_0001
Preparation of Intermediate (2S)-2- {3'-[(5"-Chloroisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyridin-l'-yl} butyric Acid tert-Butyl Ester (V4)
Intermediate F3 from above (0.781 g, 3.26 mmol, 1.1 equiv) was suspended in TΗF (10 mL). Sodium hydride (60% dispersion in mineral oil, 0.119 g, 2.98 mmol, 1 equiv) was added in one portion. After stirring 30 min, a solution of intermediate U3 (2.96 mmol, 1 equiv, based on T3) in TΗF (15 mL) was by syringe. The resulting mixture was stirred 2 hours, then diluted with EtOAc (500 mL) and washed with brine (2 x 100 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (20% EtOAc in hexanes) to provide the title intermediate (0.834 g, 74%): Ry = 0.33 (20% EtOAc in hexanes); IR (cm"1) 3334, 1732, 1694, 1650, 1603, 1537; *Η NMR (CDC13) δ 0.93 (t, 3H, 7= 7.4), 1.46 (s, 9H), 1.85-2.00 (m, IH), 2.17-2.32 (m, IH), 5.44-5.50 (m, IH), 6.33 (t, IH, 7= 7.3), 6.68 (s, IH), 7.12 (dd, IH, 7= 7.3, 1.7), 8.43 (dd, IH, 7= 7.3, 1.7), 9.61 (s, IH); Anal. C17H20CIN3O5: C, H, N.
Preparation of Intermediate (2S)-2- {3'-[(5"-Chloroisoxazole-3"-carbonyl)amino]-2'-oxo-2'H- pyridin-l'-yl} butyric Acid (W4)
Intermediate V4 from above (0.544 g, 1.42 mmol, 1 equiv) was stirred for 1 h in a mixture of TFA (9 mL) and CΗ2CI2 (9 mL). The volatiles were evaporated. The residue was suspended in CC14 and concentrated to dryness (2 x 10 mL), providing the title intermediate which was used without further purification. Preparation of Intermediate tr<3«_--(3'S,4S)-4-tert-Butoxycarbonylamino-5-[r-(2",4"- dimethoxybenzyl)-2'-oxopyrrolidin-3'-yl]pent-2-enoicAcid Cyclopentyl Ester (X3)
Intermediate Kl (prepared according to Dragovich, et al., 7 Med. Chem. 1999, 42, 1213) (2.00 g, 4.90 mmol, 1 equiv) and commercially available Dess-Martin periodinane (2.30 g, 5.38 mmol, 1.1 equiv) were combined in CH2CI2 (50 mL) and stirred 2 h. The solvent was evaporated and the residue was suspended in and then evaporated from toluene (2 x 25 mL). The resulting residue was dissolved in THF (100 mL). (Triphenyl-l5-phosphanylidene)acetic acid cyclopentyl ester (prepared in a manner analogous to that described in Baldwin, et al. 7 Org. Chem. 1971, 36, 1441) (2.28 g, 5.87 mmol, 1.2 equiv) was added. The reaction mixture was heated to reflux for 100 min, allowed to cool, diluted with EtOAc (300 mL) and washed with a mixture of brine and NaHCO3 (1 :1, 100 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 44→50% EtOAc in hexanes) to provide the title intermediate (1.58 g, 62%) as a foam: IR cm-1) 3307, 1708, 1678, 1508; Η NMR CCDCb) δ 1.43 (s, 9H), 1.50-1.93 (m, 10H), 1.97-2.08 (m, IH), 2.21-2.33 (m, IH), 2.48-2.60 (m, IH), 3.17-3.23 (m, 2H), 3.80 (s, 6H), 4.26-4.40 (m, IH), 4.41 (s, 2H), 5.18-5.24 (m, IH), 5.26-5.33 (m, IH), 5.92 (dd, 1H, 7= 15.6, 1.6), 6.41-6.47 (m, 2H), 6.82 (dd, 1H, 7= 15.6, 5.2), 7.08-7.13 (m, IH); Anal. C28H4oN2O7*0.75H2O: C, H, N.
Preparation of Intermediate tra775-(3'S,4S)-4-tert-Butoxycarbonylamino-5-(2'-oxopyrrolidin-3'- yl)pent-2-enoic Acid Cyclopentyl Ester (X4)
Intermediate X3 from above (1.54 g, 2.98 mmol, 1 equiv) was dissolved in a mixture of H O (5 mL) and CHCI3 (50 mL). DDQ (0.947 g, 4.17 mmol, 1.4 equiv) was added and the reaction mixture was heated in a 60 °C oil bath for 1.5 h. More DDQ (0.90 g, 3.96 mmol, 1.3 equiv) was added and heating at 60 °C was continued for 2 h more. After cooling to 23 °C, the reaction mixture was diluted with CH2CI2 (600 mL) and washed with a mixture of brine and 10% KHSO4 (1 :1, 150 mL) and a mixture of brine and NaHCO3 (2 x 200 mL). The organic phase was dried over MgSO4 and evaporated. The residue was purified by flash column chromatography (gradient elution, 2→3% CH3OH in CH2CI2) to provide the title intermediate (0.850 g, 78%) as a tan foam: IR (cm"1) 3305, 1698, 1522, 1279, 1165; JH NMR (CDCI3) δ 1.44 (s, 9H), 1.52-2.05 (m, IH), 2.38-2.54 (m, 2H), 3.29-3.40 (m, 2H), 4.29-4.41 (m, IH), 5.17-5.27 (m, 2H), 5.93 (dd, 1H, 7= 15.7, 1.6), 5.96-6.40 (m, IH), 6.82 (dd, 1H, 7 = 15.7, 5.3); Anal. Cι9H3oN2θ5*0.30H2O: C, H, N.
Preparation of Intermediate tra«5-(3'S,4S)-4-Amino-5-(2'-oxo-pyrrolidin-3'-yl)pent-2-enoic acid Cyclopentyl Ester (Y3)
Intermediate X4 (0.522 g, 1.42 mmol, 1 equiv) was stirred for 40 min in a solution of TFA (7 mL) in CH2CI2 (10 mL). The volatiles were evaporated. The residue was suspended in CC14 and concentrated to dryness (2 x 20 mL), providing the title intermediate which was used without further purification.
Preparation of Product R4 (Compound 29)
Intermediates W4 and Y3 (as prepared above) were combined in CH2CI2 (25 mL) and cooled in an ice bath. HOBt (0.269 g, 1.99 mmol, 1.4 equiv), iP^NEt (1.74 mL, 9.99 mmol, 7 equiv) and EDC (0.341 g, 0.1.78 mmol, 1.25 equiv) were added sequentially. The reaction mixture was allowed to warm to 23 °C over the weekend. It was then diluted with EtOAc (500 mL) and washed with a mixture of brine and 10% KHSO (1 :1, 100 mL) and brine (100 mL). The organic phase was dried over Na2SO4 and evaporated. The residue was purified by flash column chromatography (2% CH3OH in CH2CI2) to provide the title intermediate (0.590 g, 72%) as a white foam: IR (cm"1) 3295, 1690, 1649, 1590, 1531; lH NMR (CDCI3) δ 0.92 (t, 3H, J = 7.3), 1.50-2.02 (m, 10H), 2.11-2.37 (m, 4H), 3.25-3.44 (m, 2H), 3.64-3.74 (m, IH), 4.41-4.52 (m, IH), 5.18-5.24 (m, IH), 5.69 (dd, IH, J = 9.0, 6.6), 5.94 (dd, IH, J = 15.6, 1.4), 6.36 (t, IH, J = 7.3), 6.67 (s, IH), 6.82 (dd, IH, J = 15.6, 5.9), 7.15 (s, IH), 7.58 (dd, IH, J = 7.3, 1.7), 8.42 (dd, IH, J = 7.3, 1.7), 8.49 (m, IH), 9.52 (s, IH); Anal. C27H32CIN5O7O.5OH2O: C, H, N.
EXAMPLE 28 Preparation of Compound 30: trα«5-(2'S,4S)-6-Carbamoyl-4-(2'-{3"-[(5'"-chloroisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}-3'-phenylpropionylamino)hex-2-enoic Acid Ethyl Ester
Figure imgf000123_0001
Preparation of Intermediate ( S,4"S)-5-Chloroisoxazole-3-carboxylic Acid { l'-[l "-Benzyl -4"- (tert-butyldimethylsilanyloxymethyl)-2"-oxo-6"-(tritylcarbamoyl)hexyl]-2'-oxo- ,2'- dihydropyridin- 3 '-yl } amide
10 To solution of 5 -chloroisoxazole-3 -carboxylic acid (Example 22, 0.253 g, 1.71 mmol,
2.0 equiv) in CHCI3 (21 mL) was added SOCI2 (9 mL). The reaction mixture was heated to reflux for 26 h. The volatiles were removed under reduced pressure to give crude 5- chloroisoxazole-3-carbonyl chloride.
A sample of 10% Pd on C (0.070 g) was added to a solution of (l'S,4"S)-5-
15 chloroisoxazole-3-carboxylic acid { 1 '-[ 1 "-Benzyl-4"-(tert-butyldimethylsilanyloxy-methyl)-2"- oxo-6"-(tritylcarbamoyl)hexyl]-2'-oxo-r,2'-dihydropyridin-3'-yl}amide (Example 1, 0.74 g, 0.855 mmol, 1 equiv) in EtOAc (15 mL). The reaction mixture was stirred at room temperature under H2 atmosphere (balloon) for 4 h, and then was filtered through Celite. The filtrate was concentrated under reduced pressure. The residue was dissolved in THF (15 mL),
20 cooled to 0 °C, and 5-chloroisoxazole-3-carbonyl chloride (1.71 mmol, 2.0 equiv) and NMM (0.188 mL, 1.71 mmol, 2.0 equiv) were added sequentially. The resulting mixture was stirred at 0 °C for 1 h, then was partitioned between water (100 mL) and EtOAc (2 x 100 mL). The organic layers were dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography (35% EtOAc in hexanes) to afford the title intermediate (0.666 g,
25 90%): IR (cm-1) 3332, 1684, 1531; *H NMR (CDC13) δ 0.01 (s, 6H), 0.86 (s, 9H), 1.60-1.71 (m, IH), 1.80-1.83 (m, IH), 2.12-2.16 (m, 2H), 3.16-3.23 (m, IH), 3.36-3.41 (m, IH), 3.47- 3.55 (m, 2H), 3.81-3.90 (m, IH), 5.64-5.70 (m, IH), 6.26 (t, IH, 7= 7.2), 6.47 (d, IH, 7= 8.7), 6.70 (s, IH), 7.02 (s, br. IH), 7.17-7.33 (m, 20H), 7.37 (d, IH, 7= 7.2), 8.40 (d, 7= 7.8), 9.56 (s, br. IH).
Preparation of Compound 30 The preceding intermediate was converted into compound 30 by a process that was analogous to that described in Example 7 for the conversion of intermediate Cl to compound 7: mp = 163-165 °C; IR (cm"1) 3335, 1648, 1533, 1179; *H NMR (CDC13) δ 0.34 (t, 3H, 7 = 7.2), 1.76-1.86 (m, IH), 1.94-2.00 (m, IH), 2.23 (t, 2H, 7= 6.9), 3.20-3.27 (m, IH), 3.35-3.42 (m, 2H), 3.51-3.58 (m, IH), 4.22 (q, 2H, 7= 7.2), 4.52-4.57 (m, IH), 5.69 (dd, IH, 7= 15.6, 1.5), 6.70 (s, br. IH), 6.14 (s, br. IH), 6.32 (s, br. IH), 6.39 (t, IH, J = 7.2), 6.69 (dd, 1H, 7= 15.6, 5.4), 7.21-7.33 (m, 4H), 7.47 (d, IH, 7= 7.2), 8.44 (d, IH, 7= 7.5), 9.47 (s, br. IH); Anal. C27H28C1N5O7*0.6H2O: C, H, N.
EXAMPLE 29 Preparation of Compound 31 : trα«5-(2'S,3""S,4S)-4-(2'- {3"-[(5"'-Methyl-isoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}pent-4'-ynoyl-amino)-5-(2""-oxopyrrolidin-3""- yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000124_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and Y2 and an alternate deprotection of the intermediate corresponding to VI (Lil in pyridine) where appropriate: IR (cm"1) 3295, 1684, 1649, 1596, 1531 ; JΗ NMR (CDCI3) δ 1.27 (t, 3H, 7= 7.1), 1.52-1.62 (m, IH), 1.64-1.80 (m, IH), 2.04 (t, IH, 7= 2.6), 2.08-2.38 (m, 3H), 2.49 (s, 3H), 2.91 (ddd, IH, 7= 17.0, 8.4, 2.6), 3.01 (ddd, IH, 7= 17.0, 6.8, 2.6), 3.22-3.39 (m, 2H), 4.18 (q, 2H, 7= 7.1), 4.44-4.55 (m, IH), 5.71-5.78 (m, IH), 6.03 (dd, 1H, 7= 15.6, 1.5), 6.32 (t, 1H, 7= 7.2), 6.46 (s, IH), 6.85 (dd, 1H, 7= 15.6, 5.4), 6.89 (s, IH), 7.49 (dd, IH, 7= 7.2, 1.7), 8.41 (dd, IH, 7= 7.2, 1.7), 8.68 (d, IH, 7= 6.8), 9.52 (s, IH); Anal. C26H29N5O7O.75H2O: C, H, N.
