EP1196436A2 - Peptidboronsäure inhibitore des hepatitis c virus proteases - Google Patents

Peptidboronsäure inhibitore des hepatitis c virus proteases

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Publication number
EP1196436A2
EP1196436A2 EP00943413A EP00943413A EP1196436A2 EP 1196436 A2 EP1196436 A2 EP 1196436A2 EP 00943413 A EP00943413 A EP 00943413A EP 00943413 A EP00943413 A EP 00943413A EP 1196436 A2 EP1196436 A2 EP 1196436A2
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EP
European Patent Office
Prior art keywords
substituted
amino
alkyl
acid
phenyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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EP00943413A
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English (en)
French (fr)
Inventor
Charles A. Kettner
Sharada Jagannathan
Timothy Patrick Forsyth
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Bristol Myers Squibb Pharma Co
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Bristol Myers Squibb Pharma Co
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Publication of EP1196436A2 publication Critical patent/EP1196436A2/de
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/69—Boron compounds
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
    • C07F5/02—Boron compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
    • C07F5/02—Boron compounds
    • C07F5/025—Boronic and borinic acid compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0827—Tripeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1027—Tetrapeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
    • C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
    • C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates generally to novel ⁇ -aminoboronic acids and corresponding peptide analogs represented by structural Formula (I) :
  • R 1 , R 2 , R 3 , Y 1 , Y 2 , and A are described herein.
  • the invention is also concerned with pharmaceutical formulations comprising these novel compounds as active ingredients and the use of the novel compounds and their formulations in the treatment of hepatitis C viral infections .
  • the compounds of the invention are inhibitors of hepatitis C viral protease.
  • Hepatitis C virus is the major cause of transfusion and community-acquired non-A, non-B hepatitis worldwide. Approximately 2% of the world's population are infected with the virus. In the Unites States, hepatitis C represents approximately 20% of cases of acute hepatitis. Unfortunately, self-limited hepatitis is not the most common course of acute HCV infection. In the majority of patients, symptoms of acute hepatitis resolve, but ALT levels (a liver enzyme diagnostic for liver damage) often remain elevated and HCV RNA persists. Indeed, a propensity to chronicity is the most distinguishing characteristic of hepatitis C, occurring in at least 85% of patients with acute HCV infection.
  • HCV is a positive-stranded RNA virus. Based on comparison of deduced amino acid sequence and the extensive similarity in the 5' untranslated region, HCV has been classified as a separate genus in the Flaviviridae family, which also includes flaviviruses (such as yellow fever virus (YF) ) , and animal pestiviruses (like bovine viral diarrhea virus (BVDV) and swine fever virus (CSFV) ) .
  • flaviviruses such as yellow fever virus (YF)
  • BVDV bovine viral diarrhea virus
  • CSFV swine fever virus
  • Flaviviridae family All members of the Flaviviridae family have enveloped virions that contain a positive stranded RNA genome encoding all known virus-specific proteins via translation of a single, long uninterrupted, open reading frame.
  • Considerable heterogeneity is found within the nucleotide and encoded amino acid sequence throughout the HCV genome. At least 6 major genotypes have been characterized, and more than 50 subtypes have been described. The major genotypes of HCV differ in their distribution worldwide; the clinical significance of the genetic heterogeneity of HCV remains elusive despite numerous studies of the possible effect of genotypes on pathogenesis and therapy.
  • RNA genome is about 9.6 Kb in length, and encodes a single polypeptide of about 3000 amino acids.
  • the 5' and 3 ' ends are of critical importance for the replicative life cycle.
  • the 5' end contains an Internal Ribosome Entry Site or IRES, which directs cellular ribosomes to the correct AUG for initiation of translation.
  • IRES Internal Ribosome Entry Site
  • the precursor protein is cotranslationally and posttranslationally processed into at least 10 viral structural and nonstructural proteins by the action of a host signal peptidase and by two distinct viral proteinase activities.
  • the translated product contains the following proteins: core-El-E2-p7-NS2-NS3-NS4A-NS4B-NS5A- NS5B.
  • the N-terminal portion of NS3 functions as a proteolytic enzyme that is responsible for the cleavage of sites liberating the nonstructural proteins NS4A, NS4B, etc. Agents that block this protease are expected to be new antiviral agents.
  • This protease has been classified as a "serine protease" based on the catalytic residues in the active site, Eckart et al . Biochem . Biophys . Res . Commun . 192, 399-406 (1993) . It is known in the art that peptide analogs corresponding to sequences of peptide substrate and containing an electrophilic group provide good inhibitors of serine proteases.
  • Enzyme susceptibility to inhibition differs significantly by choice of the electrophilic group.
  • inhibitors of HCV protease corresponding to the sequence of the NS5A/B cleavage site have been prepared with an electrophilic boronic acid group incorporated into the sequence.
  • Boronic acids have a distinct advantage over other peptide inhibitors of HCV protease.
  • the concept of using boronic acids as serine protease inhibitors was introduced in the early 70's Antonov et al . FEBS Lett 7, 23 (1970); Koelhler and Lienhard Biochemistry 10, 2477-2483 (1971) .
  • boronic acid inhibitors specially designed as inhibitors of trypsin-like serine proteases such as thrombin, plasma kallikrein and plasmin, wherein the ⁇ -aminoboronic acid side chain is an aklyl group substituted by -NH 2 , -NH-C (NH) -NH 2 or -S-C (NH) -NH 2 .
  • boronic acid dipeptide inhibitors which are inhibitors of trypsin-like serine proteases wherein the ⁇ -aminoboronic acid side chain is a substituted alkyl or substituted alkylphenyl group.
  • boronic acid peptide inhibitors which are inhibitors of thrombosis and anticoagulants wherein the ⁇ -aminoboronic acid side chain is an aklyl, substituted alkyl, substituted phenylaklyl, or substituted cycloalkylalkyl group.
  • boronic acid inhibitors which are thrombin inhibitors wherein the ⁇ - aminoboronic acid side chain is a monosubstituted alkyl, monosubstituted alkenyl, or a monosubstituted phenylalkyl group .
  • hexapeptide boronic acid inhibitors which are HCV protease inhibitors wherein the ⁇ - aminoboronic acid side chain is an alkyl or an alkenyl group .
  • ⁇ -aminoboronic acid compounds as inhibitors of serine proteases, it is still desirable to develop more efficacious inhibitors which are enzyme specific to HCV protease.
  • the present invention discloses ⁇ -aminoboronic acid compounds as efficacious inhibitors of the NS3 protease of the hepatitis C virus.
  • One object of the presenbt invention is to provide compounds, or pharmaceutically acceptable salt forms or prodrugs thereof, which are useful as inhibitors of hepatitis C virus protease, more specifically, the NS3 protease.
  • compositions comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of Formula (I), or pharmaceutically acceptable salt form or prodrug thereof.
  • FIG. 1 illustrates plasmid construction maps for expression in cultured cells of HCV NS3 protease (pCMV NS3 PR) and substrate (pCMVNS5A/5B) .
  • FIG. 2 illustrates detection by western blotting of NS3 protease-inhibitory compound in a cell-based assay.
  • Human 293 cells were electroporated with expression plasmids described in FIG. 1. The cells were placed in tissue culture medium containing the indicated concentration of Example 10, an inhibitor of NS3 protease. The cells were allowed to synthesize proteins for 24 additional hours. Then the contents of the cells were analyzed using polyacrylamide gel electrophoresis and western blotting. The full length substrate (NS5A/5B) and cleavage product NS5A were detected with specific antiserum. Activity of the tested compound was measured by the accumulation of uncleaved NS5A/5B.
