+

WO1996019483A1 - Low molecular weight bicyclic thrombin inhibitors - Google Patents

Low molecular weight bicyclic thrombin inhibitors Download PDF

Info

Publication number
WO1996019483A1
WO1996019483A1 PCT/CA1995/000708 CA9500708W WO9619483A1 WO 1996019483 A1 WO1996019483 A1 WO 1996019483A1 CA 9500708 W CA9500708 W CA 9500708W WO 9619483 A1 WO9619483 A1 WO 9619483A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
compound according
aryl
oxo
cycloalkyl
Prior art date
Application number
PCT/CA1995/000708
Other languages
French (fr)
Inventor
John Dimaio
Annette Marian Doherty
M. Arshad Siddiqui
Jeremy John Edmonds
John W. Gillard
Yves St-Denis
Micheline Tarazi
Patrice Preville
Sophie Levesque
Benoit Bachand
Original Assignee
Biochem Pharma Inc.
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority claimed from GBGB9426038.7A external-priority patent/GB9426038D0/en
Priority claimed from GBGB9510265.3A external-priority patent/GB9510265D0/en
Priority claimed from GBGB9510267.9A external-priority patent/GB9510267D0/en
Priority claimed from GBGB9510266.1A external-priority patent/GB9510266D0/en
Priority to AU42505/96A priority Critical patent/AU4250596A/en
Priority to MD97-0253A priority patent/MD970253A/en
Priority to EP95940923A priority patent/EP0802916A1/en
Priority to SK838-97A priority patent/SK83897A3/en
Priority to BR9510433A priority patent/BR9510433A/en
Application filed by Biochem Pharma Inc. filed Critical Biochem Pharma Inc.
Priority to NZ297360A priority patent/NZ297360A/en
Priority to EE9700113A priority patent/EE9700113A/en
Priority to JP8519383A priority patent/JPH11508535A/en
Priority to APAP/P/1997/001004A priority patent/AP9701004A0/en
Publication of WO1996019483A1 publication Critical patent/WO1996019483A1/en
Priority to IS4504A priority patent/IS4504A/en
Priority to FI972466A priority patent/FI972466L/en
Priority to NO972892A priority patent/NO972892L/en
Priority to BG101647A priority patent/BG101647A/en
Priority to US08/880,885 priority patent/US6057314A/en
Priority to LVP-97-141A priority patent/LV12019B/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/08—Vasodilators for multiple indications
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
    • 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
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • 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
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • 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
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • 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
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to compounds useful for the
  • Plasma proteins such as fibrinogen, proteases and cellular receptors participating in hemostasis have emerged as important factors that play a role in acute and chronic coronary disease as well as cerebral artery disease by contributing to the formation of thrombus or blood clots that effectively diminish normal blood flow and supply.
  • Vascular aberrations stemming from primary pathologic states such as hypertension, rupture of
  • Thrombin is a key regulatory enzyme in the coagulation cascade; it serves a pluralistic role as both a positive and negative feedback regulator. However, in pathologic conditions the former is amplified through catalytic activation of cofactors required for thrombin generation as well as activation of factor XIII necessary for fibrin cross-linking and stabilization.
  • thrombin In addition to its direct effect on hemostasis, thrombin exerts direct effects on diverse cell types that support and amplify pathogenesis of arterial thrombus disease.
  • the enzyme is the strongest activator of platelets causing them to aggregate and release substances (eg. ADP TXA NE) that further propagate the thrombotic cycle.
  • Platelets in a fibrin mesh comprise the principal framework of a white thrombus.
  • Thrombin also exerts direct effects on
  • thrombin activity constitutes a viable therapeutic approach towards the attenuation of proliferative events associated with thrombosis.
  • ATIII antithrombin III
  • Heparin exerts clinical efficacy in venous thrombosis by enhancing ATIII/thrombin binding through catalysis. However, heparin also catalyzes inhibition of other proteases in the coagulation cascade and its
  • thrombocytopenia, osteoporosis and triglyceridemia have been observed following prolonged treatment with heparin.
  • Hirudin derived from the glandular secretions of the leech hirid ⁇ medicinalis is one of the high molecular weight natural anticoagulant protein inhibitors of
  • Hirudin as a therapeutic agent is likely antigenicity and lack of an effective method of neutralization, especially in view of its extremely tight binding characteristics toward
  • thrombin The exceedingly high affinity for thrombin is unique and is attributed to a simultaneous interaction with the catalytic site as well as a distal "anion binding exosite" on the enzyme.
  • Thrombin activity can also be abrogated by Hirudin-like molecules such as hirulog (Maraganore, J.M. et al.,
  • Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for
  • Blomback et al first designed a thrombin inhibitor that was modeled upon the partial sequence of the fibrinogen A(LBi)0c chain comprising its proteolytically susceptible region (Blomback, et al., J. Clin. Lab. Invest., 24 , 59, 1969). This region of fibrinogen minimally includes the residues commencing with phenylalanine: Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly
  • aldehyde group is presumed to contribute strongly to inhibitory activity in view of its chemical reactivity toward thrombin's catalytic Ser residue, generating a hemiacetal intermediate.
  • thrombin inhibitors bearing the (D)Phe-Pro-Arg general motif are those incorporating COOH- terminal boroarginine variants such as boronic acids or boronates (Kettner, C. et al., J. Biol. Chem., 268, 4734, 1993). Still other congeners of this motif are those bearing phosphonates (Wang, C-L J., Tetrahedron Letters, ll, 7667, 1992) and ⁇ -Keto esters (Iwanowicz, E.J. et al., Bioorganic and Medicinal Chemistry Letters, 12, 1607, 1992).
  • Neises, B. et al. have described a trichloromethyl ketone thrombin inhibitor (MDL-73756) and Attenburger, J.M. et al . have revealed a related difluoro alkyl amide ketone (Tetrahedron Letters, 32, 7255, 1991).
  • Maraganore et al. disclose a series of thrombin inhibitors that incorporate the D-Phe-Pro- moiety and hypothesize that this preferred structure fits well within the groove adjacent to the active site of thrombin. Variations on these inhibitors are essentially linear or cyclic peptides built upon the D-Phe-Pro moiety.
  • One object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme, thrombin.
  • a further object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme thrombin and are provided for in s pnarmacologically acceptable state.
  • Still a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof as anticoagulant and thrombin inhibitory agents.
  • Yet a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof for therapeutic treatment of various thrombotic maladies .
  • a further object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors.
  • the enzyme inhibitors of the present invention are encompassed by the structure of general Formula I.
  • the present invention provides for novel compounds that display thrombin inhibitory activity as reflected in formula I :
  • A is selected from (CH-R 8 ) , S, SO, SO , O and NR wherein R 8 is hydrogen, C alkyl optionally interupted with 1 or 2 heteroatoms; C aryl, C _ cycloalkyl or heterocyclic ring or a hydrophobic group;
  • R 6 and R 7 are independently selected from
  • D is selected from (CH-R 5 ) wherein R is hydrogen, C
  • E is selected from CH and CH substituted with the -C ( O ) F. provided that only one of D and E is substituted with with -C (O)R ;
  • X is selected from O, N-R , or CH-R ;
  • Y is selected from O, S, SO, SO , N-R and CH-R provided that when X is N-R then Y is CH-R or O, and when X is O then Y is CH-R ;
  • Z is selected from O, S and H ;
  • R is a polar amino acid residuearginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle ;
  • R 2 is selected from H and C alkyl optionally substituted with C aryl, a 6 member heterocycle or a C _ cycloalkyl ring;
  • R 3 is selected from H, NRR- and C alkyl
  • R 4 and R 5 are independently selected from H; NR,R, ; C, , aryl or C cycloalkyl optionally substituted with C . alkyl; C alkyl optionally interrupted by one or more
  • heteroatom or carbonyl group and optionally substituted with OH, SH, NRR or a C aryl, heterocycle or C cycloalkyl group optionally substituted with halogen, hydroxyl , C alkyl; an amino acid side chain; and a hydrophobic group.
  • the molecules, compositions and methods of this invention are useful as anti-coagulants, or in the treatment and
  • the present invention relates to molecules which inhibit the enzyme, thrombin. These molecules are characterized by a heterobicyclic moiety as illustrated in Formula I :
  • hydrophobic group refers to any group which lacks affinity for, or displaces water. Hydrophobic groups include but are not limited to C alkyl, C alkenyl (e.g. vinyl, allyl) or C alkynyl (e.g. propargyl) optionally interrupted by a carbonyl group, (e.g. forming an acyl group); C aryl, C
  • hydrophobic groups include cyclohexyl, benzyl, benzoyl, phenylmethyl ,
  • arginyl moiety represents an arginine amino acid residue or an analogue or derivative thereof.
  • an analogue or derivative of the natural residue may incorporate a longer or shorter methylene chain from the alpha carbon (i.e. ethylene or butylene chain);
  • ns trained group i.e. an aryl, cycloalkyl or
  • alkyl represents a straight or branched, saturated or unsaturated chain having a specified total number of carbon atoms.
  • aromatic or aryl represents an unsaturated carbocyclic ring(s) of 6 to 16 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i . e. NR R ) halogen or C alkyl.
  • Aromatic rings include benzene, napththalene, phenanthrene and anthracene.
  • Preferred aromatic rings are benzene and naphthalene.
  • cycloalkyl represents a saturated carbocyclic ring of 3 to 7 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NR.R ) halogen or C alkyl.
  • Cycloalkyl groups include cyclo- propyl , butyl, pentyl, hexyl and heptyl .
  • a preferred cycloalkyl group is cyclohexyl .
  • aralkyl represents a substituent comprising an aryl moiety attached via an alkyl chain (e.g. benzyl, phenethyl) wherein the sum total of carbon atoms for the aryl moiety and the alkyl chain is as specified.
  • the aryl or chain portion of the group is optionally mono- or disubstituted with OH, SH, amino (i.e. NR R ) halogen or C alkyl
  • heteroatom represents oxygen, nitrogen or sulfur (O, N or S) as well as sulfoxyl or sulfonyl (SO or SO ) unless otherwise indicated. It is understood that alkyl chains interrupted by one or more heteroatoms means that a carbon atom of the chain is replaced with a heteroatom having the appropriate valency Freferrably, an alkyl chain is interrupted by 0 to 4 heteroatoms and that two adjacent carbon atoms are not both replaced.
  • heterocycle represents a saturated or
  • a heterocycle is optionally mono- or di-substituted with OH, SH, amino (i.e. NR R- ) , halogen, CF , oxo or C alkyl.
  • suitable monocyclic heterocycles include but are not limited to pyridine, piperidine, pyrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran and thiophene .
  • suitable bicyclic heterocycles include but are not limited to indole, quinoline,
  • hydrophobic amino acid represents an amino acid residue that bears an alkyl or aryl group attached to the ⁇ -carbon atom.
  • glycine which has no such group attached to the ⁇ -carbon atom is not a hydrophobic amino acid.
  • the alkyl or aryl group can be substituted, provided that the substituent or substituents do not detract from the overall hydrophobic character of the amino acid.
  • hydrophobic amino acids include natural amino acid residues such as alanine; isoleucine; leucine;
  • Vellaccio Suitable non-naturally ocurring amino acids include cyclohexylalanine and 1-aminocyclohexanecarboxylic.
  • amino acid side chain is meant the substituent attached to the carbon which is ⁇ to the amino group.
  • R 2 is H or C alkyl. More preferably R is H methyl or ethyl and most preferably R is H.
  • R 3 is H or C alkyl. More preferably, R is K methyl or ethyl, and most preferably R is H.
  • one of R 4 or R 5 is a hydrophobic group such as a saturated or unsaturated carbocycle of 5 or 6 members optionally fused to another carbocyclic group while the other is H, C alkyl optionally substituted by NP. P. or carboxy.
  • the hydrophobic moiety may be linked via a spacer such as a C . alkyl chain optionally interrupted with 1 or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO ) groups.
  • one of R 4 and R 5 is phenyl , cyclohexyl, indole, thienyl, quinoline,
  • A is absent or CH .
  • B is S or CH .
  • D is CH .
  • E is CH substituted with -C(O)R wherein R is as previously defined.
  • X is CH-R or N-R .
  • Y is CH-R or S.
  • R 1 is represented by one of formula Via to VId:
  • R 11 is hydrogen or C alkyl
  • K is a bond or -NH- ;
  • G is C alkoxy; cyano ; -NH ; -CH -NH ; -CINH.-NH; -NHC(NH)-NH; -CH -NH-C (NH) -NH ; a C cycloalkyl or aryl substituted with cyano, -NH , -CH -NH , -C(NH)-NH , -NH C(NH)-NH or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally
  • U is cyano, -NH , -C(NH)-NH or -NH-C (NH) -NH ;
  • P is a bond, -C(O)- or a bivalent group:
  • J is C alkylene optionally substituted with OH, NH and C alkyl and optionally interrupted by a heteroatom selected from O, S and N;
  • n 0 or 1 ;
  • T is H, OH, amino, a peptide chain, C alkyl, C alkoxy, C aralkyl, or heterocycle optionally substituted.
  • R 11 is H or methyl and most preferably H.
  • K is a bond
  • G is -NH-C(NH)-NH attached via a methylene chain of 3-7 carbons or phenyl substituted with -C(NH)-NH attached via a methylene chain of 0 to 3 carbons. More preferably G -NH-C(NH)-NH attached via a methylene chain of 3 atoms .
  • P is -C(O)-.
  • J is selected from: -CH -S-CH -CH - ; -CH -O-CH CH - ; -CH -NH-CH -CH - ; and a bond when n is 0. More preferably, J is a bond while n is 0.
  • R is selected from the following amino acid derivatives prepared
  • T is a peptide of 1 to 4 amino acid residues in length and preferably fibrinogen' s A or B chain or fragment or derivative thereof .
  • T is a heterocycle selected from the group consisting of:
  • R' is hydrogen, C _ alkyl optionally carboxyl substituted, carboxyl, -C . alkyl-CO -C alkyl, C aralkyl, C
  • T is selected from the group consisting of
  • R' is as defined above.
  • T is selected from the group consisting of:
  • R' is as defined above.
  • T is selected from the group consisting of:
  • R' is as defined above.
  • T is or
  • R' is H or C alkyl such as methyl, ethyl, propyl or butyl and most preferably wherein R' is hydrogen, .
  • T is a 1,2 thiazole optionally
  • compounds of the invention are represented by formulas II, III, IV and V, wherein X, Y, B, R to R and R are as previously defined.
  • compounds of the invention are represented by one of formulas VII, VIII, IX and X:
  • B is O, S, -CH-, or -NH- ;
  • Y is selected from O, S, SO, SO , N-R and CH-R tenu;
  • R j is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
  • R 2 is H or C alkyl
  • R 3 is selected from H, NR R 7 and C alkyl
  • R 4 and R 5 are independently selected from H; NRR ; C aryl or C cycloalkyl optionally substituted with C alkyl C alkyl optionally interrupted by one or more
  • heteroatom or carbonyl group and optionally substitute:! with OH, SH, NRR, or a C aryl, heterocycle or C cycloalkyl group optionally substituted with halogen, hydroxyl , C . alkyl; an amino acid side chain; and a hydrophobic group;
  • R ⁇ is hydrogen, C . alkyl optionally interupted with 1 or 2 heteroatoms; C aryl, C, - cycloalkyl or heterocyclic ring or a hydrophobic group; and
  • n 1 or 2.
  • Preferred compounds according to formula VII include: 0005 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a] pyridine-3R- carboxamido (propyl
  • 0275 8a-Methyl-5-oxo-6-(3- phenyl-propyl)-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
  • 0280 8a-Methyl-5-oxo-6-(3- phenyl-propyl)-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
  • More preferred compounds according to formula (VII) include:
  • 0220 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-amide; 0240 8a-Methyl-5- oxo-6-(2-trifluoromethyl-quinolin-6-ylmethyl)- hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide; 0245 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3- carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)butyl]-amide; 0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- a]pyridine
  • thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]-amide.
  • Most preferred compounds according to formula VII include: 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido (propylcarbo methoxy
  • Preferred compounds according to formula VIII include:
  • 0670 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(2-amino-6- chloro-pyrimidin-4-yl)-1- (thiazole-2-carbonyl)-propyl]- amide
  • 0675 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(6-amino- pyridin-2-yl)-1-(thiazole-2- carbonyl)-propyl]-amide
  • 0745 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(3guanidino-cyclohexylmethyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
  • 0750 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4- guanidino-cyclohexylmethyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
  • Preferred compounds according to formula (VIII) include: 0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo[1,2- a]pyridine-6-carboxylic acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-amide
  • 0750 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-guanidinocyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide
  • 0760 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2- carbonyl)-4-guanidino-butyl]-amide
  • Preferred compounds according to formula IX include:
  • Preferred compounds according to formula X include
  • More preferred compounds according to formula X include: 925 7-Benzyl-6-oxo-octahydro-pyrido[2,1-c][1,4]thiazine-4- carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)butyl]-amide; and940 6-Oxo-7-phenethyloctahydro-pyrido[2,1-c][1,4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl)-amide .
  • Preferred compounds according to formula III include:
  • Step 2
  • Step 3
  • adduct e. from aldehyde c with d is done by stirring the reactant in aromatic solvents e.g. benzene or toluene in presence of catalytic amount of suitable acid e.g, p-toluenesulfonic acid.
  • aldehyde c to aldehyde g is readily achieved by appropriate protection deprotection protocals found in T. Greene, Protective Groups In Organic Synthesis, (John Wiley & Sons, 1981).
  • Step 4
  • Tne cylization of adduct e to f may readily be achieved by appropriate Lewis acids e.g, trimethyl aluminum in suitable solvents e.g. dichloromethane, the methodology found in T. Greene, supra. Step 4':
  • the compound f can be derived from the treatment of aldehyde g with d in presence of suitable aromatic solvents e.g, benzene.
  • Pg is a nitrogen protecting group
  • each of R 20 ; and R 21 is independently a C alkyl; and X, R 1 , R 3 , R 4 and R 5 are as previously defined.
  • the amino and carboxylic functions of the unsaturated compound of formula (a) are protected with appropriate protecting groups.
  • a variety of protecting groups known for reactive functional groups and suitable protection and deprotection protocols may be found in T. Greene,
  • the appropriate protecting group to use in a particular synthetic scheme will depend on many factors, including the presence of other reactive functional groups and the reaction conditions desired for removal .
  • the unsaturated compound of formula is easily obtained by methods and protocols known to chemist skill on the art.
  • the protected unsaturated compound of formula (a) is subjected to appropriate conditions to allow cyclisation using an appropriate reagent such as mercuric acetate in an inert solvent such as tetrahydrofuran (THF) to yield to a protected amino alcohol of formula (b) .
  • the protected amino alcohol of formula (b) is oxidized using an appropriate oxidizing agent such as sulfur trioxide pyridine complex in an appropriate solvent such as diclhoromethane or dimethylformamide to yield to a protected amino aldehyde of formula (c) .
  • an appropriate oxidizing agent such as sulfur trioxide pyridine complex
  • an appropriate solvent such as diclhoromethane or dimethylformamide
  • intermediate (C) can be made by the ozonolysis of a compound of formula (a') prepared according to Collado et al, J. Org. Chem.,1995, 60:5011.
  • the protected amino aldehyde of formula (c) is coupled with an amino acid alkyl ester of formula (d) by first forming the imine followed by contacting the obtained imine with an appropriate reagent such as sodium
  • the cyclic intermediate of formula (e) is functionalized at the amino position to yield to the amino substituted cyclic intermediate of formula (f).
  • Conditions appropriate for such reactions are well known in the art and will depend on the nature of the R 5 substituent.
  • the amino protecting group of the cyclic intermediate of formula (f) is removed under appropriate conditions and the resulting compound is then subjected to appropriate condition for internal ring closure such as low heat in an inert solvent or as a raw compound to yield to a bicyclic intermediate of formula (g).
  • the bicyclic intermediate of formula (g) can also be obtained by hydrolysing the ester function (-C(O)O-R 20 ) of the cyclic intermediate of formula (g) to the free carboxylic acid followed by standard peptide coupling using an appropriate coupling reagent such as benzotriazole-1-yloxy-tris- (dimethylamino)phosphonium hexafluorophosphate (BOP) in an inert solvent such as dimethyl formamide (DMF).
  • an appropriate coupling reagent such as benzotriazole-1-yloxy-tris- (dimethylamino)phosphonium hexafluorophosphate (BOP) in an inert solvent such as dimethyl formamide (DMF).
  • Pg is a sulfur or amino protecting group
  • L is a leaving group
  • each of R 20 ; and R 21 is independently a C alkyl; and R 1 , R,
  • R 4 and R 5 are as previously defined.
  • the carboxylic acid compound (a) is coupled to the cyclic amine compound (b) with a peptide coupling agent such as benzotriazol-1-yloxy-tris- (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in an appropriate solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to yield to an amido compound of formula (c) .
  • a peptide coupling agent such as benzotriazol-1-yloxy-tris- (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in an appropriate solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to yield to an amido compound of formula (c) .
  • a peptide coupling agent such as benzotriazol
  • the compound of formula (c) is subjected to appropriate conditions to allow internal cyclisation to yield to a bicyclic intermediate of formula (d).
  • appropriate conditions for example, acid mediated cyclisation using p-toluenesulfonic acid or TFA in an appropriate solvent such as dichloroethane.
  • each of R 20 and R 21 is independently a C alkyl; and B, R 1 , R 3 , R 4 , and R 5 are as previously defined.
  • the process depicted in scheme 4 is briefly described as follows:
  • the halogenated compound of formula (a) is converted to a halomethyl ketone of formula (b) using an appropriate reagent, such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25°C to about 0°C.
  • an appropriate reagent such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25°C to about 0°C.
  • the resulting mixture is then treated under acidic
  • halomethyl ketone of formula (b) is coupled with an amino acid alkyl ester of formula (c) with an appropriate base such as sodium cyanoborohydride in an organic solvent such as methanol (MeOH) to yield to a cyclic intermediate of formula (d).
  • an appropriate base such as sodium cyanoborohydride in an organic solvent such as methanol (MeOH)
  • the cyclic intermediate of formula (d) is treated under acidic conditions using an appropriate acid such as camphorsulfonic acid in an appropriate solvent such as toluene ti yield to a bicyclic intermediate of formula (e).
  • an appropriate acid such as camphorsulfonic acid
  • an appropriate solvent such as toluene ti yield to a bicyclic intermediate of formula (e).
  • Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Peptide Synthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.
  • Compounds of the present invention may be prepared for assay by dissolving them in buffer to give solutions ranging in concentrations from 1 to 100 ⁇ M. In an assay to determine the inhibitory dissociation
  • a chromogenic or fluorogenic substrate of thrombin would be added to a solution containing a test compound and thrombin; the resulting catalytic activity of the enzyme would be spectrophotometrically determined.
  • This type of assay is well known to those skilled in the art.
  • the compounds of the present invention may be used as anti-coagulants in vi tro or ex vivo as in the case of contact activation with foreign thrombogenic surfaces such as is found in tubing used in extracorporeal shunts.
  • the compounds of the invention may also be used to coat the surface of such thrombogenic conduits.
  • the compounds of the invention are obtained as lyophilized powders, redissolved in isotonic saline and added in an amount sufficient to maintain blood in an anticoagulated state.
  • the therapeutic agents of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers.
  • the proportion of each carrier is determined by the solubility and chemical nature of the compound, the route of administration, and standard pharmaceutical practice.
  • the compounds may be injected parenterally; this being intramuscularly, intravenously, or subcutaneously.
  • parenteral for parenteral
  • the compound may be used in the form of sterile solutions containing other solutes, for example, sufficient saline or glucose to make the solution
  • the compounds may be administered orally in the form of tablets, capsules, or granules containing suitable excipients such as starch, lactose, white sugar and the like.
  • suitable excipients such as starch, lactose, white sugar and the like.
  • the compounds may also be administered sublingually in the form of troches or lozenges in which each active ingredient is mixed with sugar or corn syrups, flavouring agents and dyes, and then dehydrated sufficiently to make the mixture suitable for pressing into solid form.
  • the compounds may be administered orally in the form of solutions which may contain colouring and/ or flavouring agents .
  • Physicians will determine the dosage of the present therapeutic agents which will be most suitable. Dosages may vary with the mode of administration and the
  • the dosage may vary with the particular patient under treatment.
  • composition When the composition is administered orally, a larger quantity cf the active agent will typically be required to produce the same effect as caused with a smaller quantity-given parenterally.
  • the preferred compounds as of the present invention are synthesized using conventional preparative steps and recovery methods known to those skilled in the art of organic and bio-organic synthesis, while providing a new a unique combination for the overall synthesis of each compound.
  • Pd/C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture with warming The hydrogenation was continued until no starting material could be detected as judged by TLC .
  • the catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
  • Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC.
  • the catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
  • Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC.
  • the catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
  • tert-Butyloxycarbonyl-para-cyano-phenylalanine-N,O- dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure toe yield a clear residue.
  • tert-Butyloxycarbonyl-ortho-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg added. The solution was shaken under H at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.
  • tert-Butyloxycarbonyl-para-aminomethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
  • tert-Butyloxycarbonyl-meta-aminomethyl-phenylalanine-N,O- dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N ' -bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.! mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
  • tert-Butyloxycarbonyl-ortho-aminomethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified on silica gel (ethyl acetate/hexane), and
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with magnesium sulfate, filtered, and concentrated under reduced pressure.
  • the crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
  • the reaction was quenched with saturated aqueous ammonium chlcride.
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
  • the mixture was diluted with ethyl acetate ( 150 mL ) , and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure.
  • the crude material was purified on silica gel ethyl acetate/hexane), and concentrated under reduced pressure .
  • tert-Butyloxycarbonyl-3-(4-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide.
  • tert-Butyloxycarbonyl-3-(3-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(3-piperidyl)alanine-N,O-dimethylamide.
  • tert-Butyloxycarbonyl-3-(2-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide.
  • tert-Butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
  • tert-Butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 filtered, and concentrated under reduced pressure.
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 ⁇ 50 mL), brine (50 mL), dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • tert-Butyloxycarbonyl-para-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in HO (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 ⁇ 50 mL) and brine (3 ⁇ 50 mL). The solution was dried with MgSI filtered, and concentrated under reduced pressure yielding the title compound.
  • tert-Butyloxycarbonyl-meta-nitro-phenylalanine-N,O-dimethylamide 13.88 g, 39.3 mmol was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in HO (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 ⁇ 50 mL), and brine (3 ⁇ 50 mL). The solution was dried with MgSO 4 , filtered, and concentrated under reduced pressure yielding the title compound.
  • tert-Butyloxycarbonyl-ortho-nitro-phenylalanine-N,O-dimethylamide 13.88 g, 39.3 mmol was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in H 2 O (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 ⁇ 50 mL), and brine (3 ⁇ 50 mL). The solution was dried with MgSO 4 , filtered, and concentrated under reduced pressure yielding the title compound.
  • tert-Butyloxycarbonyl-3-(cis/trans-4-aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqvieous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly.
  • the aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 ⁇ 50 mL), 4 N aqueous sodium bisulfate (2 ⁇ 50 mL), and brine (2 ⁇ 50 mL).
  • the solution was dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the semi-solid was chromatographed on silica gel (ethyl acetate/ hexane).
  • tert-Butyloxycarbonyl-3-(cis/ trans-3 - aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly.
  • the aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 ⁇ 50 mL), 4 N aqueous sodium bisulfate (2 ⁇ 50 mL), and brine (2 ⁇ 50 mL).
  • the solution was dried with MgSO 4 , filtered, and concentrated under reduced pressure.
  • the semi-solid was chromatographed on silica gel (ethyl acetate hexane).
  • the semi-solid was chromatographed on silica gel (ethyl acetate' hexane).
  • guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
  • guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
  • guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL ; was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
  • Oxidation of the organoborane was achieved by the addition of 4.7 g (2.0 equiv., 27.2 mmol) of 3-chloroperoxybenzoic acid at 0°C, with warming to ambient temperature and stirring for an additional hour.
  • the organic phase was washed with 5% Na 2 CO 3 , ddH 2 O, and dried over sodium sulfate. Due to the instability of the alcohol, a quick column was performed to remove the extreme polar and nonpolar material which originated from the 3-chloroperoxybenzoic acid.
  • the alcohol (4) was obtained in a yield of 65%.
  • the aldehyde, (5), (2.6 g, 7.10 mmol) was dissolved in benzene (70 mL) and a catalytic amount of ptoluenesulfonic acid was added, followed by 1.58 g (1.2 equiv., 8.52 mmol) of L-cysteine ethyl ester and 4 A molecular sieves .
  • the reaction was allowed to stir overnight at ambient temperature followed be removal of solvent in vacuo .
  • the residue was dissolved in
  • Oxalyl chloride (9) (25 g, 0.197 mol) was cooled to 0oC and cyclohexane propionic acid (20 ml, 0.14 mol) was added. This was left to stir overnight. The resultant mixture was distilled to give an 84% yield of the
  • the chiral auxiliary (11) (13.6g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78oC. Then n-BuLi ⁇ 52.8 mL, 84.4 mmol, 1.2 equiv.) was added and left for 2 mins (dark orange solution). The acid chloride (10) (13.4 g, 76.6 mmol, 1 eq) was then added and left to stir overnight. Work-up was done by quenching with saturated NH 4 Cl extracting with ethyl acetate, washing the extracts with water and brine, drying over sodium sulphate and concentration.
  • the starting material (12)(9.13g, 29 mmol, leq) was dissolved in dry THF and cooled to -78oC, after which LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise over 40 mins. Then, 30 minutes later, allyl bromide (7.5 mL, 86.9 mmol, 3 eq) was added slowly over 10 mins. The mixture was left to warm overnight. Work-up included quenching with sat. ammonium chloride, extraction with ethyl acetate, washing with 10% sodium thiosulphate, decolourising with charcoal, drying over sodium sulphate and concentration in vacuo. The product was obtained as a yellow oil (13) in 96% yield.
  • the starting material (1.97 g, 3.9 mmol, 1 equiv.) was dissolved in 20 mL of dry dichloromethane and cooled to 0oC. Trimethylaluminum (5.9 mL, 11.8 mmol, 3 equiv.), was added dropwise and the mixture was left stirring overnight. After complete reaction as evidenced by HPLC, methanol was added until a yellow t>olid mass was formed. Dichloromethane was added to dissolve the solid and the whole mixture was stirred for 15-30 minutes and then filtered.
  • the starting material (16) (0.95 g, 2.9 mmol, 1 equiv.) was dissolved in 10 mL of dioxane. The solution was cooled to 10oC, and to it was added LiOH H 2 O(0.123 g, 2.9 mmol, 1 eq.) dissolved in 10 mL of water. The bath was removed and the mixture was stirred at room temperature for 1 hour. TLC showed complete reaction and the solvent was evaporated under vacuum. The remaining aqueous layer was washed with ether (2X), acidified with 10% citric acid, and extracted with dichloromethane (3X). The combined extracts were dried over sodium sulphate and concentrated to give a white solid which was
  • BOC-DiCbz Arg (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0oC. Triethylamine (2. mL) was added followed by 14.5 mmol of a 1M toluene solution of isopropyl chloroformate via a syringe. The reaction was allowed to stir at 0oC for 30 minutes then quickly filtered. The white solid was discarded. To the filtrate was bubbled freshly prepared diazomethane until the color of the solution turned yellow. The reaction mixture was allowed to stand overnight in a well
  • the products of the reactions described above can be isolated in the free form or in the form of salts.
  • the products can be obtained as pharmaceutically acceptable acid addition salts by reacting one of the free bases with an acid.
  • the product can be obtained as pharmaceutically acceptable salts by reacting one of the free bases with an acid.
  • the pyrrolidine-aldehyde (6) is coupled with the protected diamino-propionic acid (7) by first forming the imine (8) (MgSO 4 , CH 2 Cl 2 ). Isolation of the imine (8) is done by filtration of the MgSO ⁇ and evaporation of the solvent. The crude imine is then treated with NaBH(OAc) and actic acid (AcOH) in THF for 15 hours to obtain the amine (8) after extrative work-up .
  • imine (8) MgSO 4 , CH 2 Cl 2
  • the CB7 (7) protecting group of the amine (8) is removed by hydrogenation with palladium on charcoal 10% as a catalyst in methanol (MeOH).
  • the catalyst is filtered and the MeOH evaporated to give the crude diamine (9) that can be used without any purification.
  • the cyclisation is done by heating the crude oil (9) from step 7, neat slightly above the boiling point of methanol.
  • the bicyclic lactam (10) is purified by flash
  • the carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF using BOP as the coupling agent in the presence of diisopropylethylamine (EtNiPr ).
  • the two CBZ(Z) protecting groups of compound (15) are removed by catalytic hydrogenation with Pd/C 10% as a catalyst.
  • the catalyst is filtered and the solvent is evaporated to give the amino-guanidine (16).
  • the enamine (5) (1.0eq) is treated with mercuric acetate (1.1 eq) in THF. The solvent is evaporated to dryness and the residue dissolved in methanol. The resulting organomercurial is reductivly cleaved with sodium borohydride (1.3eq). The resulting crude lactam thioether is purified by flash chromatography on silica gel affording compound (6).
  • the isolated bicyclic lactam (8) is hydrolysed with one equivalent of lithium hydroxyde in a 1:1 mixture of THF and water. The mixture is stirred at room temperature for 1 hour. The crude mixture is extracted with ether and the resulting solution is poured into 10% citric acic aqueous solution and extracted with dichloromethane to yield the corresponding carboxylic acid (9).
  • STEP 6
  • the crude carboxylic acid (9) is coupled with
  • Cyclic compound (3) (913mg, 3.32 mmol) was disolved in 5' ml of dry Toluene.
  • ( IS) - (+ ) -10-Camphorsulfonic acid 91 mg, 0.39 mmol was added and the mixture was left to reflux for 4 days.
  • the mixture was worked up byevaporation of solvent, dissolving residue in ethyl acetate and washing with 2 x 5 % NaHCO 3 .
  • the Ethyl acetate layer was dried over Na 2 SO4 and evaporated.
  • the crude residue was purified by silica gel flash column chromatography using 60 % ETOAC / 40 %Hexane followed by 70 % ETOAC / 30 % Hexane giving 62.5% of Bicyclic compound (4).
  • Benzyl bromide (0.26 ml, 2.22 mmol) is then added and the mixture is allowed to reach room temperature and stirred for 15 hours. The mixture is then poured into 10% HCl (50 ml and extracted wiht diclhoromethane (4 ⁇ 60ml). The combined organic phases are dried over MgSO and the solvent remove by evaporation to yield to the crude alkylated amide (6).
  • the steady-state was achieved within 3 min and measured for a few min.
  • the kinetic data (the steady-state velocity at various
  • the fibrin clotting assay was performed in 50 mM Tris HCl buffer (pH 7.52 at 37 °C) containing 0.1 M NaCl and 0.1% poly (ethylene glycol) 8000 with 9.0 ⁇ 10-10 M (0.1 NIH unit/mL) and 0.03 % (w/v) of the final concentrations of human thrombin and bovine fibrinogen, respectively, as reported elsewhere (Szewczuk et al., supra).
  • the clotting time was plotted against the inhibitor concentrations and the IC 50 was estimated as the inhibitor concentration required to double the clotting time relative to the control. Results are summarized in Tables 1 and 2 below.
  • the fibrin clot assay was performed essentially as
  • a serial dilution of the inhibitor was prepared in 50 mM tris HCl buffer (pH7.8 at 23 oC) containing 0. IM NaCl and 0.1% (w/v) polyethylene glycol 8000.
  • Human plasma 60uL, collected in 3.8% sodium citrate, blood/anticoagulant 9:1 was added to microtiter wells (microtiter plate, Falcon) containing 100 ⁇ L of various inhibitor dilutions. The solution was mixed after which 50 ⁇ L of human thrombin (InM final cone.) was added and mixed for 15 seconds. The turbidity of the clot was immediately monitored by
  • microplate autoreader (Dynateck MR 5000) at 405nm and recorded every 3 min. The maximal turbidity in the absence of inhibitors was reached within a 60 min. IC values were calculated at 30 minutes as the inhibitor concentration that gave half the optical density of the control.
  • Rat blood was collected into ACD (6/1 v/v) by cardiac puncture.
  • Suspensions of washed platelets were prepared as described by Ardlie et al, (Br. J. Haematol. 1970, 19:7 and Proc. Soc. Exp. Biol. Med., 1971, 136:1021).
  • the final suspending medium was a modified Tyrode solution (NaCl 138mM, KCl 2.9mM, HEPES 20mM, NaH.PO. 0.42mM, NaHCO 12mM, CaCl ImM, MgCl 2mM, 0.1% glucose, 0.35% albumin, apyrase l ⁇ L/mL pH 7.4). Platelet counts were adjusted to 5000,000/uL.
  • IC 50 values represent the concentration that was necessary to inhibit platelet aggregation or secretion to 50% of the control
  • the FeCl induced injury to the carotid artery in rats was induced according to the method described by Kurz, K.D., Main, R.W., Sandusky, G.E., Thrombosis Research 60; 269-280, 1990 and Schumacher, W.A. et al. J. Pharmacology and Experimental Therapeutics 267; 1237-1242, 1993.
  • Male, Sprague-Dawley rats ( 375-410 g) were anesthetized with urethane ( 1500 mg ⁇ kg ip). Animals were laid on a 37°C heating pad.
  • the carotid artery was exposed through a midline cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid sheath. Using forceps, the artery was lifted to provide sufficient clearance to insert two small pieces of polyethylene tubing (PE-205 ) underneath it.
  • a temperature probe PE-205
  • Inhibitor compounds were given as an iv bolus (mg/kg) followed immediately by an iv infusion ( ⁇ g/kg/min. via femoral vein) .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Pulmonology (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Peptides Or Proteins (AREA)