EXAMPLE 30 Preparation of Compound 32: tr «5-(2'S,3""S,4S)-4-(2'-{3"-[(5"'-Methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}-pent-4'-ynoylamino)-5-(2""-oxopyrrolidin-3""- yl)pent-2-enoic acid 2,2-Dimethylpropyl Ester
Figure imgf000125_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)4-tert-butoxycarbonylamino-5-(2'-oxopyrrolidin-
3'-yl)pent-2-enoic acid 2,2-dimethylpropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) and an alternate deprotection of the intermediate corresponding to VI (Lil in pyridine) where appropriate: IR (cm-1) 3295, 1690, 1649, 1596, 1531; iH NMR (CDCI3) δ 0.95 (s, 9H), 1.54-1.80 (m, 2H), 2.05 (t, 1H, 7= 2.6), 2.10-2.39 (m, 3H), 2.49 (s, 3H), 2.91 (ddd, 1H, 7= 17.0, 8.2, 2.6), 3.01 (ddd, 1H, 7= 17.0, 6.8, 2.6), 3.22-3.38 (m, 2H), 3.82 (s, 2H), 4.45-4.56 (m, IH), 5.72-5.79 (m, IH), 6.06 (dd, IH, 7= 15.7, 1.5), 6.33 (t, IH, 7= 7.2), 6.46 (s, IH), 6.86 (dd, IH, 7= 15.7, 5.4), 6.93 (s, IH), 7.51 (dd, 1H, 7= 7.2, 1.7), 8.41 (dd, 1H, 7= 7.2, 1.7), 8.69 (d, 1H, 7= 6.6), 9.52 (s, IH); Anal. C29H35N5O7O.25H2O: C, H, N. EXAMPLE 31
Preparation of Compound 33 : trans-(2'S,3 S,4S)-4-(3'-(3",4"-Difluorophenyl)-2'- {3"'-[(5""- methylisoxazole-3""-carbonyl)amino]-2"'-oxo-2"'H-pyridin-r"-yl}propionylamino)-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000126_0001
The title compound was prepared by a method analogous to that described in Example 19 for the preparation of specific intermediate Rl utilizing (2R)-3-(3',4'-difluorophenyl)-2- hydroxypropionic acid (synthesized from (2R)-2-tert-butoxycarbonylamino-3-(3',4'- difluorophenyl)propionic acid in analogy to the preparation of intermediate SI described in Example 22) and triethylphosphonoacetate where appropriate: IR (cm-1) 3331, 1690, 1649, 1596, 1531, 1455, 1278; ΪH NMR ^DC ) δ 1.30 (t, 3H, 7= 7.1), 1.44-1.54 (m, IH), 1.63- 1.78 (m, IH), 2.08-2.29 (m, 3H), 2.49 (d, 3H, 7= 0.9), 3.05 (dd, IH, 7= 13.6, 7.5), 3.20-3.42 (m, 3H), 4.19 (dq, 2H, 7= 7.1, 1.7), 4.34-4.45 (m, IH), 5.64 (dd, IH, 7= 15.7, 1.4), 6.00 (t, IH, 7= 7.8), 6.32 (t, IH, 7= 7.3), 6.45 (s, IH), 6.71 (dd, IH, 7= 15.7, 5.6), 6.86-6.91 (m, IH), 6.98-7.08 (m, 2H), 7.15 (s, IH), 7.68 (dd, IH, 7= 7.3, 1.7), 8.39 (dd, 1H, 7= 7.3, 1.7), 8.65 (d, IH, 7= 6.8), 9.46 (s, IH); Anal. C30H31F2N5O7: C, H, N.
EXAMPLE 32 Preparation of Compound 34: trans -(2'S,3""S,4S)-4-(2'- {3"-[(5'"-Methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}butyrylamino)-5-(2""-oxopyrrolidin-3""-yl)pent-2- enoic Acid tert-Butyl Ester
Figure imgf000126_0002
The title compound was prepared by a method analogous to that described for the synthesis of compound R3 in Example 25: IR (cm"1) 3331, 3295, 1690, 1649, 1590, 1531, 1455, 1155; !H NMR (CDC13) δ 0.93 (t, 3H, 7= 7.3), 1.40-1.60 (m, IH), 1.49 (s, 9H), 1.62- 1.78 (m, IH), 1.87-2.02 (m, IH), 2.09-2.36 (m, 4H), 2.50 (s, 3H), 3.26-3.43 (m, 2H), 4.39- 4.50 (m, IH), 5.78 (dd, IH, 7= 8.9, 6.7), 5.89 (dd, IH, 7= 15.7, 1.3), 6.35 (t, IH, 7= 7.3), 6.47 (s, IH), 6.76 (dd, IH, 7= 15.7, 5.9), 7.33 (s, IH), 7.64 (dd, IH, 7= 7.3, 1.8), 8.43 (dd, 1H, 7= 7.3, 1.8), 8.59 (d, 1H, 7= 6.8), 9.57 (s, IH); Anal. C27H35N5O7O.5H2O: C, H, N.
EXAMPLE 33 Preparation of Compound 35: tra«5-(2'S,3""S,4S)-4-(2'-{3"-[(5'"-Methylisoxazole-3'"- carbonyl)amino]-2"-oxo-2"H-pyridin-l"-yl}butyrylamino)-5-(2""-oxopyrrolidin-3""-yl)pent-2- enoic Acid
Figure imgf000127_0001
The title compound was prepared from compound 34 by acidic deprotection of the tert-butyl ester moiety present in that compound (following standard literature procedures): IR (cm-1) 3319, 1678, 1643, 1590, 1531; lU NMR (CDCI3) δ 0.89 (t, 3H, 7= 7.2), 1.61-1.99 (m, 3H), 2.08-2.32 (m, 3H), 2.40-2.54 (m, IH), 2.49 (s, 3H), 3.25-3.41 (m, 2H), 4.58-4.69 (m, IH), 5.56-5.64 (m, IH), 5.98 (d, IH, 7= 15.6), 6.21 (br, IH), 6.37 (t, 1H, 7= 7.3), 6.47 (s, IH), 6.96 (dd, IH, 7= 15.6, 6.6), 7.50 (dd, IH, 7= 7.3, 1.6), 7.52 (s, IH), 8.26 (d, IH, 7= 7.7), 8.44 (dd, IH, 7= 7.3, 1.6), 8.62 (s, IH); Anal. C23H27N5O7'0.75H2O: C, H, N.
EXAMPLE 34 Preparation of Compound 36: tr_.«s-(3'S,4S)-4-[(6'-Benzyloxycarbonylamino-5'-oxo-r,2',3',5'- tetrahydroindolizine-3'-carbonyl)amino]-6-carbamoylhex-2-enoicAcid Ethyl Ester
Figure imgf000128_0001
Preparation of Intermediate 6-But-3-enyl-2-hydroxynicotinonitrile (al) w-Butyllithium (100 mL of a 1.6 M solution in hexanes, 160 mmol, 2.5 equiv) was added via cannula over 10 min to a solution of diisopropylamine (22.4 mL, 160 mmol, 2.5 equiv) in THF (600 mL) at -78 °C. The resulting pale yellow solution was stirred at -78 °C for 5 min, then was warmed to 0 °C for an additional 5 min. 2-Hydroxy-6- methylnicotinonitrile (8) (8.58 g, 64.0 mmol, 1 equiv) was added as a solid in small portions over 15 min and the deep orange solution thus obtained was stirred for 1 h at 0 °C. Allyl bromide (8.31 mL, 96.0 mmol, 1.5 equiv) was then added and the reaction mixture was warmed to 23 °C, maintained at that temperature for 30 min, and was partitioned between 1.0 M HCl (300 mL) and EtOAc (2 x 250 mL). The combined organic layers were dried over Na2SO4 and were concentrated. The resulting orange solid was triturated with boiling Et2θ (100 mL) and subsequently cooled to 23 °C, then was filtered through a medium frit, washed with Et2θ (2 x 50 mL) and air-dried to give al (6.42 g, 58%) as a tan solid: mp = 122-125 °C; IR (KBr pellet, cm-1) 2223, 1654; *H NMR (DMSO- 6) δ 2.32-2.37 (m, 2H), 2.62 (t, 2H, 7= 7.6), 4.96-5.06 (m, 2H), 5.69-5.83 (m, IH), 6.23 (d, IH, 7= 7.3), 8.03 (d, IH, 7= 7.3), 12.55 (s, br, IH); Anal. CιoHιoN2O«0.10H2O: C, H, N.
Preparation of Intermediate 6-But-3-enyl-2-hydroxynicotinamide (bl)
Hydrogen peroxide (30 wt. % solution in water, 45 mL) was added to a solution of al (12.13 g, 70.2 mmol) in a mixture of EtOH (150 mL) and 10% aqueous NaOH (280 mL) at 23 °C. The reaction mixture was heated to 50 °C for 18 h, then was cooled to 23 °C and the volatiles were removed under reduced pressure. The residue was acidified with 12 M HCl to pH 2-3, and the resulting precipitate was filtered, washed with water (2 x 50 mL), and air- dried to afford bl as a yellow solid (13.48 g, 100%): mp = 195-198 °C; IR (cm"1) 3329, 3134, 1688, 1642; H NMR (DMSO-76) δ 2.31-2.38 (m, 2H), 2.64 (t, 2H, 7= 7.6), 4.96-5.05 (m, 2H), 5.71-5.84 (m, IH), 6.29 (d, IH, 7= 7.3), 7.45 (s, br, IH), 8.21 (d, IH, 7= 7.3), 9.00 (s, br, IH), 12.37 (s, br, IH); Anal. Cι0Hi2N2θ2»0!5H2θ: C, H, N.
5 Preparation of Intermediate 6-But-3-enyl-2-hydroxynicotinic Acid (cl)
A solution of bl (12.38 g, 70.1 mmol) in 10% aqueous KOH (350 mL) was heated to reflux for 20 h and subsequently cooled to room temperature. The reaction mixture was acidified with 12 M HCl to pH 2-3, and the resulting precipitate was filtered, washed with water (2 x 50 mL), and dried under vacuum to afford cl as a yellow solid (12.46 g, 92%): mp 10 = 151-155 °C; ER (cm"1) 2905 (br), 1736, 1652; !H NMR (DMSO-76) δ 2.34-2.29 (m, 2H), 2.73 (t, 2H, 7= 7.6), 4.96-5.07 (m, 2H), 5.72-5.85 (m, IH), 6.56 (d, IH, 7= 7.5), 8.28 (d, IH, 7- 7.5), 13.26 (s, br, IH), 14.64 (s, br, IH); Anal. C10H11NO3O.IOH2O: C, H, N.
Preparation of Intermediate (6-But-3-enyl-2-hydroxypyridin-3-yl)carbamic Acid Benzyl Ester
15 (dl)
Triethylamine (13.9 mL, 99.7 mmol, 2.0 equiv) and diphenylphosphoryl azide (16.1 mL, 74.7 mmol, 1.5 equiv) were added sequentially to a suspension of cl (9.63 g, 49.8 mmol, 1 equiv) in 1,4-dioxane (450 mL) at 23 °C. The resulting solution was heated to reflux for 7.5 h, then benzyl alcohol (10.3 mL, 99.5 mmol, 2.0 equiv) was added and reflux was continued
20 for an additional 16 h. The dark brown reaction mixture was cooled to 23 °C and the volatiles were removed under reduced pressure. The resulting dark brown oil was partitioned between water (300 mL) and EtOAc (2 x 250 mL) and the combined organic layers were dried over Na2SO4 and concentrated. The solid thus obtained was triturated with Et2θ (150 mL) and was filtered through a medium frit, washed with Et2θ (2 x 50 mL) and air-dried to give dl (7.34 g,
25 49%) as an off-white powder: mp = 179-180 °C; IR (cm"1) 3386, 1727, 1645; ]H NMR
(CDCI3) δ 2.39-2.46 (m, 2H), 2.65 (t, 2H, 7= 7.5), 4.97-5.07 (m, 2H), 5.21 (s, 2H), 5.73-5.87 (m, IH), 6.10 (d, IH, 7= 7.5), 7.32-7.44 (m, 5H), 7.68 (s, br, IH), 8.06 (s, br, IH), 12.74 (s, br, IH); Anal. C17H18N2O3: C, H, N. Preparation of Intermediate (6-But-3-enyl-2-methoxypyridin-3-yl)carbamic Acid Benzyl Ester (el)
Trimethyloxonium tetrafluoroborate (2.0 g, 13.5 mmol, 1.2 equiv) and 2,6-di-tert- butylpyridine (1.52 mL, 6.76 mmol, 0.6 equiv) were added to a solution of dl (3.36 g, 11.26 mmol, 1 equiv) in CH2CI2 (80 mL) at 23 °C. The reaction mixture was stirred at that temperature for 65 h, then was partitioned between water (2 x 50 mL) and CH2CI2 (2 x 200 mL) and combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography (5% EtOAc in hexanes) to afford el (3.19 g, 91%) as colorless oil: IR (cm"1) 3432, 1733; ]H NMR (CDCI3) δ 2.42-2.49 (m, 2H), 2.73 (t, 2H, 7 = 7.6), 3.96 (s, 3H), 4.94-5.07 (m, 2H), 5.20 (s, 2H), 5.80-5.94 (m, IH), 6.71 (d, IH, 7= 7.9), 7.11 (s, br, IH), 7.32-7.43 (m, 5H), 8.16 (s, br, IH); Anal. C18H20N2O3: C, H, N.
Preparation of Intermediate (3'R)-[6-(3',4'-Dihydroxybutyl)-2-methoxypyridin-3-yl]carbamic Acid Benzyl Ester (fl) To a mixture of 1 : 1 t-BuOH and water (300 mL) at 0 °C was added (DHQD)2AQN
(0.148 g, 0.164 mmol, 0.01 equiv), K3Fe(CN)6 (16.2 g, 49.2 mmol, 3 equiv), K2CO3 (6.8 g, 49.2 mmol, 3 equiv), potassium osmate dihydrate (0.024 g, 0.066 mmol, 0.004 equiv), and then el in t-BuOH (25 mL), sequentially. The resulting mixture was stirred at 0 ° C for 20 h, then warmed to room temperature and Na2SO3 (30 g) was added carefully. The mixture was stirred at room temperature for 2 h, then the volatiles were removed under reduced pressure. The residue was partitioned between water (200 mL) and EtOAc (3 x 200 mL) and the combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography (2% CH3OH in CH2C12) to afford fl (5.57 g, 98%) as pale yellow oil: ER (cm"1) 3427 (br), 1731; JH NMR (CDCI3) δ 1.76-1.87 (m, 2H), 2.07-2.11 (m, IH), 2.80-2.89 (m, 2H), 3.46-3.53 (m, IH), 3.60-3.67 (m, IH), 3.71-3.77 (m, IH), 3.96 (s, 3H), 4.64 (d, IH, 7= 3.2), 5.21 (s, 2H), 6.76 (d, IH, 7= 7.8), 7.12 (s, br, IH), 7.34-7.43 (m, 5H), 8.22 (s, br, IH); Anal. C18H22N2O5: C, H, N. Preparation of Intermediate (3V?)- {6-[4'-(tert-Butyl-dimethyl-silanyloxy)-3'-hydroxybutyl]-2- methoxypyridin-3-yl}carbamic Acid Benzyl Ester (gl)
Triethylamine (1.55 mL, 1 1.1 mmol, 2.5 equiv), tert-butyldimethylsilyl chloride (1.07 g, 7.10 mmol, 1.6 equiv), and 4-dimethylaminopyridine (0.025 g, 0.20 mmol, 0.045 equiv) were added sequentially to a solution of fl (1.54 g, 4.45 mmol, 1 equiv) in CH2CI2 (50 mL) at 23 °C. The reaction mixture was stirred for 19 h at 23 °C, then was partitioned between 0.5 M HCl (150 mL) and a 1 :1 mixture of EtOAc and hexanes (2 x 150 mL). The combined organic layers were dried over Na2SO4 and were concentrated. Purification of the residue by flash column chromatography (20% EtOAc in hexanes) provided gl (1.69 g, 82%) as a colorless oil: IR (cm"1) 3436, 1734; *H NMR (CDCI3) δ 0.07 (s, 6H), 0.90 (s, 9H), 1.72- 1.94 (m, 2H), 2.72-2.88 m, 2H), 3.26 (d, IH, 7= 3.1), 3.50 (dd, IH, 7= 9.8, 6.9), 3.58-3.63 (m, IH), 3.65-3.70 (m, IH), 3.95 (s, 3H), 5.20 (s, 2H), 6.75 (d, IH, 7= 8.1), 7.11 (s, br, IH), 7.33-7.43 (m, 5H), 8.20 (s, br, IH); Anal. C24H36N2O5Si: C, H, N.
