OA10493A

OA10493A - Low molecular weight bicyclic thrombin inhibitors

Info

Publication number: OA10493A
Application number: OA70029A
Authority: OA
Inventors: John Dimaio; Arshad M Siddiqui; John W Gillard; Yves St-Denis; Micheline Tarazi; Patrice Preville; Sophie Levesque; Benoit Bachand; Annette Marian Doherty; Jeremy John Edmunds
Original assignee: Iaf Biochem Int
Priority date: 1994-12-22
Filing date: 1997-06-18
Publication date: 2002-04-10
Also published as: IL116503A0; NO972892L; AU4062895A; CN1175259A; CA2208772A1; WO1996019483A1; MD970253A; AP9701004A0; AU715378B2; EE9700113A; BG101647A; HUT77651A; IS4504A; SK83897A3; MX9704718A; PL320965A1; LV12019B; AU4250596A; FI972466A; EP0802916A1

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 BICYCLICTHROMBIN INHIBITORS 010493

5 PIELD OP THE INVENTION

This invention relates to.compounds useful for thetreatment of thrombotic disorders, and more particularlyto novel heterocyclic inhibitors of the enzyme thrombin. 10

BACRGRODND

J

Inondinate thrombus formation on blood vessel wallsprécipitâtes acute cardiovascular disease stutes that are 15 the chief cause of death in economically developed societies. Plasma proteins such as fibrinogen, proteasesand cellular receptors participating in hemostasis hâveemerged as important factors that play a rôle in acute andchronic coronary disease as well as cérébral artery 20 -disease by contributing to the formation of thrombus orblood clots that effectively diminish normal blood flowand supply. Vascular aberrations stemming from primarypathologie States such as hypertension, rupture ofatherosclerotic plaques or denuded endothélium, activate 25 biocbemical cascades that serve to respond and repair thein jury site. Thrombin is a key regulatory enzyme in thecoagulation cascade; it serves a pluralistic rôle as botha positive and négative feedback regulator. However, inpathologie conditions the former is amplified through 30 catalytic activation of cofactors required for thrombin génération as well as activation of factor XIII necessaryror fibrin cross-linking and stabilization.

In addition to its direct effect on hemostasis, thrombin 35 eyert-.s direct effects on diverse cell types t haf-'supportand acnplify pathogenesis of arterial thrombus disease.

The enzyme is the strongest activator of platelets causing 010493 them to aggregate and release substance;; (eg. ADP TXA, NE)that further propagate the thrombotic cycle, Platelcts ina fibrin mesh comprise the principal framowork of a whitethrombus. Thrombin also exerts direct effects on 5 endothélial cells causing release of vasoconstrictor substances and translocation of adhesion molécules thatbecome sites for attachment of immune cells. In addition,the enzyme causes mitogenesis of smoo'th muscle cells andprolifération of fibroblasts. From this analysis, it is 10 apparent that inhibition of thrombin activity constitutesa viable therapeutic approach towards the atténuation ofproliférative events associated with thrombosis.

The principal endogenous neutralizing factor for thrombin 15 activity in mammals is antithrombin III (ATIII) , a circulating plasma macroglobulin having low affinity forthe enzyme. Heparin exerts clinical efficacy in venonsthrombosis by enhancing ATIII/thrombin binding throughcatalysis. However, heparin also catalyzes inhibition of 20 other proteases in the coagulation cascade and its efficacy in platelet-dependent thrombosis is largelyreduced or abrogated due to inaccessibility of thrombus-bourrd encyme. Adverse side effects such as thrcmbocytopenia, osteoporosis and triglyceridemia hâve 25 been observed following prolonged treatment with heparin.

Hirudin, derived from the glandular sécrétions of theleech hi^ido medicinalis is one of the high molecularweight natural anticoagulant protein inhibitors of 30 throcnbin activity (Markwardt F. Cardiovascular Drug