  • the present invention provides a method of treating Hepatitis C virus in a mammal comprising administering to said mammal in need of such treatment an effective amount of a compound of Formula (I) :
  • Y 1 and Y 2 are independently selected from: a) -OH, b)-F, c)-NR 18 R 19 , d) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: e) a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0, f) a cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or O, g) a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally,
  • R 1 is selected from:
  • R 1A is H, C1-C4 alkyl, phenyl, or -CH 2 phenyl, wherein phenyl of R 1A is substituted with 0-3 substituents selected from -CH 3 , -CF 3 , -N0 2 , -CN, -OH, -SH, -OCH3 , -OCF3 , -Cl, -Br, -I, and F;
  • R 1B is C1-C4 alkyl, phenyl, or -CH 2 phenyl, wherein phenyl of
  • R 1B is substituted with 0-3 substituents selected from - CH 3 , -CF 3 , -N0 2 , -CN, -OH, -SH, -0CH 3 , -OCF 3 , -Cl, -Br, - I, and F;
  • A is a bond, A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A 4 -
  • a 5 , A 1 -A 2 -A 3 -A 4 -A 5 -A 6 , A 1 -A 2 -A 3 -A -A 5 -A 6 -A 7 , A 1 -A 2 -A -A 4 - A 5 -A 6 -A 7 -A 8 , A 1 -A 2 -A 3 -A -A 5 -A 6 -A 7 -A 8 -A 9 ; or A 1 -A 2 -A 3 -A 4 - A 5 -A 6 -A 7 -A 8 -A 9 -A 10 ; A 1 , A 2 , A 3 , A 4 , A 5 , A 6 , A 7 , A 8 , A 9 , and A 10 are independently selected from an amino acid residue, wherein said amino acid residue comprises a natural amino acid, a modified amino acid or an unnatural amino acid;
  • R 2 is H, C 1 -C 4 alkyl, aryl, aryl (C 1 -C 4 alkyl)-, or C3-C 6 cycloalkyl,
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R 4B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R 4B ; or a 5-6 membered heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4C is selected at each occurrence from the group:
  • two independent R 11 groups may optionally be taken together to form -(CH 2 ) p -;
  • n 0, 1, 2, 3, or 4;
  • p 1, 2, 3, or 4;
  • Z is selected from:
  • R 12 is H
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , CF 3 , -Cl, -Br, -I, F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -N ( CH 2 CH 3 ) 2 , and C3.-C4 alkyl ;
  • R 18 and R 19 at each occurrence are independently selected from H, C 1 -C 4 alkyl, aryl (C 1 -C 4 alkyl)-, and C 3 -C 7 cycloalkyl;
  • R 20 is C 1 -C 4 alkyl
  • R 21 is, at each occurrence, independently H or C 1 -C 4 alkyl
  • R 1a is, at each occurrence, independently H, C 1 -C 4 alkyl, aryl, or C 1 -C 4 haloalkyl;
  • A is not -Asp-Glu- (2-methyl-Phe) - (3 -methyl-Val) -Leu-, -Asp-Glu- (2-methyl-Phe) - (3 -methyl-Val) - (cyclopentyl-Ala) -, -Asp-Glu- (2-methyl-Phe) - (cyclohexyl-Ala) -Leu-, -Asp-Glu- (2-methyl-Phe) - (phenyl-Gly) -Leu-, -Asp-Glu- (2-methyl-Phe) - (cyclohexyl-Ala) -Leu-, -Asp-Glu- (2-methyl-Phe) - (3-methyl-Val) - (Pro) -, -Asp-Glu- (2-methyl-Phe) -Phe-Leu-, or -Asp-Glu- ( 4-chlor
  • a 1 , A 2 , A 3 , A 4 , A 5 , A 6 , A 7 , A 8 , A 9 , and A 10 are independently selected from an amino acid residue wherein said amino acid residue comprises a natural amino acid selected from the group: Ala, Arg, Ash, Asp, Aze, Cha, Cys, Dpa, Gin, Glu, Gly, His, Hyp, lie, Irg, Leu, Lys, Met, Orn, Phe, Phe (4-fluoro) , Pro, Sar, Ser, Thr, Trp, Tyr, and Val; a modified amino acid selected from the group: Asp(OMe), Glu(OMe), Hyp(OMe), Asp(O Bu), Glu(O fc Bu), Hyp(O t Bu), Thr(O t Bu), Asp(OBzl), Glu(OBzl), Hyp(OBzl), Thr(OBzl); and
  • 2-amino-3-carboxypentanedioic acid 2-amino-4-ethylpentanedioic acid, -amino-4-propylpentanedioic acid, -amino-4-isoamylpentanedioic acid, -amino-4-phenylpentanedioic acid, 2-amino-hexanedioic acid, 2-amino-heptanedioic acid,
  • 2-amino-4-aminooxybutanoic acid 2-amino-3- (N-nitrosohydroxyamino) propanoic acid, 2-amino-3-ureidopropanoic acid, 2-amino-4-ureidobutanoic acid, 2-amino-3-phosphopropanoic acid, 2-amino-3 - thiophosphopropanoic acid, 2-amino-4-methanephosphonylbutanoic acid, 2-amino-3- (trimethylsilyl) propanoic acid, 2-amino-3 - (dimethyl (trimethylsilylmethylsilyl) propanoic acid, 2-amino-2-phenylacetic acid, 2-amino-2- (3-chlorophenyl) acetic acid, 2-amino-2- (4-chlorophenyl) acetic acid, 2-amino-2- (3 -fluorophenyl) acetic acid, 2-amino-2- (3-methylphenyl) ace
  • 2-amino-3 - (2-pyrryl) propanoic acid 2-amino-3- (1-pyrryl) propanoic acid, 2-amino-4- (1-pyrryl) butanoic acid, 2-amino-5- (1-pyrryl) pentanoic acid, 2-amino-3 - (5-imidazolyl) -3-methylpropanoic acid, 2-amino-3- (5-imidazolyl) -3-ethylpropanoic acid, 2-amino-3-hexyl-3- (5-imidazolyl) propanoic acid, 2-amino-3 -hydroxy-3- (5-imidazolyl) propanoic acid,
  • A is A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A 4 -A 5 , A 1 -A 2 -A 3 -A 4 -A 5 -A 6 , or A 1 -A 2 -A 3 -A 4 -A 5 -A 6 -A 7 ; and A 1 , A 2 , A 3 , A 4 , A 5 , A 6 , and A 7 are independently selected from Ala, Arg, Asn, Asp, Aze, Cha, Cys, Dpa, Gin, Glu, Gly, His, Hyp, lie, Irg, Leu, Lys, Met, Orn, Phe, Phe (4-fluoro) , Pro, Sar, Ser, Thr, Trp, Tyr, Val, Asp(OMe), Glu (OMe), Hyp(OMe), Asp(O t
  • Y 1 and Y 2 are independently selected from: a) -OH, b)-F, c)-NR 18 R 19 , d) C ⁇ -C 8 alkoxy, or when taken together, Y 1 and Y 2 form: e) a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0, f) a cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0, g) a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally,
  • R 1 is selected from:
  • R 1A is H, methyl , ethyl, propyl, phenyl, or -CH 2 phenyl, wherein phenyl of R 1A is substituted with 0-3 substituents selected from -CH 3 , -CF 3 , -N0 2 , -CN, -OH, - SH, -OCH 3 , -OCF 3 , -Cl, -Br, -I, and F;
  • A is A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A 4 -A 5 , or A 1 -A 2 -A 3 -A -A 5 -A 6 ;
  • a 1 , A 2 , A 3 , A 4 , A 5 , and A 6 are independently selected from
  • R 2 is H, methyl, ethyl, propyl, or butyl
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4B is selected at each occurrence from the group:
  • heterocyclic ring system consisting of carbon atoms and 1-3 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-4 R 4C ;
  • R C is selected at each occurrence from the group: H, F, Cl, Br, I, -N0 2 , -CN, -NCS, -CF 3 , -OCF 3 ,
  • two independent R 11 groups may optionally be taken together to form -(CH 2 ) p -;
  • n 0, 1, 2, 3, or 4;
  • p 1, 2, 3, or 4;
  • Z is selected from:
  • R 12 is H
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , CF 3 , -Cl, -Br, -I, F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -N(CH 2 CH 3 ) 2 , and C 1 -C 4 alkyl;
  • R 18 and R 19 at each occurrence are independently selected from H, C 1 -C 4 alkyl, aryl (C 1 -C 4 alkyl)-, and C 3 -C 7 cycloalkyl;
  • R 20 is methyl, ethyl, propyl or butyl
  • R 21 is, at each occurrence, independently H or methyl, ethyl, propyl or butyl;
  • R 21a is, at each occurrence, independently H, methyl, ethyl, propyl or butyl, phenyl, or C 1 -C 4 haloalkyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, c) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: d) a cyclic boron ester where said chain or ring contains from 2 to 16 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0,
  • A is A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , or A 1 -A 2 -A 3 -A 4 -A 5 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala,
  • R 2 is H, methyl, or ethyl
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • R 4A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R B ; naphthyl substituted with 0-3 R B ; benzyl substituted with 0-3 R 4B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • heterocyclic ring system consisting of carbon atoms and 1-3 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-4 R 4C ;
  • R C is selected at each occurrence from the group:
  • X is a bond, C 1 -C 4 alkyl substituted with 0-3 R 11 ,
  • R 11 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 ,
  • two independent R 11 groups may optionally be taken together to form -(CH ) p -;
  • n 0, 1, 2, or 3;
  • p 1, 2, 3, or 4;
  • Z is selected from: -H, -R 12 , -halo, -NHS0 2 R 12 , -S0 2 NHR 12 , -S0 2 R 12 ,
  • R 12 is H, C 1 -C 4 alkyl substituted with 0-3 R 13 ,
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , CF 3 , -Cl, -Br, -I, F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -N(CH 2 CH 3 ) 2 , and C 1 -C 4 alkyl;
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and
  • R 20 is methyl, ethyl, propyl or butyl
  • R 21 is, at each occurrence, independently H or methyl, ethyl, propyl or butyl;
  • R 21a is, at each occurrence, independently H, methyl, ethyl, propyl or butyl, phenyl, or C 1 -C 4 haloalkyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, b) C 1 -C 6 alkoxy, or when taken together, Y 1 and Y 2 form: c) a cyclic boron ester where said chain or ring contains from 2 to 12 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or O,
  • R 1 is selected from -CH 2 CH 2 CF 3 , -CH 2 CHF 2 , and -CH 2 CH 2 F,
  • A is A 1 -A 2 , A 1 -A 2 -A 3 , or A 1 -A 2 -A 3 -A 4 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala, Arg, Asn, Asp, Aze, Cha, Cys, Dpa, Gin, Glu, Gly, His, Hyp, lie, Irg, Leu, Lys, Met, Orn, Phe, Phe (4-fluoro) , Pro, Sar, Ser, Thr, Trp, Tyr, Val, Asp(OMe), Glu (OMe), Hyp (OMe), Asp(O t Bu), Glu(O Bu), Hyp(O t Bu), Thr(O fc Bu), Asp(OBzl), Glu(OBzl), Hyp(OBzl), and Thr (OBzl) ;
  • R 2 is H
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-2 R 4B ;
  • R A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R 4B ; nap thyl substituted with 0-3 R B ; benzyl substituted with 0-3 R B ; or a 5-6 membered heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4B is selected at each occurrence from the group:
  • R 4C is selected at each occurrence from the group:
  • C3-C10 carbocycle substituted with 0-2 R 11 wherein the carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantanyl, norbornanyl, norbornenyl, and fluorenyl, phenyl substituted with 0-3 R 11 , naphthyl substituted with 0-3 R 11 , C5-C1 0 heterocycle substituted with 0-2 R 11 , wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, benzthiophenyl, pyrrolidinyl, pyrrolyl, carbazolyl, pyridinyl, thiophenyl, triazolyl, thiadiazolyl, benzodioxanyl, benzodioxolyl, quinazolinyl, quinoxalinyl, and quinolinyl;
  • R 11 at each occurrence is independently selected from H,
  • n 0, 1, or 2;
  • p 2, 3, or 4;
  • Z is selected from:
  • R 12 is H
  • R 13 at each occurrence is independently selected from H,
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl ,- and
  • R 20 is methyl, ethyl, propyl or butyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, b) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: c) a cyclic boron ester where said chain or ring contains from 2 to 12 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0,
  • R 1 is -CH 2 CHF 2 ;
  • A is A 1 -A 2 , A 1 -A 2 -A 3 , or A 1 -A 2 -A 3 -A 4 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala,
  • R 2 is H
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-2 R B ;