Abstract

This invention relates to the discovery of heterocyclic competitive inhibitors of the enzyme thrombin having formula (I), their preparation, and pharmaceutical compositions thereof. As well, this invention relates to the use of such compounds and compositions in vitro as anticoagulants and in vivo as agents for the treatment and prophylaxis of thrombotic disorders such as venous thrombosis, pulmonary embolism and arterial thrombosis resulting in acute ischemic events such as myocardial infarction or cerebral infarction. Moreover, these compounds and compositions have therapeutic utility for the prevention and treatment of coagulopathies associated with coronary bypass operations as well as restenotic events following transluminal angioplasty.

Description

LOW MOLECULAR WEIGHT BICYCLIC
THROMBIN INHIBITORS
FIELD OF THE INVENTION
This invention relates to compounds useful for the
treatment of thrombotic disorders, and more particularly to novel heterocyclic inhibitors of the enzyme thrombin.
BACKGROUND
Inordinate thrombus formation on blood vessel walls precipitates acute cardiovascular disease states that are the chief cause of death in economically developed
societies. Plasma proteins such as fibrinogen, proteases and cellular receptors participating in hemostasis have emerged as important factors that play a role in acute and chronic coronary disease as well as cerebral artery disease by contributing to the formation of thrombus or blood clots that effectively diminish normal blood flow and supply. Vascular aberrations stemming from primary pathologic states such as hypertension, rupture of
atherosclerotic plaques or denuded endothelium, activate cijcnemicai cascades that serve to respond and repair the injury site. Thrombin is a key regulatory enzyme in the coagulation cascade; it serves a pluralistic role as both a positive and negative feedback regulator. However, in pathologic conditions the former is amplified through catalytic activation of cofactors required for thrombin generation as well as activation of factor XIII necessary for fibrin cross-linking and stabilization.
In addition to its direct effect on hemostasis, thrombin exerts direct effects on diverse cell types that support and amplify pathogenesis of arterial thrombus disease. The enzyme is the strongest activator of platelets causing them to aggregate and release substances (eg. ADP TXA NE) that further propagate the thrombotic cycle. Platelets in a fibrin mesh comprise the principal framework of a white thrombus. Thrombin also exerts direct effects on
endothelial cells causing release of vasoconstrictor substances and translocation of adhesion molecules that become sites for attachment of immune cells. In addition, the enzyme causes mitogenesis of smooth muscle cells and proliferation of fibroblasts. From this analysis, it is apparent that inhibition of thrombin activity constitutes a viable therapeutic approach towards the attenuation of proliferative events associated with thrombosis.
The principal endogenous neutralizing factor for thrombin activity in mammals is antithrombin III (ATIII), a
circulating plasma macroglobulin having low affinity for the enzyme. Heparin exerts clinical efficacy in venous thrombosis by enhancing ATIII/thrombin binding through catalysis. However, heparin also catalyzes inhibition of other proteases in the coagulation cascade and its
efficacy in platelet-dependent thrombosis is largely reduced or abrogated due to inaccessibility of thrombus-bound enzyme. Adverse side effects such as
thrombocytopenia, osteoporosis and triglyceridemia have been observed following prolonged treatment with heparin.
Hirudin, derived from the glandular secretions of the leech hiridσ medicinalis is one of the high molecular weight natural anticoagulant protein inhibitors of
thrombin activity (Markwardt F. Cardiovascular Drug
Reviews, 10, 211, 1992). It is a biopharmaceutical that has demonstrated efficacy in experimental and clinical thrombosis. A potential drawback to the use of Hirudin as a therapeutic agent is likely antigenicity and lack of an effective method of neutralization, especially in view of its extremely tight binding characteristics toward
thrombin. The exceedingly high affinity for thrombin is unique and is attributed to a simultaneous interaction with the catalytic site as well as a distal "anion binding exosite" on the enzyme. Thrombin activity can also be abrogated by Hirudin-like molecules such as hirulog (Maraganore, J.M. et al.,
Biochemistry, 23, 7095, 1990) or hirutonin peptides
(DiMaio, J. et al., J. Med. Chem., 35, 3331, 1992). Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for
thrombin's catalytic site, thereby inhibiting proteolysis of that protein or other protein substrates such as the thrombin receptor. A common strategy for designing enzyme inhibitory compounds relies on mimicking the specificity inherent in the primary and secondary structure of the enzyme's natural substrate. Thus, Blomback et al . first designed a thrombin inhibitor that was modeled upon the partial sequence of the fibrinogen A(LBi)0c chain comprising its proteolytically susceptible region (Blomback, et al., J. Clin. Lab. Invest., 24 , 59, 1969). This region of fibrinogen minimally includes the residues commencing with phenylalanine: Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly
-Gly-Gly-Val-Arg-Gly-Pro-Arg
↑ scissile bond
Systematic replacement of amino acids within this region has led to optimization of the tripeptidyl inhibitory sequence exemplified by the peptide (D)-Phe-Pro-Arg which corresponds to interactions within the P -P -P local binding sites on
thrombin (Bajusz S. et al. in Peptides: Chemistry
Structure and Biology: Proceedings of the Fourth American Peptide Symposium, Walter R., Meienhofer J. Eds. Ann Arbor Science Publishers Inc., Ann Arbor MI, 1975, pp 603).
Bajusz et al. have also reported related compounds such as (D)Phe-Fro-Arg-(CO)H (GYKI-14166) and (D)MePhe-Pro-Arg-(CO)H (GYKI-14766) (Peptides-Synthesis, Structure and Function: Proceedings of the Seventh American Peptide Symposium, Rich, D.H. & Gross, E. eds., Pierce Chemical Company , 1981, pp. 417). These tripeptidyl aldehydes are effective thrombin inhibitors both in vi tro and in vivo . In the case of both GYKI-14166 and GYKI-14766, the
aldehyde group is presumed to contribute strongly to inhibitory activity in view of its chemical reactivity toward thrombin's catalytic Ser residue, generating a hemiacetal intermediate.
Related work in the area of thrombin inhibitory activity has exploited the basic recognition binding motif
engendered by the tripeptide (D)Phe-Pro-Arg while
incorporating various functional or reactive groups in the locus corresponding to the putative scissile bond (i.e. P1-P1').
In U.S. Patent 4,318,904, Shaw reports chloromethyl-ketones (PPACK) that are reactive towards Ser and His . These two residues comprise part of thrombin's catalytic triad (Bode, W. et al., EMBO Journal 8 , 3467, 1989) .
Other examples of thrombin inhibitors bearing the (D)Phe-Pro-Arg general motif are those incorporating COOH- terminal boroarginine variants such as boronic acids or boronates (Kettner, C. et al., J. Biol. Chem., 268, 4734, 1993). Still other congeners of this motif are those bearing phosphonates (Wang, C-L J., Tetrahedron Letters, ll, 7667, 1992) and α-Keto esters (Iwanowicz, E.J. et al., Bioorganic and Medicinal Chemistry Letters, 12, 1607, 1992).
Neises, B. et al. have described a trichloromethyl ketone thrombin inhibitor (MDL-73756) and Attenburger, J.M. et al . have revealed a related difluoro alkyl amide ketone (Tetrahedron Letters, 32, 7255, 1991).
Maraganore et al. (European 0,333,356; WO 91/02750; U.S. 5,196,404) disclose a series of thrombin inhibitors that incorporate the D-Phe-Pro- moiety and hypothesize that this preferred structure fits well within the groove adjacent to the active site of thrombin. Variations on these inhibitors are essentially linear or cyclic peptides built upon the D-Phe-Pro moiety.
Another series of patents and patent applications have described attempts to develop effective inhibitors against thrombosis by using alpha-ketoamides and peptide aldehyde analogs (EP 0333356;WO 93/15756; WO 93/22344; WO 94/08941; WO 94/17817).
Still others have focused their attention on peptides, peptide derivatives, peptidic alcohols, or cyclic peptide; as anti-thrombotic agents (WO 93/22344, EP 0276014; EP C341607; EP 0291982). Others have examined amidine sulfonic acid moieties to achieve this same end (U.S.
4,781,866), while yet others have examined para or meta substituted phenlyalanine derivatives (WO 92/08709; WO 92/6549) .
A series of Mitsubishi patents and patent applications have disclosed apparently effective argininamide compounds for use as antithrombotic agents. The chemical structures described in these documents represent variations of side groups on the argininamide compound (U.S. 4,173,630; U.S. 4,097,591; CA 1,131,621; U.S. 4,096,255; U.S. 4,046,876; U.S. 4,097,472; CA 2,114,153) .
Canadian patent applications 2,076,311 and 2,055,850 disclose cyclic imino derivatives that exhibit inhibitory effects on cellular aggregation.
Many of the examples cited above are convergent by
maintaining at least a linear acyclic tripeptidyl motif consisting of an arginyl unit whose basic side chain is required for interaction with a carboxylate group located at the base of the P specificity cleft in thrombin. Two adjacent hydrophobic groups provide additional binding through favourable Van der Waals interactions within a contiguous hydrophobic cleft on the enzyme surface
designated the P -P site.
One object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme, thrombin.
A further object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme thrombin and are provided for in s pnarmacologically acceptable state.
Still a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof as anticoagulant and thrombin inhibitory agents.
Yet a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof for therapeutic treatment of various thrombotic maladies . A further object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The enzyme inhibitors of the present invention are encompassed by the structure of general Formula I.
SUMMARY OF THE INVENTION
The present invention provides for novel compounds that display thrombin inhibitory activity as reflected in formula I :
Figure imgf000010_0001
wherein:
A is selected from (CH-R8) , S, SO, SO , O and NR wherein R8 is hydrogen, C alkyl optionally interupted with 1 or 2 heteroatoms; C aryl, C _ cycloalkyl or heterocyclic ring or a hydrophobic group;
B is selected from S, SO , O, -N= , NH, -CH= and CR R
wherein R6 and R7 are independently selected from
hydrogen and C alkyl provided that when A is Ξ, SO, SO , O, or NR , then B is CR R ;
D is selected from (CH-R5) wherein R is hydrogen, C
alkyl or -C(O)R ; and CH with a double bond to B when B is -N= or -CH=;
E is selected from CH and CH substituted with the -C ( O ) F. provided that only one of D and E is substituted with with -C (O)R ;
X is selected from O, N-R , or CH-R ;
Y is selected from O, S, SO, SO , N-R and CH-R provided that when X is N-R then Y is CH-R or O, and when X is O then Y is CH-R ;
Z is selected from O, S and H ;
R is a polar amino acid residuearginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle ; R2 is selected from H and C alkyl optionally substituted with C aryl, a 6 member heterocycle or a C _ cycloalkyl ring;
R3 is selected from H, NRR- and C alkyl; and
R4 and R5 are independently selected from H; NR,R, ; C, , aryl or C cycloalkyl optionally substituted with C. alkyl; C alkyl optionally interrupted by one or more
heteroatom or carbonyl group and optionally substituted with OH, SH, NRR or a C aryl, heterocycle or C cycloalkyl group optionally substituted with halogen, hydroxyl , C alkyl; an amino acid side chain; and a hydrophobic group.
As will be appreciated from the disclosure to follow, the molecules, compositions and methods of this invention are useful as anti-coagulants, or in the treatment and
prevention of various diseases attributed to the
undesirable effects of thrombin, as well as for diagnostic purposes .
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to molecules which inhibit the enzyme, thrombin. These molecules are characterized by a heterobicyclic moiety as illustrated in Formula I :
Figure imgf000011_0001
wherein X, Y, Z, A, B, D, E and R to R are as previously defined. The term "hydrophobic group" (HG) as used hereinafter, refers to any group which lacks affinity for, or displaces water. Hydrophobic groups include but are not limited to C alkyl, C alkenyl (e.g. vinyl, allyl) or C alkynyl (e.g. propargyl) optionally interrupted by a carbonyl group, (e.g. forming an acyl group); C aryl, C
cycloalkyl, C aralkyl , C cycloalkyl substituted C alkyl, wherein the aliphatic portion is optionally
interrupted by a carbonyl group (e.g. forming an acyl group) and the ring portion is optionally substituted with C alkyl such as methyl ethyl or t-butyl; or a hydrophobic amino acid side chain. Preferred hydrophobic groups include cyclohexyl, benzyl, benzoyl, phenylmethyl ,
phenethyl and para-t-butyl-phenylmethyl .
The term "arginyl moiety" represents an arginine amino acid residue or an analogue or derivative thereof. For example, an analogue or derivative of the natural residue may incorporate a longer or shorter methylene chain from the alpha carbon (i.e. ethylene or butylene chain);
replacement of the guanidino group with a hydrogen bond donating or accepting group (i.e. amino, amidino or methoxv; ; replacement of the methylene chain with a
ns trained group (i.e. an aryl, cycloalkyl or
Heterocyclic ring) ; elimination of the terminal carboxyl ,i.e. des-carboxy) or hydroxyl (i.e. an aldehyde) ; or a combination thereof . The term "alkyl" represents a straight or branched, saturated or unsaturated chain having a specified total number of carbon atoms.
The term "aromatic" or "aryl" represents an unsaturated carbocyclic ring(s) of 6 to 16 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NR R ) halogen or C alkyl. Aromatic rings include benzene, napththalene, phenanthrene and anthracene.
Preferred aromatic rings are benzene and naphthalene.
The term "cycloalkyl" represents a saturated carbocyclic ring of 3 to 7 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NR.R ) halogen or C alkyl. Cycloalkyl groups include cyclo- propyl , butyl, pentyl, hexyl and heptyl . A preferred cycloalkyl group is cyclohexyl .
The term "aralkyl" represents a substituent comprising an aryl moiety attached via an alkyl chain (e.g. benzyl, phenethyl) wherein the sum total of carbon atoms for the aryl moiety and the alkyl chain is as specified. The aryl or chain portion of the group is optionally mono- or disubstituted with OH, SH, amino (i.e. NR R ) halogen or C alkyl
The term "heteroatom" as used herein represents oxygen, nitrogen or sulfur (O, N or S) as well as sulfoxyl or sulfonyl (SO or SO ) unless otherwise indicated. It is understood that alkyl chains interrupted by one or more heteroatoms means that a carbon atom of the chain is replaced with a heteroatom having the appropriate valency Freferrably, an alkyl chain is interrupted by 0 to 4 heteroatoms and that two adjacent carbon atoms are not both replaced.
The term "heterocycle" represents a saturated or
unsaturated mono- or polycyclic (i.e. bicyclic) ring incorporating 1 or more (i.e. 1-4) heteroatoms selected from N, O and S. It is understood that a heterocycle is optionally mono- or di-substituted with OH, SH, amino (i.e. NR R- ) , halogen, CF , oxo or C alkyl. Examples of suitable monocyclic heterocycles include but are not limited to pyridine, piperidine, pyrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran and thiophene . Examples of suitable bicyclic heterocycles include but are not limited to indole, quinoline,
isoquinoline, purine, and carbazole. The term "hydrophobic amino acid" represents an amino acid residue that bears an alkyl or aryl group attached to the α-carbon atom. Thus glycine, which has no such group attached to the α-carbon atom is not a hydrophobic amino acid. The alkyl or aryl group can be substituted, provided that the substituent or substituents do not detract from the overall hydrophobic character of the amino acid.
Examples of hydrophobic amino acids include natural amino acid residues such as alanine; isoleucine; leucine;
phenylalanine; and non-naturally ocurring amino acids such as those described in "The Peptides", vol. 5, 1983,
Academic Press, Chapter 6 by D.C. Roberts and F.
Vellaccio. Suitable non-naturally ocurring amino acids include cyclohexylalanine and 1-aminocyclohexanecarboxylic.
Ey "amino acid side chain" is meant the substituent attached to the carbon which is α to the amino group. F;r example, the side chain of the amino acid alanine is a methyl group and while benzyl is the side chain for phenylalanine..
Preferably R2 is H or C alkyl. More preferably R is H methyl or ethyl and most preferably R is H. Preferably, R3 is H or C alkyl. More preferably, R is K methyl or ethyl, and most preferably R is H.
Preferably, one of R4 or R5 is a hydrophobic group such as a saturated or unsaturated carbocycle of 5 or 6 members optionally fused to another carbocyclic group while the other is H, C alkyl optionally substituted by NP. P. or carboxy. The hydrophobic moiety may be linked via a spacer such as a C . alkyl chain optionally interrupted with 1 or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO ) groups. More preferably, one of R4 and R5 is phenyl , cyclohexyl, indole, thienyl, quinoline,
tetrahydroisoquinoline, naphthyl or benzodioxolane linked via C alkyl optionally interupted with a heteroatom or a carbonyl while the other is H, carboxymethyl or
carboxyethyl .
Preferably, A is absent or CH .
Preferably, B is S or CH .
Preferably, D is CH .
Preferably, E is CH substituted with -C(O)R wherein R is as previously defined.
Preferably, X is CH-R or N-R .
Preferably, Y is CH-R or S.
Preferably, Z is O. In a preferred embodiment, R1 is represented by one of formula Via to VId:
Figure imgf000015_0001
wherein :
R11 is hydrogen or C alkyl;
K is a bond or -NH- ;
G is C alkoxy; cyano ; -NH ; -CH -NH ; -CINH.-NH; -NHC(NH)-NH; -CH -NH-C (NH) -NH ; a C cycloalkyl or aryl substituted with cyano, -NH , -CH -NH , -C(NH)-NH , -NH C(NH)-NH or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally
substituted with cyano, -NH_, -CH -NH , -C(NH)-NH , -NHC(NH)-NH or -CH -NH-C (NH) -NH ;
U is cyano, -NH , -C(NH)-NH or -NH-C (NH) -NH ;
P is a bond, -C(O)- or a bivalent group:
Figure imgf000016_0001
J is C alkylene optionally substituted with OH, NH and C alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1 ; and
T is H, OH, amino, a peptide chain, C alkyl, C alkoxy, C aralkyl, or heterocycle optionally substituted. Preferably R11 is H or methyl and most preferably H.
Preferably K is a bond.
Preferably G is -NH-C(NH)-NH attached via a methylene chain of 3-7 carbons or phenyl substituted with -C(NH)-NH attached via a methylene chain of 0 to 3 carbons. More preferably G -NH-C(NH)-NH attached via a methylene chain of 3 atoms .
Preferably P is -C(O)-.
Preferably J is selected from: -CH -S-CH -CH - ; -CH -O-CH CH - ; -CH -NH-CH -CH - ; and a bond when n is 0. More preferably, J is a bond while n is 0.
In particular embodiments of the invention, R is selected from the following amino acid derivatives prepared
according to the procedures described in Bioorg . Med .
Chem., 1995, 3:1145 :
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001
wherein n=l-6, nl=l-2, n2=0-7 and T is as previously defined .
In a preferred embodiment, T is a peptide of 1 to 4 amino acid residues in length and preferably fibrinogen' s A or B chain or fragment or derivative thereof . In another preferred embodiment, T is a heterocycle selected from the group consisting of:
Figure imgf000020_0002
wherein
X5 , X10, X11 and X12 are each independently selected from the group consisting of N, or C-X where X7 is hydrogen, C alkyl, or C aryl; X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X-, or CH-X. ;
R' is hydrogen, C _ alkyl optionally carboxyl substituted, carboxyl, -C . alkyl-CO -C alkyl, C aralkyl, C
cycloalkyl, aryl or an aromatic heterocycle.
Preferably T is selected from the group consisting of
Figure imgf000021_0001
wherein R' is as defined above.
More preferably T is selected from the group consisting
Of :
Figure imgf000021_0002
wherein R' is as defined above.
More preferably T is selected from the group consisting of:
Figure imgf000022_0001
wherein R' is as defined above.
Most preferably T is or
Figure imgf000022_0002
Figure imgf000022_0003
wherein R' is H or C alkyl such as methyl, ethyl, propyl or butyl and most preferably wherein R' is hydrogen, . In another embodiment, T is a 1,2 thiazole optionally
substituted with R' and\or is attached to J at the 2, 3, 4 or 5 position of the ring.
In particular embodiments, compounds of the invention are represented by formulas II, III, IV and V, wherein X, Y, B, R to R and R are as previously defined.
Figure imgf000022_0004
Figure imgf000022_0005
Figure imgf000022_0007
Figure imgf000022_0006
In a particularly preferred embodiment, compounds of the invention are represented by one of formulas VII, VIII, IX and X:
Figure imgf000023_0001
Figure imgf000023_0004
Figure imgf000023_0002
Figure imgf000023_0003
wherein
B is O, S, -CH-, or -NH- ;
Y is selected from O, S, SO, SO , N-R and CH-R„;
Rj is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or C alkyl;
R3 is selected from H, NR R7 and C alkyl; and
R4 and R5 are independently selected from H; NRR ; C aryl or C cycloalkyl optionally substituted with C alkyl C alkyl optionally interrupted by one or more
heteroatom or carbonyl group and optionally substitute:! with OH, SH, NRR, or a C aryl, heterocycle or C cycloalkyl group optionally substituted with halogen, hydroxyl , C . alkyl; an amino acid side chain; and a hydrophobic group;
Rβ is hydrogen, C . alkyl optionally interupted with 1 or 2 heteroatoms; C aryl, C, - cycloalkyl or heterocyclic ring or a hydrophobic group; and
n is 1 or 2.
Preferred compounds according to formula VII include: 0005 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a] pyridine-3R- carboxamido (propyl
ketoarginine)
Figure imgf000024_0001
0010 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a] pyridine-3R- carboxamido (butyl
ketoarginine)
Figure imgf000024_0002
0015 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a] pyridine-3R- carboxamido(propylcarbmethoxy ketoarginine)
Figure imgf000024_0003
0020 6S-cyclohexylmethyl
hexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido(benzylketo
arginine)
Figure imgf000024_0004
0G25 6S-cyclohexyl methyl
hexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine -3R- carboxamido(carbmethoxy
propyl
Figure imgf000024_0005
cyclodithioketalarginine)
0030 6S-cyclohexylmethyl
hexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido((S)-Arg-(R)- pipecolilic acid)
Figure imgf000024_0006
0035 6S-benzylhexa hydro-5-oxo-5H- thiazolo [3,2-a]pyridine-3R- carboxamido(carboxamidopropyl cyclodithioketal arginine)
Figure imgf000025_0006
0040 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R- carboxamido((S)-Arg
nipecotamide)
Figure imgf000025_0005
0045 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2a]pyridine-3R- carboxamido((S)Arg
isonipecotamide)
Figure imgf000025_0004
0050 6S-benzylhexahydro-5-oxo-5Hthiazolo[3,2-a]pyridine-3R- carboxamido(carboxamidopentyl cyclodithioketal arginine)
Figure imgf000025_0003
0055 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido(carbmethoxy
propyl cyclodithioketal
arginine)
Figure imgf000025_0002
0060 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido(1- carboxy-3-thiobutyl
ketoarginine)
Figure imgf000025_0001
0065 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido(1- carboxy-3-thiobutyl
ketoarginine)
Figure imgf000026_0001
0070 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyrιdme-3R-carboxamιdo(1- carboxy-2-methyl-3-thiobutyl ketoarginine)
Figure imgf000026_0002
0075 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido((3- thiobutyl sulfonic acid)
ketoarginine)
Figure imgf000026_0003
0080 6S-cyclohexylmethylhexahydro¬
5-oxo-5H-thiazolo[3,2- a]pyridine-3R- carboxamido(iso-quinolinium methyl ketoarginine)
Figure imgf000026_0004
0085 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R- carboxamido(propylcarbmethoxy ketoarginine)
Figure imgf000026_0005
0090 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2- a]pyridine-3R- carboxamido( (propylketo)Arg- Phe-Arg-NH )
Figure imgf000026_0006
0095 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido((propanoic acid) ketoarginine)
Figure imgf000027_0004
0100 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido(propyl
carbmethoxy ketoarginine)