Reviews, AO, 211, 1992). It is a biopharmaceutical thathas demonstrated efficacy in experimental and clinicalthrombosis. A potential drawback to the use of Hirudin asa therapeutic agent is likely antigenicity and lack of an 35 effective method of neutralization, especial.iy iiï view of its extremely tight binding characteristics toward thrombin. The exceedingly high affinity for thrombin i;; 3 0 1 0 4 0 3 unique and is attributed to a simultaneou:; interactionwith the catalytic site as well as a distal "anion bindingexosite" on the enzyme. 5 Thrombin activity can also be abrogated by Hirudin-likemolécules such as hirulog·(Maraganore, û.M. et al.,Biochemistry, 29., 7 095, 1990) or hirutonin peptides(DiMaio, J. et al-, J. Med. Chem., 3 5'. 3331, 1992). 10 Thrombin activity can also be inhibited by low moleeularweight compounds that compete with fibrinogen forthrombin's catalytic site, thereby inhibiting proteolysisof that protein or other protein substrates such as thethrombin receptor. A common strategy for designing enzyme 15 inhibitory compounds relies on mimicking the spécificitéinhérent in the primary and secondary structure of theenzyme's natural substrate. Thus, Blomback et al. firstdesigned a thrombin inhibitor that was modeled upon thepartial sequence of the fibrinogen A(LBi)a chain comprising 20 its proteolytically susceptible région (Blomback, et al..,J. Clin. Lab. Invest., 24, 59, 1969). This région offibrinogen minimally includes the residues commencing withphenylalanine: 25 Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg-Gly-Pro-Arg T scissile bond

Systematic replacement of amino acids within this région 30 has led to optimization of the tripeptidyl inhibitory sequence exemplified by the peptide (D)-Phe-Pro-Arg whichcorresponds to interactions within the Ρ,-Ρ.,-Ρ, localbinding sites on thrombin (Bajusz S. et al. in Peptides: Chemistry 35 Structure and Biology: Proceedings of the Fourth American ο J 0498

Pepcide Symposium, Walter R., Meienhofer J. Eds. Ann ArborScience Publishers Inc., Ann Arbor MI, 1975, pp 603).

Bajusz et al. hâve also reported related compounds such as(D)Phe-Pro-Arg-(CO)H (GYKI-14166) and (D)Mel’he-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 ChemicalCompany , 1981, pp. 417). These tripeptidyl aldéhydes areeffective thrombin inhibitors both in vitro and in vivo.

In the case of both GYKI-14166 and GYKI-14766, thealdéhyde group is presumed to contribute strongly toinhibi tory activity in view of its Chemical reactivitytoward thrombin's catalytic Ser195 residue, générât ing ahemiacetal intermediate.

Related work in the area of thrombin inhibitory activityhas exploited the basic récognition binding motifengendered by the tripeptide (D)Phe-Pro-Arg whileincorporating various functional or reactive groups in thelocus corresponding to the putative scissile bond (i.e,Pl-Fl')·

In U.S. Patent 4,318,904, Shaw reports chloromethyl-ketones (PPACK) that are reactive towards Ser19!i and Hisi7.These twc residues comprise part of thrombin1 s catalytictriad (Bcde, VJ. et al., EMBO Journal .8, 3467, 1989).

Other exemples of thrombin inhibitors bearing the (D)Phe-Pro-Arg general motif are those incorporating COOH-remuLnal boroarginine variants such as boronic acids orboronates (Kettner, C. et al., J. Biol. Chem., 268. 4734, 1993

Stili other congeners of this motif are those bearing phosphonates (Wang, C-L J., Tetrahedron betters, 33 , 7667, Oî 0493 1992) and α-Keto esters (Iwanowicz, E.J. et al.,Bioorganicand Médicinal Chemistry Letters, 12, 1607, 1992).

Neises, B. et al. hâve described a trichloromethyl ketone 5 thrombin inhibitor (MDL-73756) and Attenburger, û.M. etal. hâve revealed a related difluoro alkyl timide ketone(Tetrahedron Letters, 32., 7255, 1991) .

Maraganore et al. (Européen 0,333,356; WO 91/02750; U. S. 10 5,196,404) disclose a sériés of thrombin inhibitors that incorporate the D-Phe-Pro- moiety and hypothesize thatthis preferred structure fits well within the grooveadjacent to the active site of thrombin. Variations onthese inhibitors are essentially linear or cyclic peptides 15 built upon the D-Phe-Pro moiety.

Another sériés of patents and patent applications hâvedescribed attempts to develop effective inhibitors againstthrosmbosis by using alpha-ketoamides and peptide aldéhyde 20 analogs (EP 0333356;WO 93/15756; WO 93/22344; WO 34/08941;WO 94/17817).

Still others hâve focused their attention on peptides,peptide dérivatives, peptidic alcohols, or cyclic peptides

25 as anti-thrombotic agents (WO 93/22344, EP 0276014; EP0341607; EP 0291982). Others hâve examined amidinesulfonic acid moieties to achieve this same end (U.S.4,781,866), while yet others hâve examined para or metasubstituted phenlyalanine dérivatives (WO 92/08709; WO 30 92/6549). A sériés of Mitsubishi patents and patent applicationshâve disclosed apparently effective argininamide compoundsfor use as antithrombotic agents. The Chemical structures 35 described in these documents represent variations' of sidegroups on the argininamide compound (U.S. 4,173,63 0; U. S.

r-r·»’ vr·"» |F οî0493 4,097,591; CA 1,131,621; U.S. 4,096,255; U.;;. 4,046,876; U.S. 4,097,472; CA 2,114,153).