  • R A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R C is selected at each occurrence from the group:
  • C 3 -C 10 carbocycle substituted with 0-2 R 11 wherein the carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantanyl, norbornanyl, norbornenyl, and fluorenyl, phenyl substituted with 0-3 R 11 , naphthyl substituted with 0-3 R 11 , 5 -C 10 heterocycle substituted with 0-2 R 11 , wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, benzthiophenyl, pyrrolidinyl, pyrrolyl, carbazolyl, pyridinyl, thiophenyl, triazolyl, thiadiazolyl, benzodioxanyl, benzodioxolyl, quinazolinyl, quinoxalinyl, and quinolinyl;
  • R 11 at each occurrence is independently selected from H,
  • two independent R 11 groups may optionally be taken together to form -(CH 2 ) p -;
  • n 0, 1, or 2;
  • p 2, 3, or 4 ;
  • Z is selected from:
  • R 12 is H
  • heterocycle substituted with 0-3 R 13 ; wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, pyrrolidinyl, pyrrolyl, pyridinyl, thiophenyl, triazolyl, and thiadiazolyl ; R 13 at each occurrence is independently selected from H,
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and
  • R 20 is methyl, ethyl, propyl or butyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b)-F, c)-NR 18 R 19 , d) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: e) a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0, f) a cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0, g) a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or 0;
  • R 1 is selected from:
  • R 1A is H, methyl , ethyl, propyl, phenyl, or -CH 2 phenyl, wherein phenyl of R 1A is substituted with 0-3 substituents selected from -CH 3 , -CF 3 , -N0 2 , -CN, -OH, - SH, -OCH 3 , -OCF 3 , -Cl, -Br, -I, and F;
  • A is A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A 4 -A 5 , or A 1 -A 2 -A 3 -A 4 -A 5 -A 6 ;
  • a 1 , A 2 , A 3 , A 4 , A 5 , and A 6 are independently selected from
  • R 2 is H, methyl, ethyl, propyl, or butyl
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A ,
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: O, S, and N, and said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4A is C1-C4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R 4B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4B is selected at each occurrence from the group:
  • R C is selected at each occurrence from the group:
  • X is a bond, C1-C4 alkyl substituted with 0-3 R 11 ,
  • R 11 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 ,
  • two independent R 11 groups may optionally be taken together to form -(CH2) p -;
  • n 0, 1, 2, 3, or 4;
  • p 1, 2, 3, or 4;
  • R 12 is H, C 1 -C 4 alkyl substituted with 0-3 R 13 ,
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , CF 3 , -Cl, -Br, -I, F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -N(CH 2 CH 3 ) 2 , and C 1 -C 4 alkyl;
  • R 18 and R 19 at each occurrence are independently selected from H, C 1 -C 4 alkyl, aryl(C ⁇ -C 4 alkyl)-, and C 3 -C 7 cycloalkyl;
  • R 20 is methyl, ethyl, propyl or butyl
  • R 21 is, at each occurrence, independently H or methyl, ethyl, propyl or butyl;
  • R 21a is, at each occurrence, independently H, methyl, ethyl, propyl or butyl, phenyl, or C 1 -C 4 haloalkyl;
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, c) C]_-C6 alkoxy, or when taken together, Y 1 and Y 2 form: d) a cyclic boron ester where said chain or ring contains from 2 to 16 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or O,
  • R 1 is selected from:
  • A is A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , or A 1 -A 2 -A 3 -A 4 -A 5 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala,
  • R 2 is H, methyl, or ethyl
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A , C 3 -C 6 cycloalkyl substituted with 0-3 R 4B and aryl substituted with 0-3 R 4B and
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-3 R B ;
  • R 4A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R 4B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4B is selected at each occurrence from the group:
  • R 4C is selected at each occurrence from the group: H, F, Cl, Br, I, -N0 2 , -CN, -NCS, -CF 3 , -OCF 3 ,
  • n 0, 1, 2, or 3;
  • p 1, 2, 3, or 4;
  • Z is selected from:
  • R 12 is H
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , CF 3 , -Cl, -Br, -I, F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), -N(CH 2 CH 3 ) 2 , and C 1 -C 4 alkyl;
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and
  • R 20 is methyl, ethyl, propyl or butyl
  • R 21 is, at each occurrence, independently H or methyl, ethyl, propyl or butyl; and 2 la ⁇ S/ t each occurrence, independently H, methyl, ethyl, propyl or butyl, phenyl, or C 1 -C 4 haloalkyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, b) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: c) a cyclic boron ester where said chain or ring contains from 2 to 12 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N, S, or O,
  • R 1 is selected from -CH 2 CH 2 CF 3 , -CH 2 CHF 2 , and -CH 2 CH 2 F,
  • A is A 1 -A 2 , A 1 -A 2 -A 3 , or A 1 -A 2 -A 3 -A 4 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala,
  • R 2 is H
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A , C 3 -C 6 cycloalkyl substituted with 0-3 R 4B and aryl substituted with 0-2 R 4B and 5-10 membered heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-2 R 4B ;
  • R A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R B ; or a 5-6 membered heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R B is selected at each occurrence from the group:
  • heterocyclic ring system consisting of carbon atoms and 1-3 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-3 R 4C ;
  • R C is selected at each occurrence from the group:
  • C 1 -C 4 alkyl substituted with 0-3 R 11 C 2 -C 4 alkenyl substituted with 0-2 R 11 , C 3 -C 10 carbocycle substituted with 0-2 R 11 , wherein the carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantanyl, norbornanyl, norbornenyl, and fluorenyl, phenyl substituted with 0-3 R 11 , naphthyl substituted with 0-3 R 11 , C 5 -C 10 heterocycle substituted with 0-2 R 11 , wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, benzthiophenyl, pyrrolidinyl, pyrrolyl, carbazolyl, pyridinyl, thiophenyl, triazolyl, thiadiazolyl, benzodioxanyl,
  • R 11 at each occurrence is independently selected from H,
  • two independent R 11 groups may optionally be taken together to form -(CH ) p -;
  • n 0, 1, or 2;
  • p 2, 3, or 4;
  • Z is selected from: -H, -R 12 , -halo, -NHS0 2 R 12 , -S0 2 NHR 12 , -S0 2 R 12 ,
  • R 12 is H, C 1 -C 4 alkyl substituted with 0-3 R 13 ,
  • R 13 at each occurrence is independently selected from H,
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and
  • R 20 is methyl, ethyl, propyl or butyl.
  • Y 1 and Y 2 are independently selected from: a) -OH, b) -F, b) Ci-C ⁇ alkoxy, or when taken together, Y 1 and Y 2 form: c) a cyclic boron ester where said chain or ring contains from 2 to 12 carbon atoms, and, optionally, 1, 2, or 3 heteroatoms which can be N,
  • R 1 is -CH 2 CHF 2 ;
  • A is A 1 -A 2 , A 1 -A 2 -A 3 , or A 1 -A 2 -A 3 -A 4 ;
  • a 1 , A 2 , A 3 , and A 4 are independently selected from Ala,
  • R 2 is H
  • R 4 is C 1 -C 4 alkyl substituted with 0-1 R 4A , C 3 -C 6 cycloalkyl substituted with 0-3 R 4B and aryl substituted with 0-2 R 4B and
  • heterocyclic ring system consisting of carbon atoms and 1-4 heteroatoms selected from the group: O, S, and N, and said heterocyclic ring system is substituted with 0-2 R B ;
  • R A is C 1 -C 4 alkyl, halo, -OR 20 , -SR 20 , -NR 18 R 19 , phenyl substituted with 0-3 R 4B ; naphthyl substituted with 0-3 R 4B ; benzyl substituted with 0-3 R B ; or a
  • heterocyclic ring system containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur; said heterocyclic ring system is substituted with 0-3 R 4B ;
  • R 4B is selected at each occurrence from the group:
  • heterocyclic ring system consisting of carbon atoms and 1-3 heteroatoms selected from the group: 0, S, and N, and said heterocyclic ring system is substituted with 0-3 R C ;
  • R 4C is selected at each occurrence from the group:
  • C 1 -C 4 alkyl substituted with 0-3 R 11 C 2 -C 4 alkenyl substituted with 0-2 R 11 , C 3 -C 10 carbocycle substituted with 0-2 R 11 , wherein the carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantanyl, norbornanyl, norbornenyl, and fluorenyl, phenyl substituted with 0-3 R 11 , naphthyl substituted with 0-3 R 11 ,
  • heterocycle substituted with 0-2 R 11 , wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, benzthiophenyl, pyrrolidinyl, pyrrolyl, carbazolyl, pyridinyl, thiophenyl, triazolyl, thiadiazolyl, benzodioxanyl, benzodioxolyl, quinazolinyl, quinoxalinyl, and quinolinyl;
  • R 11 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -NH 2 , -SO 3 H, -S0 2 CH 3 , -C0 2 H, -CF 3 ,
  • n 0, 1, or 2;
  • p 2, 3, or 4;
  • Z is selected from: -H, -R 12 , -halo, -NHS0 2 R 12 , -S0 2 NHR 12 , -S0 2 R 12 ,
  • R 12 is H, C 1 -C 4 alkyl substituted with 0-3 R 13 ,
  • heterocycle substituted with 0-3 R 13 ; wherein the heterocycle is selected from furanyl, oxazolyl, isoxazolyl, pyrrolidinyl, pyrrolyl, pyridinyl, thiophenyl, triazolyl, and thiadiazolyl ;
  • R 13 at each occurrence is independently selected from H, -CH 3 , -CH 2 CH 3 , -N0 2 , -S0 2 OH, -S0 2 CH 3 , -CF 3 , -Cl, -Br, - I, -F, -NH 2 , -NH(CH 3 ), -N(CH 3 ) 2 , -NH(CH 2 CH 3 ), - N(CH CH 3 ) 2 , methyl, ethyl, propyl, and butyl;
  • R 18 and R 19 are independently selected from H, methyl, ethyl, propyl, butyl, benzyl, phenylethyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; and
  • R 20 is methyl, ethyl, propyl or butyl
  • compounds of Formula (I) selected from Examples 7-17, 19-22, 27-41, 43-53, 54a-54f, 59a-59bj , and 60a-60bc.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula (I) and a pharmaceutically acceptable carrier.
  • the present invention provides a method for the treatment of HCV comprising administering to a host in need of such treatment a therapeutically effective amount of a compound of Formula (I) , or a pharmaceutically acceptable salt thereof.
  • This invention also provides compositions comprising one or more of the foregoing compounds and methods of using such compositions in the treatment of hepatitis C virus, such as inhibition of hepatitis C virus protease, in mammals or as reagents used as inhibitors of hepatitis C virus protease in the processing of blood to plasma for diagnostic and other commercial purposes.
  • the present invention provides novel compounds of Formula (I) or pharmaceutically acceptable salt forms thereof for use in therapy.
  • the present invention provides the use of novel compounds of Formula (I) or pharmaceutically acceptable salt forms thereof for the manufacture of a medicament for the treatment of HCV.
  • substituent R 1 is -CH 2 CHF 2 .
  • substituent R 1 is -CH CH 2 CF 3 .