Figure imgf000027_0003
0105 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo [3,2- a]pyridine-3R-carboxamido (α- benzothiazolo keto arginine); and
Figure imgf000027_0002
0110 6S-cyclohexylpropylhexahydro- 5-oxo-5H-thiazolo [3,2- a]pyridine-3R- carboxamido(propylcarbmethoxy ketoarginine)
Figure imgf000027_0001
0205 6-Benzyl-5-oxo-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [1- (benzothiazole-2- carbonyl)-4-guanidino
Figure imgf000028_0001
butyl]-amide
0210 6-Benzyl-5-oxo-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [1- (benzothiazole-2- carbonyl)-4-guanidino
Figure imgf000028_0002
butyl]-amide
0215 6-Benzyl-5-oxo-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [1- (benzothiazole-2- carbonyl)-4-guanidino
Figure imgf000028_0003
butyl]-amide
0220 6-Benzyl-8a-methyl-5-oxo- hexahydro-thiazolo[3,2- alpyridine-3-carboxylic
acid [1-(benzothiazole-2-
Figure imgf000028_0004
carbonyl)-4-guanidinobutyl]-amide 0225 8a-Methyl-5-oxo-6- phenethyl-hexahydrothiazolo[3,2-a]pyridine-3' carboxylic acid [1-
Figure imgf000029_0004
(benzothiazole-2- carbonyl)-4-guanidinobutyl]-amide
0230 8a-Methyl-5-oxo-6- phenethyl-hexahydrothiazolo|3,2-a]pyridine-3- carboxylic acid [1-
Figure imgf000029_0003
(benzothiazole-2- carbonyl)-4-guanidinobutyl]-amide
0240 8a-Methyl-5-oxo-6-(2- trifluoro methyl-quinolin- 6-ylmethyl)-hexahydrothiazolo[3,2-a] pyridine-3-
Figure imgf000029_0002
carboxylic acid [1- (benzothiazole-2- carbonyl)-4-guanidinobutyl]-amide
0245 6-Benzyl-5-oxo-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)butyl]-amide
Figure imgf000029_0001
0250 6-Benzyl-5-oxo-hexahydro- thiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)butyl]-amide
Figure imgf000030_0001
0255 6-Benzyl-5-oxo-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(1-methyl-1H imidazole-2-
Figure imgf000030_0002
carbonyl)butyl]-amide
0260 6-Benzyl-8a-methyl-5-oxo- hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic
acid [4-guanidino-1-
Figure imgf000030_0003
(thiazole-2-carbonyl)- butyl]-amide
0265 5-Oxo-6-(3-cyclohexylpropyl)-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid 14- guanidino-1-(thiazole-2-
Figure imgf000030_0004
carbonyl)butyl]-amide
0275 8a-Methyl-5-oxo-6-(3- phenyl-propyl)-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2-
Figure imgf000030_0005
carbonyl)-butyl]-amide 0280 8a-Methyl-5-oxo-6-(3- phenyl-propyl)-hexahydrothiazolo[3,2-a]pyridine-3- carboxylic acid [4- guanidino-1-(thiazole-2-
Figure imgf000031_0004
carbonyl)-butyl]-amide
0285 8a-Methyl-5-oxo-6-(2- trifluoromethyl-quinolin6-ylmethyl)-hexahydrothiazolo[3,2-a]pyridine-3-
Figure imgf000031_0003
carboxylic acid [4- guanidino-1-{thiazole-2- carbonyl)-butyl]-amide
0295 6-(1,3-Dioxo-1,3-dihydroisoindol-2-yl)-5-oxo- hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic
Figure imgf000031_0002
acid [4-guanidino-1- (thiazole-2-carbonyl)- butyl]-amide
0305 5-Oxo-6-(3-phenylpropionyl amino)-hexahydro thiazolo[3,2-a]pyridine-3- carboxylic acid [4-
Figure imgf000031_0001
guanidino-1-(thiazole-2- carbonyl)-butyl]-amide 0315 5-Oxo- 6 - ( 3 -phenylpropionyl amino ) -hexahydro
thiazolo[3 , 2 -a]pyridine-3 - carboxylic acid [4 -
Figure imgf000032_0001
guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
More preferred compounds according to formula (VII) include:
0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido(propylcarbo
methoxyketoarginine);
0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido((propylketo)Arg-Phe-Arg- NH);
0095 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine- 3R-carboxamido((propanoic acid) ketoarginine);
0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2- a]pyridine-3R-carboxamido (α-benzothiozolo keto arginine);
0210 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3- carboxylic acid [1-(benzothiazole-2-carbonyl)-4- guanidino-butyl]-amide;
0220 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-amide; 0240 8a-Methyl-5- oxo-6-(2-trifluoromethyl-quinolin-6-ylmethyl)- hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide; 0245 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3- carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)butyl]-amide; 0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole- 2-carbonyl)-butyl]-amide;
0265 5-Oxo-6-(3-cyclohexyl-propyl)-hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole- 2-carbonyl)butyl]-amide;
0285 8a-Methyl-5-oxo-6-(2-trifluoromethyl-quinolin-6- ylmethyl)-hexahydro-thiazolo[3,2-a]pyridine-3- carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)- butyl]-amide; and
0315 5-Oxo-6-(3-phenyl-propionylamino)-hexahydro
thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]-amide. Most preferred compounds according to formula VII include: 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2- a]pyridine-3R-carboxamido (propylcarbo methoxy
ketoarginine); and
0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2- a]pyridine-3R-carboxamido (α-benzothiozolo keto arginine).
Preferred compounds according to formula VIII include:
0325 3-Aminomethyl-2-benzoyl-4-oxo- octahydro-pyrrolo[1,2- a]pyridine-6-carboxylic acid
[1-(benzothiazole-2-carbonyl)- 4-guanidino-butyl]-amide
Figure imgf000034_0001
0330 3-Aminomethyl-4-oxo-2- phenylacetyl-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [1- (benzothiazole-2-carbonyl)-4-
Figure imgf000034_0002
guanidino-butyl]-amide
0335 2-Benzoyl-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000034_0003
butyl]-amide
0340 4-Oxo-2-(3-phenyl-propionyl)
octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid
[4-guanidino-1-(thiazole-2-
Figure imgf000034_0004
carbonyl)-butyl]-amide 0345 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid
[4-guanidino-1-(5-methyl
Figure imgf000035_0005
thiazole-2-carbonyl)-butyl]- amide
0350 2-(3-Cyclohexyl-propionyl)-4- oxo- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid
[4-guanidino-1-(2-thiazole
Figure imgf000035_0004
carbonyl)-butyl]-amide
0355 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid
[4-guanidino-1-(thiazole-2-
Figure imgf000035_0003
carbonyl)-butyl]-amide
0365 4-Oxo-2-(4-phenyl-butyryl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid
[4-guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000035_0002
0370 4-Oxo-2-phenylacetyloctahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid
[4-guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000035_0001
0375 2-(2-Amino-3-phenylpropionyl)-4-oxo-octahydropyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000036_0001
butyl]-amide
0380 2-[2-Amino-3-(4-hydroxyphenyl)-propionyl]-4-oxo- octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2-
Figure imgf000036_0002
carbonyl)-butyl]-amide
0385 2-[2-Amino-3-(4-fluorophenyl)-propionyl]-4-oxooctahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4-
Figure imgf000036_0003
guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
0390 4-Oxo-2-(3-phenyl-propyl)- octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2-
Figure imgf000036_0004
carbonyl)-butyl]-amide
0395 2-[2-Amino-3-(1H-indol-3-yl)- propionyl]-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000036_0005
butyl]-amide 0400 4-Oxo-2-(3-thiophen-3-yl- propionyl)-octahydropyrrolo[1 ,2-a] pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000037_0001
butyl]-amide
0405 4-Oxo-2-(3-thiophen-2-yl- propionyl)-octahydro- pyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000037_0002
butyl]-amide
0410 2-(3-1 H-Imidazol-4-yl- propionyl)-4-oxo-octahydro- pyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000037_0003
butyl]-amide
0415
2-(2-Amino-3-thiophen-3-yl- propionyl)-4-oxo-octahydropyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino-
Figure imgf000037_0004
1-(thiazole-2-carbonyl)- butyl]-amide
0420 4-Oxo-2-(1,2,3,4-tetrahydroisoquinoline-3-carbonyl)octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2-
Figure imgf000037_0005
carbonyl)-butyl]-amide 0425 2-(Hydroxy-phenyl-acetyl)-4- oxo-octahydro-pyrrolo[1,2-a] pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000038_0003
0430 2-(2-Hydroxy-3-phenylpropionyl)-4-oxo-octahydropyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000038_0002
butyl]-amide
0435 4-Oxo-2-phenoxyacetyloctahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000038_0001
0440 4-Oxo-2-(3-phenoxy-propionyl)- ocatahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000039_0001
0445 4-Oxo-2-(2-phenylethanesulfonyl)-octahydropyrrolo[1,2-a] pyrazine-6- carboxylic acid [4-guanidino-1- (thiazle-2-carbonyl)-butyl]-
Figure imgf000039_0002
amide
0450 2-(Naphthalene-2-sulfonyl)-4- oxo-octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000039_0003
0455 4-(6-[4-Guanidino-1-(thiazole-2- carbonyl)-butylcarbamoyl]-4-oxo- hexahydro-pyrrolo[1,2-a]
pyrazin-2yl)-4-oxo-3-(2 propyl
-pentanoylamino)-butyric acid
methyl ester
Figure imgf000039_0004
0460 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- guanidino-1)-butyl]-amide
Figure imgf000039_0005
0465 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [3- guanidino-propyl) -amide
Figure imgf000039_0006
0470 4-(6-[4-Guanidino-1-(thiazole-2- carbonyl)-butylcarbamoyl]-4-oxo- hexahydro-pyrrolo[1,2-a]pyrazin
2-yl)-4-oxo-butyric acid
Figure imgf000040_0006
0475 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (5-ethyl-thiazole-2-carbonyl)-
Figure imgf000040_0005
4-guanidino-butyl]-amide
0480 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(5-methyl-thiazole-
2-carbonyl)-butyl]-amide
Figure imgf000040_0004
0485 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(4-methyl-thiazole- 2-carbonyl)-butyl]-amide
Figure imgf000040_0003
0490 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid[1- (4-ethyl-thiazole-2-carbonyl)-
Figure imgf000040_0002
4-guanidino-butyl]-amide
0495 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (4- carbamimidoyl-pheny)-amide
Figure imgf000040_0001
0500 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(5-phenyl-thiazole- 2-carbonyl)-butyl]-amide
Figure imgf000041_0001
0505 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (5-benzyl-thiazole-2-carbonyl)- 4-guanidino-butyl]-amide
Figure imgf000041_0002
0510 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (4-carbarnimidoyl-benzyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
Figure imgf000041_0003
0515 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (3-carbamimidoyl-benzyl)-2-oxo- 2-thiazol-2-yl-ethyl]-amide
Figure imgf000041_0004
0520 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4- ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide
Figure imgf000041_0005
0525 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000041_0006
ethyl]-amide 0530 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-2- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000042_0005
ethyl]-amide
0535 [6-[4-Guanidino-1-(thiazole-2- carbonyl)-butylcarbamoyl]-4-oxo- 2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-3-yl]-acetic acid
Figure imgf000042_0004
0540 3-[6-[4-Guanidino-1-(thiazole-2- carbonyl)-butylcarbamoyl]-4-oxo- 2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazin- 3-yl]-propionic acid
Figure imgf000042_0003
0545 [6-[1-(1-Carbamimidoyl-piperin- 4-ylmethyl)-2-oxo-2-thiazol-2- yl-ethylcarbamoyl]-4-oxo-2-(3- phenyl-propionyl)-octahydro- pyrrolo[1,2-a]pyrazin-3-yl)-
Figure imgf000042_0002
acetic acid
0550 3-[6-[1-(1-Carbamimidoylpiperidin-4-ylmethyl)-2-oxo-2- thiazol-2-yl-ethylcarbamoyl]-4- oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazin- 3-yl)-acetic acid
Figure imgf000042_0001
0555 [6-[1-(1-Carbamimidoylpiperidin-3-ylmethyl)-2-oxo-2- thiazol-2-yl-ethylcarbamoyl]-4- oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazin-
Figure imgf000043_0001
3-yl)-acetic acid
0560 [6-(3-Guanidinopropylcarbamoyl)-4-oxo-2-(3- phenyl-propionyl)-octahydropyrrolo[1,2-a]pyrazin-3-yl)- acetic acid
Figure imgf000043_0002
0565 3-[6-(3-Guanidinopropylcarbamoyl)-4-oxo-2-(3- phenyl-propionyl)-octahydropyrrolo[1,2-a]pyrazin-3-yl)- propionic acid
Figure imgf000043_0003
0570 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazolc-2- carbonyl)-butyl]-methyl-amide
Figure imgf000043_0004
0575 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4- ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-methyl-amide
Figure imgf000043_0005
0580 [6-([1-Carbamimidoyl-piperidin- 4-ylmethyl)-2-oxo-2-thiazol-2- yl-ethyl]-methyl-carbamoyl)-4- oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazin- 3-yl]-acetic-acid
Figure imgf000044_0005
0585 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000044_0004
ethyl]-methyl-amide
0590 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (3- guanidino-propyl)-methyl-amide
Figure imgf000044_0003
0595 2-(Naphthalene-2-carbonyl)-4- oxo-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000044_0002
0600 2-(Naphthalene-1-carbonyl)-4- oxo-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000044_0001
0605 2-(3-Naphthalen-1-yl- proplonyl)4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]- amide
Figure imgf000045_0001
0610 2-(4-tert-Butyl-benzoyl)-4-oxo- octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000045_0002
0615 2-(Benzo[1,3]dioxole-5- carbonyl)-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]-
Figure imgf000045_0003
amide
0620 2-(3-Benzo[1,3jdioxol-5-yl- propionyl)-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]-
Figure imgf000045_0004
amide
0625 2-[2-(2-Methyl-benzylidene)-but- 3-enoyl]-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [1-(1- carbamimidoyl-piperidin-3-
Figure imgf000045_0005
ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide 0630 2-12-(2-Methyl-benzylidene)-but- 3-enoyl]-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [1-(1- carbamimidoyl-piperidin-4-
Figure imgf000046_0002
ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide
0635 2-(2-Benzylidene-pent-3-enoyl)-
4-oxo-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (3- guanidino-propyl)-amide
Figure imgf000046_0001
0640 4-Oxo-2-(3-phenyl-propionyl)octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid 4- carbamimidoyl-benzylamide
Figure imgf000047_0001
0645 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrauine- 6-carboxylic acid [4-imidazol-1- yl-1-(thiazole-2-carbonyl)-
Figure imgf000047_0002
butyl]-amide
0650 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-(2-aminoimidazol-1-yl)-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000047_0003
0655 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(2-amino-6- methyl-pyrimidin-4-yl)-1- (thiazole-2-carbonyl)-propyl]-
Figure imgf000047_0004
amide
0670 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(2-amino-6- chloro-pyrimidin-4-yl)-1- (thiazole-2-carbonyl)-propyl]-
Figure imgf000047_0005
amide 0675 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(6-amino- pyridin-2-yl)-1-(thiazole-2- carbonyl)-propyl]-amide
Figure imgf000048_0005
0680 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [3-(2-aminopyridin-4-yl)-1-(thiazole-2- carbonyl)-propyl]-amide
Figure imgf000048_0004
0685 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [2-(2-aminopyridin-4-yl)-1-(thiazole-2- carbonyl)-ethyl]-amide
Figure imgf000048_0003
0690 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine-
6-carboxylic acid [2-(6-aminopyridin-2-yl)-1-(thiazole-2- carbonyl)-ethyl]-amide
Figure imgf000048_0002
0695 2-[4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carbonyl]-3-(thiazole-2- carbonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamidine
Figure imgf000048_0001
0700 2-[4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carbonyl]-3-(thiazole-2- carbonyl)-1,2,3,4-tetrahydro
Figure imgf000049_0001
isoquinoline-7-carboxamidine
0705 N-[1-[4-Oxo-2-(3-phenylpropionyl)-octahydropyrrolo[1,2-a]pyrazine-6- carbonyl]-5-(thiazole-2- carbonyl)-pyrrolidin-3-yl]-
Figure imgf000049_0002
guanidine
0710 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4-aminocyclohexyl)-2-oxo-2-thiazol-2- yl-ethyl]-amide
Figure imgf000049_0003
0715 4-Oxo-2-(3-phenyl-propionyl)octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4-amino- cyclohexylmethyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide
Figure imgf000049_0004
0720 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4-aminobenzyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide
Figure imgf000049_0005
0725 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4- aminomethyl-benzyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide
Figure imgf000050_0005
0730 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(3- aminomethyl-benzyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide
Figure imgf000050_0004
0735 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid (2-oxo-1- piperidin-4-ylmethyl-2-thiazol- 2-yl-ethyl)-amide
Figure imgf000050_0003
0740 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid (2-oxo-1- piperidin-3-yl-2-thiazol-2-yl- ethyl)-amide
Figure imgf000050_0002
0745 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(3guanidino-cyclohexylmethyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
Figure imgf000050_0001
0750 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(4- guanidino-cyclohexylmethyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
Figure imgf000051_0001
0755 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(2- guanidino-cyclohexylmethyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
Figure imgf000051_0002
0760 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [1-(5-benzylthiazole-2-carbonyl)-4- guanidino-butyl]-amide
Figure imgf000051_0003
0765 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-guanidino1-(5-phenyl-thiazole-2- carbonyl)-butyl]-amide
Figure imgf000051_0004
0770 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrido[1,2-a]pyrazine- 6-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]- amide
Figure imgf000051_0005
0775 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid
[4-guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000051_0006
0780 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid [1-(4-carbamimidoyl-benzyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
Figure imgf000052_0001
0785 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid
[1-(3-carbamimidoyl-benzyl)-2- oxo-2-thiazo1-2-yl-ethyl]-amide
Figure imgf000052_0002
0790 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid
[1-(1-carbamimidoyl-piperidin-3- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000052_0003
ethyl]-amide
0795 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza- naphthalene-4-carboxylic acid [1-(1-carbamimidoyl-piperidin-4- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000052_0004
ethyl]-amide
0800 [4-[4-Guanidino-1-(thiazole-2- carbonyl)-butylcarbamoyl]-5-oxo- 7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalen-6-yl]-acetic acid
Figure imgf000052_0005
0805 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000053_0001
0810 3-[4-[4-Guanidino-1-(thiazoie-2- carbonyl-butylcarbamoyl]-5-oxo- 7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalen-6-yl]-propionic acid
Figure imgf000053_0002
0815 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diazanaphthalene-4-carboxylic acid
[3-guanidino-propyl]-amide
Figure imgf000053_0003
0820 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza- naphthalene-4-carboxylic acid
[1-(1-carbamimidoyl-piperidin-3- ylmethyl)-2-oxo-2-thiazol-2-yl-
Figure imgf000053_0004
ethyl]-amide
0825 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-guanidino- 1-(hydroxy-thiazol-2-yl-methyl)- butyl]-amide
Figure imgf000053_0005
0830 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine-6- carboxylic acid (4-guanidino-1- thiazol-2-ylmethyl-butyl)-amide
Figure imgf000054_0006
0835 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine-6- carboxylic acid (4-guanidino-1- thiazol-2-yl-butyl)-amide
Figure imgf000054_0005
0840 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-methoxy-1- (thiazole-2-carbonyl)-butyl]-
Figure imgf000054_0004
amide
0845 [6-[4-Methoxy-1-(thiazole-2- carbonyl)-butylcarbamoyl]-4-oxo- 2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazin- 3-yl]-acetic acid
Figure imgf000054_0003
085C [2-(5-Methoxy-2-([4-oxo-2-(3- phenyl-propionyl)-octahydro- pyrrolo[1,2-a]pyrazine-6- carbonyl]-amino)-pentanoyl)-
Figure imgf000054_0002
thiazol-5-yl]-acetic acid
0855 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-amino-1- (thiazole-2-carbonyl)-butyl]- amide
Figure imgf000054_0001
0860 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [5-amino-1- (thiazole-2-carbonyl)-pentyl]- amide
Figure imgf000055_0001
0865 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [5-guanidino-
1-(thiazole-2-carbonyl)-pentyl]- amide
Figure imgf000055_0002
0870 2-(3-Naphthalen-2-yl-propionyl)- 4-oxo-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000055_0003
0875 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-guanidino- 1-(1-methyl-1H-imidazole-2- carbonyl)-butyl]-amide
Figure imgf000055_0004
0880 4-Oxo-2-(3-phenyl-propionyl)-
Figure imgf000055_0005
octahydro-pyrrolo[1,2-alpyrazine- 6-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]amide
Figure imgf000055_0006
0885 8,8-Dimethyl-4-oxo-2-(3-phenylpropionyl)-octahydropyrrolo[1,2-a]pyrazine-6- carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]-
Figure imgf000056_0001
amide
Preferred compounds according to formula (VIII) include: 0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo[1,2- a]pyridine-6-carboxylic acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-amide
0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydropyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0515 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (1-(3-carbamimidoylbenzyl)-2-oxo-2-thiazol-2-yl-ethyl)-amide
0530 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (1-(1-carbamimidoylpiperidin-2-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]- amide
0545 [6-[1-(1-Carbamimidoyl-piperin-4-ylmethyl)-2-oxo-2- thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenylpropionyl)-octahydro-pyrrolo]1,2-a]pyrazin-3-yl)- acetic acid
0550 3-[6-[1-(1-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxo- 2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenylpropionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)- acetic acid
0555 [6-[1-(1-Carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2- thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenylpropionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)- acetic acid 0560 [6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenylpropionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)- acetic acid
0565 3-[6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenylpropionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)- propionic acid 0575 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(1-carbamimidoylpiperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]- methyl-amide 0580 [6- ((1-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2- thiazol-2-yl-ethyl]-methyl-carbamoyl)-4-oxo-2-(3- phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3- yl]-acetic-acid 0585 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(1-carbamimidoylpiperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]- methyl-amide 0590 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (3-guanidino-propyl)- methyl-amide
0595 2-(Naphthalene-2-carbonyl)-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6-carboxylic acid (4-guanidino- 1-(thiazole-2-carbonyl)-butyl]-amide 0625 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo- octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide
0630 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo- octahydro-pyrrolo[1 ,2-a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide
0635 2-(2-Benzylidene-pent-3-enoyl)-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6-carboxylic acid (3-guanidinopropyl)-amide 0625 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo- octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2- thiazol-2-yl-ethyl)-amide 0630 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxooctahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2- thiazol-2-yl-ethyl]-amide 0635 2-(2-Benzylidene-pent-3-enoyl)-4-oxo-octahydropyrrolo[1,2-a]pyrazine-6-carboxylic acid (3-guanidinopropyl)-amide
0645 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo]1,2- a]pyrazine-6-carboxylic acid [4-imidazol-1-yl-1- (thiazole-2-carbonyl)-butyl]-amide 0670 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid 13-(2-amino-6-chloropyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide 0675 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [3-(6-amino-pyridin-2- yl)-1-(thiazole-2-carbonyl)-propyl]-amide
0680 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (3-(2-amino-pyridin-4- yl)-1-(thiazole-2-carbonyl)-propyl]-amide
0685 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [2-(2-amino-pyridin-4- yl)-1-(thiazole-2-carbonyl)-ethyl]-amide
0690 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [2-(6-amino-pyridin-2- yl)-1-(thiazole-2-carbonyl)-ethyl]-amide