Preparation of Intermediate (3S)-(3-Hydroxymethyl-5-oxo-l,2,3,5-tetrahydroindolizin-6-yl)- Carbamic Acid Benzyl Ester (hi)
2,6-Lutidine (2.43 mL, 20.84 mmol, 4.0 equiv) and trifluoromethanesulfonic anhydride (1.31 mL, 7.82 mmol, 1.5 equiv) were added sequentially to a solution of gl (2.4 g, 5.21 mmol, 1 equiv) in CH2CI2 (100 mL) at -78 °C. The colorless reaction mixture was stirred at -78 °C for 45 min, warmed to 23 °C for an additional 15 min, then was partitioned between 0.5 M HCl (150 mL) and CH2CI2 (2 x 150 mL). The organic layer was dried over Na2SO4 and was concentrated. The residue thus obtained was dissolved in THF (120 mL) at 23 °C and tetrabutylammonium fluoride (15.63 mL of a 1.0 M solution in THF, 15.63 mmol, 3.0 equiv) was added. The reaction mixture was stirred at that temperature for 1 h, then was partitioned between 0.5 M HCl (150 mL) and EtOAc (2 x 150 mL). The organic layers were dried over Na2SO and were concentrated. The residue thus obtained was purified by flash column chromatography (80% EtOAc in hexanes) to give hi (0.953 g, 58%) as a colorless oil: IR (cm-1) 3379 (br), 1727, 1649; H NMR (CDCI3) δ 1.86-1.97 (m, IH), 2.29-2.41 (m, IH), 2.90-3.15 (m, 2H), 3.80-3.93 (m, 2H), 4.78-4.86 (m, IH), 5.11-5.15 (m, IH), 5.20 (s, 2H), 6.20 (d, IH, 7= 7.5), 7.30-7.41 (m, 5H), 7.75 (s, br, IH), 8.07 (d, IH, 7= 7.5); Anal. C7Hi8N2θ4'0.75H2O: C, H, N. Preparation of Intermediate (3S)-6-Benzyloxycarbonylamino-5-oxo- 1,2,3, 5- tetrahydroindolizine-3-carboxylic Acid (il)
Dimethylsulfoxide (0.522 mL, 7.36 mmol, 2.6 equiv) was added dropwise to a solution of oxalyl chloride (0.321 mL, 3.68 mmol, 1.3 equiv) in CH2CI2 (80 mL) at -78 °C. The reaction mixture was stirred for 20 min at that temperature, then a solution of hi (0.890 g, 2.83 mmol, 1 equiv) in CH2CI2 (20 mL) was added via cannula. After stirring an additional 20 min at -78 °C, triethylamine (1.97 mL, 14.15 mmol, 5.0 equiv) was added dropwise. The reaction mixture was maintained at -78 °C for 1.5 h, then acetic acid (15.57 mmol, 0.891 mL, 5.5 equiv) was added. The reaction mixture was warmed to 0 °C for 5 min, then was washed with water (50 mL), saturated NaHCO3 (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated to afford the crude aldehyde product as off-white foam. This material was utilized without further purification.
A solution of NaClO2 (2.88 g, 25.47 mmol, 9.0 equiv) and NaH2PO (2.73 g, 19.81 mmol, 7.0 equiv) in water (60 mL) was added dropwise to a solution of the proceeding crude aldehyde (2.83 mmol, 1 equiv) in a mixture of tBuOH (60 mL) and 2-methyl-2-butene (10 mL) over 15 min at 23 °C. The resulting mixture was stirred at room temperature for 1 h, then, the volatiles were removed under reduced pressure. The residue thus obtained was partitioned between 0.5 M HCl (150 mL) and 10% CH3OH in CH2CI2 (2 x 150 mL). The organic layers were dried over Na2SO4 and were concentrated. The resulting residue was purified by flash column chromatography (10% CH3OH in CH2CI2) to give il (0.346 g, 37%) as off-white solid: mp = 204-206 °C; ER (cm"1) 3298, 1722, 1564, 1208; !H NMR (DMSO-<.6) δ 2.17-2.55 (m, 2H), 2.45-2.59 (m, 2H), 4.98 (dd, IH, 7= 9.6, 2.7), 5.16 (s, 2H), 6.23 (d, IH, 7= 7.5), 7.34-7.45 (m, 5H), 7.83 (d, IH, 7= 7.5), 8.34 (s, IH); Anal. C17H16N2O5: C, H, N.
Preparation of Product kl (Compound 36)
Intermediate il was transformed into compound 36 (via coupling with intermediate AAl) by a process that was analogous to that described in Example 23 for the transformation of W2 to product J2: Η NMR (OMSO-d6) δ 1.20 (t, 3H, 7= 7.1), 1.64-1.84 (m, 2H), 2.01- 2.16 (m, 2H), 2.41-2.50 (m, IH), 3.01-3.06 (m, 2H), 4.11 (q, 2H, 7= 7.1), 4.41 (m, br, IH), 5.01 (dd, IH, 7= 9.2, 2.7), 5.14 (s, 2H), 5.87 (dd, 1H, 7= 15.8, 1.4), 6.20 (d, IH, 7= 7.6), 6.75 (s, br, IH), 6.82 (dd, IH, 7= 15.8, 5.0), 7.09-7.42 (m, 7H), 7.81 (d, IH, 7= 7.6), 8.38 (s, IH), 8.55 (d, 1H, 7= 8.4).
EXAMPLE 35 Preparation of Compound 37: trαws-(3'S,3mS,4S)-4-({6'-[(5''-Methylisoxazole-3"- carbonyl)amino] -5 '-oxo- 1 ',2',3 ',5 '-tetrahydroindolizine-3 '-carbonyl } amino)-5 -(2'"- oxopyrrolidin-3'"-yl)-pent-2-enoic Acid Ethyl Ester
Figure imgf000133_0001
Preparation of Intermediate (3"R)-(5-Methylisoxazole-3-carboxylic Acid {6'-[4"-(tert- butyldimethylsilanyloxy)-3"-hydroxybutyl]-2'-methox pyridin-3'-yl} amide (g2)
A suspension of intermediate gl (prepared as described in Example 34) (4.21 g, 9.14 mmol, 1 equiv) and Pd on C (10%, 0.375 g) in EtOAc (120 mL) was stirred under an H2 atmosphere (balloon) at 23 °C for 1 h. The reaction mixture was filtered through Celite and the filtrate was concentrated under reduced pressure. The residue was dissolved in CH3CN (120 mL), cooled to 0 °C, and NMM (1.00 mL, 9.10 mmol, 1.0 equiv) and 5-methylisoxazole- 3-carbonyl chloride (1.33 g, 9.14 mmol, 1.0 equiv) were added sequentially. The reaction mixture was stirred at 0 °C for 1 h, then was partitioned between 0.5 M HCl (150 mL) and EtOAc (2 x 150 mL). The organic layers were dried over Na2SO4 and were concentrated. Trituration of the residue with a 1 :3 mixture of Et2θ and hexanes (130 mL) provided a white solid which was collected by filtration, washed with hexanes (25 mL), and air-dried (2.61 g, 66%): mp = 100-102 °C; IR (cm-1) 3390, 1696, 1593, 1536; iH NMR (CDCI3) δ 0.07 (s, 6H), 0.90 (s, 9H), 1.78-1.95 (m, 2H), 2.52 (s, 3H), 2.78-2.90 (m, 2H), 3.45-3.53 (m, 2H), 3.60- 3.74 (m, 2H), 4.03 (s, 3H), 6.51 (s, IH), 6.79 (d, IH, 7= 7.9), 8.55 (d, IH, 7= 7.9), 8.95 (s, lH); Anal. C2iH33N3O5Si: C, H, N. Preparation of Intermediates (3S)-6-[(5'-Methylisoxazole-3'-carbonyl)amino]- 5-oxo-l,2, 3, 5 -tetrahydroindolizine-3 -carboxylic Acid (i2) and (3S)-8-Chloro-6-[(5'- methylisoxazole-3'-carbonyl)amino]-5-oxo-l,2,3,5-tetrahydroindolizine-3-carboxylic Acid (i3) 5 The preceding intermediate (g2) was transformed into a 1 :4 mixture of intermediates i2 and i3 by a process that was analogous to that described in Example 34 for the transformation of gl to il: (i2) IR (cm-1) 3343, 2977 (br), 1741, 1688, 1648; lR NMR (CDC13) δ 2.28-2.74 (m, 2H), 2.50 (s, 3H), 3.04-3.32 (m, 2H), 5.31 (d, IH, 7= 7.9), 6.36 (d, IH, 7= 7.7), 6.48 (s, IH), 8.58 (d, IH, 7= 7.7), 9.40 (s, IH); (i3) ER (cm"1) 2240, 1652, 1532, 10 1260; iH NMR (DMSO-76) δ 1.53-1.63 (m, 2H), 3.11-3.20 (m, 2H), 5.14 (dd, IH, J = 6.9, 2.7), 6.75 (s, IH), 8.32 (s, IH), 9.41 (s, br. IH).
Preparation of Product 11 (Compound 37)
Intermediate i2 was transformed into compound 37 (via coupling with intermediate
15 Y2) by a process that was analogous to that described in Example 25 for the transformation of W3 to product R3: IR (cm"1) 3336, 1684, 1651, 1596, 1536; Η NMR (CDCI3) δ 1.28 (t, 3H, 7= 7.1), 1.63-1.89 (m, 2H), 2.04-2.15 (m, IH), 2.38-2.64 (m, 3H), 2.50 (s, 3H), 2.98-3.06 (m, IH), 3.26-3.42 (m, 4H), 4.19 (q, 2H, 7= 7.1), 4.63-4.65 (m, IH), 5.15 (dd, IH, 7= 8.3, 2.7), 5.97-6.05 (m, 2H), 6.21 (d, IH, 7= 7.3), 6.48 (s, IH), 6.89 (dd, IH, 7= 15.6, 5.5), 8.44-8.52 0 (m, 2H), 9.40 (s, IH); Anal. C25H29N5O7*0.6TFA: C, H, N.
EXAMPLE 36 Preparation of Compound 38: trα«5-(2"S,3S,3""S)-6-[(5'-Methylisoxazole-3'-carbonyl)amino]- 5-oxo-l ,2, 3, 5-tetrahydroindolizine-3-carboxylic Acid [2"-(2"'-oxodihydrofuran-3"'-ylidene)-l"- 5 (2""-oxoρyrrolidin-3""-ylmethyl)ethyl]amide
Figure imgf000134_0001
The title compound was prepared from intermediate i2 (via coupling with intermediate Yl) by a process that was analogous to that described in Example 22 for the transformation of Wl to product R2: IR (cm"1) 3336, 1752, 1686, 1651, 1594, 1535; JH NMR (CDCI3) δ 1.59-1.68 (m, IH), 1.77-1.81 (m, IH), 2.09-1.81 (m, IH), 2.37-2.45 (m, 2H), 2.50 (s, 3H), 2.58-2.66 (m, IH), 2.91-3.05 (m, 2H), 3.23-3.44 (m, 4H), 4.39 (t, 2H, 7= 7.2), 4.60- 4.67 (m, IH), 5.10-5.13 (m, IH), 6.09 (s, IH), 6.21 (d, IH, 7= 7.9), 6.41-6.48 (m, 2H), 8.45- 8.50 (m, 2H), 8.77 (d, IH, 7= 6.2), 9.39 (s, IH).
EXAMPLE 37 Preparation of Compound 39: trα^-(3'S,3"'S,4S)-4-({8'-Chloro-6'-[(5"-methylisoxazole-3"- carbonyl)amino]-5'-oxo- ,2,,3',5'-tetrahydroindolizine-3'-carbonyl}amino)-5-(2'"- oxopyrrolidin-3"'-yl)-pent-2-enoic Acid Ethyl Ester
Figure imgf000135_0001
The title compound was prepared from intermediate i3 (via coupling with intermediate Y2) by a process that was analogous to that described in Example 25 for the transformation of W3 to product R3: IR (cm"1) 3334, 1683, 1532; JH NMR (CDCI3) δ 1.31 (t, 3H, 7= 7.2), 1.66-1.89 (m, 4H), 2.33-2.50 (m, IH), 2.53 (s, 3H), 2.67-2.72 (m, IH), 3.04-3.20 (m, 2H), 3.27-3.48 (m, 2H), 4.21 (q, 2H, 7= 7.2), 4.62-4.64 (m, IH), 5.16-5.20 (m, IH), 5.94 (s, br. IH), 5.99 (dd, IH, 7= 15.6, 1.5), 6.51 (s, IH), 6.89 (dd, IH, 7= 15.6, 5.7), 8.52 (s, IH), 8.56 (s, br. IH), 9.41 (s, br. IH); Anal. C25H28C1N5O7*0.50H2O: C, H, N. EXAMPLE 38 Compound 40: trα«s-(2'S,3,""S,4S)-4-[2'-(3"- {[1 '"-(5""-Methylisoxazol-3""- yl)methanoyl]amino}-2"'-oxo-2"'H-pyridin- "-yl)pent-4"-ynoylamino]-5-(2,""-oxopyrrolidin- 3"'"-yl)pent-2-enoic Acid Cyclopentyl Ester
Figure imgf000136_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and Y3 and an alternate deprotection of the intermediate corresponding to VI (Lil in pyridine) where appropriate: IR (cm-1) 3302, 1690, 1649, 1632; ]Η NMR (CDC13) δ 1.62-1.80 (m, 7H), 1.87-1.92 (m, 2H), 2.01-2.12 (m, 2H), 2.26-2.40 (m, 2H), 2.53 (s, 3H), 2.95-3.12 (m, 2H), 3.26-3.36 (m, 2H), 4.50-4.53 (m, IH), 5.20-5.25 (m, IH), 5.54-5.60 (m, IH), 6.01 (dd, 1H, 7= 15.6, 1.5), 6.31 (t, 2H, 7= 7.2), 6.40 (s, br, IH), 6.49 (s, IH), 6.83 (dd, IH, 7= 15.6, 5.4), 7.43 (dd, IH, 7= 6.9, 1.8), 8.44 (dd, IH, 7= 6.9, 1.5), 8.58 (d, IH, 7= 6.6), 9.56 (s, IH); Anal. (C29H33N5O7O.75H2O) C, H, N.