  • substituent R 1 is allyl
  • Ala is L-alanine
  • Alg is L-2-amino-4-pentenoic acid
  • Ape is L-2-aminopentanoic acid; Arg is L-arginine;
  • Asn is L-asparagine
  • Asp is L-aspartic acid
  • Aze is azedine-2-carboxlic acid
  • Cha is L-2-amino-3 -cyclohexylpropionic acid
  • Cpa is L-2-amino-3-cyclopropylpropionic acid
  • Cpg is L-2-amino-2-cyclopropylacetic acid
  • Cys is L-cysteine
  • Dfb is L-4, 4 ' -difluoro-1-amino-butyric acid
  • Dpa is L-2-amino-3 , 3 -diphenylpropionic acid
  • Gin is L-glutamine
  • Glu is L-glutamic acid; Gly is glycine;
  • HomoLys is L-homolysine
  • Hyp is L-4-hydroxyproline; lie is L-isoleucine; Irg is isothiouronium analog of L-Arg;
  • Leu is L-leucine
  • Lys is L-lysine
  • Met is L-methionine
  • Orn is L-ornithine; Phe is L-phenylalanine;
  • Phe (4-fluoro) is para-fluorophenylalanine
  • Pro is L-proline
  • Ser is L-serine; Thr is L-threonine;
  • Tpa is L-2-amino-5, 5, 5-trifluoropentanoic acid
  • Trp is L-tryptophan
  • Tyr is L-tyrosine
  • Val is L-valine.
  • boroVal-OH where "-OH” indicates the boronic acid is in the form of the free acid.
  • the pinanediol boronic acid ester and the pinacol boronic acid ester are abbreviated “- 10 H 16 " and “-C 6 H ⁇ ", respectively.
  • Examples of other useful diols for esterification with the boronic acids are 1, 2-ethanediol, 1, 3-propanediol, 1 , 2-propanediol, 2,3- butanediol, 1 , 2-diisopropylethanediol, 5 , 6-decanediol, and 1, 2-dicyclohexylethanediol .
  • DIBAL diisobutylalummum hydride.
  • RaNi means
  • the abbreviation "LAH” means lithium aluminum hydride .
  • the abbreviation “1,1' -CDI” means 1,1'- carbonyldiimidazole.
  • the abbreviation “Bn” means benzyl.
  • the abbreviation “BOC” means t-butyl carbamate .
  • the abbreviation “CBZ” means benzyl carbamate.
  • BSA benzene sulfonic acid
  • THF tetrahydrofuran
  • Boc- t-butoxycarbonyl-
  • Ac- acetyl
  • pNA p-nitro-aniline
  • DMAP 4-N,N-dimethylaminopyridine
  • Tris Tris (hydroxymethyl) aminomethane
  • MS mass spectrometry
  • FAB/MS fast atom bombardment mass spectrometry.
  • LRMS (NH 3 - CI)and HRMS (NH 3 -CI)are low and high resolution mass spectrometry, respectively, using NH 3 as an ion source.
  • the reactions of the synthetic methods claimed herein are carried out in suitable solvents which may be readily selected by one of skill in the art of organic synthesis, said suitable solvents generally being any solvent which is substantially nonreactive with the starting materials (reactants) , the intermediates, or products at the temperatures at which the reactions are carried out .
  • a given reaction may be carried out in one solvent or a mixture of more than one solvent.
  • suitable solvents for a particular reaction step may be selected.
  • stable compound or stable structure it is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • substituted means that any one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound.
  • 2 hydrogens on the atom are replaced.
  • any variable e.g., R 11 or R 13
  • its definition at each occurrence is independent of its definition at every other occurrence.
  • a group is shown to be substituted with 0-2 R 11 , then said group may optionally be substituted with up to two R 11 groups and R 11 at each occurrence is selected independently from the definition of R 11 .
  • R 11 at each occurrence is selected independently from the definition of R 11 .
  • combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
  • stable compound it is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture.
  • the substituent A is intended to be absent (i.e. a bond), a single amino acid residue, or a peptide of 2 to 10 amino acid residues.
  • the scope of A can be described as a bond, A 1 , A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A 4 -A 5 , A 1 -A 2 -A 3 -A 4 -A 5 -A 6 , A 1 -A 2 -A 3 -A 4 - A 5 -A 6 -A 7 , A 1 -A 2 -A 3 -A 4 -A 5 -A 6 -A 7 -A 8 , A 1 -A 2 -A 3 -A -A 5 -A 6 -A 7 -A 8 , A 1 -A 2 -A 3 -A -A 5 -A 6 -A 7
  • A can be described as (A") n wherein n is O, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • each amino acid residue of A is independently selected apart from each other amino acid residue.
  • a 1 , A 2 , A 3 , A 4 , A 5 , A 6 , A 7 , A 8 , A 9 and A 10 are independently selected from the defined list of possible amino acid residues, including modified or unnatural amino acid residues, disclosed herein.
  • each A" when n is 2 or greater, is independently selected from the defined list of possible amino acid residues, including modified or unnatural amino acid residues, disclosed herein. Therefore, A is intended to be absent, a single amino acid residue, a homopeptide, or a heteropeptide .
  • a preferred scope of substituent A is A 1 , A 1 -A 2 , A 1 -A 2 - A 3 , A 1 -A 2 -A 3 -A 4 , A 1 -A 2 -A 3 -A -A 5 , and A 1 -A 2 -A 3 -A 4 -A 5 -A 6 .
  • a more preferred scope of substituent A is A 1 , A ⁇ A , A 1 -A 2 - A 3 , A 1 -A 2 -A -A 4 , and A 1 -A 2 -A 3 -A 4 -A 5 .
  • substituent A is A 1 -A 2 , A 1 -A 2 -A 3 , A 1 -A 2 -A 3 -A 4 , and A 1 -A 2 -A 3 -A -A 5 .
  • a most preferred scope of substituent A is A 1 -A 2 , A 1 -A 2 -A 3 , and A 1 -A 2 -A 3 -A 4 .
  • substituent A 1 is Pro, 3- hydroxyproline, 4-hydroxyproline, Hyp (OMe), Hyp(O t Bu), and Hyp(OBzl) .
  • amino acid residue refers to natural, modified or unnatural amino acids of either D- or L-configuration and means an organic compound containing both a basic amino group and an acidic carboxyl group.
  • Natural amino acids residues are Ala, Arg, Asn, Asp, Aze, Cys, Gin, Glu, Gly, His, Hyp, lie, Irg Leu, Lys, Met, Orn, Phe, Phe(4-fluoro) , Pro, Sar, Ser, Thr, Trp, Tyr, and Val. Roberts and Vellaccio, The Peptides, Vol 5; 341-449 (1983), Academic Press, New York, discloses numerous suitable unnatural amino acids and is incorporated herein by reference for that purpose. Additionally, said reference describes, but does not extensively list, acylic N-alkyl and acyclic ⁇ , ⁇ -disubstituted amino acids.
  • N-alkyl, aryl, and alkylaryl analogs of both in chain and N-terminal amino acid residues.
  • alkyl, aryl, and alkylaryl maybe substituted for the alpha hydrogen.
  • Illustrated below are examples of N-alkyl and alpha alkyl amino acid residues, respectively.
  • Modified amino acids which can be used to practice the invention include, but are not limited to, D-amino acids, hydroxylysine, 4-hydroxyproline, 3-hydroxyproline, an N-CBZ-protected amino acid, 2 , 4-diaminobutyric acid, homoarginine, norleucine, N-methylaminobutyric acid, naphthylalanine, phenylglycine, ⁇ -phenylproline, tert-leucine, 4-aminocyclohexylalanine, N-methyl-norleucine, 3 , 4-dehydroproline,
  • Unnatural amino acids that fall within the scope of this invention are by way of example and without limitation: 2-aminobutanoic acid, 2-aminopentanoic acid, 2- aminohexanoic acid, 2-aminoheptanoic acid, 2-aminooctanoic acid, 2-aminononanoic acid, 2-aminodecanoic acid, 2- aminoundecanoic acid, 2-amino-3 , 3-dimethylbutanoic acid, 2- amino-4, 4-dimethylpentanoic acid, 2-amino-3 -methylhexanoic acid, 2-amino-3 -methylheptanoic acid, 2-amino-3- methyloctanoic acid, 2-amino-3 -methylnonanoic acid, 2- amino-4-methylhexanoic acid, 2-amino-3 -ethylpentanoic acid, 2-amino-3 , 4-dimethylpentanoic acid, 2-amino-3,5- dimethylhe
  • amino acids residue also refers to various amino acids where sidechain functional groups are modified with appropriate protecting groups known to those skilled in the art.
  • the Peptides Vol 3, 3-88 (1981) discloses numerous suitable protecting groups and is incorporated herein by reference for that purpose.
  • amino acids where sidechain functional groups are modified with appropriate protecting groups include, but are not limited to, Asp(OMe), Glu (OMe), Hyp(OMe), Asp(O t Bu), Glu(O Bu), Hyp(O t Bu), Thr(O t Bu), Asp(OBzl), Glu(OBzl), Hyp(OBzl), and Thr(OBzl); wherein OMe is methoxy, O t Bu is tert-butoxy, and OBzl is benzyloxy.
  • alkyl or “alkylene” is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms; for example, "Ci-C ⁇ alkyl” denotes alkyl having 1 to 6 carbon atoms.
  • alkyl examples include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, n-hexyl, 2- methylbutyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl .
  • alkenyl or “alkenylene” is intended to include hydrocarbon chains of either a straight or branched configuration having the specified number of carbon atoms and one or more unsaturated carbon-carbon bonds which may occur in any stable point along the chain.
  • alkenyl examples include, but are not limited to, ethenyl, 1- propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3, pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5- hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, and the like.
  • Alkynyl or “alkynylene” is intended to include hydrocarbon chains of either a straight or branched configuration and one or more carbon-carbon triple bonds which may occur in any stable point along the chain, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like.
  • Cycloalkyl is intended to include saturated ring groups, having the specified number of carbon atoms.
  • C 3 -C 6 cycloalkyl denotes such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
  • Alkoxy or "alkyloxy” represents an alkyl group as defined above with the indicated number of carbon atoms attached through an oxygen bridge.
  • alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, n-pentoxy, and s-pentoxy.
  • alkylthio or “thioalkoxy” represents an alkyl group as defined above with the indicated number of carbon atoms attached through a sulpher bridge .
  • Halo or "halogen” as used herein refers to fluoro, chloro, bromo, and iodo; and "counterion” is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate, and the like.
  • haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2 , 2 , 2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl .
  • haloalkyl also include “fluoroalkyl” which is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with 1 or more fluorine atoms.
  • fluoroalkyl is intended to mean any stable 3- to 7-membered monocyclic or bicyclic or 7- to 13 -membered bicyclic or tricyclic, any of which may be saturated, partially unsaturated, or aromatic.
  • carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4. O]bicyclodecane (decalin) , [2.2.2]bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl, or tetrahydronaphthyl (tetralin) .
  • heterocycle or “heterocyclic ring” is intended to mean a stable 5- to 7- membered monocyclic or bicyclic or 7- to 14-membered bicyclic heterocyclic ring which is saturated partially unsaturated or unsaturated (aromatic) , and which consists of carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O and S and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring.