0695 2-[4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carbonyl]-3-(thiazole-2-carbonyl)- 1,2,3,4-tetrahydro-isoquinoline-6-carboxamidine 0700 2-[4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carbonyl]-3-(thiazole-2-carbonyl)- 1,2,3,4-tetrahydro-isoquinoline-7-carboxamidine
0705 N-[1-[4-Oxo-2-(3-phenyl-propionyl)-octahydropyrrolo[1,2-a]pyrazine-6-carbonyl]-5-(thiazole-2- carbonyl)-pyrrolidin-3-yl]-guanidine 0710 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-amino-cyclohexyl)- 2-oxo-2-thiazol-2-yl-ethyl]-amide 0730 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(3-aminomethylbenzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide
0745 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(3-guanidinocyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide
0755 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(2-guanidinocyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide
0795 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [1-(1- carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol- 2-yl-ethyl]-amide
0800 [4-[4-Guanidino-1-(thiazole-2-carbonyl)- butylcarbamoyl]-5-oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza-naphthalen-6-yl]-acetic acid
0810 3-[4-[4-Guanidino-1-(thiazole-2-carbonylbutylcarbamoyl]-5-oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza-naphthalen-6-yl]-propionic acid 0815 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [3-guanidinopropyl]-amide 0820 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [1-(1- carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol- 2-yl-ethyl]-amide 0830 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2a]pyrazine-6-carboxylic acid (4-guanidino-1-thiazol-2- ylmethyl-butyl)-amide
0835 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2a]pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2- yl-butyl)-amide
More preferred compounds according to formula VIII
include:
0335 2-Benzoyl-4-oxo-octahydro-pyrrolo[1,2-alpyrazine-6- carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)- butyl]-amide
0650 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4-(2-amino-imidazol-1- yl)-1-(thiazole-2-carbonyl)-butyl]-amide 0655 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [3-(2-amino-6-methyl- pyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide 0715 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-aminocyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0720 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-amino-benzyl)-2- oxo-2-thiazol-2-yl-ethyl]-amide
0725 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-aminomethylbenzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide
0735 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (2-oxo-1-piperidin-4- ylmethyl-2-thiazol-2-yl-ethyl)-amide
0740 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (2-oxo-1-piperidin-3-yl- 2-thiazol-2-yl-ethyl)-amide
0750 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(4-guanidinocyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0760 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2- carbonyl)-4-guanidino-butyl]-amide
0765 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (4-guanidino-1-(5-phenylthiazole-2-carbonyl)-butyl]-amide 0770 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrido[1,2- a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole- 2-carbonyl)-butyl]-amide 0775 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]-amide
0780 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [1-(4- carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]- amide
0785 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [1-(3- carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]- amide
0790 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [1-(1- carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol- 2-yl-ethyl[-amide
0805 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)-butyl]-amide
0825 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a)pyrazine-6-carboxylic acid [4-guanidino-1-(hydroxythiazol-2-y1-methyl)-butyl]-amide 0840 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4-methoxy-1-(thiazole-2- carbonyl)-butyl]-amide 0845 [6-(4-Methoxy-1-(thiazole-2-carbonyl)-butylcarbamoyl]- 4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazin-3-yl]-acetic acid
0850 [2-(5-Methoxy-2-([4-oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[1,2-a]pyrazine-6-carbonyl]-amino)- pentanoyl)-thiazol-5-yl]-acetic acid
0855 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [4-amino-1-(thiazole-2- carbonyl)-butyl]-amide
0860 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid [5-amino-1-(thiazole-2- carbonyl)-pentyl]-amide
0865 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (5-guanidino-1-(thiazole- 2-carbonyl)-pentyl]-amide Most preferred compounds according to formula VIII
include:
0345 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- a]pyrazine-6-carboxylic acid (4-guanidino-1-(5-methyl- thiazole-2-carbonyl)-butyl]-amide; and0340 4-Oxo-2- (3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine- 6-carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)-butyl]-amide.
Preferred compounds according to formula IX include:
0890 3-Amino-4-oxo-2-phenylhexahydro-pyrrolo[2,1- b][1,3]thiazine-6-carboxylic
acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-
Figure imgf000066_0001
amide
0895 3-Amino-2-benzyl-4-oxo- hexahydro-pyrrolo[2,1b][1,3]thiazine-6-carboxylic
acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]- amide
Figure imgf000066_0002
0900 3-Amino-2-cyclohexyl-4-oxo- hexahydro-pyrrolo(2,1- bll1,3]thiazine-6-carboxylic
acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]- amide
Figure imgf000066_0003
Preferred compounds according to formula X include
0905 7-Benzyl-6-oxo-octahydropyrido[2,1-c][1,4]thiazine-4- carboxylic acid [1- (benzothiazole-2-carbonyl)-4- guanidino-butyl]-amide
Figure imgf000067_0005
0910 7-(4-tert-Butyl-benzyl)-6-oxo- octahydro-pyrido[2,1- c][1,4]thiazine-4-carboxylic
acid [1-(benzothiazole-2- carbonyl)-4-guanidino-butyl]-
Figure imgf000067_0004
amide
0915 6-Oxo-octahydro-pyrido[2,1- c][1,4jthiazine-4-carboxylic
acid [4-guanidino-1-(thiazole- 2-carbonyl)-butyl]-amide
Figure imgf000067_0003
0925 7-Benzyl-6-oxo-octahydropyrido[2,1-c][1,4]thiazine-4- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)butyl]-
Figure imgf000067_0002
amide
0935 7-Benzyl-6-oxo-octahydropyrido(2,1-c][1,4]thiazine-4- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)butyl]-
Figure imgf000067_0001
amide 0940 6-Oxo-7-phenethyl-octahydro- pyrido[2,1-c][1,4jthiazine-4- carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-
Figure imgf000068_0004
butyl]-amide
0950 7-Benzyl-2,2,6-trioxo- octahydro-21>6_pyrido[2,1- c][1,4]thiazine-4-carboxylic
acid [4-guanidino-1-(thiazole-
Figure imgf000068_0003
2-carbonyl)-butyl]-amide
More preferred compounds according to formula X include: 925 7-Benzyl-6-oxo-octahydro-pyrido[2,1-c][1,4]thiazine-4- carboxylic acid [4-guanidino-1-(thiazole-2- carbonyl)butyl]-amide; and940 6-Oxo-7-phenethyloctahydro-pyrido[2,1-c][1,4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl)-amide .
Preferred compounds according to formula III include:
096C 4-oxo-1-(3-phenyl-propionyl)- octahydropyrrolo[1,2]pyrimidin
e-6-carboxylic acid[4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000068_0002
0965 4-oxo-1-(phenethylsulfonyl)- octahydropyrrolo[1,2]pyrimidin
e-6-carboxylic acid[4- guanidino-1-(thiazole-2- carbonyl)-butyl]-amide
Figure imgf000068_0001
For preparation of the compounds of formula (VII) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme is one particular method of
Figure imgf000070_0001
Step 1:
The alkylation of a is done with appropriate bases
according to the procedures described in Evans et al (J. Am. Chem. Soc., 1981, 103, 2127; ibid, 1982, 104, 1737; Aldrichimica Acta, 1982, 15, 23) to give b.
Step 2 :
Compound b upon hydroboration and oxidation following conditions available in the literature (Synthesis, 1980, 151) results in the aldehyde c.
Step 3 :
The formation of adduct e. from aldehyde c with d is done by stirring the reactant in aromatic solvents e.g. benzene or toluene in presence of catalytic amount of suitable acid e.g, p-toluenesulfonic acid.
Step 3':
The inter conversion of aldehyde c to aldehyde g is readily achieved by appropriate protection deprotection protocals found in T. Greene, Protective Groups In Organic Synthesis, (John Wiley & Sons, 1981).
Step 4 :
Tne cylization of adduct e to f may readily be achieved by appropriate Lewis acids e.g, trimethyl aluminum in suitable solvents e.g. dichloromethane, the methodology found in T. Greene, supra. Step 4':
Alternatively, the compound f can be derived from the treatment of aldehyde g with d in presence of suitable aromatic solvents e.g, benzene. Step 5:
The ester function (-C(O)O-R ) of the bicyclic
intermediate of formula f is then subjected to hydrolysation using an appropriate agent such as HCl in an appropriate solvent such as ethyl ether to yield to the free carboxylic acid . The resulting compound is then coupled to R1H with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to yield to a bicyclic coupled compound of formula (VIII). Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin,
Heidelberg, New York, Tokyo 1984; and The Peptides,
Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academic Press , New York, San
Francisco, London, 1979. For preparation of the compounds of formula (VIII) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme is one particular method of preparation.
Figure imgf000073_0001
wherein;
Pg is a nitrogen protecting group;
each of R20; and R21 is independently a C alkyl; and X, R1, R3, R4 and R5 are as previously defined.
The process in scheme 2 is briefly described as follows:
STEP 1:
The amino and carboxylic functions of the unsaturated compound of formula (a) are protected with appropriate protecting groups. A variety of protecting groups known for reactive functional groups and suitable protection and deprotection protocols may be found in T. Greene,
Protective Groups In Organic Synthesis. (John Wiley & Sons, 1981) . The appropriate protecting group to use in a particular synthetic scheme will depend on many factors, including the presence of other reactive functional groups and the reaction conditions desired for removal . The unsaturated compound of formula is easily obtained by methods and protocols known to chemist skill on the art. The protected unsaturated compound of formula (a) is subjected to appropriate conditions to allow cyclisation using an appropriate reagent such as mercuric acetate in an inert solvent such as tetrahydrofuran (THF) to yield to a protected amino alcohol of formula (b) .
STEP 2
The protected amino alcohol of formula (b) is oxidized using an appropriate oxidizing agent such as sulfur trioxide pyridine complex in an appropriate solvent such as diclhoromethane or dimethylformamide to yield to a protected amino aldehyde of formula (c) . Alternatively, intermediate (C) can be made by the ozonolysis of a compound of formula (a') prepared according to Collado et al, J. Org. Chem.,1995, 60:5011. STEP 3
The protected amino aldehyde of formula (c) is coupled with an amino acid alkyl ester of formula (d) by first forming the imine followed by contacting the obtained imine with an appropriate reagent such as sodium
triacetoxy borohydride NaBH(OAc) to yield to a cyclic intermediate of formula (e). STEP 4
The cyclic intermediate of formula (e) is functionalized at the amino position to yield to the amino substituted cyclic intermediate of formula (f). Conditions appropriate for such reactions are well known in the art and will depend on the nature of the R5 substituent.
STEP 5
The amino protecting group of the cyclic intermediate of formula (f) is removed under appropriate conditions and the resulting compound is then subjected to appropriate condition for internal ring closure such as low heat in an inert solvent or as a raw compound to yield to a bicyclic intermediate of formula (g). The bicyclic intermediate of formula (g) can also be obtained by hydrolysing the ester function (-C(O)O-R20) of the cyclic intermediate of formula (g) to the free carboxylic acid followed by standard peptide coupling using an appropriate coupling reagent such as benzotriazole-1-yloxy-tris- (dimethylamino)phosphonium hexafluorophosphate (BOP) in an inert solvent such as dimethyl formamide (DMF).
STEP 6
The ester function (-C(O)O-R21) of the bicyclic
intermediate of formula (g) is then subjected to
hydrolysation using an appropriate agent such as HCl in an appropriate solvent such as ethyl ether to yield to the free carboxylic acid. The resulting compound is then coupled to R1H with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to yield to a bicyclic coupled compound of formula (VIII). Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin,
Heidelberg, New York, Tokyo 1984; and The Peptides,
Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academic Press , New York, San
Francisco, London, 1979.
For preparation of the compounds of formula (IX) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme is one particular method of preparation.
SCHEME 3
Figure imgf000077_0001
wherein -.
Pg is a sulfur or amino protecting group;
L is a leaving group;
each of R20; and R21 is independently a C alkyl; and R1, R,
R4 and R5 are as previously defined.
The process depicted in scheme 3 is briefly described as follows :
STEP 1 :
The carboxylic acid compound (a) is coupled to the cyclic amine compound (b) with a peptide coupling agent such as benzotriazol-1-yloxy-tris- (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in an appropriate solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to yield to an amido compound of formula (c) . Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides,
Analysis. Synthesis, Biology, Vol, 1. edited by Gross E., and Meienhofer J., Academic Press , New York, San
Francisco, London, 1979.
STEP 2
The compound of formula (c) is subjected to appropriate conditions to allow internal cyclisation to yield to a bicyclic intermediate of formula (d). For example, acid mediated cyclisation using p-toluenesulfonic acid or TFA in an appropriate solvent such as dichloroethane.
STEP 3
The ester function (-C(O)O-R ) of the bicyclic
intermediate of formula (d) is subjected to hydrolysis using an appropriate agent such as lithium hydroxide
(LiOH) in an appropriate solvent such as tetrahydrofuran (THF) to yield to the free carboxylic acid. The resulting compound is then coupled to R H with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to give compound (e). Suitable conditions for peptide bond formation are well known in the art of peptide chemistry. For example see Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol. 1.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.
For preparation of the compounds of formula (X) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme 4 is one particular method of
preparation. SCHEME 4
Figure imgf000079_0001
wherein :
each of R20 and R21 is independently a C alkyl; and B, R1, R3, R4, and R5 are as previously defined. The process depicted in scheme 4 is briefly described as follows:
STEP 1 :
The halogenated compound of formula (a) is converted to a halomethyl ketone of formula (b) using an appropriate reagent, such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25°C to about 0°C. The resulting mixture is then treated under acidic
conditions to yield to the halomethyl ketone of formula (b).
STEP 2
The halomethyl ketone of formula (b) is coupled with an amino acid alkyl ester of formula (c) with an appropriate base such as sodium cyanoborohydride in an organic solvent such as methanol (MeOH) to yield to a cyclic intermediate of formula (d). STEP 3
The cyclic intermediate of formula (d) is treated under acidic conditions using an appropriate acid such as camphorsulfonic acid in an appropriate solvent such as toluene ti yield to a bicyclic intermediate of formula (e).
STEP 4
The ester function (-C(O)O-R20) of the bicyclic
intermediate of formula (e) is subjected to hydrolysation using an appropriate reagent such as LiOH to yield to the free carboxylic acid. The resulting compound is then coupled to R1H with a peptide coupling agent such as BOP in an appropriate solvent such as dimethylformamide to yield to a coupled bicyclic compound of formula (X).
Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Peptide Synthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.
Compounds of the present invention are further
characterized by their ability to inhibit the catalytic activity of thrombin, which is demonstrated in the assay as follows. Compounds of the present invention may be prepared for assay by dissolving them in buffer to give solutions ranging in concentrations from 1 to 100μM. In an assay to determine the inhibitory dissociation
constant, K, for a given compound, a chromogenic or fluorogenic substrate of thrombin would be added to a solution containing a test compound and thrombin; the resulting catalytic activity of the enzyme would be spectrophotometrically determined. This type of assay is well known to those skilled in the art.
The compounds of the present invention may be used as anti-coagulants in vi tro or ex vivo as in the case of contact activation with foreign thrombogenic surfaces such as is found in tubing used in extracorporeal shunts. The compounds of the invention may also be used to coat the surface of such thrombogenic conduits. To this end, the compounds of the invention are obtained as lyophilized powders, redissolved in isotonic saline and added in an amount sufficient to maintain blood in an anticoagulated state.
The therapeutic agents of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers. The proportion of each carrier is determined by the solubility and chemical nature of the compound, the route of administration, and standard pharmaceutical practice. For example, the compounds may be injected parenterally; this being intramuscularly, intravenously, or subcutaneously. For parenteral
administration, the compound may be used in the form of sterile solutions containing other solutes, for example, sufficient saline or glucose to make the solution
isotonic . The compounds may be administered orally in the form of tablets, capsules, or granules containing suitable excipients such as starch, lactose, white sugar and the like. The compounds may also be administered sublingually in the form of troches or lozenges in which each active ingredient is mixed with sugar or corn syrups, flavouring agents and dyes, and then dehydrated sufficiently to make the mixture suitable for pressing into solid form. The compounds may be administered orally in the form of solutions which may contain colouring and/ or flavouring agents .
Physicians will determine the dosage of the present therapeutic agents which will be most suitable. Dosages may vary with the mode of administration and the
particular compound chosen. In addition, the dosage may vary with the particular patient under treatment.
When the composition is administered orally, a larger quantity cf the active agent will typically be required to produce the same effect as caused with a smaller quantity-given parenterally.
To further assist in understanding the present invention, the following non-limiting examples of such thrombin inhibitory compounds are provided. The following
examples, of course, should not be construed as
specifically limiting the present invention, variations presently known or later developed, which would be within the purview of one skilled in the art and considered to fall within the scope of the present invention as
described herein. The preferred compounds as of the present invention are synthesized using conventional preparative steps and recovery methods known to those skilled in the art of organic and bio-organic synthesis, while providing a new a unique combination for the overall synthesis of each compound. Preferred synthetic routes for intermediates involved in the synthesis as well as the resulting anti-thrombotic compounds of the present
invention follow.
EXAMPLE 1
Figure imgf000084_0001
A solution of tert-butyloxycarbonyl-iodo-alanine-N,O- dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N- dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o- tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) was added followed by 4-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.1N), distilled H2O (3 × 50 mL), drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash
chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.
Figure imgf000084_0002
A solution of tert-butyloxycarbonyl-iodo-alanine-N,O- dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o- tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 3-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.1N), distilled HO (3 × 50 mL), drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash
chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.
Figure imgf000085_0001
A solution of tert-butyloxycarbonyl-iodo-alanine-N,O- dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N- dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o- tolylphosphine(palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.IN), distilled HO (3 × 50 mL), drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash
chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.
Figure imgf000086_0001
To a solution of tert-butyloxycarbonyl-para-cyanophenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture with warming The hydrogenation was continued until no starting material could be detected as judged by TLC . The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
Figure imgf000087_0001
To a solution of tert-butyloxycarbonyl-meta-cyanophenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
Figure imgf000087_0002
To a solution of tert-butyloxycarbonyl-ortho-cyanophenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
Figure imgf000088_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78 C, and the solution stirred. tert-Butyloxycarbonyl-para-amidino-phenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000089_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C, and the solution stirred. tert-Butyloxycarbonyl-meta-amidino-phenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel
(ethyl acetate/hexane), and concentrated under reduced
pressure .
Figure imgf000089_0002
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78 C, and the solution stirred. tert- Butyloxycarbonyl-ortho-amidino-phenylalanine-N,O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel
(ethyl acetate/hexane), and concentrated under reduced
pressure.
Figure imgf000090_0001
tert-Butyloxycarbonyl-para-cyano-phenylalanine-N,O- dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure toe yield a clear residue.
The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated unde: reduced pressure.
Figure imgf000091_0001
tert-Butyloxycarbonyl-meta-cyano-phenylalanine-N,O- dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H2 at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.
The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure.
Figure imgf000091_0002
tert-Butyloxycarbonyl-ortho-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg added. The solution was shaken under H at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.
The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure
Figure imgf000092_0002
tert-Butyloxycarbonyl-para-aminomethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified compound.
Figure imgf000092_0001
tert-Butyloxycarbonyl-meta-aminomethyl-phenylalanine-N,O- dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.! mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified compound.
Figure imgf000093_0001
tert-Butyloxycarbonyl-ortho-aminomethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified compound.
Figure imgf000093_0002
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C, and the solution stirred. The protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and
concentrated under reduced pressure.
r
Figure imgf000094_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C, and the solution stirred. The protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride ( 2. × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and
concentrated under reduced pressure.
Figure imgf000095_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C and the solution stirred. The protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000095_0002
A solution of tert-butyloxycarbonyl-iodo-alanine-N,O- dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3357-3404) in dry benzene (30 mL), and dry N,N- dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol) . The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.1N), distilled H2O (3 × 50 mL), drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash
chromatography over silica gel (light petroleum/ethyl acetate gradient) afforded the purified compound.
Figure imgf000096_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C, and the solution stirred. The amino acid-N,O-dimethylamide (1.07 g, 3.3 mmol) in anhydrous THF (15 mL ) was then added dropwise and the resulting mixture stirred.
The reaction was quenched with saturated aqueous ammonium chlcride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000097_0001
A solution of tert-butyloxycarbonyl-iodo-alanine-N,O-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404 ) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g) . The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.1N), distilled H2O (3 × 50 mL), drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash
chromatography over silica gel (light petroleum/ethyl acetate gradient, afforded the purified compound.
Figure imgf000097_0002
To a solution o f tert-butyloxycarbonyl - ( 4 -- cyano ) 3 ¬pyr idylalanine-N , O-dimethylamide ( 1 . 34 g , 4 . 0 mmo l ) in dry ethanol (20 mL) was added N,O-hydroxlyamine hydrochloride
(0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The
precipitate was filtered, washed with cold ethanol,
diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, and the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL . The solution was chilled overnight yielding the title compound.
Figure imgf000098_0001
To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous TKF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78º C, and the solution stirred. The amino acid-N,O-dimethylamide (1.16 g, 3.3 mmol) in anhydrous THF (15 mLi was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate ( 150 mL ) , and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel ethyl acetate/hexane), and concentrated under reduced pressure .
Figure imgf000099_0001
tert-Butyloxycarbonyl-3-(4-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (2 × 50 mL), brine (2 × 50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure to yield the title compound.
Figure imgf000099_0002
tert-Butyloxycarbonyl-3-(3-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(3-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (2 × 50 mL), brine (2 × 50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure to yield the title compound.
Figure imgf000100_0002
tert-Butyloxycarbonyl-3-(2-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (2 × 50 mL), brine (2 × 50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure to yield the title compound.
Figure imgf000100_0001
tert-Butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound.
Figure imgf000101_0001
tert-Butyloxycarbonyl-3-(3-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was
concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound.
Figure imgf000102_0001
tert-Butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was
concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound.
Figure imgf000102_0002
To a solution of thiazole in anhydrous THF (1.23 g, 14.4 mmol) was added n-BuLi (1.6 M/hexane, 8.4 mL, 13.4 mmol) dropwise at -78º C and the solution stirred. The guanidylated 4- piperidylalanine derivative (2.00 g, 3.2 mmol) in anhydrous THF (15 mL) was added dropwise , and the resulting mixture stirred . The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4 filtered, and concentrated under reduced pressure.
Figure imgf000103_0002
To a solution of thiazole in anhydrous THF (1.23 g, 14.4 mmol) was added n-BuLi (1.6 M/hexane, 8.4 mL, 13.4 mmol) dropwise at -78º C with stirring. The mixture was stirred at -78º C for 1 h. The guanidylated 3-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure.
Figure imgf000103_0001
To a solution of thiazole in anhydrous THF (1.23 g, 14.4 mmol) was added n-BuLi (1.6 M/hexane, 8.4 mL, 13.4 mmol) dropwise at -78 C with stirring. The mixture was stirred at -78º C for 1 h. The guanidylated 2-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4 , filtered, and concentrated under reduced pressure.
Figure imgf000104_0001
tert-Butyloxycarbonyl-para-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in HO (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 × 50 mL) and brine (3 × 50 mL). The solution was dried with MgSI filtered, and concentrated under reduced pressure yielding the title compound.
Figure imgf000104_0002
tert-Butyloxycarbonyl-meta-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in HO (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 × 50 mL), and brine (3 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure yielding the title compound.
Figure imgf000105_0002
tert-Butyloxycarbonyl-ortho-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in H2O (150 mL), and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 × 50 mL), and brine (3 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure yielding the title compound.
o
Figure imgf000105_0001
1. tert-Butyloxycarbonyl-3-(cis/trans-4-aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqvieous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 × 50 mL), 4 N aqueous sodium bisulfate (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure.
The semi-solid was chromatographed on silica gel (ethyl acetate/ hexane).
2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)
dropwise at -78º C and the solution stirred. The above
protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000107_0001
1. tert-Butyloxycarbonyl-3-(cis/ trans-3 - aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 × 50 mL), 4 N aqueous sodium bisulfate (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure.
The semi-solid was chromatographed on silica gel (ethyl acetate hexane).
2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)
dropwise at -78º C and the solution stirred. The above
protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000108_0001
1. tert-Butyloxycarbonyl-3-(cis/ trans-2-aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF
[60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF
(10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 × 50 mL), 4 N aqueous sodium bisulfate (2 × 50 mL), and brine (2 × 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure.
The semi-solid was chromatographed on silica gel (ethyl acetate' hexane).
2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)
dropwise at -78º C and the solution stirred. The above
protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000109_0001
1. tert-Butyloxycarbonyl-3-(cis/ trans-4 -aminocyclohexyl)alanine-N,O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring. The solution was cooled to 0 C, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified product.
2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -78º C and the solution stirred. The above
guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000110_0001
1. tert-Butyloxycarbonyl-3-(cis/ trans-3 -aminocyclohexyl)alanine-N,O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring.
The solution was cooled to 0 C, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate
(300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified product.
2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -78 ºC and the solution stirred. The above
guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.
Figure imgf000111_0001
1. tert-Butyloxycarbonyl-3-(cis/ trans-2 - aminocyclohexyl)alanine-N,O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring. The solution was cooled to 0 C, N,N'-bis-(benzyloxycarbonyl)- S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified product.
2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -78º C and the solution stirred. The above
guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL ; was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium
chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 × 50 mL), brine (50 mL), dried with MgSO4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. Example 2
Synthesis of Intermediates
Figure imgf000112_0001
(4S, 5R)-3-(1-oxo-3-phenylpropyl)-4-(phenyl)-5-(methyl)-2- oxazolidone (2). A solution of 10.0 g (1.0 equiv., 56.4 mmol) of (4S, 5R)-4-phenyl-5-methyl-2-oxazolidone (1) in 250 mL of dry THF, stirred at -78°C under argon, was treated dropwise with n-butyllithium (1.6 M in hexane, 1.1 eq., 38.8 mL). After stirring for 30 min., 8.4 mL (1.0 equiv., 56.4 mmol) of hydrocinnamoyl chloride was
introduced dropwise over a 10 minute period. The
resulting mixture was warmed to 0°C, stirred for an additional hour, and quenched with saturated ammonium chloride. The solvent was removed in vacuo and the resulting white solid dissolved in ethyl acetate and ddH2O . The aqueous phase was removed and extracted with two additional portions of ethyl acetate. The extracts were combined, washed with saturated sodium chloride, dried over sodium sulfate, and the solvent was removed ir. vacuo to afford a white crystalline solid (2) (in 91% yield). m. p.95-96.5; [α]D-35 c=1, CH2Cl2) ; [α]D-26.6 (c=1.01c CH2Cl2) .
1HNMR (CDCI3) δ 0.89(d, 3H, CH^, J=6.6 Hz) , 3.00-3.05 im, 2H), 3.26-3.34 (m, 2H), 4.73-4.78 (m, 1H), 5.64, d, 1H, J=7.4 Hz) , 7.22-7.46 (m, 10H) .
Figure imgf000113_0001
A solution of 5.0 g (1.00 equiv., 16.2 mmol) of (2) in 100 mL of dry THF, was cooled to -78°C. Enolization was achieved with 17.8 mL (1.1 equiv., 17.8 mmol) of lithium bis-trimethylsilylamide, which was added dropwise via syringe. The solution was stirred for 30 min . before 4.45 mL (3.0 equiv., 48.5 mmol) of allyl iodide was introduced and the reaction warmed to -15°C. After 1 h the reaction was quenched with saturated ammonium chloride and
extracted (3X) with ethyl acetate. The organic phase was washed with sodium metabisulfite, dried over sodium sulfate and the solvent removed in vacuo to afford an off- colour oil. Purification was achieved by flash
chromatography on silica gel using a stepwise gradient (15:1, 12-.1, 10:1) to yield (3), a colourless oil (95%).
[α]D 47.5 (c=3.12, CH2C l2). 1HNMR (CDCI3) δ 0.82 (d, 3H, CH3, J=6.6), 2.31-2.40 (m, 1H, RCH=CHCH2), 2.49-2.57 (m, 1H, RCH=CHCH2), 2.84-3.00 (m, 2H, Ph-CH2), 4.32-4.37 (m, 1H, CH-(N)CO), 4.53-4.58 (m, 1H, CH3-CH-), 5.03-5.13 (m, 2H, ABX, CH=CH2), 5.21 (d, 1H, Ph-CH, J=7.1 Hz), 5.81-5.89 (m, 1H, CH=CH2), 7.20-7.42 (m, 10H, ArH);
13C (CDCI3) δ 14.4, 36.2, 38.2, 43.9, 54.7, 78.4, 117.1, 125.4, 126.3, 128.2, 128.5, 129.0, 133.1, 134.8, 138.9, 152.4, 174.9.
Figure imgf000114_0001
A sample of the allyl compound, (3), (4.75 g, 13.6 mmol) in THF (100 mL) was treated with 13.6 mL (1.0 equiv., 13.6 mmol) of a 1.0 M solution of borane-tetrahydrofuran complex at 0°C and stirred for 2h. The solvent was evaporated and chloroform (100 mL) added via syringe.
Oxidation of the organoborane was achieved by the addition of 4.7 g (2.0 equiv., 27.2 mmol) of 3-chloroperoxybenzoic acid at 0°C, with warming to ambient temperature and stirring for an additional hour. The organic phase was washed with 5% Na2CO3 , ddH2O, and dried over sodium sulfate. Due to the instability of the alcohol, a quick column was performed to remove the extreme polar and nonpolar material which originated from the 3-chloroperoxybenzoic acid. The alcohol (4) was obtained in a yield of 65%.
[α]D 39.3 (c=1.038, CH2C l2).
1HNMR (CDCI3) δ 1.07 (d, 3H, CH3-CH, J=6.5 Hz), 1.81-1.93 (m, 3H, CH2-CH-H), 2.10-2.19 (m, 1H, CH2-CH-H), 3.10-3.17 (m, 2H, Ph-CH2), 3.873.90 (m, 2H, CH2OH), 4.43-4.49 (m, 1H, CH-CO), 4.70-4.75 (m, 1H, CH3-CH), 5.36 (d, 1H, Ph-CH, J=7.l Hz), 7.41-7.63 (m, 10H, ArH);
Figure imgf000115_0001
To a solution of the alcohol (4) (1.0 g, 2.7 mmol), dissolved in dichloromethane (27 mL), 876 mg (1.5 equiv., 4.1 mmol) of pyridinium chlorochromate and 1.0 g of 4 angstrom molecular sieves were introduced and the mixture changed from bright orange to a black colour. The reaction was monitored by TLC and after 30 min., if starting material remained, additional molecular sieves were added. The solution was filtered through celite and the solvent was evaporated. The residue was dissolved in ethyl acetate and washed with saturated sodium chloride . If the orange colour persisted in the organic phase, additional filterings through celite pads were performed. The aldehyde was obtained in a quantitative yield as a transparent, colourless oil (5).
1HNMR (CDCI3) δ 0.84 (d, 3H, CH3-CH, J=6.6Hz), 1.87-1.94
(m, 1H, CH2-CH(H) -CHO), 2.04-2.13 (m, 1H, CH2-CH(H)-CHO),
2.45-2.50 (m, 2H, Ph-CH2), 2.79-2.85 (dd, 1H, CH2-CHO,
J=13.3 & J=6.6), 2.92-2.99 (dd, 1H, CH2-CHO, J=13.2 &
J=8.8), 4.19-4.22 (m, 1H, CH-CO), 4.46-4.51 (m, 1H, CH3- CH), 5.13-5.25 (m, 1H, Ph-CH), 7.20-7.39 (m, 10H, ArH), 9.69 (s, 1H, CHO ) ;
13C (CDCI3) δ 14.2, 23.8, 39.0, 41.2, 43.8, 54.9, 78.6,
125.3, 126.4, 128.2, 128 4, 128.5, 128.9, 132.8, 138.4,
152.4, 174.9, 201.1.
Figure imgf000116_0001
The aldehyde, (5), (2.6 g, 7.10 mmol) was dissolved in benzene (70 mL) and a catalytic amount of ptoluenesulfonic acid was added, followed by 1.58 g (1.2 equiv., 8.52 mmol) of L-cysteine ethyl ester and 4 A molecular sieves . The reaction was allowed to stir overnight at ambient temperature followed be removal of solvent in vacuo . The residue was dissolved in
chloroform, washed with saturated sodium chloride, ddH2O, and dried over sodium sulfate. The solvent was removed in vacuo to afford a gummy solid (6).
Figure imgf000116_0002
2.0M trimethylaluminum in hexane (2.4mL, 4.8mmol, 3 equivalents) was added slowly to starting material (6) (800mg, 1.61mmol) stirring in anhydrous dichloromethane under argon, using ovendried equipment. After stirring overnight, HPLC indicated that the reaction had gone to completion. The mixture was quenched with excess methanol, then filtered on a short silica gel column (washing through
with excess 10% methanol in ethylacetate . Evaporation
gave 784 mg of crude material that was purified using 2:1 hexane : EtOAc on a silica gel column affording 258mg
(0.δlmmol being a 50% yield) of pure compound (7), a 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-ethyl ester as a white/yellow solid. 1H NMR (CDCl3) d 1.28-1.31 (m, 3H), 1.72-1.81 (m, 3H),
2.10-2.13 (m, 1H), 2.66 (dd, 1H, J= 11.5 and 6.0 Hz),
3.29-3.34 (m, 2H), 4.19-4.29 (m, 2H), 4.88 (dd, 1H, J= 9.0 and 5.0 Hz), 5.22 (dd, 1H, J= 8.0 and 6.0 Hz), 7.18-7.23
(m, 3H), 7.28-7.31 (m, 2H) .
Figure imgf000117_0001
LiOH.H2O (48mg, 1.12mmol) in 10mL of water was added to starting material (7) (240mg, 0.76mmol) dissolved in 10mL :f dioxane . After 1 hour, TLC in 1:1 Hexane:EtOAc showed no starting material. The reaction was quenched with 10% citricacid, then extracted twice with dichloromethane. Drying and evaporating the combined organic layers gave 354mg of crude product. This was redissolved in dichloromethane, then precipitated by adding excess hexane. The product was filtered to give 200mg (0.68mmol being a 90% yield)) of an off-white solid, (8), also known as 6S-benzylhexahydro-5-oxo- 5H-thiazolo[3,2-a]pyridine-3R-carboxylic acid. 1 H NMR (CD3OD) d 1.71-1.82 (m, 3H), 2.12-2.17 (m, 1H), 2.67 (dd, 1H, J= 14 and 11 Hz), 2.77-2.81 (m, 1H), 3.30-3.40 (m, 3H), 4.81 (dd, 1H, J= 8.5 and 4.9 Hz), 5.16 (t, 1H, J= 7.5 Hz), 7.18-7.31 (m, 5H).
Figure imgf000118_0001
Oxalyl chloride (9) (25 g, 0.197 mol) was cooled to 0ºC and cyclohexane propionic acid (20 ml, 0.14 mol) was added. This was left to stir overnight. The resultant mixture was distilled to give an 84% yield of the
colorless liquid (10), cyclohexyl propionic acid chloride
Figure imgf000118_0002
The chiral auxiliary (11) (13.6g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78ºC. Then n-BuLi \52.8 mL, 84.4 mmol, 1.2 equiv.) was added and left for 2 mins (dark orange solution). The acid chloride (10) (13.4 g, 76.6 mmol, 1 eq) was then added and left to stir overnight. Work-up was done by quenching with saturated NH4Cl extracting with ethyl acetate, washing the extracts with water and brine, drying over sodium sulphate and concentration. A fast column, with dry loading, (6:1 hexane ethyl acetate) was run to purify the product. This afforded in a white solid (12) which was recrystallized from ether and hexane to give the title compound in 78% yield.
[α]D=-20.1(c=1,EtOH); MP/BP mp=90.5-91.5oC
1H NMR (CDCl3) d 0.86-1.10 (m, 5H), 1.18-1.30 (m, 4H), 1.54-1.75 (m, 7H), 2.86-2.97 (m, 2HO, 4.70-4.76 (m, 1H), 5.65 (d, 1H, J= 7.2 Hz) , 7.28-7.42 (m, 5H) .
Figure imgf000119_0001
The starting material (12)(9.13g, 29 mmol, leq) was dissolved in dry THF and cooled to -78ºC, after which LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise over 40 mins. Then, 30 minutes later, allyl bromide (7.5 mL, 86.9 mmol, 3 eq) was added slowly over 10 mins. The mixture was left to warm overnight. Work-up included quenching with sat. ammonium chloride, extraction with ethyl acetate, washing with 10% sodium thiosulphate, decolourising with charcoal, drying over sodium sulphate and concentration in vacuo. The product was obtained as a yellow oil (13) in 96% yield.
[α]D=+9.5 (c=1.0, EtOH)
H NMR (CDC13) d 0.92-1.10 (m, 5H), 1.10-1.39 (m, 5H), 1.63-1.75 (m, 6H), 2.27-2.42 (m, 2H), 4.01-4.14 (m, 1H) 4.76-4.85 (m, 1H), 5.00-5.07 (m, 2H0 , 5.65 (d, 1H, J= 7 Hz), 5.64-5.88 (m, 1HO , 7.27-7.46 (m, 5H) .
Figure imgf000120_0001
2-methyl-2-butene was added dropwise to borane
dimethylsulphide complex at -12ºC. The reaction was maintained at this temperature for 15 minutes and then it was warmed to 0ºC, after which it was stirred for 2 hours. The disiamyl borane was then added to a mixture of the starting material 13 in THF using a double-ended needle at 0ºC. The mixture was then stirred for 2 hours after which the solvents were removed and the residue dissolved in dichloromethane. It was carefully added to a suspension of pyridinium chlorochromate in dichloromethane contained in a flask equipped with a reflux condenser. After the initial exothermic reaction had subsided, the mixture was refluxed at 50ºC for 1 hour. The dark brown liquid was dissolved m ethyl acetate and filtered through Florisil. The black residue of PCC was extracted with ethyl acetate and also filtered through the same Florisil pad.
Concentration of the filtrates resulted in a 78% yield of a yellow gummy product (14).
[α]D=-17.8(c=1.245,EtOH) 1H NMR (CDCl3) d 0.89-1.18 (m, 5H), 1.20-1.47 (m, 8H),
1.60-1.74 (m, 6H), 1.83-2.00 (m, 1H), 2.48-2.53 (m, 2H), 3.90-4.10 (m, 1HO, 4.12-4.16 (m, 1H), 4.76-4.80 (m, 1HO, 5.67 (d, 1H, J= 7Hz), 7.27-7.46 (m, 5H), 9.77 (s, 1H).
Figure imgf000121_0002
The resulting aldehyde (14) (7.7g crude, 20.8 mmol 1 equiv.) was dissolved in 75 mL of toluene. To the
solution was added a catalytic amount of p- toluenesulphonic acid (50 mg), 10 g of 4A molecular sieves, and L-cysteine ethyl ester (3.87 g, 20.8 mmol, 1 equiv.). The mixture was stirred overnight, filtered and concentrated. The residue was then purified by silica gel chromatography (6:1 hexane: ethyl acetate) afforded 6.36 g of the product (15) in 61% yield.
[α]D=-48.3(c=1.095,EtOH)
Η NMR (CDC13) d 0.84-0.98 (m, 4H), 1.11-1.38 (m, 7H), 1.50-1.90 (m, 10H), 2.80-2.99 (m, 1H), 3.24-3.34 (m, 1H) 3.77-4.29 (m, 4H) , 4.46-4.81 (m, 2H), 5.66 (d, 1H, J=
7Hz), 7.27-7.46 (m, 5H) .
Figure imgf000121_0001
The starting material (15) (1.97 g, 3.9 mmol, 1 equiv.) was dissolved in 20 mL of dry dichloromethane and cooled to 0ºC. Trimethylaluminum (5.9 mL, 11.8 mmol, 3 equiv.), was added dropwise and the mixture was left stirring overnight. After complete reaction as evidenced by HPLC, methanol was added until a yellow t>olid mass was formed. Dichloromethane was added to dissolve the solid and the whole mixture was stirred for 15-30 minutes and then filtered. The residue after concentration in vacuo was run through a quick column (6:1 hexane: ethyl acetate), to remove auxilary and as many of the polar decomposition products as possible, affording in a 50% yield of a yellow oil (16). 1H NMR (CDCl3) d 0.83-0.98 (m, 2H), 1.09-1.38 (m, 10H), 1.57-2.00 (m, 11H), 2.12-2.18 (m, 1H), 2.49-2.54 ( m, 1H), 3.10 (dd, 1H, J= 11 and 6 Hz), 3.27 (dd, 1H, J= 11.5 and 8.0 Hz), 4.11-4.25 (m, 2H), 4.88 (dd, 1H, J= 11.0 and 5.o Hz), 5.14 (dd, 1H, J= 10 and 6 Hz).
Figure imgf000122_0001
The starting material (16) (0.95 g, 2.9 mmol, 1 equiv.) was dissolved in 10 mL of dioxane. The solution was cooled to 10ºC, and to it was added LiOH H2O(0.123 g, 2.9 mmol, 1 eq.) dissolved in 10 mL of water. The bath was removed and the mixture was stirred at room temperature for 1 hour. TLC showed complete reaction and the solvent was evaporated under vacuum. The remaining aqueous layer was washed with ether (2X), acidified with 10% citric acid, and extracted with dichloromethane (3X). The combined extracts were dried over sodium sulphate and concentrated to give a white solid which was
recrystallized from ether. Concentration of the filtrate and purification by silica gel column chromatography (2:1 hexane: ethyl acetate) resulted in more product (17) with a m.p. of 198.2-199C.
1H NMR (DMSO-d6) d 0.78-0,93 (m, 2H), 1.11-1.27 (m, 5H), 1.34-1.36 (m, 1H), 1.51-1.56 (m, 1H), 1.60-1.75 (m, 1H), 1.82-1.87 (m, 1H), 2.15-2.18 (m, 1H), 2.37-2.41 (m, 1H),
3.03 (dd, 1H, J= 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.82 (dd, 1H, J= 9 and 4 Hz), 4.95 (dd, 1H, J= 8 and 5.5 Hz).
Figure imgf000123_0001
BOC-DiCbz Arg (18) (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0ºC. Triethylamine (2. mL) was added followed by 14.5 mmol of a 1M toluene solution of isopropyl chloroformate via a syringe. The reaction was allowed to stir at 0ºC for 30 minutes then quickly filtered. The white solid was discarded. To the filtrate was bubbled freshly prepared diazomethane until the color of the solution turned yellow. The reaction mixture was allowed to stand overnight in a well
ventilated fumehood which facilitated the discharge of excess diazomethane. Dry ether was added to precipitate the diazoketone. The product was filtered and dried under vacuum to give light yellow fluffy solid (4.6 g, 58%). Diazoketone (19) (lg, 1.77 mmol) was dissolved in THF (20 mL) and to this solution was added 1M HCl in ether (20 mL) at 0ºC. The reaction was allowed to stir at ambient temperature overnight during which time a white
precipitate was formed. Further precipitation was achieved by adding ether. Filtration and drying the solid afforded the product (20) (1.02 g , 100%). 1 H NMR (DMSO-d6) d 1.65-1.77 (m, 3H), 2.06-2.50 (m, 1H), 3.86-3.90 (m, 2H), 4.29 (m, 1H), 4.76 (d, 1H, J= 18Hz),
4.95 (d, 1H, J= 18 Hz), 7.35 (s, 2H), 7.36 (s, 2H), 7.35- 7.41 (m, 10H), 8.71 (br s, 3H), 10.1 (br s, 2H). 13 C NMR (DMSO-d6) d 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3, 128.6, 128.7, 128.8, 128.9, 135.2, 135.9, 153.4, 157.4, 198.9.
Figure imgf000124_0001
To mimetic (17) (0.422 g, 1.42 mmol) in THF (50 mL ) at 0ºC and in presence of N-methyl morpholine (0.19 mL), was slowly added 1M toluene solution of isopropylchloroformate (1.71 mL) . The reaction was allowed to stir at 0ºC for 30 minutes then treated with aminochloromethylketone (20) in small portions. Once the addition was complete the reaction was further stirred for 15 minutes followed by addition of N-methyl morpholine (0.19 mL). The reaction was stirred at ambient temperature for 3 hours, then extracted with ethyl acetate followed by washing with brine and 10% aqueous citric acid. Removal of organic solvent gave a white foam (21) (1.03 g, 96%) which was used further without purification.
Η NMR (CDCl3) d 0.07-0.97 (m, 1H), 1.15-1.41 (m, 7H), 1.62-1.91 (m, 10H), 2.10-2.16 (m, 1H), 2.43-2.48 (m, 1H)
2.74-2.80 (m, 1H), 3.01-3.07 (m, 1H), 3.87-3.94 (m, 1H),
4.11-4.19 (m, 2H), 4.60-4.66 (m, 1H), 4.74-4.86 (m, 2H),
5.09-5.24 (m, 4H), 7.30-7.39 (m, 10H), 7.95 (d, 1H, J= 8 Hz), 9.4 (br s, 1H), 9.56 (br s, 1H).
EXAMPLE 3
Figure imgf000126_0001
(N-t-BOC-N-tosyl)butyrylketoarginine (240mg, 0.515 mmol), was deprotected using 30% TFA in dichloromethane. The deprotected arginine derivative, was coupled with the mimetic (8) (100 mg, 0.343 mmol) in DMF under basic
conditions (Et.N, pH = 8-9), using BOP reagent (228 mg, 0.52 mmol) as the dehydrating agent. The reaction was typically complete within 2-4 hours. Extraction with ethyl acetate followed by successive washing with brine and 10% aqueous citric acid yielded the crude product. The crude product was purified by column chromatography affording IF; mg (76%) of pure product. This product was then treated with HF to remove the tosyl group. Purification of the isolated deprotected product by HPLC afforded BCH-2737.
EXAMPLE 4
Figure imgf000126_0002
Chloromethylketone (21) (0.188 g, 0.245 mmol) was
dissolved in THF (10 mL) treated with NMM (0.036 mL) followed by mercapto acetic acid (0.02 mL, 0.299 mmol). The reaction was stirred at ambient temperature overnight. Extraction of the reaction mixture with ethyl acetate followed by successive washing with brine and 10 % aqueous citric acid and evaporation of organic solvent gave the crude product which was purified by column chromatography to give foamy solid as the product (0.125 g, 62%).