EXAMPLE 39 Preparation of Compound 41 : trα«s-(2,S,3"'"S,4S)-4-[2'-(3"-{[l,"-(5""-Methylisoxazol-3""- yl)methanoyl]amino}-2'"-oxo-2'"H-pyridin-r"-yl)pent-4"-ynoylamino]-5-(2'""-oxopyπOlidin- 3"'"-yl)pent-2-enoic Acid tert-Butyl Ester
Figure imgf000136_0002
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2, Kl, and (triphenyl-l5-phosphanylidene)-acetic acid tert-butyl ester and an alternate deprotection of the intermediate corresponding to VI (Lil in pyridine) where appropriate: mp = 195 °C, dec; IR (cm"1) 3295, 1690, 1649; !H NMR (CDC13) δ 1.47 (s, 9H), 1.50-1.79 (m, 2H), 2.04 (t, IH, 7= 2.6), 2.05-2.37 (m, 3H), 2.49 (s, 3H), 2.92 (ddd, IH, 7= 5 17.0, 8.6, 2.6), 3.02 (ddd, IH, 7= 17.0, 6.7, 2.6), 3.20-3.37 (m, 2H), 4.41-4.52 (m, IH), 5.65- 5.73 (m, IH), 5.94 (dd, 1H, 7= 15.6, 1.4), 6.33 (t, 1H, 7= 7.3), 6.46 (s, 1 H), 6.70 (s, 1 H), 6.73 (dd, 1 H, 7= 15.6, 5.3), 7.48 (dd, 1 H, 7= 7.3, 1.7), 8.41 (dd, 1 H, 7= 7.3, 1.7), 8.62 (d, 1 H, 7= 6.6), 9.53 (s, IH); Anal. (C28H33N5O7) C, H, N.
10 EXAMPLE 40
Preparation of Compound 42: trans-(2'S,3 S,4S)-4-[2'-(3"-{[l"'-(5""-Methylisoxazol-3""- yl)methanoyl]amino}-2'"-oxo-2'"H-pyridin-l'"-yl)butanoylamino]-5-(2"'"-oxopyrrolidin-3'""- yl)pent-2-enoic Acid 2,2-Dimethylpropyl Ester
Figure imgf000137_0001
15
The title compound was prepared by a method analogous to that described in Example
25 for the preparation of specific intermediate R3 utilizing trans-(3'S,4S)4-tert- butoxycarbonylamino-5-(2'-oxopyιτolidin-3'-yl)pent-2-enoic acid 2,2-dimethylpropyl ester
(prepared from Kl in a manner similar to that described for the preparation of X4 in Example
20 27) where appropriate: IR (cm"1) 3331, 3295, 1690, 1649; !H NMR (CDCI3) δ 0.87-0.99 (m, 12H), 1.51-1.61 (m, IH), 1.64-1.79 (m, IH), 1.83-2.00 (m, IH), 2.10-2.37 (m, 4H), 2.49 (s, 3H), 3.26-3.43 (m, 2H), 3.83 (s, 2H), 4.43-4.54 (m, IH), 5.71 (dd, IH, 7= 8.7, 6.9), 6.00 (dd, IH, 7= 15.7, 1.3), 6.35 (t, IH, 7= 7.2), 6.46 (s, IH), 6.86 (dd, IH, 7= 15.7, 5.9), 7.39 (s, IH), 7.58 (dd, 1H, 7= 7.2, 1.7), 8.42 (dd, 1H, 7= 7.2, 1.7), 8.53 (d, 1H, 7= 7.0), 9.55 (s, IH); Anal.
25 (C28H37N5O7*0.5H2O) C, H, N. EXAMPLE 41 Preparation of Compound 43: tr_.«5-(2'S,3'""S,4S)-4-[2'-(3"-{[l'"-(5""-Methylisoxazol-3""- yl)methanoyl]amino}-2"'-oxo-2"'H-pyridin- "-yl)pent-4"-ynoylamino]-5-(2""'-oxopyrrolidin- 3"'"-yl)pent-2-enoic Acid
Figure imgf000138_0001
The title compound was prepared from compound 41 by acidic deprotection of the tert-butyl ester moiety present in that compound (following standard literature procedures): Ry = 0.37 (10% CH3OH in CHCI3); IR (cm"1) 3295, 2950 (br), 1696, 1649; Η NMR (CDCI3) δ 1.62-1.85 (m, 2H), 2.05-2.16 (m, 2H), 2.24-2.36 (m, IH), 2.43-2.56 (m, IH), 2.52 (s, 3H), 2.92-3.11 (m, 2H), 3.30-3.42 (m, 2H), 4.60-4.71 (m, IH), 5.65-5.73 (m, IH), 6.03 (d, 1H, 7= 15.6), 6.37 (t, IH, 7= 7.2), 6.50 (s, IH), 6.94 (dd, IH, 7= 15.6, 6.0), 7.24 (s, IH), 7.47 (dd, IH, 7= 7.2, 1.3), 8.31 (d, IH, 7= 7.5), 8.45 (dd, 1H, 7= 7.2, 1.3), 9.53 (s, IH); Anal. (C24H25N5θ7O.5H2O) C, H, N.
EXAMPLE 42 Compound 44: tra«5-(2'S,3""'S,4S)-2-Methyl-4-[2'-(3"-{[l"'-(5""-methylisoxazol-3""- yl)methanoyl] amino} -2"'-oxo-2'"H-pyridin- 1 "'-yl)pent-4"-ynoylamino]-5-(2"",-oxopyrrolidin- 3"'"-yl)pent-2-enoic Acid Ethyl Ester
Figure imgf000138_0002
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid ethyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) and an alternate deprotection of the intermediate corresponding to VI (Lil in pyridine) where appropriate: ER (cm-1) 3307, 1690, 1649; JH NMR (CDC13) δ 1.29 (t, 3H, 7= 7.1), 1.36-1.47 (m, IH), 1.65-1.80 (m, IH), 1.95- 2.00 (m, 4H), 2.14-2.40 (m, 3H), 2.50 (s, 3H), 2.87-3.03 (m, 2H), 3.23-3.38 (m, 2H), 4.18 (q, 2H, 7= 7.1), 4.56-4.68 (m, IH), 5.63-5.72 (m, IH), 6.34 (t, IH, 7= 7.3), 6.47 (s, IH), 6.52- 6.58 (m, IH), 6.81 (s, IH), 7.46 (dd, 1H, 7= 7.3, 1.6), 8.42 (dd, 1H, 7= 7.3, 1.6), 8.65 (d, IH, 7= 6.4), 9.54 (s, IH).
EXAMPLE 43 Preparation of Compound 45: 4S-[2S-(3-Methyl-7-oxo-l,4,5,7-tetrahydro-pyrrolo[2,3- c]pyridin-6-yl)3-phenyl-propionylamino]-5-(2-oxo-pyrrolidin-3-yl)pent-2(trans)-enoic acid ethyl ester.
Figure imgf000139_0001
3-(2-Hydroxy-ethyl)-5-iodo-4-methyl-lH-pyrrole-2-carboxylic acid tert-butyl ester (0.34 mmol, 114 mg) in DMSO (2 mL) was treated with o-iodoxybenzoic acid (EBX, 0.51 mmol, 148 mg), then held at room temperature for 3h. The mixture was diluted with ethyl acetate (50 mL), washed with brine (10 mL), then concentrated under reduced pressure to provide 84 mg (75%) of 5-iodo-4-methyl-3-(2-oxo-ethyl)-lH-pyrrole-2-carboxylic acid tert- butyl ester. 'H NMR (CDC13) δ 9.63 (IH, s), 8.91 (IH, br s), 4.09 (2H, s), 2.62 (3H, s), 1.52 (9H, s).
5-Iodo-4-methyl-3-(2-oxo-ethyl)-lH-pyrrole-2-carboxylic acid tert-butyl ester (0.25 mmol, 84 mg) in MeOH (2 mL) was treated with phenylalanine methyl ester (0.25 mmol, 45 mg), then treated with sodium cyanoborohydride (0.25 mmol, 16 mg). The mixture was held at room temperature for 3h, then concentrated under reduced pressure. Purification of the residue by silica gel chromatography gave 80 mg (46%) of 5-iodo-3-[2S-(l-methoxycarbonyl- 2-phenyl-ethylamino)-ethyl]-4-methyl-lH-pyrrole-2-carboxylic acid tert-butyl ester. 'H NMR (CDC13) δ 8.81 (IH, br s), 7.30-7.13 (5H, m), 3.61 (3H, s), 3.55 (IH, t, J = 6.8), 2.94 (2H, d, J = 6.3), 2.90-2.78 (2H, m), 2.76-2.70 (IH, m), 2.62-2.56 (IH, m), 1.94 (3H, s), 1.51 (9H, s). 5-Iodo-3-[2S-(l-methoxycarbonyl-2-phenyl-ethylamino)-ethyl]-4-methyl-lH-pyrrole-
2-carboxylic acid tert-butyl ester (0.37 mmol, 0.18 g) in dioxane (2 mL) at 0°C was treated with HCl (0.37 mmol, 1 mL of 4M in dioxane). The mixture was held at 0°C for 15 minutes, then concentrated under reduced pressure. The residue was diluted with DMF (3 mL), then treated with HOBT (0.37 mmol, 50 mg), DCC (0.37 mmol, 77 mg), and K2CO3 (0.74 mmol, 0.10 g). The mixture was stirred at room temperature for 30 minutes, then diluted with ethyl acetate (50 mL), washed with brine (10 mL), then concentrated under reduced pressure. The residue was purified by silica gel chromatography to provide 21 mg (13%) of 2S-(3-methyl-7- oxo-l,4,5,7-tetrahydro-pyrrolo[2,3-c]pyridin-6-yl)-3-phenyl propionic acid methyl ester. It was noticed that this material had inadvertently de-iodinated. Η NMR (CDC13) δ 9.28 (IH, br s), 7.31-7.18 (5H, m), 6.64 (IH, s), 5.33 (IH, dd, J = 10.4, 5.6), 3.79-3.74 (IH, m), 3.75 (3H, s), 3.57-3.42 (4H, m), 3.14 (IH, dd, J = 14.5, 10.4), 2.59-2.49 (2H, m), 2.00 (3H, s). MS (FAB) 311 (MH+).
2S-(3-methyl-7-oxo-l,4,5,7-tetrahydro-pyrrolo[2,3-c]pyridin-6-yl)-3-phenyl propionic acid methyl ester (0.07 mmol, 21 mg) in 1 :1 dioxane-water (3 mL) was treated with lithium hydroxide (0.2 mmol, 5 mg), then heated to reflux for 30 minutes. The solution was acidified with saturated aqueous citric acid (3 mL), then extracted with ethyl acetate (15 mL), washed with brine (5 mL), and concentrated under reduced pressure to provide 15 mg (75%) of 2S-(3- methyl-7-oxo-l,4,5,7-tetrahydro-pyrrolo[2,3-c]pyridin-6-yl)-3-phenyl propionic acid. This material (15 mg, 0.05 mmol) was coupled to 4S-amino-5-(2-oxo-pyrrolidin-3S-yl)-pent- 2(trans)-enoic acid ethyl ester (11 mg, 0.05 mmol) in DMF (1 mL) by treatment with diisopropylethyl amine (0.02 mL, 0.01 mmol) and HATU (19 mg, 0.05 mmol) at room temperature for 2 h. The resulting solution was washed with brine (10 mL), and extracted with EtOAc (30 mL). The organics were concentrated under reduced pressure and the residue purified by preparative reverse phase chromatography (CH3CN-H2O) to provide 10 mg (40 %) of 4S-[2S-(3-methyl-7-oxo-l,4,5,7-tetrahydro-ρyrrolo[2,3-c]ρyridin-6-yl)-3-ρhenyl- propionylamino]-5-(2-oxo-pyrrolidin-3-yl)pent-2(trans)-enoic acid ethyl ester. Η NMR (CDCI3) δ 10.21 (IH, br s), 7.54 (IH, d, J = 7.7), 7.26-7.11 (4H, m), 6.80 (IH, d, J = 15.6), 6.62 (IH, br s), 6.23 (IH, br s), 5.82 (IH, dd, J = 15.5, 5.4), 5.28 (IH, t, J = 8.0), 4.68-4.50 (IH, m), 4.19 (2H, q, J = 7.1), 3.67-2.96 (6H, m), 2.56-2.51 (3H, m), 2.21-2.09 (2H, m), 1.96 (3H, s), 1.75-1.64 (IH, m), 1.53-1.43 (3H, m), 1.29 (3H, t, J = 7.1). HRMS (FAB) 639.1577 (MCs+, calcd. 639.1584).
EXAMPLE 44 Preparation of Compound 46: tra«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l ""-(5'""-methylisoxazol-3'""-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid tert-Butyl Ester
Figure imgf000141_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 and Example 19 for the conversion of intermediate LI to product Rl utilizing tert- butyl diethylphosphonoacetate where appropriate: TR (cm"1) 3337, 1693, 1531, 1286, 1155; !Η NMR (CDC13) δ 1.51.(s, 9H), 1.70-1.82 (m, 2H), 2.13-2.28 (m, 2H), 2.51 (s, 3H), 3.07-3.14 (m, IH), 3.25-3.47 (m, 3H), 4.23 (m, IH), 5.71 (d, 1H, 7= 15.9), 5.89-5.98 (m, IH), 6.33 (t, IH, 7= 7.2), 6.48 (s, IH), 6.66 (dd, IH, 7= 15.6, 5.7), 6.87 (m, 2H), 7.01-7.10 (m, 2H), 7.29 (s, IH), 7.58 (d, IH, 7= 5.7), 8.40(dd, IH, 7= 7.5, 1.8), 8.55 (d, IH, 7= 6.9), 9.48 (s, IH); Anal. (C32H35F2N5O7Η2O) C, H, N. EXAMPLE 45 Preparation of Compound 47: trfl«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l ""-(5'""-methylisoxazol-3'""-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2" ' ' "-oxopyrrolidin-3 " " "-yl)pent-2-enoic Acid 2,2-Dimethylpropyl Ester.
Figure imgf000142_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 and trα«5-(3'S,4S)4-tert-butoxycarbonylamino-5-(2'-oxopyrrolidin-3'-yl)pent-2- enoic acid 2,2-dimethylpropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) utilizing intermediate F2 where appropriate: IR (cnr >) 3335, 1693, 1531, 1248; *H NMR (CDC13) δ 0.99 (s, 9H), 1.72-1.83 (m, 2H), 2.11-2.29 (m, 2H), 2.52 (s, 3H), 3.09-3.16 (m, IH), 3.26-3.48 (m, 3H), 3.82-3.92 (m, 2H), 4.48 (m, IH), 5.86 (d, IH, 7= 15.6), 5.91-5.94 (m, IH), 6.33 (t, IH, 7= 8.1), 6.48 (s, IH), 6.76 (dd, IH, 7= 15.6, 6.0), 6.85 (m, 2H), 7.00-7.09 (m, 2H), 7.29 (s, IH), 7.55 (d, IH, 7= 6.9), 8.41 (d, IH, 7= 7.5), 8.60 (d, IH, 7= 6.9), 9.48 (s, IH); Anal. (C33H37F2N5O7*0.5H2O) C, H, N.