  • the nitrogen and sulfur heteroatoms may optionally be oxidized.
  • the heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom which results in a stable structure.
  • heterocyclic rings described herein may be substituted on carbon or on a nitrogen atom if the resulting compound is stable. If specifically noted, a nitrogen in the heterocycle may optionally be quaternized. It is preferred that when the total number of S and O atoms in the heterocycle exceeds 1, then these heteroatoms are not adjacent to one another. It is preferred that the total number of S and O atoms in the heterocycle is not more than 1.
  • heterocycles include, but are not limited to, lH-indazole, 2-pyrrolidonyl, 2H, 6H-1, 5, 2-dithiazinyl, 2H-pyrrolyl, 3H-indolyl, 4-piperidonyl, 4aH-carbazole, 4H-quinolizinyl, 6H-1, 2 , 5-thiadiazinyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl , benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalonyl, carbazolyl, 4aH-carbazolyl, b-carbolinyl, chromanyl, chromenyl, cinnolin
  • Preferred heterocycles include, but are not limited to, pyridinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrazinyl, piperazinyl, imidazolyl, indolyl, benzimidazolyl, lH-indazolyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, benzoxazolyl, oxindolyl, benzoxazolinyl, benzthiazolyl, benzisothiazolyl, isatinoyl, isoxazolopyridinyl, isothiazolopyridinyl, thiazolopyridinyl, oxazolopyridinyl, imidazolopyridinyl, and pyrazolopyridinyl .
  • Preferred 5 to 6 membered heterocycles include, but are not limited to, pyridinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrazinyl, piperazinyl, imidazolyl, and oxazolidinyl. Also included are fused ring and spiro compounds containing, for example, the above heterocycles .
  • aryl or aromatic residue, is intended to mean an aromatic moiety containing the specified number of carbon atoms, such as phenyl, pyridinyl and naphthyl.
  • NH 2 -blocking group refers to various acyl, thioacyl, alkyl, sulfonyl, phosphoryl, and phosphinyl groups comprised of 1 to 20 carbon atoms. Substitutes on these groups maybe either alkyl, aryl, alkylaryl which may contain the heteroatoms, 0, S, and N as a substituent or in-chain component.
  • a number of NH 2 -blocking groups are recognized by those skilled in the art of organic synthesis. By definition, an NH 2 -blocking group may be removable or may remain permanently bound to the NH 2 .
  • Suitable groups include formyl, acetyl, benzoyl, trifluoroacetyl, and methoxysuccinyl; aromatic urethane protecting groups, such as, benzyloxycarbonyl; and aliphatic urethane protecting groups, such as t- butoxycarbonyl or adamantyloxycarbonyl .
  • aromatic urethane protecting groups such as, benzyloxycarbonyl
  • aliphatic urethane protecting groups such as t- butoxycarbonyl or adamantyloxycarbonyl .
  • Amine protecting groups may include, but are not limited to the following: 2,7-di-t- butyl- [9- (10, 10-dioxo-lO, 10, 10, 10-tetrahydrothio- xanthyl) ]methylo xycarbonyl; 2- trimethylsilylethyloxycarbonyl; 2-phenylethyloxycarbonyl; 1, l-dimethyl-2 , 2-dibromoethyloxycarbonyl; 1-methyl-l- (4- biphenylyl) ethyloxycarbonyl; benzyloxycarbonyl; p- nitrobenzyloxycarbonyl; 2-(p- toluenesulfonyl ) ethyloxycarbonyl
  • phrases "pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salts refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof.
  • pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • the pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
  • inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like
  • organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic,
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.
  • Lists of suitable salts are found in Remington ' s Pharmaceutical Sciences, 17th ed. , Mack Publishing Company, Easton, PA, 1985, p. 1418, the disclosure of which is hereby incorporated by reference.
  • Prodrugs are intended to include any covalently bonded carriers which release the active parent drug according to formula (I) in vivo when such prodrug is administered to a mammalian subject.
  • Prodrugs of a compound of formula (I) are prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound.
  • Prodrugs include compounds of formula (I) wherein a hydroxy, amino, or sulfhydryl group is bonded to any group that, when the prodrug or compound of formula (I) is administered to a mammalian subject, cleaves to form a free hydroxyl, free amino, or free sulfhydryl group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol and amine functional groups in the compounds of Formula (I), and the like.
  • Solid compound and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent .
  • Scheme 1 shows the synthesis of ⁇ -aminoboronic acids containing sidechains where R is ethyl, allyl, vinyl, and cyclopropyl.
  • a Grignard reagent is added to a trialkyl boronate to give a substituted dialkyl boronate .
  • Li + CHC1 2 ⁇ is prepared in situ by the addition of LDA to a -78°C solution of the alkyl boronic acid ester in methylene chloride.
  • CHCl ⁇ Li + is prepared by reacting n-butyl lithium with methylene chloride at -100°C followed by the addition of the alkyl boronic acid 2.
  • ZnCl 2 is added to more hindered alkyl boronic acid.
  • 3 is treated with the lithium salt of hexamethyldisilazane to give the jis-silane protected amine 4.
  • Compound 4 is treated with either anhydrous HCl or trifluoroacetic acid to give the amine 5 as a hydrochloride salt or trifluoroacetate salt.
  • Scheme 2a outlines a novel method of preparing ⁇ - aminoboronic acids suitable for incorporation in to peptide and applied as enzyme inhibitors.
  • Matteson (Matteson and Majumdar J “ . Organometallic Chem. 170 , 259- 264, 1979; Matteson and Arne O ganomet allies 1, 280-288, 1982) prepared ⁇ -haloboronic acids by this method, but did not expand this method to the preparation of ⁇ -aminoboronic acids with primary or secondary amino groups required for the preparation of peptides.
  • Compound 6 is prepared by the method described by Sadhu and Matteson Organometallics 4 . , 1687-1689, 1985.
  • Compound 6 is allowed to react with thiophenol in presence of tertiary base to give the thiol ether 7.
  • 7 can be prepared by reacting the lithium salt of thioanisole with a trialkyl boronate as described by Matteson and Arne Organometallics 1 , 280-288 (1982) . 7 is treated with LDA followed by a hydrocarbon containing an electrophilic center. For this reaction 1- bromo-2, 2-difluoroethane was used to give an a 2,2- difluoroethyl substituent 8 .
  • the ⁇ -aminoboronic acid 9 was obtained by treating 8 with methyl iodide or other suitable alkylating agent in the presence of iodide ion followed by lithium hexamethyldisilazane and HCl.
  • the sidechain substituent is an electrophile.
  • This provides a method of preparing 2-amino-3 , 3-difluoropropyl boronic acid where conventional methods have failed. For example, hydroboration of 1, 1' -difluoroethene to give difluoroethyl boronate failed.
  • H-boroAsp(OMe) - C 10 H 16 can be synthesized from methyl bromoacetate and the final product is obtained by treating the sidechain methyl ester with potassium trimethylsilanolate (Laganis and Chenard Tetrahedron Letters 25, 5831-5834, 1984).
  • H-boroGlu-C ⁇ oHi 6 is also readily prepared by the sequence of reactions shown in Scheme 2a.
  • ⁇ -Aminoacids containing a sidechain carboxylate are novel. Attempts to make boronic acid analog of aspartic acid and glutamic acid following the reaction scheme shown in Scheme 1 have failed.
  • compounds containing a carboxylate, R -CH 2 -C (O) O fc Bu or -CH 2 -CH 2 -C (0) -O t Bu, failed to react to give 3.
  • the methylene adjacent to the carboxylate is of sufficient acidity that it reacts with CHCl 2 "Li + required for the generation of 3.
  • R -CH 2 CHF 2 or alkyl or -CH 2 CH 2 C0 2 CH 3
  • Scheme 2b illustrates the preparation of ⁇ - aminoboronic acids with hydroxy substituted side chains, boroSerine and boroThreonine . Both are synthesized as their benzyl protected form and incorporated into peptides , The benzyl protecting groups are removed by catalytic hydrogenation to give the final product.
  • the synthesis of 2-benzyloxy-l-chloroethane boronic acids esters has been described previously (Matteson et al . Organometallics 3_, 1284-1288, 1984), but this chemistry has not been extended to the preparation of ⁇ -aminoboronic acids.
  • R H- or CH 3 -
  • Scheme 2c describes the novel synthesis of boronic acid analogs of cysteine.
  • Vinylmagnisium bromide is allowed to react with triethyl boronate to give vinylboronate diethyl ester.
  • Transesterification with pinanediol gives the corresponding ester 16.
  • the ⁇ -chloro group is readily converted to the amine using chemistry previously described (Scheme 1) .
  • Final deprotection of the thiol is achieved after incorporation of the amine in peptides.
  • acyl group or N-protected peptide with suitable side chain protection is coupled to 5.
  • This method is sufficiently versatile to allow the synthesis of any peptide within the limits normally encountered during peptide synthesis such as insufficient solubility. Acid chlorides or other active forms of acyl groups can be coupled.
  • the preferred method of coupling of protected amino acids and peptides to the ⁇ -aminoboronic acids is either the mixed anhydride procedure (Anderson et al . , J “ . Am . Chem . Soc . 89, 5012, 1967) or procedures using PyAOP or a related coupling agent (Albericio et al . Tetrahedron Lett. 38, 4853-4856, 1997). This is illustrated in Scheme 3 for the preparation of Ac-Asp-Glu-Val-Val-Pro-boroAlg-OH.
  • the mixed anhydride of Ac-As (O t Bu) - GlufO t Bu) -Val-Val-Pro-OH 10 is prepared in THF or DMF by allowing it to react with isobutyl chloroformate in the presence of a N-methylmorpholine or other stericly hindered base. After allowing the reaction to proceed for 5 min at -20°C, 5 is added as a cold solution in either THF or chloroform followed by the addition of a second equivalent of base. The reaction mixture is routinely stirred one hour at -20°C followed by 1-2 h of'Stirring at room temperature.
  • Insoluble material is removed by filtration, the solvent removed by evaporation, and the residue dissolved in ethyl acetate.
  • the organic solution is washed with 0.20 N hydrochloric acid, 5% aqueous sodium bicarbonate, and saturated aqueous sodium chloride.
  • the organic phase is then dried over anhydrous sodium sulfate, filtered, and subjected to evaporation.
  • 11 is further purified by techniques known to those skilled the art. These include silica gel chromatography, reverse phase HPLC, and size exclusion chromatography using SephedexTM LH-20 in methanol.