This protected precursor (0.125 g, 0.154 mmol) was
dissolved in DCM (5 mL) and cooled to -78ºC. A 1M DCM solution of BBr (1.54 mL, 1.54 mmol) was slowly added. The reaction was stirred at ambient temperature for 5 hours, then cooled to -78ºC again and treated with anhydrous methanol (2mL). The reaction was brought to room
temperature and stirred for 2 additional hours. The
solvents were removed under reduced pressure and the residue was partitioned between ether and water. The water layer was collected, lyophilized and the final product (23) obtained as a powder after HPLC purification and
lyophilization. The products of the reactions described above can be isolated in the free form or in the form of salts. In addition, the products can be obtained as pharmaceutically acceptable acid addition salts by reacting one of the free bases with an acid. In a similar manner, the product can be obtained as pharmaceutically acceptable salts by
reacting one of the free carboxylic acids with a base. Likewise, treatment of the salts with a base or acid results in a regeneration of the free amide. EXAMPLE 5
A general method of synthesizing compound of formula II or
III:
Figure imgf000128_0001
EXAMPLE 6
Synthes is of
Figure imgf000129_0001
STEP 1
Synthesis of 2-Benzyloxycarbonylamino-4-hydroxybutyric acid tert-butyl ester
Figure imgf000129_0002
T: a solution of the protected aspartic acid (1) (Bachem, 2.50 g, 4.95 mmols) in 50 mL of dry tetrahydrofuran
(THF), at -10°C, under N2, was added N-methylmorpholine
(109 μL, 0.2 eq) and isopropyl chloroformate (1.0
M: toluene : 384 μL, 1.1 eq). The soluion was stirred at -10°C for 60 min. In another flask, NaBH4 (375 mg, 2 eq) was suspended in a dry 5:1 mixture of THF/MeOH (50 mL), at -78°C, under N2. This suspension was stirred at -78°C for 30 min. The mixted anhydride solution was then added to the NaBH4 suspension dropwise via canula , and the final solution was stirred at -78°C for 3 hr. Acetic acid (2.8 mL, 10 eq) was then added and the solution was warmed to r.t. (30 min) . The solvents were evaporated, the residue taken up in EtOAc and washed with sat.aq. NaHCO3 (2x) and brine. The organic layer was dried over MgSO4 , the solids were filtered and the solvent
evaporated to give 1.53 g (4.95 mmols, 100%) of the alcohol (2) as a clear oil.
1H NMR (CDC13, 400 MHz) : δ 7.40-7.31 (m, 5H, ArH), 5.63 (d, 1H, J=7.3, NH), 5.13 (AB system, 2H, J=12.2, CH2Ph), 4.43 (m, 1H, H-2), 3.69 (m, 2H, H-4 ), 2.17 (m, 1H, H-3), 1.63 (m, 1H, H-3), 1.48 (s, 9H, t-Bu).
STEP 2
2-Benzyloxycarbonylamino-4-iodobutyric acid tert-butyl ester
Figure imgf000130_0001
To a solution of the alcohol (2) (1.53 g, 4.95 mmols) in a 1:1 mixture of CH3CN/Et2O (50 mL), at -10ºC, under N2, were added successively imidazole (607 mg, 1.8 eq) and Ph,P (2.21 g, 1.7 eq). Iodine (2.14 g, 1.7 eq) was then added in small portions over a period of 15 min. After the addition was completed, a white precipitate formed and the solution was brown. It was stirred at -10°C for 45 min. It was then poured in Et2O and the organic phase was washed with sat.aq. Na:SO3, sat.aq. CuSO4, H2O and dried over MgSO4. The solids were filtered and the solvent evaporated to give a yellow oil that was purified by flash chromatography (silica gel, 5% to 20% EtOAc/ Hex). The iodide (3) was obtained in 83% yield (1.71 g) as a clear oil. 1H NMR (CDCI3, 400 MHz) : δ 7.41-7.31 (m, 5H, ArH), 5.35
(bd, 1H, J=7.3, NH), 5.13 (s, 2H, CH2Ph), 4.30 (m, 1H, H- 2), 3.22-3.12 (m, 2H, H-4), 2.42 (m, 1H, H-3), 2.20 (m, 1H, H-3), 1.48 (s, 9H, t-Bu).
STEP 3
Synthesis of 2-Benzyloxycarbonylamino-4-hexenoic acid tert-butyl ester
Figure imgf000131_0001
To a suspension of Cul (2.27 g, 5 eq) in dry THF (20 mL), at -78°C, under N2, was added slowly a 1.0M solution in THF of vinyl magnesium bromide (23.4 mL, 9.8 eq). The solution was then warmed up to -10°C for 30 min (it turned then black) and cooled back to -78°C. A solution of the iodide (3) (1.00g, 2.39 mmols) in dry THF (3.5 mL) was then added slowly to the cuprate solution. The reaction mixture was stirred at -78°C for 2.5 hr .
Sat.aq. NH4Cl (50 mL) was added and the mixture was brought back to room temp, with vigorous stirring. It was then poured in Et:O and stirred for 5 min. The dark suspension was filtered through a cintered funnel and the phases were separated. The aqueous phase was extracted with Et2O (2x) and the combined organic extracts were dried over MgSO4. The solids were filtered, the solvents evaporated and the crude oil purified by flash
chromatography (silica gel, 5% AcOEt/Hex) to give 0.51 g (67%) of the pure alkene (4). 1H NMR (CDCI3, 400 MHz) : δ 7.37-7.31 (m, 5H, ArH), 5.80 (m, 1H, H-5) , 5.33 (d, 1H, J=7.8, NH), 5.12 (s, 2H, CH2Ph), 5.05 (d, 1H, J=17.2, H-6) , 5.01 (d, 1H, J=10.4, H-6) , 4.30 (q, 1H, J=7.4, H-2) , 2.16-2.08 (m, 2H, H-4) , 1.92 (m, 1H, H-3) , 1.74 (m, 1H, H-3) , 1.48 (s, 9H, t-Bu)
STEP 4
Synthesis of 1-Benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinecarboxylic acid tert-butyl ester
Figure imgf000132_0001
To a solution of the alkene (4) (50 mg, 0.157 mmol) in dry THF (3.1 mL), at r.t., under N2, was added mercuric acetate (75 mg, 1.5 eq). The solution was stirred at r.t. for 18 hr after which it was cooled down to 0°C. Sat.aq. NaHCO3 (2 mL) was then added and the mixture was stirred at 0ºC for 30 min. KBr (0.11g, 6 eq) was added and the mixture was stirred at r.t. for 2 hr. It was then poured in H2O/Et2O and the phases were separated. The aqueous phase was extracted with Et2O (2x) and the combined organic extracts were dried over MgSO4. The solids were filtered and the solvents evaporated. Oxygen (O2) was bubbled into a suspension of NaBH4 (3.3 mg, 0.55 eq) in dry DMF (0.4 mL) for 1 hr, and to this was added dropwise (syringe pump, 3 mL/hr) a solution of the organomercurial bromide in DMF (3.1 mL) with continuous introduction of O2. The bubbling was continued for 1 hr and Et2O (5 mL) was added. The grey suspension was filtered through Celite and the filtrate was evaporated.
The residue was chromatographed (silica gel, 6:4
Hex/EtOAc) to give the pyrrolidinol (5) (30 mg, 57%) as a clear oil.
1H NMR (CDCl3, 400 MHz) : δ 7.37-7.28 (m, 5H, ArH), 5.22-5.09 (m, 2H, CH2Ph), 4.30 (dd, 1H, J=1.4, 8.3, H-2) , 4.24 (m, 1H, H-5) , 3.70-3.57 (m, 3H, CH2-OH), 2.25 (m, 1H), 2.13 (m, 1H), 1.92 (m, 1H), 1.70 (m, 1H), 1.34 (s, 9H, t-Bu) .
STEP 5
Synthesis of 1-Benzyloxycarbonyl-5-carboxy-2-pyrrolidinecarboxylic acid tert-butyl ester
Figure imgf000133_0001
To a solution of the alcohol (5) (50 mg, 0.149 mmol) and Et3N (62μL, 3 eq) in dry CH2Cl2 (0.8 mL) is added slowly, under N2, at 0°C, a solution of SO3-Pyridine complex (71 mg, 3 eq) in dry DMSO . The solution was stirred at 0ºC for 30 min and 10% citric acid (2 mL) is added. The pH is brought to 4 with 1M NaOH and the aqueous phase is extracted with Et2O (3x). The combined organic extract: were dried over MgSO4. The solids were filtered and the solvents evaporated to give a crude oil which was
purified by flash chromatography (silica gel, 7:3
Hex/EtOAc). The pure aldehyde (6) was obtained as a clear oil (45 mg, 90%). 1H NMR (CDCI3, 400 MHz) : δ 9.68 + 9.56 (ds, 1H, CHO) ,
7.36-7.29 (m, 5H, ArH) , 5.23-5.11 (m, 2H, CH2Ph), 4.57- 4.39 (m, 2H, H-2, H-5) , 2.30-1.97 (m, 4H, H-3, H-4) , 1.47 + 1.36 (2s, 9H, t-Bu) .
STEP 6
Figure imgf000134_0001
The pyrrolidine-aldehyde (6) is coupled with the protected diamino-propionic acid (7) by first forming the imine (8) (MgSO4, CH2Cl2). Isolation of the imine (8) is done by filtration of the MgSO^ and evaporation of the solvent. The crude imine is then treated with NaBH(OAc) and actic acid (AcOH) in THF for 15 hours to obtain the amine (8) after extrative work-up . STEP 7
Figure imgf000134_0002
The CB7 (7) protecting group of the amine (8) is removed by hydrogenation with palladium on charcoal 10% as a catalyst in methanol (MeOH). The catalyst is filtered and the MeOH evaporated to give the crude diamine (9) that can be used without any purification.
STEP 8
Figure imgf000135_0001
The cyclisation is done by heating the crude oil (9) from step 7, neat slightly above the boiling point of methanol. The bicyclic lactam (10) is purified by flash
chromatography.
STEP 9
Figure imgf000136_0001
The secondary amine of the bicyclic lactam (10) is
protected as an amide using benzoyl chloride in pyridine Evaporation of the pyridine and extractive work-up give the bicyclic lactam-amide (11).
STEP 10
Figure imgf000136_0002
The BOC and t-butyl ester protecting groups of bicyclic lactam amide (11) are removed under acidic conditions (HCl m ethyl ether (Et 0)). The amine salt (12) precipitates out of solution and is collected by filtration.
STEP 11
Figure imgf000137_0001
The primary amine of compound (12) is protected with a CBZ group by reacting it with benzyl chloroformate in
acetonitrile (CH2CN) with K2CO as a base. Extrative work-up gives fully protected carboxylic acid (13) which can be use for step 12 without further purification..
STEP 12
Figure imgf000137_0002
The carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF using BOP as the coupling agent in the presence of diisopropylethylamine (EtNiPr ).
Extraction with ethyl acetate (EtOAC) gives compound (15 as a solid which is purified by chromatography. STEP 13
Figure imgf000138_0001
The two CBZ(Z) protecting groups of compound (15) are removed by catalytic hydrogenation with Pd/C 10% as a catalyst. The catalyst is filtered and the solvent is evaporated to give the amino-guanidine (16).
EXAMPLE 7
Figure imgf000139_0001
STEP 1
Figure imgf000139_0002
4-methylmorpholine (NMM) was added to a solution of the carboxylic acid (2) (1.7g, 4.9mmol, 1.0eq), 4- hydroxyproline (3) (5.39mmol, 1.1eq), and BOP reagent (2.17g, 4.9mmol, 1.0eq) in anhydrous DMF (10mL) at room temperature. The reaction mixture was stirred at room tenperature over night, quenched with brine (50mL) and ethyl acetate (100mL). The organic layer washed with aqueous citric acid (10%, 2×50mL), sodium bicarbonate (10%, 2x50mL) and brine (50mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered and the solvent evaporated. The crude residue was purified by flash chromatography (5:4:1, ethyl acetate-hexane-methanol). 1. lg of pure product (4) was recovered 48% yield.
STEP 2
Figure imgf000140_0001
To a solution of 4-hydroxyproline derivative (4 ) (115mg 240umol, 1.0eq) in dichloromethane (10 mL, anhydrous) at 0°C is added triethylamine (72mg, 720umol, 3.0eq) and methanesulfonyl chloride (28mg, 240umol, 1.0eq) and the reaction mixture is stirred at room temperature. The mixture is then quenched with an aqueous solution of ammonium chloride and extracted with ethyl acetate. The organic layer is washed with 10% citric acid and brine, dried, filtered and the solvent is evaporated to dryness yielding compound (5). STEP 3
Figure imgf000141_0001
The enamine (5) (1.0eq) is treated with mercuric acetate (1.1 eq) in THF. The solvent is evaporated to dryness and the residue dissolved in methanol. The resulting organomercurial is reductivly cleaved with sodium borohydride (1.3eq). The resulting crude lactam thioether is purified by flash chromatography on silica gel affording compound (6).
STEP 4
Figure imgf000141_0002
To a solution of the lactam thioether (6) (1.0eq) in dry dichloromethane N-chlorosuccinimide (1.0eq) is added at 0°C. The reaction mixture is warmed to room temperature. When the reaction shows no more starting material the solid is filtered and the solvent evaporated to dryness . The crude material (7) is use without any further
purification for step 5. STEP 5
Figure imgf000142_0002
To a solution of the alpha-chlorothioether (7) (1.0eq) in THF (anhydrous) a solution of phenylcuprate (1.0eq)
(prepared according to litterature procedure) is added at low temperature. When the reaction mixture shows no starting chlorothioether, brine and ethyl acetate is added. The organic layer is dried, filtered and evaporated to dryness to afford the desired product (8) .
STEP 6
Figure imgf000142_0001
The isolated bicyclic lactam (8) is hydrolysed with one equivalent of lithium hydroxyde in a 1:1 mixture of THF and water. The mixture is stirred at room temperature for 1 hour. The crude mixture is extracted with ether and the resulting solution is poured into 10% citric acic aqueous solution and extracted with dichloromethane to yield the corresponding carboxylic acid (9). STEP 6
Figure imgf000143_0001
The crude carboxylic acid (9) is coupled with
benzythiazole keto arginine in DMF using BOP as the coupling reagent in the presence of diisopropylethylamme . Extraction with EtOAc gives a solid that is purified on silica gel to give the protected amide. The CBZ protecting group is removed with BBr in dichloromethane at room temperature finally gives the bicyclic benzothiazole keto arginine inhibitors (10).
The following compounds are produced accordingly with the execption that the appropriate substitution of products were made in order to obtain the final compounds.
Figure imgf000143_0002
Figure imgf000144_0002
EXAMPLE 8
STEP 1
Figure imgf000144_0001
Commercially available glutaric acid monomethyl ester chloride (1) (20 ml, 0.144 mol) was disolved in 40 ml of dry tetrahydrofuran (THF) and cooled to -15°C. Excess diazomethane freshly prepared in 300 ml of Ether was introduced via cannula at -15°C to the solution. The mixture was left to warm up to room temperature overnight Excess diazomethane was evacuated from the flask with a current of argon. To bring the reaction to completion, 75 ml of 1 N HCl in Ether was added at 0°C and left to warm up to room temperature for 5 hours. The volume of the solvent was reduced and then washed with 2x 5% NaHCO3 dried over Na2CO3 and evaporated to give crude
chloromethylketone (20.46 g , 79%) wich was used in the next step without further purification. 1H NMR (CDCL3, 400 MHz) d 1.16-1.2 (t,1H), 1.83-1.9
(m,2H), 2.27-2.35 (m,2H), 2.6-2.64 ( t, 1H),
3.6 (s,3H), 4.04 (s,2H).
STEP 2
Figure imgf000145_0001
Crude chloromethylketone (2) (10.04 g, 56.15 mmol) was disolved in 300 ml of dry MeOH. Sodium acetate (2 eq, 9.21 g, 112.3 mmol) was added followed by L-Cysteine ethyl ester hydrochloride salt (1.3 eq, 13.55g, 72.98 mmol) and sodium cyanoborohydride (1.4 eq, 4.9 g, 78.59 mmol). The heterogeneous mixture was left to stir at room temperature for 2h30 min. 200 ml of methanol (MeOH) was then added to disolve all the solid and the pH was brought to 2 with 1N HCl . The mixture was then basified with saturated NaHCO3 until pH= 8. MeOH was evaporated and the remaining aqueous solvent was washed with ethyl acetate and dichloromethane. Solvents were combined, dried over Na2SO4 and evaporated. The crude residue was purified by silica gel flash column chromatography using a gradient. of eluents ethyl acetate / hexane in the following ratios: (3:7, 5:5, 6:4, 7:3) giving cyclic compound (3 ) .
J-H NMR (CDCI3 , 400MHz) of compound (3) d 1.21-1.27 (t, 3H, J=7.06 Hz), 1.41-1.48 (m, 2H), 1.65-1.73 (m, 2H), 2.28-2.39 (m, 4H), 2.57-2.63 (t, 1H, J=10.9), 2.722.76 (dd, 1H, J=10.7 Hz), 2.8-2 86 (m, 1H), 3.6-3.64 (d, 4H, .J=2.55 Hz), 3.63 (s, 3H), 4.13-4.2 (m, 2H) 13C NMR (CDCI3, 400 MHz) 13.078, 19.888, 28.326,
31.133, 32.741, 35.277, 50.462, 56.394, 59.149, 60.188, 69.713, 170.182, 172.52
STEP 4
Figure imgf000146_0001
Cyclic compound (3) (913mg, 3.32 mmol) was disolved in 5' ml of dry Toluene. ( IS) - (+ ) -10-Camphorsulfonic acid ( 91 mg, 0.39 mmol) was added and the mixture was left to reflux for 4 days. When all starting material was shown to be consumed (by TLC), the mixture was worked up byevaporation of solvent, dissolving residue in ethyl acetate and washing with 2 x 5 % NaHCO3. The Ethyl acetate layer was dried over Na2SO4 and evaporated. The crude residue was purified by silica gel flash column chromatography using 60 % ETOAC / 40 %Hexane followed by 70 % ETOAC / 30 % Hexane giving 62.5% of Bicyclic compound (4).
1H NMR (CDCI3, 400MHz) of compound (4) d 1.27-1.31 (t, 3H, J=7 Hz), 1.5-1.6 (m, 1H), 1.72-1.87 (m, 2H), 2.02- 2.1 (m, 1H), 2.33-2.46 (m, 2H), 2.52-2.59 (m, 2H), 2.83-2.88 (dd, 1H,J=14 , 4 Hz), 3.14-3.18 (d, 1H), 3.78-3.85 (m, 1H), 4.2-4.27 (q, 2H, J= 3.9 Hz), 5.9- 5.92 (t, 1H, J=3.4 Hz).
STEP 5
Figure imgf000147_0001
Bicyclic (4) (366mg,1.5 mmol) was dissolved in 25 ml of THF and 5 ml H2O Lihium hydroxide, monohydrate (1.1 eq, 7.05 mg, 1.68 mmol) was added in 2.3 ml of H2O, at 0°C and the mixture was left to stirr at 0°C for 1 hr and at room temperature for 3 hrs. THF was then evaporated and the remaining aqueous mixture was acidified by addition of Citric acid until pH=2. Extraction of aqueous mixture with 2 × CH2CL2 and 2 × ETOAC, drying of combined organic layers with Na2SO4 and evaporation gave a crude residue wich was purified by silica gel flash column chromatography using 70 % ETOAC / 30 %Hexane followed by 4.7 % HOAC / Ethyl acetate giving the pure acid (5) in 54 % yield . 16 % of starting material (4) was recovered.
1H NMR (MeOD, 400 MHz) of compound (5) d 1.57-1.69 (m, 1H), 1.70-1.80 (m, 1H), 1.81-1.89 (m, 1H), 2.05-2.12 (m, 1H), 2.35-2.5 (m, 2H), 2.51-2.66 (m, 2H), 2.86- 2.91 (dd, 1H, J=13.8, 4 Hz) , 3.12-3.17 (d, 1H) , 3.3- 3.32 (m, 1H) , 3.78-3.84 (m, 1H) , 5.76-5.78(t, 1H, J=3.53 Hz) . 13C NMR (MeOD, 400 MHz) d 17.052, 27.07, 28.928,
31.382, 32.096, 51.016, 55.138, 170,088, 171.24
STEP 6
Figure imgf000148_0001
To a solution of lithium bis (trimethylsilyl) amide (5ml of 1M THF solution, 5mmol) in THF (10 ml) is added at -78°C a solution of the carboxylic acid (5) (500mg, 2.32 mmol). the resulting solution is stirred at -78°C for 1 hour.
Benzyl bromide (0.26 ml, 2.22 mmol) is then added and the mixture is allowed to reach room temperature and stirred for 15 hours. The mixture is then poured into 10% HCl (50 ml and extracted wiht diclhoromethane (4× 60ml). The combined organic phases are dried over MgSO and the solvent remove by evaporation to yield to the crude alkylated amide (6).
STEP 7
Figure imgf000149_0002
The crude aklylated amide (6) is coupled with
benzythiazole keto arginine in DMF using BOP as the coupling reagent in the presence of diisopropylethylamme . Extraction with EtOAc gives a solid that is purified on silica gel to give the protected amide. The CBZ protecting group is removed with BBr in dichloromethane at room temperature finally gives the bicyclic benzothiazole keto arginine inhibitors (7).
The following compound is produced accordingly with the exeption that the appropriate substitution of products were made in order to obtain the final compounds .
COMPOUND #8
Figure imgf000149_0001
EXAMPLE 9
Determination of K Values for Heterocyclics
The affinity cf inhibitors for thrombin was measured
according to the procedures described in (DiMaio et al , J. Bio. Chem., 1990, 265:21698) Inhibition of amidolytic activity of human thrombin was measured fluorometrically using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50 mM Tris-HCl buffer (pH 7.52 at 37°C) containing 0.1 M NaCl and 0.1% poly (ethylene glycol) 8000 at room temperature, and (Szewczuk et al . , Biochemistry, 1992 31:9132). The hydrolysis of the substrate by thrombin was monitored on a Varian-Cary 2000™ spectrophotometer in the
fluorescence mode (λeX = 383 nm, λem = 455 nm) or on a Hitachi F2000™ fluorescence spectrophotometer (λer = 383 nm, λer = 455 nm), and the fluorescent intensity was calibrated using AMC . The reaction reached a steady-state within 3 minutes after mixing thrombin with the substrate and an inhibitor. The steady-state velocity was then measured for a few minutes . The compounds of this invention were also pre- incubated with thrombin for 20 minutes at room
temperature before adding the substrate. The steady-state was achieved within 3 min and measured for a few min. The kinetic data (the steady-state velocity at various
concentrations of the substrate and the inhibitors) of the competitive inhibition was analyzed using the methods described by Segel (1975). A non-linear regression program, RNLIN in the IMSL library (IMSL, 1987), LMDER in MINPACK library (More et al., 1980) or Microsoft™ Excell™ , was used to estimate the kinetic parameters (Kn Vn, and K ) . dTT assay The fibrin clotting assay was performed in 50 mM Tris HCl buffer (pH 7.52 at 37 °C) containing 0.1 M NaCl and 0.1% poly (ethylene glycol) 8000 with 9.0 × 10-10 M (0.1 NIH unit/mL) and 0.03 % (w/v) of the final concentrations of human thrombin and bovine fibrinogen, respectively, as reported elsewhere (Szewczuk et al., supra). The clotting time was plotted against the inhibitor concentrations and the IC50 was estimated as the inhibitor concentration required to double the clotting time relative to the control. Results are summarized in Tables 1 and 2 below.
Fibrin Clot Assay
The fibrin clot assay was performed essentially as
described by Krtenansky et al, FEBS, 1987, 211:10. A serial dilution of the inhibitor was prepared in 50 mM tris HCl buffer (pH7.8 at 23 ºC) containing 0. IM NaCl and 0.1% (w/v) polyethylene glycol 8000. Human plasma (60uL, collected in 3.8% sodium citrate, blood/anticoagulant 9:1) was added to microtiter wells (microtiter plate, Falcon) containing 100μL of various inhibitor dilutions. The solution was mixed after which 50μL of human thrombin (InM final cone.) was added and mixed for 15 seconds. The turbidity of the clot was immediately monitored by
microplate autoreader (Dynateck MR 5000) at 405nm and recorded every 3 min. The maximal turbidity in the absence of inhibitors was reached within a 60 min. IC values were calculated at 30 minutes as the inhibitor concentration that gave half the optical density of the control.
Platelet Aggregation and Secretion
Rat blood was collected into ACD (6/1 v/v) by cardiac puncture. Suspensions of washed platelets were prepared as described by Ardlie et al, (Br. J. Haematol. 1970, 19:7 and Proc. Soc. Exp. Biol. Med., 1971, 136:1021). The final suspending medium was a modified Tyrode solution (NaCl 138mM, KCl 2.9mM, HEPES 20mM, NaH.PO. 0.42mM, NaHCO 12mM, CaCl ImM, MgCl 2mM, 0.1% glucose, 0.35% albumin, apyrase lμL/mL pH 7.4). Platelet counts were adjusted to 5000,000/uL.
To permit measurement of the extent of release of the contents of the dense granules, the platelets were
labelled in the first washing solution with 14C-serotonin (5-HT) (lμCi/10mL of washing fluid) and release of 14C-serotonin was determined as described in Holmsen et al, (Enzymology, 1989, 169:206). Inipramine (5μM final cone.) was added to present the reuptake of released serotonin.
Platelet aggregation was recorded at 37ºC in an
aggregometer (BioData PAP-4) at a stirring speed of 1,100 rpm by measuring the variations of light transmission.
Percentage of aggregation was determined 3 min. after the addition of the stimulating agent (human thrombin 0. lIU/mL final cone.). Inhibitors were preincubated 1 minute at 37ºC before addition of stimulating agent. IC50 values represent the concentration that was necessary to inhibit platelet aggregation or secretion to 50% of the control
Arterial Thrombosis Model
FeCl Induced Carotid Arterial Injury Model
The FeCl induced injury to the carotid artery in rats was induced according to the method described by Kurz, K.D., Main, R.W., Sandusky, G.E., Thrombosis Research 60; 269-280, 1990 and Schumacher, W.A. et al. J. Pharmacology and Experimental Therapeutics 267; 1237-1242, 1993. Male, Sprague-Dawley rats ( 375-410 g) were anesthetized with urethane ( 1500 mg\kg ip). Animals were laid on a 37°C heating pad. The carotid artery was exposed through a midline cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid sheath. Using forceps, the artery was lifted to provide sufficient clearance to insert two small pieces of polyethylene tubing (PE-205 ) underneath it. A temperature probe
(Physitemp MT23/3) was placed between one of the pieces of tubing and the artery. Injury was induced by topical application on the carotid artery above the temperature probe of a small disc (3 mm dia.) of Whatman No .1 filter paper previously dipped in a 35% solution of FeCl3. The incision area was covered with aluminum foil in order to protect the FeCl from degradation by light. The vessel temperature was monitored for 60 minutes after application of FeCl3 as an indication of blood flow. Vessel
temperature changes were recorded on a thermister (Cole-Palmer Model 08533-41).
The time between the FeCl application and the time at which the vessel temperature decreased abruptly (>2.4°C) was recorded as the time to occlusion of the vessel.
Inhibitor compounds were given as an iv bolus (mg/kg) followed immediately by an iv infusion (μg/kg/min. via femoral vein) . The dose of inhibitor needed to double the time to occlusion in comparison to control animals in which injury was induced in the absence of inhibitor was determined .
Figure imgf000154_0001
Figure imgf000155_0001
Figure imgf000156_0001