Canadian patent applications 2,076,313 êind 2,055,850 5 disclose cyclic imino dérivatives thaï exhibit inhibitoryeffects on cellular aggregation.

Many of the examples cited above are convergent bymaintaining at least a linear acyclic tripeptidyl motif 10 consisting of an arginyl unit whose basic side chain is required for interaction with a carboxylate group locatedat the base of the Pj specificity cleft in thrombin. Twoadjacent hydrophobie groups provide additional bindingthrough favourable Van der Waals interactions within a 15 contiguous hydrophobie cleft on the enzyme surfacedesignated the P3-P2 site.

One object of the présent invention is to provide thrombininhibitors that display inhibitory activity towards the 20 target enzyme, thrombin. A further object of the présent invention is to providethrombin inhibitors that display inhibitory activitytowards the target enzyme thrombin and are provided for in 25 a pharmacologically acceptable State.

Still a further object of the présent invention is toprovide for the use of heterocyclic thrombin inhibitorsand formulations thereof as anticoagulant and thrombin 30 inhibitory agents.

Yet a further object of the présent invention is toprovide for the use of heterocyclic thrombin inhibitorsand formulations thereof for therapeutic treatment of 35 various thrombotic maladies. ~ 010493 A further object of the présent invention in a process forthe synthesis of these low molecular weight thrombininhibitors. The enzyme inhibitors of the présent inventionare encompassed by the structure of general Formula I.

G 010493

SDMMARY OF THE INVENTION

The présent invention provides for novel compounds thaïdisplay thrombin inhibitory activity as rcflected in 5 formula I:

wherein: A is selected from (CH-Re)„., , S, SO, SOs, O and NR„ wherein 10 R, is hydrogen, C,_6 alkyl optionally interupted with 1 or2 heteroatoms; C6.16 aryl, C3.7 cycloalkyl or heterocyclicring or a hydrophobie group; B is selected from S, SO2, O, -N=, NH, -CH= und CRcR.,wherein R, and R7 are independently selected from 15 hydrogen and C,_6 alkyl provided that when A is S, SO, SO,, O, or NRe, then B is CR6R7; D is selected from <CH-R9)0_2 wherein R, is hydrogen, C,.calkyl or -C(O)R,; and CH with a double bond to B when Bis -N= or -CH=; 20 E is selected from CH2 and CH substituted with the -C(O)R,,provided that only one of D and E is substituted withwith -C(O)R,; X is selected from O, N-Rs, or CH-R, ,· Y is selected from O, S, SO, SO?, N-R,, and CH-RU provided

25 that when X is N-Rs then Y is CH-R„ or O, and when X is O then Y is CH-R„; Z is selected from O, S and H2;

Rj is a polar amino acid residuearginyl moiety or an analogor dérivative thereof optionally substituted with an 30 amino acid, a peptide or a heterocyclo; "•ai 9 0 1 0493 R2 is selected from H and Cj_6 alkyl optionally substitutedwith Ct. aryl, a 6 member heterocycle or a C3.7 cycloalkylring ; R3 is selected from H, NR6R, and C,.c alkyl; and R4 and R5 are independently selected from II; NR6R7; C6.„, arylor C3.7 cycloalkyl optionally substituted with C,.6 alkyl;C..u alkyl optionally interrupted by one or moreheteroatom or carbonyl group and optionally substitutedwith OH, SH, NR^ or a C6_H. aryl, heterocycle or C,..,cycloalkyl group optionally substituted with halogen,hydroxyl, C,.(. alkyl; an amino acid side chain; and ahydrophobie group.

As will be appreciated from the disclosure to follow, themolécules, compositions and methods of this invention areusefu.1 as anti-coagulants, or in the treatment andprévention of various diseases attributed to the undesirable effects of thrombin, as well as for diagnosticpurposes.