EXAMPLE 46 Preparation of Compound 48: tr-z«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l "".(5""'.methylisoxazol-3'""-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid Isopropyl Ester.
Figure imgf000143_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 and tr_z«5-(3'S,4S)4-tert-butoxycarbonylamino-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid isopropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) utilizing intermediate F2 where appropriate: JΗ NMR (CDC13) δ 1.27 (d, 3H, 7= 6.2), 1.28 (d, 3H, 7= 6.2), 1.45-1.56 (m, IH), 1.62-1.78 (m, IH), 2.08-2.29 (m, 3H), 2.49 (s, 3H), 3.06 (dd, IH, 7= 13.8, 7.8), 3.21-3.44 (m, 3H), 4.36-4.47 (m, IH), 4.99-5.12 (m, IH), 5.66 (dd, 1H, 7= 15.6, 1.5), 5.96-6.04 (m, IH), 6.32 (t, IH, 7=7.3), 6.45 (s, IH), 6.71 (dd, 1H, 7= 15.6, 5.6), 6.84-6.91 (m, IH), 6.98-7.17 (m, 3H), 7.66 (dd, 1H, 7= 7.3, 1.6), 8.39 (dd, IH, 7= 7.3, 1.6), 8.64 (d, IH, 7= 6.6), 9.45 (s, IH).
EXAMPLE 47 Preparation of Compound 49: trα«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2,-(3'"- {[l ""-(5'""-methylisoxazol-3'""-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid Cyclopentyl Ester.
Figure imgf000144_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates Y3 (Example 27) and F2 where appropriate: ^H NMR (CDC13) δ 1.45-1.98 (m, 10Η), 2.08-2.30 (m, 3Η), 2.49 (s, 3H), 3.07 (dd, IH, 7= 13.7, 8.1), 3.20-3.44 (m, 3H), 4.35-4.47 (m, IH), 5.18-5.23 (m, IH), 5.71 (d, 1H, 7= 15.7), 5.93-6.02 (m, IH), 6.31 (t, 1H, 7= 7.2), 6.45 (s, IH), 6.70 (dd, 1H, 7= 15.7, 5.8), 6.83-6.89 (m, IH), 6.97- 7.10 (m, 3H), 7.63 (dd, IH, 7= 7.2, 1.6), 8.38 (dd, IH, 7= 7.2, 1.6), 8.62 (d, IH, 7= 6.8), 9.45 (s, IH).
EXAMPLE 48 Preparation of Compound 50: trα«5-(2'S,3'""S,4S)-4-[2,-(3"-{[l '"-(5""-Methylisoxazol- 3 " "-yl)methanoyl]amino} -2" '-oxo-2" 'H-pyridin- 1 " '-yl)pent-4"-ynoylamino]-5-(2" " '- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Isopropyl Ester
Figure imgf000144_0002
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid isopropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: JR (cm-1) 3304, 1692, 1650; JH NMR (CDC13) δ 1.25 (d, 6H, 7= 6.2), 1.54-1.78 (m, 2H), 2.03 (t, 1H, 7= 2.5), 2.06-2.33 (m, 3H), 2.49 (s, 3H), 2.88-3.07 (m, 2H), 3.21-3.35 (m, 2H), 4.47-4.49 (m, IH), 5.00-5.08 (m, IH), 5.66-5.71 (m, IH), 6.00 (dd, 1H, 7= 15.7, 1.4), 6.33 (t, 1H, 7= 7.2), 6.46 (s, 1H), 6.73 (br s, IH), 6.83 (dd, 1H, 7= 15.7, 5.4), 7.48 (dd, 1H, 7= 7.2, 1.7), 8.41 (dd, 1H, 7 = 7.2, 1.7), 8.65 (d, IH, 7= 6.6), 9.53 (s, IH); Anal. (C27H3ιN5θ7*0.50H2θ) C, H, N.
EXAMPLE 49 Preparation of Compound 51 : trα«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l "".(5""'.methylisoxazol-3""'-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2 " " "-oxopyrrolidin-3 " " "-yl)pent-2-enoic Acid.
Figure imgf000145_0001
The title compound was prepared by deprotection of the tert-butyl ester present in compound 46 (Example 44) under acidic conditions using standard literature procedures: ER (cm-1) 3413, 1684, 1590, 1519; iH NMR (DMSO) δ 1.42-1.54 (m, IH), 1.59-1.68 (m, IH), 1.84-1.94 (m, IH), 2.10-2.16 (m, IH), 2.51 (s, 3H), 3.05-3.13 (m, IH), 3.32-3.45 (m, 3H), 4.52 (m, IH), 5.74 (d, IH, 7= 15.9), 5.80-5.82 (m, IH), 6.36 (t, IH, 7= 7.2), 6.71 (s, IH), 6.77 (dd, IH, 7= 15.6, 5.4), 7.04 (m, IH), 7.28-7.34 (m, 3H), 7.06-7.77 (m, 2H), 8.23 (d, IH, 7= 7.5), 8.61 (d, IH, 7= 7.2), 9.42 (s, IH); Anal. (C28H27F2N5O7Η2O) C, H, N. EXAMPLE 50 Preparation of Compound 52: tran5-(2'S,3'""S,4S)-4-[2,-(3"-{[l '"-(5""-Methylisoxazol- 3 " "-yl)methanoyl]amino} -2" Oxo-2" 'H-pyridin- 1 " '-yl)butanoylamino]-5-(2" " '- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Isopropyl Ester.
Figure imgf000146_0001
The title compound was prepared by a process analogous to that described in Example 25 for the conversion of intermediate V3 to product R3 utilizing intermediates F2 and trans- (3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid isopropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: JR (cm"1) 3333, 1681, 1531, 1276; JH NMR (CDC13) δ 0.98 (t, 3H, 7= 7.2), 1.29 (d, 6H, 7= 6.6), 1.46-1.73 (m, IH), 1.89-2.07 (m, IH), 2.13-2.37 (m, 4H), 2.52 (s, 3H), 3.30-3.43 (m, 3H), 4.48 (m, IH), 5.02-5.14 (m, IH), 5.69 (t, IH, 7= 6.9), 5.96 (d, IH, 7= 15.6), 6.39 (t, IH, 7= 7.5), 6.49 (s, IH), 6.86 (dd, IH, 7= 15.6, 6.0), 6.91 (s, br. IH), 7.56 (d, IH, 7= 7.2), 8.44 (d, IH, 7= 7.5), 8.53 (d, IH, 7= 6.3), 9.59 (s, IH); Anal. (C26H33N5O7O.5H2O) C, H, N.
EXAMPLE 51 Preparation of Compound 53: tra«5-(2,S,3'""S,4S)-4-[2,-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2'"-oxo-2'"H-pyridin-l '"-yl)pent-4"-ynoylamino]-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Cyclobutyl Ester.
Figure imgf000146_0002
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid cyclobutyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: ΪH NMR (CDC13) δ 1.52-1.87 (m, 4H), 2.01-2.41 (m, 8H), 2.50 (s, 3H), 2.92 (ddd, 1H, 7= 17.0, 8.5, 2.6), 3.02 (ddd, IH, 7= 17.0, 6.8, 2.6), 3.21-3.37 (m, 2H), 4.43-4.54 (m, IH), 4.96-5.08 (m, IH), 5.68-5.76 (m IH), 6.00 (dd, IH, 7= 15.7, 1.5), 6.33 (t, IH, 7= 7.3), 6.45-6.48 (m, IH), 6.83 (s, IH), 6.84 (dd, 1H, 7= 15.7, 5.4), 7.49 (dd, 1H, 7= 7.3, 1.7), 8.41 (dd, 1H, 7= 7.3, 1.7), 8.68 (d, IH, 7= 6.6), 9.52 (s, IH).
EXAMPLE 52 Preparation of Compound 54: tra«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l "".(5'"''.methylisoxazol-3""'-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2" ' ' "-oxopyrrolidin-3 " " "-yl)pent-2-enoic Acid Cyclohexyl Ester.
Figure imgf000147_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2- methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid cyclohexyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: Η NMR CDCy 1.20-1.61 (m, 7Η), 1.62-1.79 (m, 3H), 1.82-1.93 (m, 2H), 2.07-2.31 (m, 3H), 2.49 (d, 3H, 7= 0.7), 3.08 (dd, IH, 7= 13.8, 8.2), 3.20-3.45 (m, 3H), 4.36-4.48 (m, IH), 4.75-4.85 (m, IH), 5.71 (dd, 1H, 7= 15.6, 1.4), 5.90-5.98 (m, IH), 6.31 (t, 1H, 7= 7.3), 6.44- 6.46 (m, IH), 6.72 (dd, IH, 7= 15.6, 5.7), 6.83-6.90 (m, IH), 6.95-7.08 (m, 3H), 7.59 (dd, IH, 7= 7.3, 1.6), 8.38 (dd, IH, 7= 7.3, 1.6), 8.60 (d, IH, 7= 6.6), 9.45 (s, IH).
EXAMPLE 53 Preparation of Compound 55: tran5-(2'S,3"",S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2'"-oxo-2'"H-pyridin-l '"-yl)pent-4"-ynoylamino]-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Cyclohexyl Ester.
Figure imgf000148_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid cyclohexyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: IR (cm-1) 3305, 1685, 1650, 1597, 1534; lK NMR (CDC13) δ 1.24-1.98 (m, 12Η), 2.03 (t, IH, 7= 2.5), 2.05- 2.39 (m, 3H), 2.50 (s, 3H), 2.90-3.09 (m, 2H), 3.20-3.34 (m, 2H), 4.45-4.52 (m, IH), 4.75- 4.83 (m, IH), 5.59-5.64 (m, IH), 6.01 (dd, IH, 7= 15.6, 1.2), 6.33 (t, IH, 7= 7.2), 6.46 (s, IH), 6.54 (br s, IH), 6.82 (dd, IH, 7= 15.6, 5.3), 7.44 (dd, 1H, 7= 7.2, 1.7), 8.41 (dd, 1H, 7= 7.2, 1.7), 8.59 (d, IH, 7= 6.6), 9.53 (s, IH); Anal. (C30H35N5O7O.75H2O) C, H, N.
EXAMPLE 54 Preparation of Compound 56: tra«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l ""-(5'"".methylisoxazol-3'""-yl)methanoyl]amino}-2""-oxo-2""H-pyridm-l ""- yl)propanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid Cycloheptyl Ester.
Figure imgf000149_0001
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2- methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid cycloheptyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: lli NMR (CDC13) δ 1.40-2.35 (m, 17Η), 2.48 (s, 3H), 3.09 (m, IH), 3.20-3.50 (m, 3H), 4.35- 4.50 (m, IH), 4.90-5.05 (m, IH), 5.73 (d, IH, 7= 15.7), 5.86 (t, IH, 7= 7.5), 6.21 (t, IH, 7 = 7.2), 6.44 (s, IH), 6.69 (dd, IH, 7 = 15.6, 7.4), 6.75-6.80 (m, IH), 6.82-6.89 (m, IH), 6.95- 7.08 (m, 2H), 7.52 (d, IH, 7= 6.1), 8.37 (d, IH, 7= 6.1), 8.48 (d, IH, 7= 6.6), 9.45 (s, IH); Anal. (C35H39N5O7F3O.3OH2O) C, H, N.
EXAMPLE 55 Preparation of Compound 57: tra«5-(2'S,3""'S,4S)-4-[2,-(3"-{[l "'-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2'"-oxo-2'"H-pyridin-l '"-yl)pent-4"-ynoylamino]-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Cycloheptyl Ester.
Figure imgf000150_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid cycloheptyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: lK NMR (CDC13) δ 1.40-2.00 (m, 15Η), 2.00 (s, IH), 2.19-2.42 (m, 2H), 2.49 (s, 3H), 2.90-3.13 (m, 2H), 3.14-3.33 (m, 2H), 4.43-4.58 (m, IH), 4.90-5.05 (m, IH), 5.48 (t, 1H, 7= 6.6), 5.98 (d, IH, 7= 15.6), 6.21 (s, IH), 6.33 (t, IH, 7= 7.3), 6.46 (s, IH), 6.80 (dd, IH, 7= 15.6, 5.4), 7.36 (d, 1H, 7= 6.1), 8.35-8.50 (m, 2H), 9.52 (s, IH); Anal. (C31H37N5O7) C, H, N.
EXAMPLE 56 Preparation of Compound 58: tra«5-(2'S,3"""S,4S)-4-[3'-(3",4"-Difluorophenyl)-2'-(3'"- {[l "".(5'"".methylisoxazol-3"",-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)propanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid Cyclobutyl Ester.
Figure imgf000150_0002
The title compound was prepared from D-(3,4)-difluorophenylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2- methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid cyclobutyl ester (prepared from Kl in a 5 manner similar to that described for the preparation of X4 in Example 27) where appropriate: IR cm-1) 3322, 1682, 1531, 1284; lH NMR (CDCl3) δ 1.50-1.89 (m, 5H), 2.04-2.18 (m, 3H), 2.21-2.31 (m, 2H), 2.34-2.44 (m, 2H), 2.52 (s, 3H), 3.07-3.14 (m, IH), 3.26-3.46 (m, 3H), 4.43 (m, IH), 5.00-5.11 (m, IH), 5.68 (dd, 1H, 7= 15.9, 1.2), 5.91 (t, IH, 7= 7.8), 6.33 (t, IH, 7= 7.2), 6.47 (s, IH), 6.74 (dd, IH, 7= 15.6, 5.7), 6.83 (s, br. IH), 6.89-6.92 (m, IH), 7.05 (t, 10 IH, 7= 8.7), 7.59 (d, IH, 7= 7.2), 8.41 (dd, IH, 7= 7.5, 1.8), 8.61 (dd, IH, 7= 6.6, 1.8), 9.49 (s, IH); Anal. (C32H33F2N5θ7-H2θ) C, H, N.
EXAMPLE 57 Preparation of Compound 59: tra^-(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 15 3 " "-yl)methanoyl]amino} -2" Oxo-2" 'H-pyridin- 1 ' "-yl)butanoylamino]-5-(2" ' ' '- oxopyrrolidin-3"'"-yl)pent-2-enoic Acid Cyclobutyl Ester.