  • the tert-butyl ester protecting groups on Asp and Glu are removed allowing 11 to react with anhydrous HCl to give 12.
  • the allyl side chain on the boronic acid is compatible with this procedure.
  • a broader range protecting groups can be used for compounds with other side chains. This includes protecting groups that are labile to catalytic hydrogenation. These techniques are known to those skilled in the art and are described in Stewart and Young "Solid Phase Peptide Synthesis" Pierce Chemical Company, (1984) .
  • the boronic acid ester is removed by the procedure described in Kettner US patent 5,384,410 (1995).
  • the boronic acid ester is suspended in ammonium acetate buffer, pH 6.0, and is allowed to react with an excess of phenyl boronic acid added in an equal volume of ether.
  • the product is readily separated from phenyl boronic acid and phenyl boronic acid pinanediol ester by extracting with ether.
  • the free boronic acid, 13, is obtained by lyophilizing the aqueous phase.
  • Pinanediol esters are also readily removed by treating with anhydrous boron trichloride in methylene chloride as described by Kinder et al., J " . Med. Chem.
  • the boronic acid ester is treated with a 2-3 fold excess of BCI 3 for 5 min at -78°C and the mixture is allowed to stir 15 min in a 0° ice bath. Excess BCI 3 is hydrolyzed by the slow addition of water. Less structurally rigid boronic acid esters such as pinacol esters can be prepared by transesterification with diethanolamine and by hydrolyzing the diethanolamine ester with aqueous acid (Kettner and Shenvi J. Biol . Chem . 259, 15106-15114, 1984) . Compound 13 can be converted to the difluoroborane (-BF 2 ) using a modification of the procedure of Kinder et al . , J " .
  • Scheme 4 shows the synthesis of the cyclopropyl and cyclopropylalkyl side chain inhibitors using the procedure described for the preparation of cyclopropylglycine (Hallinan et al . J. Chem. Soc. Perkin Trans 3537-3543, 1994) .
  • the peptide boronic acid containing an unsaturated alkyl sidechain 14 is treated with diazomethane in the presence of palladium acetate to give the product 15.
  • n 0 or 1
  • a diverse series of inhibitors is obtained by coupling H-boroAlg-C ⁇ oHi 6 , H-Pro-boroAlg-CioHig, H-Leu-boroAlg-C ⁇ oH 5 , and H-Val-Pro-boroAlg-C ⁇ oHi 6 to various acyl chlorides and sufonyl chlorides.
  • the acyl chloride or sulfonyl chloride (Aldrich Chem. Co., 25 ⁇ mol) was dissolved in 200 ⁇ l of ethyl acetate in a screw capped test tube.
  • R is either R 4 -S0 2 - or R 4 -CO- EXAMPLES
  • l-Chloro-3-butene boronate pinanediol ester pinanediol ester.
  • the ⁇ -chloro compound was prepared by homologation of the corresponding allyl boronate.
  • the aqueous layer was washed with hexane (600 mL) .
  • the combined organic phases were concentrated to 1 L and washed with 5% sodium bicarbonate (1 L) and saturated sodium chloride (1 L) . They were dried over sodium sulfate and filtered.
  • the filtrate was concentrated in vacuo and distilled (bp 130-132°C, 0.5 mm Hg) to give 60 g (42 %) of the ⁇ -chloroboronic acid as a clear yellow oil.
  • Cvclopropylboronate pinacol ester The pinacol cyclopropyl bornate ester was prepared by the addition of cyclopropyl magnesium bromide was added to isopropylboronate pinacol ester. The latter compound was prepared by a previously described procedure (Andersen, M. W. ; Hildebrandt, B.; Koster, G. ; Hoffmann, R. W. Chem. Ber. 122, 1989, 1777-1782) .
  • the Grignard reagent was prepared by adding cyclopropylbromide (3.0 mL, 37 mmol) to magnesium turnings (11 g, 0.46 mole) in THF (300 mL) at room temperature under nitrogen.
  • Iodomethyl boronate pinacol THF (800 mL) was placed in a 3 L, 3-necked flask equipped with two addition funnels. Triisopropyl boronate (Aldrich) (128 mL, 0.55 mol) and chloro-iodomethane (Aldrich) (100 g, 0.56 mol) were added. The mixture was cooled to -78°C and n butyl lithium (330 mL, 0.53 mol, 1.6 M in hexanes) was added dropwise. The solution was stirred for 2 hand slowly allowed to warm to - 10°C.
  • Methyl orange indicator was added and HCl (1.0 M in ether) was added until the methyl orange endpoint was reached.
  • Pinacol 65 g, 0.55 mol was added and reaction mixture was allowed to stir 12 h. It was filtered and evaporated in vacuo. The residue was dissolved in acetone (500 mL) and sodium iodide (70 g, 0.47 mol) was added.
  • Phenylthiomethane boronate pinacol ester Phenylthiomethane boronate pinacol ester. Thiophenol (11.6 mL, 113 mmol) was dissolved in DMF (40 mL) and diisopropylethylamme (19.8 mL, 113 mmol) and chloromethyl boronate pinacol ester (20 g, 113 mmol) were added sequentially. (Iodomethyl boronate pinacol can be readily substituted for the chloro compound.) After stirring for 12 hours, solvent was removed by rotary evaporation and ether (70 mL) was added.
  • the reaction mixture was washed with 0.2 N HCl (70 mL) , 5 % NaHC0 3 (70 mL) and saturated sodium chloride (70 mL) .
  • the combined organic phases were dried over sodium sulfate and filtered.
  • the filtrate was concentrated in vacuo and distilled (bp 125-127°C, 0.6 mm
  • boro-Vinylqlycine pinanediol Ester*HCl The ⁇ -chlorovinyl boronate pinanediol ester (10.6 g, 41.7 mmol) was dissolved in THF (100 mL) and added to a freshly prepared solution of lithium hexamethyldisilazide (45.9 mmol) in THF (150 mL) at -78°C. This solution was stirred for 20 h while warming to room temperature . THF was removed in vacuo and hexanes (150 mL) were added. The resulting precipitate was removed by filtration.
  • Pinacol (1-chloroethyl) boronate A 250 mL round bottom flask is charged with THF (60 mL) and CH 2 C1 2 (2.63 mL, 41.0 mmol) . The solution was cooled to - 100°C with a liquid nitrogen/methanol/H0 bath. n-BuLi (1.6 N in hexanes, 25.7 mL) was added slowly over the course of 1 h. The resulting solution was stirred for an additional 45 min at -100°C. Pinacol methyl boronate, dissolved in THF (40 mL) , was added and the solution was stirred overnight while warming to room temperature.
  • Pinanediol ( 1-benzyloxyethyl ) boronate n-BuLi (1.6 N, 13.8 mL) was added to a solution of benzyl alcohol (2.3 mL, 22 mmol) in THF (60 mL) at -78 °C followed by DMSO (1.6 mL, 22 mmol) . The solution was allowed to warm to room temperature and stir for 1 h. The solution was recooled to 0°C and a solution of Pinacol (1-chloroethyl) boronate (2.06 g, 11 mmol) in THF (60 mL) was added.
  • Pinanediol (2 -benzyloxy-1-chloropropyl ) boronate CH 2 C1 2 (0.80 mL, 12.7 mmol) was added to THF (40 mL) and cooled to -100°C. n-BuLi (1.6 N, 6.3 mL) was slowly added while maintaining a temperature of -100°C. The flask was stirred at -100°C for an additional 45 min.
  • H-boroSer (OBzl) -pinanediol HCl was prepared by adding
  • Pinanediol l-chloro-2-benzyloxy-boronate (5.0g, 14.3 mmol) in THF (60 mL) to a solution of LiHMDS (15 mmol) in THF (60 mL) at -78°C. The solution was allowed to stir while warming to room temperature over a period of 3 h. The THF was evaporated, the residue redissolved in anhydrous hexanes (200 mL) , cooled to -78°C, and a solution of HCl in dioxane (4 N, 11.3 mL) was added. The resulting mixture was allowed to stir while warming to room temperature. The solids were removed by filtration.
  • Phenylsulfenyl chloride (2.0 g, 13.8 mmol) was added to a solution of pinanediol vinyl boronate (2.85 g, 13.8 mmol) in CH 2 C1 2 (30 mL) . The solution was stirred for 30 min and then the solution was evaporated to yield 3.9 g (81%) of a pale yellow oil.
  • X H-NMR (CDC1 3 ) ⁇ 7.40 (m, 5H) , 4.40 (d,
  • Pinanediol l-amino-2-thiophenylethylboronate HCl Pinanediol l-amino-2-thiophenylethylboronate HCl. Pinanediol l-chloro-2-thio (phenyl) ethylboronate (2.0 g, 5.7 mmol) dissolved in THF (40 mL) was added to a solution LiHMDS (6.0 mmol) in THF (60 mL) at -78°C. The solution was allowed to warm to room temperature and solvent was evaporated. The residue was redissolved in hexanes, filtered and recooled to -78°C. A solution of HCl in dioxane (4 N, 5 mL) was added and the mixture was allowed to stir overnight while warming to room temperature.
  • Phenylthiosulfenyl chloride was prepared by reacting benzene thiol with sulfur dichloride at -78°C using a published procedure (Can. J. Chem. , 5_1, 3403-3412, 1973).
  • phenyl ethyl boronate pinanediol was obtained by adding phenylthiosulfenyl chloride (3.2 g, 18.2 mmol) dissolved in dichloromethane (30 mL) dropwise over a period of 10 min to a solution of pinanediol vinylboronate (3.7 g, 18.2 mmol) in CH 2 C1 2 (50 L) in the presence of CaC0 3 (30 mg) . The resulting solution was stirred for an additional 1 h at room temperature.
  • Pinanediol l-amino-2-thiolsulfenyl (phenyl) ethyl boronate pinanediol was treated with lithium hexamethyldisilane by the procedure in Example 5d to yield the alpha-amino compound. MS/ESI calculated for C ⁇ 8 H 6 BN0 2 S 2 + H: 364. Found: 364.
  • Butyllithium (12.8 mL, 32.0 mmol, 2.5M in hexanes) was added dropwise to the solution.
  • a solution of phenylthiomethane boronate pinacol ester (8.0 g, 32.0 mmol) in THF (10 mL) was added dropwise rapidly, yielding a white precipitate.
  • the reaction mixture was stirred for 1 hour at 0 °C, followed by the dropwise addition of 3,3,3- trifluoropropyl iodide (Lancaster) (15. Og, 64.0 mmol). The precipitate dissolved and the solution was allowed to warm to room temperature and stirred for 12 hours . The mixture was then treated with excess cold 10 % phosphoric acid and stirred for 5 minutes.