DETAZLED DESCRIPTION OF THE INVENTION

The présent invention relates to molécules which inhibitthe enzyme, thrombin. These molécules are characterizedby a heterobicyclic moiety as illustrated in Formula I:

vherein X, Y, Z, A, B, D, E and R, to R. are as previously cefined. 10 010493

The term "hydrophobie group" (HG) as used hereinafter,refers to any group which lacks affinity foi-, or displaceswater. Hydrophobie groups include but are not limited to 5 C,.2O alkyl, C2.20 alkenyl (e.g. vinyl, allyi) or C2„,0 alkynyl (e.g. propargyl) optionally interrupted by a carbonylgroup, (e.g. forming an acyl group); C„ aryl, C,_7cycloalkyl, C6.20 aralkyl, C6.20 cycloalk'yl substituted C,..,,,alkyl, wherein the aliphatic portion is optionally 10 interrupted by a carbonyl group (e.g. forming an acyl group) and the ring portion is optionally substituted withC,_6 alkyl such as methyl ethyl or t-butyl; or a hydrophobieamino acid side chain. Preferred hydrophobie groupsinclude cyclohexyl, benzyl, benzoyl, phenylmethyl, 15 phenethyl and para-t-butyl-phenylmethyl.

The term "arginyl moiety" represents an arginine aminoacid residue or an analogue or dérivative thereof. Forexample, an analogue or dérivative of the natural residue 20 may incorporate a longer or shorter methylene chain fromthe alpha carbon (i.e. ethylene or butylène chain);replacement of the guanidino group with a hydrogen bonddonating or accepting group (i.e. amino, amidino ormethoxy); replacement of the methylene chain with a 25 constrained group (i.e. an aryl, cycloalkyl or heterocyclic ring); élimination of the terminal carboxyl(i.e. des-carboxy) or hydroxyl (i.e. an alde?hyde) ; or acombination thereof. 30 The term "alkyl" represents a straight or branched, saturated or unsaturated chain having a specified totalnumber of carbon atoms.

The term "aromatic" or "aryl" represents an unsaturated 35 carbocyclic ring(s) of 6 to 16 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NRfR7) halogen or C,., alkyl. Aromatic rings include 11 010493 benzene, napththalene, phenanthrene and anthracene.Preferred aromatic rings are benzene and naphthalene.

The term "cycloalkyl" represents a saturated carbocyclic 5 ring of 3 to 7 carbon atoms which is optionally mono- ordi-substituted with OH, SH, amino (i.e. NRtR.,) halogen orC,.é alkyl. Cycloalkyl groups include cyclo- propyl, butyl,pentyl, hexyl and heptyl. A preferred cycloalkyl group iscyclohexyl. 10

The term “aralkyl" represents a substituent comprising anaryl moiety attached via an alkyl chain (e.g. benzyl,phenethyl) wherein the sum total of carbon atoms for thearyl moiety and the alkyl chain is as specified. The aryl 15 or chain portion of the group is optionally mono- or di-substituted with OH, SH, amino (i.e. NRtR7) halogen or C,^alkyl

The term "heteroatom" as used herein represents oxygen, 20 nitrogen or sulfur (O, N or S) as well as sulfoxyl orsulfonyl (SO or SO2) unless otherwise indicated. It isunderstood that alkyl chains interrupted by one or moreheteroatoms means that a carbon atom of the chain isreplaced with a heteroatom having the appropriate valency. 25 Preferrably, an alkyl chain is interrupted by 0 to 4 heteroatoms and that two adjacent carbon atoms are notboth replaced.

The term "heterocycle" represents a saturated or 30 unsaturated mono- or polycyclic (i.e. bicyclic) ring incorporating 1 or more (i.e. 1-4) heteroatoms selectedfrom N, O and S. It is understood that a heterocycle isoptionally mono- or di-substituted with OH, SH, amino(i.e. NRtR7) , halogen, CF3, oxo or C,.( alkyl. Examples of 35 suitable monocyclic heterocycles include but are~not limited to pyridine, piperidine, pyrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran and » » >· v 12 010493 thicphene. Examples of suitable bicyclie het.erocycler.include but are not limited to indole, quinoline,isocuinoline, purine, and carbazole. 5 The term "hydrophobie amino acid" représente. an amino acidresidue that bears an alkyl or aryl group at.tached to therc-r^-rbon atom. Thus glycine, which has no such groupattached to the α-carbon atom is not a hydrophobie aminoacid. The alkyl or aryl group can be substit.uted, provided 10 that the substituent or substituents do not detract fromthe overall hydrophobie character of the amino acid.Examples of hydrophobie amino acids include natural aminoacid residues such as alanine; isoleucine; leucine;pherrylalanine; and non-naturally ocurring amino acids such 15 as those described in “The Peptides", vol. 5, 1983,Academie Press, Chapter 6 by D.C. Roberts and F.

Vellaccio. Suitable non-naturally ocurring amino acidsinclude cyclohexylalanine and 1-aminocyclohoxane-carboxylic. 20

By "amino acid side chain" is meant the substituentattached to the carbon which is a to the amino group. Forexample, the side chain of the amino acid alanine is amethyl group and while benzyl is the side chain for 25 pherrylalanine.