Figure imgf000151_0001
The title compound was prepared by a process analogous to that described in Example 20 25 for the conversion of intermediate V3 to product R3 utilizing intermediates F2 and trans- (3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid cyclobutyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: ER (cm"1) 3334, 1690, 1632; JΗ NMR (CDC13) δ 0.95 (t, 3H, 7= 7.5), 1.55-1.85 (m, 5H), 1.88-2.01 (m, IH), 2.04-2.44 (m, 7H), 2.52 (s, 3H), 3.31- 25 3.41 (m, 2H), 4.48 (m, IH), 5.01-5.11 (m, IH), 5.63-5.71 (m, IH), 5.96 (dd, 1H, 7= 15.0, 1.5), 6.36 (t, IH, 7= 7.5), 6.49 (s, IH), 6.83-6.90 (m, 2H), 7.54 (d, IH, 7= 7.2), 8.45 (dd, IH, 7= 7.5, 1.8), 8.53 (dd, IH, 7= 6.6, 1.8), 9.59 (s, IH); Anal. (C27H33N5O7O.75H2O) C, H, N. EXAMPLE 58 Preparation of Compound 60: trfl«5-(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2"Oxo-2'"H-pyridin-l '"-yl)butanoylamino]-5-(2'""- oxopyιτolidin-3'""-yl)pent-2-enoic Acid Cyclohexyl Ester.
Figure imgf000152_0001
The title compound was prepared by a process analogous to that described in Example 25 for the conversion of intermediate V3 to product R3 utilizing intermediates F2 and trans- (3 'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3 '-yl)pent-2-enoic acid cyclohexyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: JR (cm"1) 3334, 1687, 1649, 1595, 1532; lΗ NMR (CDC13) δ 0.92 (t, 1Η, 7= 7.4), 1.24-1.99 (m, 14Η), 2.06-2.30 (m, 3H), 2.49 (s, 3H), 3.28-3.39 (m, 2H), 4.45-4.47 (m, IH), 4.77-4.84 (m, IH), 5.65-5.70 (m, IH), 5.95 (dd, 1H, 7= 15.7, 1.1), 6.34 (t, 1H, 7= 7.3), 6.46 (s, IH), 6.83 (dd, 1H, 7= 15.7, 5.8), 6.93 (br s, IH), 7.54 (dd, IH, 7= 7.3, 1.7), 8.42 (dd, IH, 7= 7.3, 1.7), 8.49 (d, IH, 7= 6.8), 9.57 (s, IH); Anal. (C29H37N5O7O.5OH2O) C, H, N.
EXAMPLE 59 Preparation of Compound 61 : tra«5-(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2"Oxo-2,"H-pyridin-l '"-yl)butanoylamino]-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Cycloheptyl Ester.
Figure imgf000152_0002
The title compound was prepared by a process analogous to that described in Example 25 for the conversion of intermediate V3 to product R3 utilizing intermediates F2 and trans- (3 'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3 '-yl)pent-2-enoic acid cycloheptyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: Η NMR (CDC13) δ 0.85-2.35 (m, 22H), 2.49 (s, 3H), 3.23-3.38 (m, 2H), 4.46-4.57 (m, IH), 4.93-5.03 (m, IH), 5.44-5.52 (m, IH), 5.93 (dd, IH, 7=15.6, 1.2), 6.12 (s, br, IH), 6.32 (t, IH, 7=7.2), 6.46 (s, IH), 6.80 (dd, IH, 7=15.7, 5.7), 7.35 (dd, IH, 7=7.2, 1.7), 8.15 (d, IH, 7=6.8), 8.41 (dd, IH, 7=7.4, 1.6), 9.58 (s, IH); Anal. (C30H39N5O7O.8OH2O) C, H, N.
EXAMPLE 60 Preparation of Compound 62: tra«_--(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2"^xo-2"'H-pyridin-l ,"-yl)butanoylamino]-5-(2"",- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Benzyl Ester.
Figure imgf000153_0001
The title compound was prepared by a process analogous to that described in Example 25 for the conversion of intermediate V3 to product R3 utilizing intermediates F2 and trans- (3 'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3 '-yl)pent-2-enoic acid benzyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: ER (cm-1) 3335, 1685, 1649, 1595, 1533; !H NMR (CDCI3) δ 0.91 (t, 3H, 7= 7.2), 1.50-1.99 (m, 3H), 2.11-2.31 (m, 4H), 2.48 (s, 3H), 3.27-3.38 (m, 2H), 4.45-4.47 (m, IH), 5.17 (s, 2H), 5.65-5.70 (m, IH), 6.01 (dd, IH, 7= 15.6, 1.5), 6.33 (t, IH, 7= 7.1), 6.46 (s, IH), 6.90 (dd, IH, 7= 15.6, 5.8), 7.00 (br s, IH), 7.30-7.39 (m, 5H), 7.54 (dd, IH, 7= 7.1, 1.8), 8.42 (dd, IH, 7= 7.4, 1.8), 8.56 (d, IH, 7= 6.8), 9.56 (s, IH); Anal. (C30H33N5O7O.5OH2O) C, H, N. EXAMPLE 61 Preparation of Compound 63: trα«5-(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol- 3""-yl)methanoyl]amino}-2,"-oxo-2'"H-pyridin-l '"-yl)pent-4"-ynoylamino]-5-(2'""- oxopyrrolidin-3'""-yl)pent-2-enoic Acid Benzyl Ester
Figure imgf000154_0001
The title compound was prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid benzyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: IR (cm-1) 3298, 1685, 1650, 1596, 1534; JH NMR (CDC13) δ 1.55-1.74 (m, 2H), 1.97 (t, IH, 7= 2.7), 2.01- 2.19 (m, IH), 2.22-2.39 (m, 2H), 2.49 (s, 3H), 2.88-3.07 (m, 2H), 3.19-3.32 (m, 2H), 4.44- 4.52 (m, IH), 5.16 (s, 2H), 5.55-5.60 (m, IH), 6.07 (dd, 1H, 7= 15.7, 1.5), 6.32 (t, 1H, 7= 7.2), 6.45 (s, IH), 6.48 (br s, IH), 6.89 (dd, IH, 7= 15.7, 5.3), 7.29-7.41 (m, 5H), 7.44 (dd, IH, 7= 6.2, 1.7), 8.40 (dd, IH, 7= 7.5, 1.7), 8.61 (d, IH, 7= 6.6), 9.52 (s, IH); Anal. (C31H31N5O7O.5OH2O) C, H, N.
EXAMPLE 62 Preparation of Compounds 64 and 65: trα«_--(2'S,3'""S,4S)-4-[2'-(3"-{[l '"-(5""- Methylisoxazol-3 ' ' "-yl)methanoyl] amino} -2" '-oxo-2" 'H-pyridin-1 ' ' '-yl)pent-4"- ynoylamino]-5-(2'""-oxopyπOlidin-3'""-yl)pent-2-enoic Acid Pyridin-2-ylmethyl Ester and trαn5-(2'R,3,""S,4S)-4-[2'-(3"-{[l '"-(5""-Methylisoxazol-3""-yl)methanoyl]amino}-2'"- oxo-2 ' ' 'H-pyridin- 1 " ' -yl)pent-4' ' -ynoylamino] -5 -(2 " " '-oxopyιτolidin-3 ' " " -yl)pent-2-enoic Acid Pyridin-2-ylmethyl Ester.
Figure imgf000154_0002
The title compounds were prepared from D-propargylalanine by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'- oxopyrrolidin-3'-yl)pent-2-enoic acid pyridin-2-ylmethyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate. They were isolated as a 2:1 mixture of inseparable diasteroemers: ER (cm-1) 3297, 1690, 1596, 1273; *H NMR (CDC13) δ 1.58-1.69 (m), 2.02-2.08 (m), 2.09-2.48 (m), 2.53 (s), 2.81-3.12 (m), 3.24-3.49 (m), 5.26 (s), 5.31 (s), 5.52 (t, 7= 7.5), 5.82 (t, 7= 7.5), 5.92 (d, 7= 15.6), 6.15 (d, 7= 15.6), 6.36 (t, 7= 7.2), 6.46 (s), 6.53 (s), 6.87-6.70 (m), 7.24-7.43 (m), 7.68-7.76 (m), 8.43-8.50 (m), 8.61-8.69 (m), 9.56 (s), 9.61 (s); Anal. (C30H30N6O7O.5H2O) C, H, N.
EXAMPLE 63 Preparation of Compound 66: tra«5-(2'S,3'""S,4S)-4-[2'-(3"-{[l'"-(5""-Methylisoxazol- 3 " " -yl)methanoyl] amino} -2 " ' -oxo-2 ' ' 'H-pyridin- 1 ' " -yl)-3 ' ' -phenylpropanoylamino] -5 - (2 " " ' -oxopyrrolidin-3 " " ' -yl)pent-2-enoic Acid Isopropyl Ester.
Figure imgf000155_0001
The title compound was prepared from (2R)-2-hydroxy-3-phenylpropionic acid by a process analogous to that described in Example 22 for the conversion of intermediate SI to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2- methyl-5-(2 '-oxopyrrolidin-3 '-yl)pent-2-enoic acid isopropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: JΗ NMR (CDCI3) δ 1.27 (d, 3H, 7= 6.2), 1.28 (d, 3H, 7= 6.2), 1.44-1.54 (m, IH), 1.61-1.75 (m, IH), 2.09-2.33 (m, 3H), 2.47 (d, 3H, 7= 0.9), 3.13 (dd, IH, 7= 13.7, 8.2), 3.20-3.38 (m, 2H), 3.44 (dd, 1H, 7= 13.7, 7.7), 4.37-4.48 (m, IH), 4.99-5.12 (m, IH), 5.71 (dd, 1H, 7= 15.6, 1.4), 5.96-6.04 (m, IH), 6.29 (t, IH, 7= 7.3), 6.43-6.45 (m, IH), 6.69 (dd, IH, 7= 15.6, 5.7), 7.19-7.28 (m, 6H), 7.65-7.69 (m, IH), 8.36 (dd, IH, 7= 7.3, 1.6), 8.49 (d, IH, 7= 6.8), 9.45 (s, IH).
EXAMPLE 64 Preparation of Compound 67: tra«5-(2'S,3"""S,4S)-4-[3'-(4"-Fluorophenyl)-2'-(3'"-{[l ""- (5""'-methylisoxazol-3""'-yl)methanoyl]amino}-2""-oxo-2""H-pyridin-l ""- yl)ρropanoylamino]-5-(2"""-oxopyrrolidin-3"""-yl)pent-2-enoic Acid Isopropyl Ester.
Figure imgf000156_0001
The title compound was prepared from intermediate Tl by a process analogous to that described in Example 22 for the conversion of Tl to product R2 utilizing intermediates F2 and trans-(3'S,4S)-4-tert-butoxycarbonylamino-2-methyl-5-(2'-oxopyrrolidin-3'-yl)pent-2-enoic acid isopropyl ester (prepared from Kl in a manner similar to that described for the preparation of X4 in Example 27) where appropriate: IR (cm-1) 3335, 1693, 1649, 1596, 1533; Η NMR (CDC13) δ 1.27 (d, 3Η, 7= 2.6), 1.29 (d, 3H, 7= 2.6), 1.59-1.78 (m, 2H), 1.81-1.99 (m, IH), 2.20-2.29 (m, IH), 2.47 (s, 3H), 3.13-3.29 (m, 3H), 3.42-3.49 (m, IH), 4.50-4.52 (m, IH), 5.02-5.09 (m, IH), 5.67-5.75 (m, 2H), 6.24 (t, IH, 7= 7.3), 6.41 (s, IH), 6.67-6.74 (m, 2H), 6.92 (t, 2H, 7= 8.5), 7.10-7.14 (m, 2H), 7.41 (d, IH, 7= 7.3), 8.15 (br s , IH), 8.33 (d, 1H, 7= 7.7), 9.32 (s, IH).
EXAMPLE 65 Preparation of Compound 68: tra«5-(2'S,3""S,4S)-4-[2'-({[l "-(5'"-Methylisoxazol-3'" yl)methanoyl]amino}-oxotrifluoromethyl-2"H-pyridin-l"-yl)butanoylamino]-5-(2""- oxopyrrolidin-3""-yl)pent-2-enoic Acid Ethyl Ester.
Figure imgf000157_0001
The title compound was prepared from intermediate T3 and by a process analogous to that described in Example 22 for the conversion of D-(4-fluorophenylalanine) to product R2 utilizing nitrotrifluoromethylpyridin-2-ol and intermediate X2 where appropriate: Η NMR (CDC13) δ 0.93 (t, 3H, 7= 7.7), 1.28 (t, 3H, 7= 7.2), 1.60-2.33 (m, 7H), 2.45 (s, 3H), 3.20- 3.38 (m, 2H), 4.18 (q, 2H, 7= 7.0), 4.42-4.53 (m, IH), 5.68 (t, IH, 7= 7.9), 5.98 (d, IH, 7= 16.0), 6.40 (s, IH), 6.60 (s, IH), 6.84 (dd, IH, 7= 16.0, 5.8), 7.92 (s, IH), 8.50 (d, IH, 7= 2.3), 8.86 (d, IH, 7= 5.7), 9.39 (s, IH); Anal. (C26H3oN5θ7F3«1.70H2O) C, H, N.
Results of tests conducted using exemplary compounds of the invention are described below.
BIOCHEMICAL AND BIOLOGICAL EVALUATION
Inhibition of Rhinovirus 3C Protease: Stock solutions (50 mM, in DMSO) of various compounds were prepared; dilutions were in the same solvent. Recombinant rhinovirus 3C proteases (see Birch et al., "Purification of recombinant human rhinovims 14 3C protease expressed in Escherichia coli," Protein Expr. Pur. (1995), vol. 6(5), 609-618) from serotypes 14, 16, and 2 were prepared by the following standard chromatographic procedures: (1) ion exchange using Q Sepharose Fast Flow from Pharmacia; (2) affinity chromatography using Affi-Gel Blue from Biorad; and (3) sizing using Sephadex G-100 from Pharmacia. Each assay sample contained 2% DMSO, 50 mM tris pH 7.6, 1 mM EDTA, a test compound at the indicated concentration, approximately 1 μM substrate, and 50-100 nM protease. The k0bs/I values were obtained from reactions initiated by addition of enzyme rather than substrate. RVP activity was measured in the fluorescence resonance energy transfer assay. The substrate was (N-terminal) DABCYL-(Gly-Arg-Ala-Val- Phe-Gln-Gly-Pro-Val-Gly)-EDANS. In the uncleaved peptide, the EDANS fluorescence was quenched by the proximal DABCYL moiety. When the peptide was cleaved, the quenching was relieved, and activity was measured as an increase in fluorescence signal. Data were analyzed using standard non-linear fitting programs (Enzfit), and are shown in the table below. The tabulated data in the column designated kobs/[I] were measured from progress curves in enzyme start experiments.