  • reaction mixture was cooled to -78 °C and 4M anhydrous hydrogen chloride in dioxane (7.2 ml, 28.7 mmol) was added dropwise. The solution was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was concentrated and chloroform was added. Insoluble material was removed by filtration. The filtrate was evaporated almost to dryness and hexanes were added. Upon standing the product crystallized. It was isolated and washed with cold hexanes to yield 1.7g (69.8 %) of a brown solid.
  • Boc-Pro-borocyclopropylmethylglycine pinanediol (Boc-Pro-NH-CH[-CH 2 -cyclopropyl] B0 2 C ⁇ oHi 6 ) Boc-Pro-boroAlq pinanediol ester.
  • Boc-Proline (1.07 g, 4.95 mmol) was dissolved in THF (15 L) and N- methylmorpholine (0.540 mL, 4.95 mmol) was added. The solution was cooled to -20°C and isobutyl chloroformate (0.640 mL, 4.95 mmol) was added.
  • the crude material was purified on silica gel.
  • the column was eluted using a stepwise gradient of ethyl acetate: hexane from a ratio of 9:1 to a ratio of 1:1.
  • TLC in 1:1 ethyl acetate / hexane indicated the product at Rp of 0.30. Fractions containing the product were concentrated in vacuo to give 1.1 g (50 %) of 9.
  • Diazomethane was prepared from Diazald (Aldrich) using the procedure provided by the manufacturer.
  • the allyl boronic acid ester (1.00 g, 2.20 mmol) was dissolved in ether (10 mL) and diazomethane (700 mg, 16.6 mmol) was added.
  • Palladium acetate 50 mg was dissolved in THF (1 mL) was added to the flask. Vigorous bubbling was observed.
  • the reaction was allowed to stir for 10 minutes and excess diazomethane was removed by evaporation using a stream of nitrogen. Ether was added and the reaction mixture was filtered using a paper filter.
  • H-Pro-boroCpa pinanediol ester hvdrochloride .
  • the free N- terminal amine was prepared by treating Boc-Pro-boroCpa pinanediol ester (Example 6, 210 mg, 0.45 mmol) with 4 N HCl in dioxane (10 mL) for 2 hours. The material was concentrated in vacuo and dried under high vacuum to give a brown oil (180 mg, 98%).
  • Boc-ASP (Q t Bu) -Glu (Q t ⁇ u) -Val-Val-OH was prepared by coupling Boc-Val-OH to H-Val-OBzl.
  • H-Val-OBzl* HCl (5.0 g, 20.5 mmol), Boc-Val-OH (4.45 g, 20.5 mmol), and l-hydroxybenzotriazole»H 2 0 (HOBT, 5.55 g, 41.1 mmol) were dissolved in 50 mL of chloroform.
  • N-Methylmorpholine (NMM, 2.24 mL, 20.5 mmol) and N, N' -dicyclohexylcarbodiimide (DCC, 4.2 g, 20.5 mmol) were added and the reaction mixture was allowed to stir overnight at room temperature. The reaction mixture was filtered and solvent was evaporated. Ethyl acetate was added and the mixture was filtered. The filtrate was washed with 0.20 N HCl, 5% NaHC03 , and saturated aqueous NaCl. It was dried over Na 2 S0 4 , filtered, and evaporated to give 7.2 g (88%) of the desired product.
  • H-Val-Val-OBzl*HCl (21.3 g, 62.1 mmol) was dissolved in 150 mL of DMF and Z-Glu (O t ⁇ u) -OH (20.9 g, 62.1 mmol), HOBT (16.8 g, 124 mmol), NMM (6.8 mL, 62.1 mmol) and DCC (12.8 g, 62.1 mmol) were added.
  • the reaction mixture was stirred overnight at room temperature. The mixture was filtered and solvent was evaporated. Ethyl acetate was added and insoluble material was removed by filtration. The filtrate was washed with 0.2 N HCl, 5% NaHC ⁇ 3 , and saturated aqueous NaCl.
  • Boc-Asp(O t Bu) -Glu- (O fc Bu) -Val-Val-OH was prepared by coupling the tripeptide to the active ester of Boc- Asp (OBzl) -OH.
  • Boc-Asp (O fc Bu) -N-hydroxysuccinimide ester was prepared by dissolving Boc-Asp (O fc Bu) -OH (3.00 g, 10.4 mmol) and N-hydroxysuccinimide (1.19 g, 10.4 mmol) in 50 mL of ethylene glycol dimethyl ether. The flask was cooled to
  • Asp (O fc Bu) -N-hydroxysuccinimide ester (4.37 g, 11.3 mmol) was dissolved in 100 mL of dioxane and was added to a solution of H- Glu- (O t Bu) -Val-Val-OH (4.92 g, 12.3 mmol) dissolved in 150 mL of water and 50 mL of dioxane.
  • Sodium bicarbonate (3.07 g, 36.7 mmol) was added and the mixture was stirred at room temperature for 5 h.
  • Dioxane was removed in vacuo and concentrated HCl was added to adjust the pH to approximately 2. The product was extracted into ethyl acetate.
  • Boc-Asp(O fc Bu) -GlutO t ⁇ u) -Val-Val-Pro-boroCpa-pinanediol was prepared by coupling the protected tetrapeptide to the dipeptidyl boronic acid.
  • H-Pro-boroCpa pinanediol ester * HCl (180 g, 0.46 mmol) and Boc-Asp (O fc Bu) -Glu (O fc Bu) -Val- Val-OH (310 mg, 0.46 mmol) were dissolved in chloroform (15 mL) and HOBT (120 mg, 0.92 mmol) and NMM (50 ⁇ L, 0.46 mmol) were added.
  • Boc-Val-Pro-OBzl was prepared by dissolving H-Pro-OBzl (20 g, 83 mmol) in 50 mL of chloroform and adding Boc-Val-OH
  • Boc-Val-Val-Pro-OBzl was prepared by dissolving Boc-Val- Pro-OBzl (14.0 g, 35.0 mmol) in 4N HCl in dioxane (20 mL) and allowing the reaction to stir for 2 h under an inert atmosphere at room temperature. The reaction mixture was concentrated by evaporation in vacuo and ether was added to yield a precipitate. It was collected by filtration under nitrogen. After drying in vacuo with P 2 Os, H-Val-Pro-OBzl was obtained as a white solid (22.6 g, 30.3 mmol, 89%). (ESI/MS calculated for C ⁇ 7 H 24 N 2 0 3 +H: 305.2.
  • H-Glu(O t Bu) -Val-Val-Pro-OH was prepared by dissolving Z- GlufO t BuJ-Val-Val-Pro-OBzl (2.90 g, 3.89 mmol) in 100 mL methanol containing 1% acetic acid. Pearlman's catalyst, Pd(OH) 2 , (lOOmg) was added and the flask was placed on the Parr hydrogenation apparatus with an initial H 2 pressure of 34 psi. After three hours, the catalyst was removed by filtration through a celite pad and the filtrate was evaporated in vacuo to yield a yellow oil (1.30 g, 2.61 mmol, 67%). ESI/MS calculated for C 2 H 2 N 4 ⁇ 7 +H: 499.3 Found: 499.4.
  • Boc-Asp(O fc Bu) -Glu(O t Bu) -Val-Val-Pro-OH was prepared by active ester coupling.
  • Boc-Asp (O fc Bu) -N-hydroxysuccinimide ester was prepared by coupling Boc-Asp (OtBu) -OH (3.00 g, 10.4 mmol) to N-hydroxysuccinimide (1.19 g, 10.4 mmol) in 50 mL of ethylene glycol dimethyl ether.
  • the reaction flask was placed in an ice bath at 0°C and DCC was added. The reaction mixture was slowly allowed to warm to room temperature and to stir overnight. The mixture was filtered and the filtrate was evaporated in vacuo .
  • Glu(O fc Bu) -Val-Val-Pro-OH (5.40 g, 10.8 mmol) was dissolved in 100 mL of water. Sodium bicarbonate (0.92 g, 11.0 mmol) was added followed by triethylamine (2.30 mL, 16.5 mmol) . The N-hydroxysuccinimide ester (3.84 g, 10.0 mmol) was dissolved in 100 mL dioxane and was added to the H- Glu(O fc Bu) -Val-Val-Pro-OH solution. The mixture stirred overnight at room temperature. Dioxane was removed in vacuo and 1.0 M HCl was added to give pH ⁇ 1.
  • Boc-Asp (O t ⁇ u) -Glu (O t Bu) -Val-Val-Pro-boroAlg-pinanediol was prepared by coupling the protected pentapeptide to H- boroAlg-pinanediol (Example 1).
  • Boc-Asp (O t ⁇ u) -Glu (O t ⁇ u) - Val-Val-Pro-OH (1.8 g, 2.3 mmol) was dissolved 10 mL THF and was cooled to -20°C. Isobutyl chloroformate (0.30 mL, 2.3 mmol) and NMM (0.25 mL, 2.3 mmol) were added.
  • Half of the crude product (1.5 g) was purified in 250 mg lots by HPLC using a 4 cm x 30 cm Rainin C-18 reverse phase column. A gradient from 60: 40 acetonitrile: water to 100% acetonitrile was run over a period of 28 minutes at a flow rate of 40 mL/min. The fractions containing the desired product were pooled and lyophilized to yield a white solid (46 mg) .
  • the hexapeptide analog, Example 9, (22.5 mg, 0.023 mmol) was treated with 2 mL of TFA: CH 2 C1 2 (1: 1) for 2 h.
  • the material was concentrated in vacuo and purified by HPLC using C-18 Vydac reverse phase (2.2 x 25 cm) column with a gradient starting at 60:40 acetonitrile/water with 0.1%TFA going to 95:5 over 25 minutes with a flow rate of 8 mL/min.
  • the product eluted at 80% acetonitrile.
  • the fractions were evaporated and dried under high vacuum to give 8.9 mg (49%) of the desired product as white amorphous solid.
  • Ac-Asp (O t ⁇ u) -Glu (O Bu) -Val-Val-Pro-OH was prepared by coupling the N-hydroxysuccinimde ester of Ac-Asp (OBzl) -OH to H-Glu(OtBu)-Val-Val-Pro-OH (See Example 9.)
  • the tetrapeptide (2.67 g, 5.36 mmol) was dissolved in 100 mL of water and NaHC ⁇ 3 (0.45g, 5.36 mmol) and triethylamine (1.12 mL, 8.00 mmol) were added.