Claims

AIM:
Figure imgf000158_0001
wherein:
A is selected from (CH-R.),., , S, SO, SO , 0 and NR, wherein R, is hydrogen, C . alkyl optionally interupted with 1 or 2 heteroatoms; C. ι aryl, C,.cycloalkyl or heterocyclic ring or a hydrophobic group ;
B is selected from S, SO2, O, -N= , NH, -CH= and CRfRwherein R6 and R7 are independently selected from hydrogen and C,.. alkyl provided that when A is S, SO, SO2, O, or NR8, then B is CR,R, ;
D is selected from (CH-R,) wherein R is hydrogen, C alkyl or -C(0)R.; and CH with a double bond to B when B is -N= or -CH=;
E is selected from CH: and CH substituted with the C(0)R , provided that only one of D and E is substituted with with -C(0)R ;
X is selected from O, N-R,, or CH-R ;
Y is selected from O, S, SO, SO , N-R and CH-R
provided that when X is N-R. then Y is CH-R or 0, and when X is 0 then Y is CH-R. ;
Z is selected from 0, S and H ;
R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is selected from H and C alkyl optionally
substituted with C. aryl, a 6 member heterocycle or a C cycloalkyl ring; R3 is selected from H, NRfR- and C,. alkyl; and
R4 and R5 are independently selected from H; NR6R, ; C6-10. aryl or C _, cycloalkyl optionally substituted with C_, alkyl; C1-16 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C6-10 aryl, heterocycle or C - cycloalkyl group optionally substituted with halogen, hydroxyl, C_. alkyl; an amino acid side chain; and a hydrophobic group.
A compound according to claim 1, wherein R is one of formula Via to VId:
Figure imgf000159_0001
wherein :
R11 is hydrogen or C,. alkyl;
K is a bond or -NH- ;
G is C, , alkoxy; cyano; -NH ; -CH -NH_ ; -C(NH)-NH ; -NHC(NH)-NH_; -CH -NH-C (NH) -NH ; a C cycloalkyl or aryl substituted with cyano, -NH , -CH -NH , -C(NH)NH, -NH-C(NH)-NH_ or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH , -CH -NH , C(NH)-NH, -NH-C(NH)-NH or -CH -NH-C (NH) -NH ;
U is cyano, -NH , -C(NH)-NH_ or -NH-C (NH) -NH ;
P is a bond, -C(0)- or a bivalent group:
Figure imgf000160_0001
J is C,.. alkylene optionally substituted with OH, NH and C , alkyl and optionally interrupted by a heteroatom selected from 0, S and N;
n is 0 or 1; and
T is H, OH, amino, a peptide chain, C. alkyl, C,_ . alkoxy, C aralkyl, or heterocycle optionally substituted.
A compound according to claim 2, wherein T is a heterocycle selected from the group consisting of:
Figure imgf000160_0002
wherein
X, , X10, X11 and X12 are each independently selected from the group consisting of N, or C-X where X. is
hydrogen, C_ alkyl, or C. aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X. , or CH-X ; and
R' is hydrogen, C .. alkyl optionally carboxyl
substituted, carboxyl, -C . alkyl-CO -C .. alkyl, C aralkyl, C cycloalkyl, aryl or an aromatic
heterocycle .
A compound according to claim 3, wherein T is
selected from the group consisting of:
Figure imgf000161_0001
wherein R' is hydrogen, C1-10 alkyl optionally carboxyl substituted, carboxyl, -C0-10 alkyl-CO -C , alkyl, C, aralkyl, C, . cycloalkyl, aryl or an aromatic
heterocycle .
A compound according to claim 4, wherein T is
selected from:
Figure imgf000161_0002
wherein R' is hydrogen, C.,., alkyl optionally carboxy substituted, carboxyl, -C, .,. alkyl-CO -C alkyl, C aralkyl, C .- cycloalkyl, aryl or an aromatic
heterocycle . A compound according to claim 1, wherein one of P^ and Rc is a hydrophobic group selected from C alkyl, C alkenyl or C _ alkynyl optionally interrupted by a carbonyl group, C._, aryl, C - cycloalkyl, C._
aralkyl, C_ cycloalkyl substituted C.. alkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group and the ring portion is optionally substituted with C alkyl; and a hydrophobic amino acid side chain.
7. A compound according to claim 6, wherein R is H.
8. A compound according to claim 1, wherein Z is O.
9. A compound according to claim 1, wherein R is H. 10. A compound of the formula (VII):
Figure imgf000162_0001
wherein
R is an arginyl moiety or an analog or derivative
thereof optionally substituted with an amino acid, a peptide or a heterocycle;
is H or C1-6 alkyl;
is selected from H, NR R- and C alkyl; and and R5 are independently selected from H; NRR ; C aryl or C cycloalkyl optionally substituted with C alkyl; C alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionallysubstituted with OH, SH, NRR. or a C aryl, heterocycle or C - cycloalkyl group optionallysubstituted with halogen, hydroxyl , C alkyl; an amino acid side chain; and a hydrophobic group.
11. A compound according to claim 10, wherein R, is one of formula Via to VId:
wherein :
R11 is hydrogen or C. alkyl;
K is a bond or -NH- ;
G is C ... alkoxy; cyano; -NH_; -CH-NH.; -C (NH) -NH_; -NHC(NH)-NH ; -CH -NH-C (NH) -NH, ; a C. cycloalkyl or aryl substituted with cyano, -NH , -CH-NH , -C(NH)NH , -NH-C(NH) -NH. or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH , -CH-NH , C(NH)-NH, -NH-C(NH)-NH or -CH -NH-C (NH) -NH ; ϋ is cyano, -NH, , -C(NH)-NH or -NH-C (NH) -NH ;
P is a bond, -C(O)- or a bivalent group:
Figure imgf000163_0002
J is C alkylene optionally substituted with OH, NH and C alkyl and optionally interrupted by a heteroatom selected from 0, S and N;
n is 0 or 1 ; and
T is H, OH, amino, a peptide chain, C... alkyl, C.
alkoxy, C. aralkyl, or heterocycle optionally substituted. A compound according to claim 11, wherein T is a heterocycle selected from the group consisting of:
Figure imgf000164_0001
wherein
X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X- where X7 is
hydrogen, C alkyl, or C aryl;
X6 and X13 are each independently selected from the group consisting of C, 0, N, S, N-X-, or CH-X,;
R' is hydrogen, C1-1 alkyl optionally carboxyl
substituted, carboxyl, -C alkyl-CO -C alkyl, C, aralkyl, C _ cycloalkyl, aryl or an aromatic
heterocycle .
A compound according to claim 12, wherein T is selected from the group consisting of :
Figure imgf000165_0001
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C alkyl, C aralkyl, C - cycloalkyl, aryl or an aromatic heterocycle .
A compound according to claim 13, wherein T is selected from:
Figure imgf000165_0002
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C . alkyl-CO -C alkyl, C aralkyl, C cycloalkyl, aryl or an aromatic heterocycle . 15. A compound according to claim 10, wherein R and R are H.
16. A compound according to claim 10, wherein R4 is C1-1 alkyl optionally interupted with a heteroatom or a carbonyl, and optionally substituted with a C....
aromatic, C .. cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF. or oxo.
17. A compound according to claim 10, wherein R. is H. 18 A compound according to claim 12, wherein:
R is H;
R, is C;. alkyl optionally interupted with a
heteroatom or a carbonyl, and optionally
substituted with a C. aromatic, C cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF or oxo; and
R is H.
19. A compound according to claim 10, selected from:
0085 6S-cyclohexylmethylhexahydro-5-oxo-5Hthiazolo [ 3 , 2-a] pyridine-3R-carboxamido (propylcarbo methoxy ketoarginine); and
0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo
[3 , 2-a] pyridine-3R-carboxamido (α-benzothiozolo keto arginine) .
20. A compound according to claim 1, of formula (VIII)
Figure imgf000166_0001
wherein R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or Cl-6 alkyl;
R3 is selected from H, NR.R- and C,_ alkyl; and
R4 and R5 are independently selected from H; NRR, ; C. aryl or C .- cycloalkyl optionally substituted with C .. alkyl; C,. alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRR. or a C. aryl, heterocycle or C - cycloalkyl group optionally substituted with halogen, hydroxyl , C alkyl; an amino acid side chain; and a hydrophobic group. 21. A compound according to claim 20, wherein R. is one of formula Via to VId:
Figure imgf000167_0001
wherein :
R11 is hydrogen or C alkyl;
K is a bond or -NH- ;
G is C .. alkoxy; cyano ; -NH ; -CH-NH; -C(NH)-NH ; -NHC(NH)-NH ; -CH -NH-C(NH) -NH ; a C cycloalkyl or aryl substituted with cyano, -NH , -CH-NH , -C(NH)NH , -NH-C(NH)-NH or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH , -CH-NH , C(NH)-NH, -NH-C(NH)-NH or -CH -NH-C (NH) -NH ; U is cyano, -NH , -C(NH)-NH or -NH-C (NH) -NH ; P is a bond, -C(O)- or a bivalent group:
Figure imgf000168_0001
J is C;., alkylene optionally substituted with OH, NH and C,. alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and
T is H, OH, amino, a peptide chain, C,. alkyl, C alkoxy, C. aralkyl, or heterocycle optionally substituted.
22. A compound according to claim 21, wherein T is a heterocycle selected from the group consisting of:
Figure imgf000168_0002
wherein
X5, X10, X.. and X12 are each independently selected : the group consisting of N, or C-X where X7 is hydrogen, C. alkyl, or C. aryl;
X6 and X13 are each independently selected from the group consisting of C, 0, N, S, N-X-, or CH-X.. ;
R' is hydrogen, C.., alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO_-C alkyl, C aralkyl, C - cycloalkyl, aryl or an aromatic heterocycle .
23 A compound according to claim 22, wherein T is
selected from the group consisting of :
Figure imgf000169_0001
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C alkyl, C aralkyl, C - cycloalkyl, aryl or an aromatic heterocycle .
24 A compound according to claim 23, wherein T is selected from:
Figure imgf000169_0002
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C alkyl, C aralkyl, C - cycloalkyl, aryl or an aromatic heterocycle . 25. A compound according to claim 20, wherein R and R are both H.
26. A compound according to claim 20, wherein R. is H or C,. alkyl substituted with COOH . 27. A compound according to claim 20, wherein R:, R and R, are H and R. is C. ; alkyl optionally interrupted by one or more heteroatom or carbonyl group and
optionally substituted with OH, SH, NRR, or a C,.,.
aryl, heterocycle or C , cycloalkyl group optionally substituted with halogen, hydroxyl or C alkyl.
28. A compound according to claim 22, wherein:
R , R and R.. are H; and
R is C . alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR,R, or a C,.,, aryl, heterocycle or C - cycloalkyl group optionally substituted with halogen, hydroxyl or C alkyl. 29. A compound according to claim 20, selected from:
0345 4-Oxo-2- (3-phenyl-propionyl ) -octahydropyrrolo]1,2-a]pyrazine-6-carboxylic acid [4guanidino-1- ( 5-methyl-thiazole-2-carbonyl ) butyl]-amide; and0340 4-Oxo-2- ( 3-phenylpropionyl) -octahydro-pyrrolo[1 , 2-a]pyrazine-6carboxylic acid |4-guanidino-1- ( thiazole-2carbonyl ) -butyl]-amide .
30. A compound according to claim 1, of formula (IX)
Figure imgf000171_0001
wherein
Y is selected from 0, S, SO, SO , N-R, and CH-R9;
R1 is an arginyl moiety or an analog or derivative
thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or Cl-6 alkyl;
R3 is selected from H, NRR- and C._. alkyl; and
R4 and R5 are independently selected from H; NR,R_; C. _ . aryl or C .- cycloalkyl optionally substituted with C.. alkyl; C._. alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRR- or a C.. aryl, heterocycle or C... cycloalkyl group optionally substituted with halogen, hydroxyl, C alkyl; an amino acid side chain; and a hydrophobic group;
Rβ is hydrogen, C_ alkyl optionally interupted with 1 or 2 heteroatoms; C0-1 aryl, C - cycloalkyl or heterocyclic ring or a hydrophobic group; and n is 1 or 2;
31. A compound according to claim 30, wherein R is one formula VIa to VId:
Figure imgf000171_0002
wherein :
R11 is hydrogen or C .. alkyl;
K is a bond or -NH- ;
G is C1_4 alkoxy; cyano; -NH ; -CH.-NH2; -C (NH) -NH2; -NHC(NH)-NH ; -CH -NH-C (NH) -NH ; a C6 cycloalkyl or aryl substituted with cyano, -NH:, -CH-NH , -C(NH)NH , -NH-C(NH)-NH or -CH.-NH-C (NH) -NH, ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH , -CH.-NH , C(NH)-NH_, -NH-C(NH)-NH or -CH -NH-C (NH) -NH ;
U is cyano, -NH , -C(NH)-NH or -NH-C (NH) -NH ;
P is a bond, -C(O)- or a bivalent group:
Figure imgf000172_0001
J is C_ alkylene optionally substituted with OH, NH and C,_ alkyl and optionally interrupted by a heteroatom selected from 0, S and N;
n is 0 or 1; and
T is H, OH, amino, a peptide chain, C alkyl, C, alkoxy, C. aralkyl, or heterocycle optionally substituted.
A compound according to claim 31, wherein T is a heterocycle selected from the group consisting of:
Figure imgf000172_0002
wherein
X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X. where X7 is
hydrogen, C1-4 alkyl, or C aryl; X6 and X13 are each independently selected from the group consisting of C, 0, N, S, N-X, , or CH-X. ;
R' is hydrogen, C1-1, alkyl optionally carboxyl substituted, carboxyl, -C0-10 alkyl-CO2-C1-16 alkyl, C6_ aralkyl, C3- cycloalkyl, aryl or an aromatic heterocycle .
33. A compound according to claim 32, wherein T is
selected from the group consisting of:
Figure imgf000173_0001
and R' is hydrogen, C, alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C .. alkyl, C aralkyl, C ._ cycloalkyl, aryl or an aromatic heterocycle .
34 A compound according to claim 33, wherein T is
selected from:
Figure imgf000174_0001
and R' is hydrogen, C1-10 alkyl optionally carboxyl substituted, carboxyl, -C1-10 alkyl-CO2-C1-10 alkyl, C6-20 aralkyl, C - cycloalkyl, aryl or an aromatic
heterocycle.
35. A compound according to claim 30, wherein R and R are both H. 36. A compound according to claim 30, wherein R. is H, NRR- or C, alkyl substituted COOH.
37. A compound according to claim 30, wherein R5 is C1-10 aryl, C6-20 aralkyl, or C1-10 alkyl substituted with C _cycloalkyl.
38. A compound according to claim 32, wherein
n is 1 ;
R , R and R. are H; and
R is C1-10 aryl, C6-20 aralkyl, or C1-10 alkyl substituted with C - cycloalkyl.
39. A compound according to claim 30, selected from:
0890 3-Amino-4-oxo-2-phenyl-hexahydro-pyrrolo[2 , 1b][1 , 3]thiazine-6-carboxylic acid [1(benzothiazole-2-carbonyl ) -4-guanidino-butyl]amide ;
0895 3-Amino-2-benzyl-4-oxo--hexahydro-pyrrolo|2 , 1b][l , 3]thiazine-6-carboxylic acid |1¬
(benzothiazole-2-carbonyl ) -4-guanidino-butyl]amide ; and 0900 3 -Amino-2-cyclohexyl-4-oxo-hexahydro-pyrrolo[2 , 1b][l , 3]thiazine-6-carboxylic acid [1¬
(benzothiazole-2-carbonyl ) -4-guanidino-butyl]amide .
0 A compound according to claim 1, of formula (X)
Figure imgf000175_0001
wherein
B is 0, S, -CH-, or -NH- ;
R1 is an arginyl moiety or an analog or derivative
thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or Cl-6 alkyl;
R3 is selected from H, NRR- and C,. alkyl; and
R4 and R5 are independently selected from H; NR.R-; C aryl or C .- cycloalkyl optionally substituted with C1-10 alkyl; C1-10 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR R5 or a C . aryl, heterocycle or C __ cycloalkyl group optionally substituted with halogen, hydroxyl, C alkyl; an amino acid side chain; and a hydrophobic group.
41. A compound according to claim 40, wherein R is one c formula Via to VId:
Figure imgf000176_0001
wherein :
Rxl is hydrogen or Cl alkyl ;
K is a bond or -NH- ;
G is C alkoxy; cyano; -NH ; -CH-NH; -C(NH)-NH; -NHC(NH)-NH; -CH2-NH-C (NH) -NH ; a C cycloalkyl or aryl substituted with cyano, -NH , -CH-NH , -C(NH)NH , -NH-C(NH)-NH2 or -CH -NH-C (NH) -NH ; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH-NH , C(NH)-NH, -NH-C(NH)-NH2 or -CH -NH-C (NH) -NH ;
U is cyano, -NH , -C(NH)-NH or -NH-C (NH) -NH ;
P is a bond, -C(O)- or a bivalent group:
Figure imgf000176_0002
J is C alkylene optionally substituted with OH, NH and C alkyl and optionally interrupted by a heteroatom selected from 0, S and N;
n is 0 or 1; and
T is H, OH, amino, a peptide chain, C alkyl, C alkoxy, C aralkyl, or heterocycle optionally substituted.
42. A compound according to claim 41, wherein T is a heterocycle selected from the group consisting of:
Figure imgf000177_0001
wherein
X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X- where X7 is
hydrogen, C alkyl, or C5-8 aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X-, or CH-X.;
R' is hydrogen, C _.. alkyl optionally carboxyl
substituted, carboxyl, -C0-10 alkyl-CO2-C1-10 alkyl, C6-20 aralkyl, C - cycloalkyl, aryl or an aromatic
heterocycle .
43. A compound according to claim 42, wherein T is
selected from the group consisting of:
Figure imgf000178_0001
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C alkyl, C aralkyl, C . cycloalkyl, aryl or an aromatic heterocycle .
4 . A compound according to claim 43 , wherein T is selected from:
Figure imgf000178_0002
and R' is hydrogen, C alkyl optionally carboxyl substituted, carboxyl, -C alkyl-CO -C alkyl, C aralkyl, C cycloalkyl, aryl or an aromatic heterocycle .
45. A compound according to claim 40, wherein R and R are both H.
46. A compound according to claim 40, wherein R. is Ci_1 alkyl substituted with C,.,, aryl optionally
substituted with C..,. alkyl. 47. A compound according to claim 40, wherein R5 is H.
48. A compound according to claim 42, wherein
B is S;
R , R and R are H; and
R is C . alkyl substituted with C aryl optionally substituted with C1-10 alkyl.
49. A compound according to claim 40, selected from:
925 7-Benzyl-6-oxo-octahydro-pyrido[2 , 1c][l , 4]thiazine-4-carboxylic acid [4-guanidino-1¬
( thiazole-2-carbonyl)butyl]-amide; and940 6-Oxo7-phenethyl-octahydro-pyrido[2, l-c][l , 4jthiazine4-carboxylic acid [4-guanidino-1-(thiazole-2carbonyl) -butyl]-amide .
50. A method for the treatment or prophylaxis of
thrombotic disorders in a mammal, comprising
administering to said mammal an effective amount of a compound according to claim 1.
51. A method according to claim 50, wherein said
thrombotic disorder is venous thrombosis.
52. A method according to claim 50, wherein said
thrombotic disorder is a pulmonary embolism.
53. A method according to claim 50, wherein said
thrombotic disorder is arterial thrombosis.
54. A method according to claim 50, wherein said
thrombotic disorder is myocardial infarction.
55. A method according to claim 50, wherein said
thrombotic disorder is cerebral infarction.
56. A process for producing a compound according to claim
1. 57. A process for producing a compound according to any one of claims 10, 20, 30 or 40.
PCT/CA1995/000708 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors WO1996019483A1 (en)

Priority Applications (15)

Application Number Priority Date Filing Date Title
APAP/P/1997/001004A AP9701004A0 (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors.
JP8519383A JPH11508535A (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitor
EE9700113A EE9700113A (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors
NZ297360A NZ297360A (en) 1994-12-22 1995-12-21 5-oxo-5H-thiazolo[3,2-a]pyridine, 5-oxo-2-thia-4a,7-diaza-naphthalene, 4-oxo-octahydro-pyrrolo[1,2-a]pyrazine or 6-oxo-octahydro-pyrido-[2,1-c][1,4]thiazine derivatives and medicaments
MD97-0253A MD970253A (en) 1994-12-22 1995-12-21 Low-molecular bicyclic thrombin inhibitors
AU42505/96A AU4250596A (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors
EP95940923A EP0802916A1 (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors
SK838-97A SK83897A3 (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors
BR9510433A BR9510433A (en) 1994-12-22 1995-12-21 Low molecular weight cyclic thrombin inhibitor compounds method for their treatment and process for their production
IS4504A IS4504A (en) 1994-12-22 1997-06-11 Double-ring thrombin inhibitors with low molecular weight
FI972466A FI972466L (en) 1994-12-22 1997-06-11 Small molecular weight bicyclic thrombin inhibitors
BG101647A BG101647A (en) 1994-12-22 1997-06-20 Low molecular weight bicyclic thrombin inhibitors
NO972892A NO972892L (en) 1994-12-22 1997-06-20 Bicyclic Low Molecular Weight Thrombin Inhibitors
US08/880,885 US6057314A (en) 1995-12-21 1997-06-23 Low molecular weight bicyclic thrombin inhibitors
LVP-97-141A LV12019B (en) 1994-12-22 1997-07-15 BULK CYCLIC FIBERS INHIBITORS