Antirhino viral H 1 -HeLa Cell Culture Assay:
In this cell protection assay, the ability of compounds to protect cells against HRV infection was measured by the XTT dye reduction method, which is described in Weislow et al., 7 Natl. Cancer Inst. (1989), vol. 81, 577-586. Hl-HeLa cells were infected with HRV-14 at a multiplicity of infection (m.o.i.) of 0.13 (vims particles/cell) or mock-infected with medium only. Infected or mock-infected cells were resuspended at 8 x 10^ cells per mL, and incubated with appropriate concentrations of the compounds to be tested. Two days later, XTT/PMS was added to test plates and the amount of formazan produced was quantified spectrophotometrically at 450/650 nm. The EC50 value was calculated as the concentration of compound that increased the percentage of formazan production in compound-treated, virus- infected cells to 50% of that produced by compound- free, mock-infected cells. The 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased the percentage of formazan produced in compound-treated, mock-infected cells to 50% of that produced by compound-free, mock-infected cells. The therapeutic index (Tl) was calculated by dividing the CC50 value by the EC50 value. All strains of human rhinovirus (HRV) for use in this assay were purchased from
American Type Culture Collection (ATCC), except for HRV serotype-14 (produced from the infectious cDNA clone constructed by Dr. Robert Rueckert, Institute for Molecular Virology, University of Wisconsin, Madison, Wisconsin). HRV stocks were propagated and viral assays were performed in Hl-HeLa cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum, available from Life Technologies (Gaithersburg, MD). Test results for the HRV assay are shown in the table below. Anticoxsackieviral Cell Culture Assay:
Coxsackievims types A-21 (CAV-21) and B3 (CVB3) were purchased from American Type Culture Collection (ATCC, Rockville, MD). Vims stocks were propagated and antiviral assays were performed in Hl-HeLa cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum (Life Technologies, Gaithersburg, MD). The ability of the compounds of this invention to protect cells against either CAV-21 or CVB3 infection was measured by the XTT dye reduction method. This method is described in Weislow et al., 7 Natl. Cancer Ins t. (1989), vol. 81, 577-586. Hl-HeLa cells were infected with CAV-21 or CVB3 at a multiplicity of infection (m.o.i.) of 0.025 or 0.075, respectively, or mock-infected with medium only. Hl-HeLa cells were plated at 4 x 10^ cells per well in a 96- well plate and incubated with appropriate concentrations of the test compound. One day (CVB3) or two days (CAV-21) later, XTT /PMS was added to test plates and the amount of formazan produced was quantified spectrophotometrically at 450/650 nm. The EC50 was calculated as the concentration of compound that increased the formazan production in compound-treated, vims-infected cells to 50% of that produced by compound- free, uninfected cells. The 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased formazan production in compound-treated, uninfected cells to 50% of that produced in compound-free, uninfected cells. The therapeutic index (Tl) was calculated by dividing the CC50 by the EC50-
Anti-Echoviral and Anti-Enteroviral Cell Culture Assays
Echovirus type 11 (ECHO 11) was purchased from ATCC (Rockville, MD). Vims stocks were propagated and antiviral assays were performed in MRC-5 cells (ATCC). Cells were grown in minimal essential medium with 10% fetal bovine serum (Life Technologies, Gaithersburg, MD). The ability of the compounds of this invention to protect cells against ECHO 11 infection was measured by the XTT dye reduction method (Weislow et al., 7. Natl. Cancer Inst. (1989), vol. 81, 577-586). MRC-5 cells were infected with ECHO 11 at an m.o.i. of 0.003 or 0.004, respectively, or mock-infected with medium only. Infected or uninfected cells were added at 1 x 10^ cells per well and incubated with appropriate concentrations of compound. Four days later, XTT/PMS was added to test plates, and the amount of formazan produced was quantified spectrophotometrically at 450/650 nm. The EC50 was calculated as the concentration of compound that increased the formazan production in compound-treated, vims-infected cells to 50% of that produced by compound- free, uninfected cells. The 50% cytotoxic dose (CC50) was calculated as the concentration of compound that decreased formazan production in compound-treated, uninfected cells to 50% of that produced in compound- free, uninfected cells. The therapeutic index (Tl) was calculated by dividing the CC50 by the ECso- Activity of the compounds against enterovims type 70 (EV 70) may be measured by the same assay as described above in this section. Enterovims type 70 (EV 70) may be obtained from the American Type Culture Collection ATCC (Rockville, MD).
Results obtained for the compounds of the invention may be compared to results obtained in the same manner for control compounds WEN 51711, WEN 52084, and WIN 54954 (obtained from Sterling- Winthrop Pharmaceuticals), Pirodavir (obtained from Janssen Pharmaceuticals), and Pleconaril (prepared according to the method described in Diana et al., 7. Med. Chem. (1995), vol. 38, 1355). Antiviral data obtained for the test compounds are shown in the table below. The designation "ND" indicates that a value was not determined for that compound, and the designation "NA" means not applicable.
TABLE
Figure imgf000160_0001
Figure imgf000161_0001
Figure imgf000162_0001
Figure imgf000163_0001
Figure imgf000164_0001
Figure imgf000165_0001
Figure imgf000166_0001
Figure imgf000167_0001
Figure imgf000168_0001
Figure imgf000169_0001
Figure imgf000170_0001
Notes: αHRV = human rhinovirus of designated serotype.
^3C protease inhibition activity.
CCAV-21 = coxsackievirus A21.
^ECHO-11 = echovirus 11. eCVB3 = coxsackievirus B3.
JENT-70 = enterovirus 70.
SCVB2 = coxsackievirus B2.
ΛCVB5 = coxsackievirus B5.
ΕCHO-6 = echovirus 6.
. ECHO-9 = echovirus 9.
*Ki value. While the invention has been described in terms of preferred embodiments and specific examples, those skilled in the art will recognize through routine experimentation that various changes and modifications can be made without departing from the spirit and scope of the invention. Thus, the invention should be understood as not being limited by the foregoing detailed description, but as being defined by the appended claims and their equivalents.

Claims

WE CLA :
1. A compound having the formula:
Figure imgf000172_0001
wherein:
Ra is substituted or unsubstituted heterocycloalkyl or heterocycloalkylalkyl; Rb is a substituent having the formula:
Figure imgf000172_0002
wherein:
Rf and R8 are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5;
A. is CH or N;
A2 is C(Rh)(R'), N(RJ), S, S(O), S(O)2, or O; where each R\ R1 and RJ is independently H or lower alkyl; each A3 present is independently C(Rh)(R'), N(RJ), S, S(O), S(O)2, or O; where each Rh, R1 and RJ is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A4 is N(Rk), C(Rh)(R"), or O; and when p is 0, A4 is N(Rk)(R'), C(Rh)(R')(RJ), and O(R'), where each Rh, R1 and RJ is independently H or lower alkyl, each Rk is H, alkyl, aryl, or acyl, and each R1 is H, alkyl, or aryl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond when A2 is present and a hydrogen atom when A2 is absent;
Rcis H, halogen or a substituted or unsubstituted lower alkyl group;
Rd is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
Re is H or a substituted or unsubstituted alkylgroup; and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)Rn, -CO2Rn, -CN, -C(O)NRnR°, -C(O)NROR°, -C(S)Rn, -C(S)ORn, -C(S)NRnR°, -NO2, -SOR°, -SO2Rn, -SO2NRnR°, -SO2(NRn)(OR°), -SONR", -SO3Rn, -PO(OR")2, -PO(ORn)(OR0)_ -PO(NRnR°)(ORp)ι -PO(NRnR°)(NRpRcl), -C(O)NRnNR°Rp, or -C(S)NRnNR°Rp, wherein Rn, R°, Rp and Rq are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the Rn, R°, Rp and Rq, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Rd, together with the atoms to which they are bonded, form a cycloalkyl or heterocycloalkyl group, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group; or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
2. A compound according to claim 1, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)Rn, -CO2Rn, -CN, -C(O)NRnR°, -C(O)NROR°, -C(S)Rn, -C(S)NRnR°, -NO2, -SOR°, -SO2Rn, -SO2NRnR°, -SO2(NRn)(OR°), -SONR", -SO3R", -PO(ORn)2, -PO(ORn)(OR°), -PO(NRnR°)(ORp) -PO(NRnR°)(NRpRq), -C(O)NRnNR°Rp, or -C(S)NRnNR°Rp, wherein Rn, R°, Rp and Rq are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the Rn, R°, Rp and Rq, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
3. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 1 or 2, wherein Ra is substituted or unsubstituted heterocycloalkylalkyl.
4. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 1 or 2, wherein the alkyl moiety of the substituted or unsubstituted heterocycloalkylalkyl group is a substituted or unsubstituted saturated alkyl moiety.
5. A compound having the formula:
Figure imgf000174_0001
wherein:
R1 is H, a substituted or unsubstituted lower alkyl group or a suitable nitrogen protecting group;
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties in the above groups may be substituted or unsubstituted, or a suitable nitrogen protecting group;
R3 is H or a suitable substituent; or
R1 together with R2 form a suitable nitrogen protecting group; or
R2 together with R3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
R4 is H or a suitable substituent; the dotted line represents an optional bond;
R5 is H or a suitable substituent;
R6 is H or a substituted or unsubstituted alkyl group; or R5 together with R6 form a heterocycloalkyl ring, which may be optionally substituted;
R7 and R10are independently H, halogen or a substituted or unsubstituted lower alkyl group;
R8 is H or a substituted or unsubstituted lower alkyl group;
R1 ' is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
R9 is a substituent having the formula:
Figure imgf000175_0001
wherein:
R12 and R13 are independently H or lower alkyl; m is O or l; p is an integer of from 0 to 5;
A, is CH or N;
A2 is C(R,4)(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; each A3 present is independently C(RI4)(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A4 is N(R17), C(R14)(R15), or O; and when p is 0, A4 is N(R17)(R18), C(R14)(R15)(R16), and O(R18), where each R14, R15 and R16 is independently H or lower alkyl, each R17 is H, alkyl, aryl, or acyl, and each R18 is H, alkyl, or aryl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond when A2 is present and a hydrogen atom when A2 is absent; and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)OR19, -C(S)NRI9R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR'9, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO NR'^XOR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R2', or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and R", together with the atoms to which they are bonded, form a cycloalkyl or heterocycloalkyl group, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Z1 are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group; or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
6. A compound according to claim 5, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R'9, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR,9)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR,9)(OR20)ι -PO(NR19R20)(OR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
7. A compound according to claim 5, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR,9R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO(NR19R20)(OR21)> -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted.
A compound having the formula:
Figure imgf000177_0001
wherein:
R1 is H, a substituted or unsubstituted lower alkyl group or a suitable nitrogen protecting group;
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties in the above groups may be substituted or unsubstituted, or a suitable nitrogen protecting group;
R3 is H or a suitable substituent; or
R1 together with R2 form a suitable nitrogen protecting group; or
R2 together with R3 form a heterocycloalkyl ring or heteroaryl ring, which may be optionally substituted;
R4 is H or a suitable substituent;
R5 is H or a suitable substituent; R6 is H or a substituted or unsubstituted alkyl group; or
R5 together with R6 form a heterocycloalkyl ring, which may be optionally substituted;
R7 and R10 are independently H, halogen or a substituted or unsubstituted lower alkyl group;
R8is H or a substituted or unsubstituted lower alkyl group;
R1 ' is H, halogen, hydroxyl, a substituted or unsubstituted alkyl, alkoxy or alkylthio group;
R9 is a substituent having the formula:
Figure imgf000178_0001
wherein:
R12 and R13 are independently H or lower alkyl; m is 0 or 1 ; p is an integer of from 0 to 5;
A, is CH or N;
A2 is C(R14)(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; each A3 present is independently C(R1 )(R15), N(R16), S, S(O), S(O)2, or O; where each R14, R15 and R16 is independently H or lower alkyl; when p is 1, 2, 3, 4, or 5, A4 is N(R17), C(R14)(R15), or O; and when p is 0, A4 is N(R,7)(R18), C(R14)(R,5)(R16), and O(R18), where each R14, R15 and R16 is independently H or lower alkyl, each R'7 is H, alkyl, aryl, or acyl, and each R18 is H, alkyl, or aryl; provided that no more than two heteroatoms occur consecutively in the above-depicted ring formed by A,, (A2)m, (A3)p, A4, and C=O, where each dotted line in the ring depicts a single bond when A2 is present and a hydrogen atom when A2 is absent; and
Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R'9, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)OR19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR,9R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20)> -PO(NR19R20)(OR21), -PO(NR19R20)(NR2IR22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and R", together with the atoms to which they are bonded, form a cycloalkyl or heterocycloalkyl group, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group, where Z and Z1 are as defined above except for moieties that cannot form the cycloalkyl or heterocycloalkyl group; or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
9. A compound according to claim 8, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20)ι -PO(NR19R20)(OR21χ -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
10. A compound according to claim 8, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO(NR19R20)(OR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted.
11. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 1, 5 or 8, wherein said compound, prodrug, salt, metabolite or solvate is a single stereoisomer, a mixture of stereoisomers or a racemic mixture of stereoisomers.
12. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 11 , wherein said compound, prodrug, salt, metabolite or solvate is substantially stereoisomerically pure.
13. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein R1, R7, R8 and R10are each H.
14. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein Z and Z1 are independently H, substituted or unsubstituted alkyl, -CO2R19, or taken together with the atom to which they are attached, form a heterocycloalkyl group, which may be optionally substituted.
15. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein at least one of Z or Z1 is selected from -CO2H, -CO2-alkyl, -CO2-cycloalkyl, -CO2-alkylaryl, and -CO2-alkylheteroaryl, or taken together with the atom to which they are attached form a heterocycloalkyl group , which may be optionally contain at least one O, N, S or P and may be substituted by one or more of keto or thioketo.
16. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 1, 5 or 8, wherein Z and Z1 are not both H.
17. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 5 or 8, wherein when R3, R4 and R5 are independently H or a suitable substituent, said suitable substituent is independently selected from alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto, thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents are optionally substituted.
18. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 17, wherein R3, R4 and R5 are independently selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, amino, cyano, halogen, haloalkyl, hydroxyl, keto, alkoxy, aryloxy, cycloalkoxy, heterocycloalkoxy, alkyloxycarbonyl, aryloxycarbonyl, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroaryloxycarbonyl, heteroaryl carbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, alkylamino, arylamino, dialkylamino, alkylaminocarbonyl, alkylsulfonyl, and arylsulfonyl, wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl moieties of any of the above substituents may be optionally substituted by one or more of haloalkyl, nitro, amino, cyano, halogen, hydroxyl, haloalkoxy, mercapto, keto or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, aryloxy, alkylamino, dialkylamino, alkylthio or arylthio groups.
19. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 8, having the formula:
Figure imgf000182_0001
wherein:
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl and heteroaryl moieties of the above groups is substituted or unsubstituted;
R6 is H or a substituted or unsubstituted alkyl groupf and R1, R3, R4, R5, R7, R8, R9, R10, Z and Z1 are as defined in claim 8.
20. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 8, having the formula:
Figure imgf000183_0001
wherein:
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, an aryloxycarbonyl group or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl and heteroaryl moieties of the above groups is substituted or unsubstituted;
Rx represents H or one or more suitable substituents; and R1, R3, R4, R9, R10, Z and Z1 are as defined in claim 8.
21. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 8, having the formula:
Figure imgf000184_0001
wherein:
R7, R8 and R'°are independently H or a substituted or unsubstituted lower alkyl group;
R6 is H or a substituted or unsubstituted alkyl group;
Rγ represents H or one or more suitable substituents; and R1, R4, R5, R9, Z and Z1 are as defined in claim 8.
22. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 5, having the formula:
Figure imgf000184_0002
wherein:
R7, R8 and R'°are independently H or a substituted or unsubstituted lower alkyl group;
Rγ represents H or one or more suitable substituents;
R4 and R5 are independently H or a suitable substituent; and
R1, R6, R9, Z and Z1 , and the variables contained therein, are as defined in claim 5.
23. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 8, having the formula:
Figure imgf000185_0001
wherein:
R7, R8 and R10are independently H or a substituted or unsubstituted lower alkyl group;
Rγ represents H or one or more suitable substituents;
R4 and R5 are independently H or a suitable substituent; and
R1, R6, R9, Z and Z1 , and the variables contained therein, are as defined in claim 8.
24. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 19 to 23, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20)( -PO(NR19R20)(OR21)ι -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
25. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 19 to 23, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NRl9)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO(NR19R20)(OR21),
-PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted.
26. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein R2 is a substituted or unsubstituted alkyloxycarbonyl group, alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocycloalkylcarbonyl or heteroarylcarbonyl group.
27. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 26, wherein R2 is a heteroarylcarbonyl group, wherein the heteroaryl moiety is a five- membered heterocycle having from one to three heteroatoms selected from O, N, and S.
28. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 27, wherein R2 is heteroarylcarbonyl, where the heteroaryl moiety is a five-membered heterocycle having at least one nitrogen heteroatom and at least one oxygen heteroatom.
29. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 27, wherein R2 is heteroarylcarbonyl, where the heteroaryl moiety is unsubstituted or substituted 1,2-oxazolyl, 1,3-oxazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, or 1,2,5- oxadiazolyl.
30. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 27, wherein R2 is heteroarylcarbonyl, where the heteroaryl moiety is selected from unsubstituted and monomethyl-substituted 1,2,4-oxadiazolyl.
31. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 28, wherein R2 is heteroarylcarbonyl, where the heteroaryl moiety is selected from 3- isoxazolyl and 5-isoxazolyl, each unsubstituted or substituted with one or two substituents selected from methyl groups and halogens.
32. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein R6 is H or a substituted or unsubstituted lower alkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl group, a straight-chain saturated hydrocarbon moiety or an unsaturated hydrocarbon moiety.
33. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 32, wherein R6 is H, ethyl, 2-propyn-l-yl, methylcyclohexyl, or substituted or unsubstituted benzyl, wherein the phenyl moiety of the substituted benzyl has one or more substituents independently selected from lower alkyl, lower alkoxy and halogen.
34. The compound, prodrug, pharmaceutically acceptable salt, pharmaceuticaly active metabolite, or pharmaceutically acceptable salt according to any one of claims 1 or 5 to 10, wherein when p is 0, m is 0.
35. The compound, prodrug, pharmaceutically acceptable salt, pharmaceuticaly active metabolite, or pharmaceutically acceptable salt according to any one of claims 1 or 5 to 10, wherein m is 1 and p is 1 or 2.
36. The compound, prodrug, pharmaceutically acceptable salt, pharmaceuticaly active metabolite, or pharmaceutically acceptable salt according to any one of claims 1 or 5 to 10, wherein m is 1 and p is 1.
37. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 5 to 10, wherein R9 is selected from -CH2CH2C(O)NH2; -CH2CH2C(O)NH- alkyl; -CH2NHC(O)CH3; and
Figure imgf000188_0001
where n is 1 or 2.
38. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 37, wherein R9 is
Figure imgf000188_0002
39. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 1 or 5 to 10, wherein Z1 is H or lower alkyl and Z is substituted or unsubstituted ethoxycarbonyl, t-butoxycarbonyl, isopropoxycarbonyl, (2,2- dimethylpropyl)-oxycarbonyl, benzyloxycarbonyl, pyridylmethyleneoxycarbonyl, cyclobutyloxycarbonyl, cyclopentyloxycarbonyl, cyclohexyloxycarbonyl,, cycloheptyloxycarbonyl, or Z taken together with Z1 and the atom to which they are
attached form
Figure imgf000189_0001
40. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 20, wherein R„ is selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto, thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, and aryloxy.
41. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 21, 22 or 23, wherein R_, is selected from H, alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, nitro, amino, cyano, halogen, haloalkyl, hydroxyl, alkoxy, alkylenedioxy, aryloxy, cycloalkoxy, heterocycloalkoxy, heteroaryloxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy, arylcarbonyl, arylcarbonyloxy, aryloxycarbonyl, cycloalkylcarbonyl, cycloalkylcarbonyloxy, cycloalkyoxycarbonyl, heteroarylcarbonyl, heteroarylcarbonyloxy, heteroaryloxycarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyloxy, heterocycloalkyloxycarbonyl, carboxyl, carbamoyl, formyl, keto, thioketo, sulfo, alkylamino, cycloalkylamino, arylamino, heterocycloalkylamino, heteroarylamino, dialkylamino, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heterocycloalkylaminocarbonyl, heteroarylaminocarbonyl, dialkylaminocarbonyl, alkylaminothiocarbonyl, cycloalkylaminothiocarbonyl, arylaminothiocarbonyl, heterocycloalkylaminothiocarbonyl, heteroarylaminothiocarbonyl, dialkylaminothiocarbonyl, alkylsulfonyl, arylsulfonyl, alkylsulfenyl, arylsulfenyl, alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heterocycloalkylcarbonylamino, heteroarylcarbonylamino, alkylthiocarbonylamino, cycloalkylthiocarbonylamino, arylthiocarbonylamino, heterocycloalkylthiocarbonylamino, heteroarylthiocarbonylamino, alkylsulfonyloxy, arylsulfonyloxy, alkylsulfonylamino, arylsulfonylamino, mercapto, alkylthio, arylthio and heteroarylthio, wherein any of the alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl moieties present in the above substituents may be further substituted with one or more substituents selected from nitro, amino, cyano, halogen, haloalkyl, hydroxyl, keto and unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, and aryloxy.
42. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 19, having the formula:
Figure imgf000191_0001
wherein R1, R2, R3, R4, R5, R6, R9, Z and Z1 are as defined in claim 19.
43. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 20, having the formula:
Figure imgf000191_0002
wherein R1, R2, R3, R4, R% Rx, Z and Z1 are as defined in claim 20.
44. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 21, having the formula:
Figure imgf000192_0001
wherein R , R , R , R9, Ry, Z and Z are as defined in claim 21.
45. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 22, having the formula:
Figure imgf000192_0002
wherein R4, R5, R6, R9, Ry, Z and Z1 are as defined in claim 22.
46. A compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 23, having the formula:
Figure imgf000193_0001
wherein R , R , R , R , Ry, Z and Z are as defined in claim 23.
47. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 42 to 46, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR,9R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO(NR19R20)(OR 1χ -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted, or Z and Z1, together with the atom to which they are bonded, form a cycloalkyl or heterocycloalkyl group.
48. The compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to any one of claims 42 to 46, wherein Z and Z1 are independently H, F, a unsubstituted or substituted alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group, -C(O)R19, -CO2R19, -CN, -C(O)NR19R20, -C(O)NR19OR20, -C(S)R19, -C(S)NR19R20, -NO2, -SOR20, -SO2R19, -SO2NR19R20, -SO2(NR19)(OR20), -SONR19, -SO3R19, -PO(OR19)2, -PO(OR19)(OR20), -PO(NR19R20)(OR21), -PO(NR19R20)(NR21R22), -C(O)NR19NR20R21, or -C(S)NR19NR20R21, wherein R19, R20, R21 and R22 are independently H, a substituted or unsubstituted alkyl group, cycloalkyl group, aryl group, heterocycloalkyl group, acyl group or thioacyl group, or wherein any two of the R19, R20, R21 and R22, taken together with the atoms to which they are bonded, form a heterocycloalkyl group, which may be optionally substituted.
49. The compound according to any one of claims 1 or 5 to 10, having antipicomaviral activity corresponding to an EC50 less than or equal to 100 μM in an Hl-HeLa cell culture assay.
50. A pharmaceutical composition comprising: a therapeutically effective amount of at least one antipicomaviral agent selected from compounds, prodrugs, pharmaceutically acceptable salts, pharmaceutically active metabolites, and pharmaceutically acceptable solvates defined in any one of claims 1 or 5 to 10; and a pharmaceutically acceptable carrier, diluent, vehicle, or excipient.
51. A method of treating a mammalian disease condition mediated by picornaviral protease activity, comprising administering to a mammal in need thereof a therapeutically effective amount of at least one compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate defined in any one of claims 1 or 5 to 10.
52. The method according to claim 51 , wherein the at least one compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate is administered orally.
53. A method of inhibiting the activity of a picornaviral 3C protease, comprising contacting the picornaviral 3C protease with an effective amount of at least one compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate defined in any one of claims 1 or 5 to 10.
54. The method according to claim 53, wherein the at least one compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate is administered orally.
55. A method of inhibiting the activity of a picornaviral 3C protease, comprising contacting the picornaviral 3C protease with an effective amount of at least one compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate defined in any one of claims 1 or 5 to 10.
56. The method as defined in claim 55, wherein the picornaviral 3C protease is a rhinoviral protease.
57. A compound having the formula:
Figure imgf000195_0001
wherein:
R3 and R4 are independently H or a suitable substituent;
Rx represents H or one or more suitable substituents;
B is H, -OR24, where R24 is a suitable protecting group for a carboxyl moiety; and R1 is H, a substituted or unsubstituted lower alkyl group or a suitable nitrogen protecting group;
R2 is an alkylcarbonyl group, an arylcarbonyl group, a cycloalkylcarbonyl group, a heterocycloalkylcarbonyl group, a heteroarylcarbonyl group, or an alkyloxycarbonyl group, wherein each of the alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl moieties in the above groups may be substituted or unsubstituted, or a suitable nitrogen protecting group; or
R1 together with R2 form a suitable nitrogen protecting group; or a pharmaceutically acceptable salt thereof.
58. The compound according to claim 57, having the formula:
Figure imgf000196_0001
or a pharmaceutically acceptable salt thereof.
59. A compound having the formula:
Figure imgf000196_0002
or a prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate thereof.
60. A stereoisomer of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59, having the formula:
Figure imgf000197_0001
61. A stereoisomer of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59, having the formula:
Figure imgf000197_0002
62. A stereoisomer of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59, having the formula:
Figure imgf000197_0003
63. A stereoisomer of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59, having the formula:
Figure imgf000198_0001
64. A pharmaceutical composition comprising: a therapeutically effective amount of an antipicomaviral agent selected from the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59 or 63; and a pharmaceutically acceptable carrier, diluent, vehicle, or excipient.
65. A method of inhibiting the activity of a picornaviral 3C protease, comprising contacting the picornaviral 3C protease with an effective amount of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate according to claim 59 or 63.
66. The method as defined in claim 65, wherein the picornaviral 3C protease is a rhinoviral protease.
67. A method of treating a mammalian disease condition mediated by picornaviral protease activity, comprising: administering to a mammal in need thereof a therapeutically effective amount of the compound, prodrug, pharmaceutically acceptable salt, pharmaceutically active metabolite, or pharmaceutically acceptable solvate as defined in claim 59 or 63.
8. A compound selected from:
Figure imgf000199_0001
Figure imgf000200_0002
Figure imgf000200_0001
Figure imgf000200_0003
Figure imgf000201_0001
Figure imgf000202_0001
Figure imgf000202_0002
Figure imgf000203_0001
Figure imgf000204_0001
Figure imgf000205_0001
and
Figure imgf000205_0002
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US6995142B2 (en) 1998-04-30 2006-02-07 Agouron Pharmaceuticals, Inc. Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
US6610730B2 (en) 2000-04-14 2003-08-26 Agouron Pharmaceuticals, Inc. Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
US6872745B2 (en) 2000-04-14 2005-03-29 Agouron Pharmaceuticals, Inc. Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
WO2001096297A3 (en) * 2000-06-14 2003-07-10 Agouron Pharma Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
US6632825B2 (en) 2000-06-14 2003-10-14 Agouron Pharmaceuticals, Inc. Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
WO2001096297A2 (en) * 2000-06-14 2001-12-20 Agouron Pharmaceuticals, Inc. Antipicornaviral compounds and compositions, their pharmaceutical uses, and materials for their synthesis
EP1357111A1 (en) * 2000-12-28 2003-10-29 Shionogi & Co., Ltd. Pyridone derivative having affinity for cannabinoid 2-type receptor
EP1357111A4 (en) * 2000-12-28 2005-10-05 Shionogi & Co Pyridone derivative having affinity for cannabinoid 2-type receptor
WO2005110988A1 (en) * 2004-05-07 2005-11-24 Janssen Pharmaceutica, N.V. Pyridone compounds as inhibitors of bacterial type iii protein secretion systems
US7087622B2 (en) 2004-05-07 2006-08-08 Janssen Pharmaceutica N.V. Pyridone compounds as inhibitors of bacterial type III protein secreation systems
US11124497B1 (en) 2020-04-17 2021-09-21 Pardes Biosciences, Inc. Inhibitors of cysteine proteases and methods of use thereof
US11312704B2 (en) 2020-04-17 2022-04-26 Pardes Biosciences, Inc. Inhibitors of cysteine proteases and methods of use thereof
US11472793B2 (en) 2020-04-17 2022-10-18 Pardes Biosciences, Inc. Inhibitors of cysteine proteases and methods of use thereof
US11174231B1 (en) 2020-06-09 2021-11-16 Pardes Biosciences, Inc. Inhibitors of cysteine proteases and methods of use thereof
US11524940B1 (en) 2020-06-09 2022-12-13 Pardes Biosciences, Inc. Inhibitors of cysteine proteases and methods of use thereof
US11351149B2 (en) 2020-09-03 2022-06-07 Pfizer Inc. Nitrile-containing antiviral compounds
US11452711B2 (en) 2020-09-03 2022-09-27 Pfizer Inc. Nitrile-containing antiviral compounds
US11541034B2 (en) 2020-09-03 2023-01-03 Pfizer Inc. Nitrile-containing antiviral compounds

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