  • Boc-Asp (O t Bu) -Glu (O fc Bu) -Val-Val-Pro-OH (See Example 9) (1.51 g, 1.95 mmol) was dissolved in THF (20 mL) and cooled to -20°C. Isobutylchloroformate (0.251 mL, 1.95 mmol) and 4-methylmorpholine (0.214 mL, 1.95 mmol) were added. After 5 min, a solution of boroVinylglycine pinanediol ester, Example 5, (0.529 g, 1.95 mmol) dissolved in DMF (20 mL) and cooled to -20°C was added. Triethylamine (0.272 mL,
  • Boc-Asp (OtBu) -Glu (OtBu) -Val-Val-Pro-OH (See Example 9.) (1.5 g, 1.9 mmol) was dissolved in THF (15 mL) and N- methyl orpholine (0.21 mL, 1.9 mmol) was added. The solution was cooled to -20°C and isobutylchloroformate
  • H-Hyp (Bzl) -boroAbu-pinanediol The mixed anhydride of Boc- Hyp(OBzl)-OH (2.48 g, 7.71 mmol) was prepared and coupled H-boroAbu-C ⁇ oHi 6 (Example 2) by the procedure described for the preparation of Example 6. The final product was purified by silica gel chromatography using ethyl acetate: hexane as a solvent. Boc-Hyp (OBzl) -boroAbu-pinanediol was obtained in a yield of 55%.
  • Pz-CO-Val-Val-Hyp (Bzl) -boroAbu-pinanediol was prepared by coupling the two dipeptide analogs.
  • the pyrazine peptide was prepared by coupling pyrazine carboxylic acid (2.14 g, 17.3 mmol) to H-Val-Val-OBzl (See Example 7.) in 50 mL of chloroform using the DCC coupling procedure described for the preparation of Example 7.
  • Pz-CO-Val-Val-OBzl was obtained in a yield of 86%.
  • Example 19 (0.025 g, 0.025 mmol) was treated with 2 mL of
  • Boc-Pro-boroAlg-C ⁇ oHi 6 The synthesis of Boc-Pro-boroAlg-C ⁇ oHi 6 is described in the preparation of Example 6.
  • the Boc peptide (1.0 g, 2.2 mmol) was dissolved in 10 mL of 1:1 TFA: CH 2 C1 and stirred at room temperature for 1 h. Solvent was evaporated to yield H-Pro-boroAlg-C ⁇ 0 H ⁇ 6 .TFA (0.76 g, 91%).
  • ESI/MS calculated for C ⁇ 9 H 3 ⁇ N 2 0 3 B +H: 347.2. Found: 347.4.
  • H- Pro-boroAlg-C ⁇ oHi 6 »TFA (50 mg, 0.10 mmol) was dissolved in methylene chloride (10 mL) .
  • Boc-Val-Pro-OH was prepared by dissolving Boc-Val-Pro-OBzl, (See the preparation of Example 9, 7.80 g, 19.3 mmol) in 100 mL methanol containing 1% acetic acid. Pearlman's catalyst, Pd(OH) 2 , (100 mg) was added and the compound was hydrogenated on a Parr apparatus. After hydrogen consumption was complete, the catalyst was removed by filtration through a celite pad and the filtrate was evaporated in vacuo to yield an oil (6.1 g, 100%). ESI/MS calculated for C ⁇ 5 H 6 2 ⁇ 5 +H: 315.2. Found: 315.3.
  • Example 24 (100 mg, 0.18 mmol) was dissolved in 2 mL of 1:1 TFA: CH 2 C1 and stirred at room temperature for 2. The reaction mixture was evaporated in vacuo and stored under vacuum with P 2 Os overnight to yielded a yellow solid (80 mg, 0.17 mmol, 94%). ESI/MS calculated for C 24 H 40 N 3 ⁇ B +H: 446.3. Found: 446.3.
  • Example 26
  • Boc-Val-Val-Pro-OBzl (See preparation of Example 9) (1.2 g, 2.4 mmol) was dissolved in 100 L of methanol with 1% acetic acid. Pearlman's catalyst, Pd(OH) 2 , (lOOmg) was added and the flask was placed on the Parr hydrogenation apparatus under an initial hydrogen pressure of 30 psi. After 3 h, the catalyst was removed by filtration through a celite pad. The filtrate was concentrated in vacuo to yield the carboxylic acid (0.83 g, 87%) .
  • ESI/MS calculated for C 20 H35N3 ⁇ 6 +H: 413.3. Found: 414.2.
  • the mixed anhydride of Boc-Val-Val-Pro-OH (0.83 g, 2.0 mmol) was prepared in THF (10 mL) and added to H-boroAlg pinanediol ester (Example 1) dissolved in CHCI3 (6 mL) using the procedure describe in Example 6.
  • the product was purified by silica gel chromatography by first eluting the column with a stepwise gradient of hexane: ethyl acetate and then eluting with ethyl acetate: methanol (9: 1). TLC run in 1 : 1 ethyl acetate: hexane indicate a single spot, Rf 0.10.
  • the desired product was obtained in a yield of 0.29 g (23%).
  • ESI/MS calculated for C 34 H 57 N 4 O 7 B 1 - H: 643.2. Found: 643.4.
  • Example 27 (0.23 g, 0.36 mmol) was dissolved in 5 mL 4N HCl in dioxane and stirred at room temperature for 3 h. Ether was added to yield a white precipitate that was isolated by filtration. After drying over P Os in vacuo the desired product was obtained as a white solid in a yield of 86 mg, 44%) .
  • ESI/MS calculated for C 29 H 49 ⁇ 5 B ⁇ + H: 544.4. Found: 545.4.
  • Glut-Val-Val-Pro-boroAlg-pinanediol H-Val-Val-Pro-boroAlg-C ⁇ oH ⁇ 6 *HCl (Example 28, 100 mg, 0.18 mmol) was dissolved in methylene chloride (10 mL) . Glutaric anhydride (19 mg, 0.17 mmol) and diisopropylethylamme (63 ⁇ L, 0.36 mmol) were added. The reaction mixture stirred at room temperature overnight. The reaction mixture was washed with 0.20 N HCl and the organic phase was dried over Na 2 S ⁇ 4 , filtered, and concentrated in vacuo to yield a oil.
  • This Example was prepared was prepared according to the procedure described for Example 9 except Z-D-Glu (O t ⁇ u) -OH was used in place of the Z-Glu (O t ⁇ u) -OH.
  • ESI/MS calculated for C 5 iH 86 N 6 0 ⁇ 3 B ⁇ +H: 1001.6. Found: 1001.8.
  • Example 6 the mixed anhydride of Ac-Glu (O t ⁇ u) -Val-Val-Pro-OH (1.0 g, 1.9 mmol) was prepared in 20 mL DMF and coupled to H- boroAlg-CioHi ⁇ (Example 1) dissolved in 10 mL of THF.
  • the product was purified by silica gel chromatography. The column was eluted using a stepwise gradient of hexane and ethyl acetate. The product was eluted with 9 : lethyl acetate: methanol.
  • Example 27 The preparation of Boc-Val-Val-Pro-OH is described in the synthesis of Example 27.
  • the mixed anhydride of Boc-Val- Val-Pro-OH (1.8 g, 4.3 mmol) was prepared in 10 mL THF by the procedure described for the preparation of Example 6. It was coupled to H-boroCpg-pinacol «HCl dissolved in 10 mL of DMF.
  • the product was purified by silica gel chromatography. The column was eluted with a stepwise gradient of ethyl acetate: hexane and final elution was achieved with 9:1 ethyl acetate: methanol. The pooled fractions were concentrated in vacuo and lyophilized to yield a white solid 0.64 g (25%).
  • ESI/MS calculated for C 30 H 53 N 4 O 7 B 1 + H: 593.4. Found: 593.5.
  • H-Val-Val-Pro-boroCpg pinacol ester (Example 36, 78 mg, 0.15 mmol) was dissolved in water (10 mL) and glutaric anhydride (17.5 mg, 0.15 mmol) was dissolved in dioxane (10 mL) and was added. Sodium bicarbonate (38 mg, 0.45 mmol) was added and the reaction mixture was allowed to stir until the amine could not be detected by TLC. Pinanediol (51 mg, 0.30 mmol) and the reaction mixture was stirred for 1 h. It was acidified with 1 M HCl prepared in saturated aqueous NaCl.
  • the product was extracted into ethyl acetate, dried over MgS0 4 , filtered and concentrated in vacuo to yield a clear oil. It was purified by HPLC using C-18 Rainin reverse phase (4 x 30 cm) column with a gradient from 95:5 water/acetonitrile to 5:95 water acetonitrile over 31 minutes (All solvents contained 0.10% TFA) . The isolated product was lyophilized from acetonitrile/water to yield a white solid (25.1 mg, 0.04 mmol, 25%) ESI/MS calculated for C 34 H 55 N0 8 B ⁇ - ⁇ -H: 659.4. Found: 659.5.
  • the mixed anhydride of Boc-Asp (O fc Bu) -Glu (O fc Bu) -Val-Val-Pro- OH (1.24 g, 1.61 mmol) was prepared in 10 mL of THF by the procedure described for Example 6 and was coupled to boroDfb-pinanediol (Example 4) dissolved in 5 mL of THF. Following purified on a 5 x 90 cm column of SephadexTM LH- 20 column using as a solvent methanol, the desired product as an amorphous solid (0.51 g, 30.9%) was obtained. TLC in 100 % ethyl acetate indicated the product as a single spot with R F of 0.45.
  • the hexapeptide analog, Example 39, (0.26 g, 0.25 mmol) was treated with 4 N HCl in dioxane (5 mL) for 3 h.
  • the material was concentrated in vacuo and a sample (40 mg) was purified by HPLC using C-8 Phenomenex reverse phase (2.1 x 25 cm) column using a water: acetonitrile gradient (All solvents were adjusted to 0.1% TFA.).
  • the product eluted at 80% acetonitrile.
  • the fractions were evaporated and dried under high vacuum to obtain 10.1 mg (25.3 %) of the desired product as a white amorphous solid.
  • the mixed anhydride of Boc-Val-Val-Pro-OH (0.16 g, 0.39 mmol) was prepared in THF (5 mL) and was coupled to H- boroDfb-pinanediol»HCl (Example 4, 0.12 g, 0.39 mmol) dissolved in CHCI 3 (10 mL) using the procedure in Example 6. After purification by silica gel chromatography using ethyl acetate as a solvent, the desired product was obtained as an amorphous solid (44 mg) .

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AU5920400A (en) 2001-01-22
WO2001002424A2 (en) 2001-01-11
WO2001002601A2 (en) 2001-01-11
WO2001002424A3 (en) 2001-07-19
WO2001002601A3 (en) 2001-07-26
CA2376965A1 (en) 2001-01-11
AU5788800A (en) 2001-01-22

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