Applications Claiming Priority (10)

Application Number Priority Date Filing Date Title
GBGB9426038.7A GB9426038D0 (en) 1994-12-22 1994-12-22 Low molecular weight bicyclic thrombin inhibitors
GBGB9503136.5A GB9503136D0 (en) 1994-12-22 1995-02-17 Low molecular weight bicyclic thrombin inhibitors
GB9510265.3 1995-05-22
GB9510267.9 1995-05-22
GBGB9510267.9A GB9510267D0 (en) 1995-05-22 1995-05-22 Low molecular weight thiobicyclic thrombin inhibitors
GBGB9510266.1A GB9510266D0 (en) 1995-05-22 1995-05-22 Low molecular weight bicyclic thrombin inhibitors
GB9510266.1 1995-05-22
GBGB9510265.3A GB9510265D0 (en) 1995-05-22 1995-05-22 Low molecular weight diaminobicyclic thrombin inhibitors
GB9503136.5 1995-05-22
GB9426038.7 1995-05-22

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US08/880,885 Continuation US6057314A (en) 1995-12-21 1997-06-23 Low molecular weight bicyclic thrombin inhibitors

Publications (1)

Publication Number Publication Date
WO1996019483A1 true WO1996019483A1 (en) 1996-06-27

Family

ID=27517273

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA1995/000708 WO1996019483A1 (en) 1994-12-22 1995-12-21 Low molecular weight bicyclic thrombin inhibitors

Country Status (22)

Country Link
EP (1) EP0802916A1 (en)
JP (1) JPH11508535A (en)
CN (1) CN1175259A (en)
AP (1) AP9701004A0 (en)
AU (2) AU4250596A (en)
BG (1) BG101647A (en)
CA (1) CA2208772A1 (en)
CZ (1) CZ189997A3 (en)
EE (1) EE9700113A (en)
FI (1) FI972466L (en)
HU (1) HUT77651A (en)
IL (1) IL116503A0 (en)
IS (1) IS4504A (en)
LV (1) LV12019B (en)
MD (1) MD970253A (en)
MX (1) MX9704718A (en)
NO (1) NO972892L (en)
NZ (1) NZ297360A (en)
OA (1) OA10493A (en)
PL (1) PL320965A1 (en)
SK (1) SK83897A3 (en)
WO (1) WO1996019483A1 (en)

Cited By (47)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997005160A1 (en) * 1995-08-01 1997-02-13 A. Menarini Industrie Farmaceutiche Riunite S.R.L. Bicyclic lactam derivatives as thrombin inhibitors
WO1997017363A1 (en) * 1995-11-03 1997-05-15 Akzo Nobel N.V. Thrombin inhibitors
WO1997030073A1 (en) * 1996-02-13 1997-08-21 Akzo Nobel N.V. Serine protease inhibitors
WO1997031937A1 (en) * 1996-03-01 1997-09-04 Akzo Nobel N.V. Serine protease inhibitors
WO1997031939A1 (en) * 1996-03-01 1997-09-04 Akzo Nobel N.V. Serine protease inhibitors
WO1997048706A1 (en) * 1996-06-18 1997-12-24 Warner-Lambert Company Pyrrolo[1,2-a]pyrazine-1,4-dione serine protease inhibitors
WO1998005333A1 (en) * 1996-08-05 1998-02-12 Molecumetics Ltd. Use of beta-sheet mimetics as protease and kinase inhibitors and as inhibitors of transcription factors
WO1998009987A1 (en) * 1996-09-06 1998-03-12 Biochem Pharma, Inc. Lactam inhibitors of thrombin
WO1998028326A1 (en) * 1996-12-23 1998-07-02 Biochem Pharma Inc. Bicyclic thrombin inhibitors
US6020331A (en) * 1995-03-24 2000-02-01 Molecumetics, Ltd. β-sheet mimetics and use thereof as protease inhibitors
EP1017383A1 (en) * 1997-09-23 2000-07-12 Merck & Co., Inc. Thrombin inhibitors
US6117896A (en) * 1997-02-10 2000-09-12 Molecumetics Ltd. Methods for regulating transcription factors
US6133256A (en) * 1997-04-14 2000-10-17 Cor Therapeutics Inc Selective factor Xa inhibitors
EP1058549A1 (en) * 1998-12-23 2000-12-13 Du Pont Pharmaceuticals Company THROMBIN OR FACTOR Xa INHIBITORS
FR2795072A1 (en) * 1999-06-15 2000-12-22 Adir New bicyclic amino-pyrazinone derivatives and their isomers and salts, useful as thrombin inhibitors for treating cardiovascular diseases such as angina pectoris, thrombosis, atherosclerosis, arteritis and venous disease
US6194435B1 (en) 1996-10-11 2001-02-27 Cor Therapeutics, Inc. Lactams as selective factor Xa inhibitors
US6204268B1 (en) 1997-04-14 2001-03-20 Cor Therapeutics, Inc Selective factor Xa inhibitors
US6218382B1 (en) 1997-08-11 2001-04-17 Cor Therapeutics, Inc Selective factor Xa inhibitors
US6228854B1 (en) 1997-08-11 2001-05-08 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6245764B1 (en) 1995-03-24 2001-06-12 Molecumetics Ltd. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
US6262047B1 (en) 1996-10-11 2001-07-17 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6323219B1 (en) 1998-04-02 2001-11-27 Ortho-Mcneil Pharmaceutical, Inc. Methods for treating immunomediated inflammatory disorders
US6333321B1 (en) 1997-08-11 2001-12-25 Cor Therapeutics, Inc. Selective factor Xa inhibitors
WO2002002519A2 (en) * 2000-06-29 2002-01-10 Bristol-Myers Squibb Pharma Company THROMBIN OR FACTOR Xa INHIBITORS
JP2002503674A (en) * 1998-02-12 2002-02-05 モレキュメティックス リミテッド Beta sheet mimics and methods for their use
US6369080B2 (en) 1996-10-11 2002-04-09 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6369063B1 (en) 1997-04-14 2002-04-09 Cor Therapeutics, Inc. Selective factor Xa inhibitors
EP1215213A2 (en) * 2000-12-14 2002-06-19 Les Laboratoires Servier Bicyclic amino-pyrazone derivatives, their preparation and pharmaceutical compositions containing them
US6469036B1 (en) 1999-01-27 2002-10-22 Ortho-Mcneil Pharmaceutical, Inc. Peptidyl heterocyclic ketones useful as tryptase inhibitors
US6525076B1 (en) 1996-10-11 2003-02-25 Millennium Pharmaceuticals, Inc. Selective factor Xa inhibitors
US6699869B1 (en) 1995-03-24 2004-03-02 Myriad Genetics Inc. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
US7030239B2 (en) * 2000-03-23 2006-04-18 Elan Pharmaceuticals, Inc. Compounds to treat Alzheimer's disease
US7053214B2 (en) 2002-02-14 2006-05-30 Myriad Genetics, Inc. β-sheet mimetics and composition and methods relating thereto
EP1661566A2 (en) * 1996-08-05 2006-05-31 Myriad Genetics, Inc. Use of beta-sheet mimetics as protease and kinase inhibitors and as inhibitors of transcription factors
US7192615B2 (en) 2001-02-28 2007-03-20 J&J Consumer Companies, Inc. Compositions containing legume products
US7309688B2 (en) 2000-10-27 2007-12-18 Johnson & Johnson Consumer Companies Topical anti-cancer compositions and methods of use thereof
US7629318B2 (en) 2002-03-22 2009-12-08 Gpc Biotech Ag Immunosuppressant compounds, methods and uses related thereto
US8093293B2 (en) 1998-07-06 2012-01-10 Johnson & Johnson Consumer Companies, Inc. Methods for treating skin conditions
US8106094B2 (en) 1998-07-06 2012-01-31 Johnson & Johnson Consumer Companies, Inc. Compositions and methods for treating skin conditions
US8431550B2 (en) 2000-10-27 2013-04-30 Johnson & Johnson Consumer Companies, Inc. Topical anti-cancer compositions and methods of use thereof
US9758473B2 (en) 2014-10-06 2017-09-12 Cortexyme, Inc. Inhibitors of lysine gingipain
US10208048B2 (en) 2015-04-28 2019-02-19 Janssen Sciences Ireland Uc RSV antiviral pyrazolo- and triazolo-pyrimidine compounds
US10730826B2 (en) 2016-09-16 2020-08-04 Cortexyme, Inc. Ketone inhibitors of lysine gingipain
US10906881B2 (en) 2015-11-09 2021-02-02 Cortexyme, Inc. Inhibitors of arginine gingipain
US11339165B2 (en) 2017-11-29 2022-05-24 Janssen Sciences Ireland Unlimited Company Pyrazolopyrimidines having activity against the respiratory syncytial virus (RSV)
US11491157B2 (en) 2018-01-31 2022-11-08 Janssen Sciences Ireland Unlimited Company Co Cork, IE Cycloalkyl substituted pyrazolopyrimidines having activity against RSV
US11708369B2 (en) 2018-04-23 2023-07-25 Janssen Sciences Ireland Unlimited Company Heteroaromatic compounds having activity against RSV

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 108, no. 9, 29 February 1988, Columbus, Ohio, US; abstract no. 75817d, UKON NAGAI ET AL: "Synthesis of an LH-RH analog with restricted conformation by incorporation of a bicyclic beta-turn dipeptide unit" page 728; *
K. KAWASAKI ET AL: "Amino acids and peptides. XIII. Synthesis studies on N-terminal tripeptide amide analogs of fibrin alpha-chain", CHEMICAL AND PHARMACEUTICAL BULLETIN., vol. 39, no. 3, TOKYO JP, pages 584 - 589 *
PEPT. CHEM., vol. 24, pages 295 - 298 *
U. NAGAI ET AL: "Bicyclic turned dipeptide (BTD) as a beta-turn mimetic; its design, synthesis and incorporation into bioactive peptides", TETRAHEDRON, vol. 49, no. 17, OXFORD GB, pages 3577 - 3592 *

Cited By (76)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6020331A (en) * 1995-03-24 2000-02-01 Molecumetics, Ltd. β-sheet mimetics and use thereof as protease inhibitors
US6245764B1 (en) 1995-03-24 2001-06-12 Molecumetics Ltd. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
US7511039B2 (en) 1995-03-24 2009-03-31 Myriad Genetics, Inc. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
US6586426B1 (en) 1995-03-24 2003-07-01 Molecumetics Ltd. β-sheet mimetics and use thereof as protease inhibitors
US6699869B1 (en) 1995-03-24 2004-03-02 Myriad Genetics Inc. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
US7125872B2 (en) 1995-03-24 2006-10-24 Myriad Genetics, Inc. β-sheet mimetics and use thereof as inhibitors of biologically active peptides or proteins
WO1997005160A1 (en) * 1995-08-01 1997-02-13 A. Menarini Industrie Farmaceutiche Riunite S.R.L. Bicyclic lactam derivatives as thrombin inhibitors
WO1997017363A1 (en) * 1995-11-03 1997-05-15 Akzo Nobel N.V. Thrombin inhibitors
US6432921B2 (en) 1995-11-03 2002-08-13 Akzo Nobel N.V. Thrombin inhibitors
US6034067A (en) * 1996-02-13 2000-03-07 Akzo Nobel, N.V. Serine protease inhibitors
WO1997030073A1 (en) * 1996-02-13 1997-08-21 Akzo Nobel N.V. Serine protease inhibitors
US6218365B1 (en) * 1996-03-01 2001-04-17 Akzo Nobel N.V. Serine protease inhibitors
WO1997031939A1 (en) * 1996-03-01 1997-09-04 Akzo Nobel N.V. Serine protease inhibitors
US6410684B1 (en) 1996-03-01 2002-06-25 Akzo Nobel N.V. Serine protease inhibitors
WO1997031937A1 (en) * 1996-03-01 1997-09-04 Akzo Nobel N.V. Serine protease inhibitors
US6124291A (en) * 1996-06-18 2000-09-26 Warner-Lambert Company Pyrrolo[1,2-a]pyrazine-1,4-dione serine protease inhibitors
WO1997048706A1 (en) * 1996-06-18 1997-12-24 Warner-Lambert Company Pyrrolo[1,2-a]pyrazine-1,4-dione serine protease inhibitors
WO1998005333A1 (en) * 1996-08-05 1998-02-12 Molecumetics Ltd. Use of beta-sheet mimetics as protease and kinase inhibitors and as inhibitors of transcription factors
EP1661566A3 (en) * 1996-08-05 2008-04-16 Myriad Genetics, Inc. Use of beta-sheet mimetics as protease and kinase inhibitors and as inhibitors of transcription factors
EP1661566A2 (en) * 1996-08-05 2006-05-31 Myriad Genetics, Inc. Use of beta-sheet mimetics as protease and kinase inhibitors and as inhibitors of transcription factors
WO1998009987A1 (en) * 1996-09-06 1998-03-12 Biochem Pharma, Inc. Lactam inhibitors of thrombin
US6262047B1 (en) 1996-10-11 2001-07-17 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6525076B1 (en) 1996-10-11 2003-02-25 Millennium Pharmaceuticals, Inc. Selective factor Xa inhibitors
US6194435B1 (en) 1996-10-11 2001-02-27 Cor Therapeutics, Inc. Lactams as selective factor Xa inhibitors
US6369080B2 (en) 1996-10-11 2002-04-09 Cor Therapeutics, Inc. Selective factor Xa inhibitors
WO1998028326A1 (en) * 1996-12-23 1998-07-02 Biochem Pharma Inc. Bicyclic thrombin inhibitors
US6117896A (en) * 1997-02-10 2000-09-12 Molecumetics Ltd. Methods for regulating transcription factors
US6372744B1 (en) 1997-02-10 2002-04-16 Molecumetics Ltd. β-sheet mimetics and methods relating to the use thereof
US6369063B1 (en) 1997-04-14 2002-04-09 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6204268B1 (en) 1997-04-14 2001-03-20 Cor Therapeutics, Inc Selective factor Xa inhibitors
US6133256A (en) * 1997-04-14 2000-10-17 Cor Therapeutics Inc Selective factor Xa inhibitors
US6228854B1 (en) 1997-08-11 2001-05-08 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6333321B1 (en) 1997-08-11 2001-12-25 Cor Therapeutics, Inc. Selective factor Xa inhibitors
US6218382B1 (en) 1997-08-11 2001-04-17 Cor Therapeutics, Inc Selective factor Xa inhibitors
EP1017383A1 (en) * 1997-09-23 2000-07-12 Merck & Co., Inc. Thrombin inhibitors
EP1017383A4 (en) * 1997-09-23 2001-11-28 Merck & Co Inc Thrombin inhibitors
JP2002503674A (en) * 1998-02-12 2002-02-05 モレキュメティックス リミテッド Beta sheet mimics and methods for their use
US6323219B1 (en) 1998-04-02 2001-11-27 Ortho-Mcneil Pharmaceutical, Inc. Methods for treating immunomediated inflammatory disorders
US8093293B2 (en) 1998-07-06 2012-01-10 Johnson & Johnson Consumer Companies, Inc. Methods for treating skin conditions
US8106094B2 (en) 1998-07-06 2012-01-31 Johnson & Johnson Consumer Companies, Inc. Compositions and methods for treating skin conditions
EP1058549A1 (en) * 1998-12-23 2000-12-13 Du Pont Pharmaceuticals Company THROMBIN OR FACTOR Xa INHIBITORS
US6602871B2 (en) 1998-12-23 2003-08-05 Bristol-Myers Squibb Pharma Company Thrombin or factor Xa inhibitors
EP1058549A4 (en) * 1998-12-23 2003-11-12 Bristol Myers Squibb Pharma Co THROMBIN OR FACTOR Xa INHIBITORS
US6403583B1 (en) 1998-12-23 2002-06-11 Patrick Y. S. Lam Thrombin or factor Xa inhibitors
US6469036B1 (en) 1999-01-27 2002-10-22 Ortho-Mcneil Pharmaceutical, Inc. Peptidyl heterocyclic ketones useful as tryptase inhibitors
US7132418B2 (en) 1999-01-27 2006-11-07 Ortho-Mcneil Pharmaceutical, Inc. Peptidyl heterocyclic ketones useful as tryptase inhibitors
FR2795072A1 (en) * 1999-06-15 2000-12-22 Adir New bicyclic amino-pyrazinone derivatives and their isomers and salts, useful as thrombin inhibitors for treating cardiovascular diseases such as angina pectoris, thrombosis, atherosclerosis, arteritis and venous disease
EP1069132A1 (en) * 1999-06-15 2001-01-17 Adir Et Compagnie Bicyclic derivatives of amino-pyrazinones, process of preparation and pharmaceutical compositions comprising them
US7030239B2 (en) * 2000-03-23 2006-04-18 Elan Pharmaceuticals, Inc. Compounds to treat Alzheimer's disease
WO2002002519A3 (en) * 2000-06-29 2002-07-25 Bristol Myers Squibb Pharma Co THROMBIN OR FACTOR Xa INHIBITORS
US6586418B2 (en) 2000-06-29 2003-07-01 Bristol-Myers Squibb Company Thrombin or factor Xa inhibitors
WO2002002519A2 (en) * 2000-06-29 2002-01-10 Bristol-Myers Squibb Pharma Company THROMBIN OR FACTOR Xa INHIBITORS
US7309688B2 (en) 2000-10-27 2007-12-18 Johnson & Johnson Consumer Companies Topical anti-cancer compositions and methods of use thereof
US8431550B2 (en) 2000-10-27 2013-04-30 Johnson & Johnson Consumer Companies, Inc. Topical anti-cancer compositions and methods of use thereof
EP1215213A3 (en) * 2000-12-14 2002-09-25 Les Laboratoires Servier Bicyclic amino-pyrazone derivatives, their preparation and pharmaceutical compositions containing them
FR2818277A1 (en) * 2000-12-14 2002-06-21 Servier Lab NOVEL BICYCLIC DERIVATIVES OF AMINO-PYRAZINONES, PROCESS FOR PREPARING THEM AND PHARMACEUTICAL COMPOSITIONS CONTAINING SAME
EP1215213A2 (en) * 2000-12-14 2002-06-19 Les Laboratoires Servier Bicyclic amino-pyrazone derivatives, their preparation and pharmaceutical compositions containing them
US7192615B2 (en) 2001-02-28 2007-03-20 J&J Consumer Companies, Inc. Compositions containing legume products
US7897144B2 (en) 2001-02-28 2011-03-01 Johnson & Johnson Comsumer Companies, Inc. Compositions containing legume products
US7053214B2 (en) 2002-02-14 2006-05-30 Myriad Genetics, Inc. β-sheet mimetics and composition and methods relating thereto
US7629318B2 (en) 2002-03-22 2009-12-08 Gpc Biotech Ag Immunosuppressant compounds, methods and uses related thereto
US9988375B2 (en) 2014-10-06 2018-06-05 Cortexyme, Inc. Inhibitors of lysine gingipain
US10676470B2 (en) 2014-10-06 2020-06-09 Cortexyme, Inc. Inhibitors of lysine gingipain
US11332464B2 (en) 2014-10-06 2022-05-17 Cortexyme, Inc. Inhibitors of lysine gingipain
US10301301B2 (en) 2014-10-06 2019-05-28 Cortexyme, Inc. Inhibitors of lysine gingipain
US9758473B2 (en) 2014-10-06 2017-09-12 Cortexyme, Inc. Inhibitors of lysine gingipain
US10611769B2 (en) 2015-04-28 2020-04-07 Janssen Sciences Ireland Unlimited Company RSV antiviral pyrazolo- and triazolo-pyrimidine compounds
US11084826B2 (en) 2015-04-28 2021-08-10 Janssen Sciences Ireland Unlimited Company RSV antiviral pyrazolo- and triazolo-pyrimidine compounds
US10208048B2 (en) 2015-04-28 2019-02-19 Janssen Sciences Ireland Uc RSV antiviral pyrazolo- and triazolo-pyrimidine compounds
US10906881B2 (en) 2015-11-09 2021-02-02 Cortexyme, Inc. Inhibitors of arginine gingipain
US10730826B2 (en) 2016-09-16 2020-08-04 Cortexyme, Inc. Ketone inhibitors of lysine gingipain
US11325884B2 (en) 2016-09-16 2022-05-10 Cortexyme, Inc. Ketone inhibitors of lysine gingipain
US12110263B2 (en) 2016-09-16 2024-10-08 Lighthouse Pharmaceuticals, Inc. Ketone inhibitors of lysine gingipain
US11339165B2 (en) 2017-11-29 2022-05-24 Janssen Sciences Ireland Unlimited Company Pyrazolopyrimidines having activity against the respiratory syncytial virus (RSV)
US11491157B2 (en) 2018-01-31 2022-11-08 Janssen Sciences Ireland Unlimited Company Co Cork, IE Cycloalkyl substituted pyrazolopyrimidines having activity against RSV
US11708369B2 (en) 2018-04-23 2023-07-25 Janssen Sciences Ireland Unlimited Company Heteroaromatic compounds having activity against RSV

Also Published As

Publication number Publication date
AU4250596A (en) 1996-07-10
FI972466A0 (en) 1997-06-11
MD970253A (en) 1999-05-31
IL116503A0 (en) 1996-03-31
NO972892L (en) 1997-08-20
NZ297360A (en) 2000-03-27
BG101647A (en) 1998-03-31
LV12019A (en) 1998-04-20
AP9701004A0 (en) 1997-07-31
FI972466L (en) 1997-08-19
LV12019B (en) 1998-07-20
SK83897A3 (en) 1998-05-06
NO972892D0 (en) 1997-06-20
AU715378B2 (en) 2000-02-03
AU4062895A (en) 1996-07-04
CN1175259A (en) 1998-03-04
CZ189997A3 (en) 1998-09-16
PL320965A1 (en) 1997-11-24
EP0802916A1 (en) 1997-10-29
CA2208772A1 (en) 1996-06-27
MX9704718A (en) 1998-06-28
EE9700113A (en) 1997-12-15
IS4504A (en) 1997-06-11
JPH11508535A (en) 1999-07-27
OA10493A (en) 2002-04-10
HUT77651A (en) 1998-07-28

Similar Documents

Publication Publication Date Title
WO1996019483A1 (en) Low molecular weight bicyclic thrombin inhibitors
WO1998009987A1 (en) Lactam inhibitors of thrombin
AU743735B2 (en) Selective factor Xa inhibitors
AU699679B2 (en) Heterocyclic keto arginine peptides as thrombin inhibitors
EP0934064B1 (en) Thrombin inhibitors
US5869487A (en) Pyrido 3,4-B!pyrazines for use as thrombin inhibitors
US6515011B2 (en) Thrombin inhibitors
NZ502877A (en) Bicyclic aryl azepinone selective factor Xa inhibitors for treating thrombosis related diseases
CA2268281A1 (en) Selective factor xa inhibitors
US6124291A (en) Pyrrolo[1,2-a]pyrazine-1,4-dione serine protease inhibitors
NZ509400A (en) Inhibitors of urokinase and blood vessel formation
JP2000516598A (en) Thrombin inhibitor
EP0846120A1 (en) Low molecular weight bicyclic-area type thrombin inhibitors
AU6962398A (en) Selective factor xa inhibitors
NZ500352A (en) Cyclic diaza selective factor Xa inhibitors
US6057314A (en) Low molecular weight bicyclic thrombin inhibitors
WO1998028326A1 (en) Bicyclic thrombin inhibitors
WO1998028326A9 (en) Bicyclic thrombin inhibitors
AU705819B2 (en) Acylated enol derivatives of alpha-ketoesters and alpha-ketoamides
JP2002504490A (en) Azole peptidomimetics as thrombin receptor antagonists
HU221310B1 (en) Acylated enol di and tripeptide derivatives, their use and pharmaceutical compositions comprising thereof
LT4368B (en) Low molecular weight bicyclic thrombin inhibitors
AU753842B2 (en) Selective factor Xa inhibitors
EP0865445A1 (en) 3-aminoethyl-n-amidino-2,5-dihydropyrrole derivatives having arginine mimetic properties
MXPA00001443A (en) SELECTIVE FACTOR Xa INHIBITORS

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 95197614.1

Country of ref document: CN

AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU BB BG BR BY CA CH CN CZ DE DK EE ES FI GB GE HU IS JP KE KG KP KR KZ LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TT UA UG US UZ VN

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 972466

Country of ref document: FI

WWE Wipo information: entry into national phase

Ref document number: PV1997-1899

Country of ref document: CZ

ENP Entry into the national phase

Ref document number: 2208772

Country of ref document: CA

Ref document number: 2208772

Country of ref document: CA

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 83897

Country of ref document: SK

Ref document number: 297360

Country of ref document: NZ

Ref document number: 97-01144

Country of ref document: RO

WWE Wipo information: entry into national phase

Ref document number: 1019970704288

Country of ref document: KR

ENP Entry into the national phase

Ref document number: 1996 519383

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: PA/a/1997/004718

Country of ref document: MX

Ref document number: 08880885

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 97132

Country of ref document: LT

Ref document number: 97-0253

Country of ref document: MD

Ref document number: 97-132

Country of ref document: LT

WWE Wipo information: entry into national phase

Ref document number: 1995940923

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 1995940923

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 97-132

Country of ref document: LT

WWP Wipo information: published in national office

Ref document number: 1019970704288

Country of ref document: KR

WWG Wipo information: grant in national office

Ref document number: 97-132

Country of ref document: LT

WWP Wipo information: published in national office

Ref document number: PV1997-1899

Country of ref document: CZ

WWW Wipo information: withdrawn in national office

Ref document number: 97-132

Country of ref document: LT

WWR Wipo information: refused in national office

Ref document number: 1019970704288

Country of ref document: KR

WWR Wipo information: refused in national office

Ref document number: 1995940923

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1995940923

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: PV1997-1899

Country of ref document: CZ

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载