Preferably Rj is H or C,.t alkyl. More preferably R2 is II,methyl or ethyl and most preferably R2 is H. 30 Preferably, Rj is H or C,.t alkyl. More preferably, R, is H,methyl or ethyl, and most preferably R, is H.

Preferably, one of R4 or Rs is a hydrophobie group such asa sarurated or unsaturated carbocycle of 5 or 6 members 35 optionally fused to another carbocyclic group while theother is H, C..,{ alkyl optionally substituted by NR,R.. or '·* 13 010493 carboxy. The hydrophobie moiety may be linked via a spacersuch as a C,_16 alkyl chain optionally interrupted with 1or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO2)groups. More preferably, one of R4 and Rt is phenyl,cyclohexyl, indole, thienyl, quinoline, tetrahydroisoquinoline, naphthyl or benzodioxolane linkedvia Cj.,* alkyl optionally interupted with a heteroatom or acarbonyl while the other is H, carbo'xyniethyl or carboxyethyl. 10 15

Preferably, A is absent or CH2.

Preferably, B is S or CH2.

Preferably, D is CH2.

Preferably, E is CH substituted with -C(O)R, wherein R, is as previously defined.

Preferably, X is CH-RS or N-Rrj.

Preferably, Y is CH-Re or S.

Preferably, Z is O. 20 In a preferred embodiment, Rx is represented by one offormula Via to VId:

wherein: R_ bs hydrogen or C,.e alkyl; 25 K is a bond or -NH-; G is C..4 alkoxy; cyano; -NH2; -CH..-NH.,; -C (NH)-NH2~-NH-C(33H)-NH2; -CH2-NH-C (NH)-NH2 ; a Ct cycloalkyl or arylsubstituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH- 0 11)493 C(NH)-NH2 or -CH2-NH-C (NH) -NH?; or a .5 or 6 meraber,saturated or unsaturated heterocycle opt.ion.allysubstituted with cyano, -NH2, -CH2-NH,, -C (NH)-NII2, -NH-C(2ÎH)-NH2 or -CH2-NH-C (NH)-NH? ; ü is cyano, -NH2, -C(NH)-NH2 or -NH-C(NH)-NH, ; P is a bond, -C(0)- or a bivalent group:

OH

J is Cj_fc alkylene optionally substituted with OH, NR, andC,„. alkyl and optionally interrupted by a lieteroatomselected from O, S and N; n is 0 or 1; and T is H, OH, amino, a peptide chain, C,.u alkyl, C,_)t alkoxy,Ct,20 aralkyl, or heterocycle optionally substituted.

Preferably Rn is H or methyl and most preferably H.Preferably K is a bond.

Preferably G is -NH-C(NH)-NH2 attached via a methylenechain of 3-7 carbons or phenyl substituted with -C(NH)-NH2attached via a methylene chain of 0 to 3 carbons. Morepreferably G -NH-C(NH)-NH2 attached via a methylene chainof 3 atoms.

Preferably P is -C(0)-.

Preferably J is selected from: -CH2-S-CH,-CH2-; -CR,-O-CH-CR-; -CH.-NH-CH2-CH2-; and a bond when n is 0. Morepreferably, J is a bond while n is 0. in perticular embodiments of the invention, R, is selectedfrom the following amino acid derivativers preparedaccording to the procedures described in Bioorg. Med.Chem., 1935, 3:1145 : 010493

010493

17 «1 0493

ry-ü'-s-.-?· τγ’·/):’*. ™·,τ 18 010403

5 wherein n=l-6, nl=l-2, n2=O-7 and T is as previouslydefined.

In a preferred embodiment, T is a peptide of 1 to 4 amino10 acid residues in length and preferably fibrinogen's A or B chain or fragment or dérivative thereof. In anotherpreferred embodiment, T is a heterocycle selected from thegroup consisting of:

wherein

Xs, X. and X12 are each independently selected from the croup consisting of N, or C-X7 where X7 is hydrogen, C,.4alkyL, or C,.u aryl; 19 θ I 0493 Χ6 and Χ13 are each independently selected irom the groupconsisting of C, O, N, S, N-X,, or CH-X, ; R' is hvdrogen, C,.u alkyl optionally carboxyl substituted,carboxyl, -C0.16 alkyl-CO2-C,.16 alkyl, C6,20 aralkyl, C3.7 5 cycloalkyl, aryl or an aromatic heterocycle.

Preferably T is selected from the group consisting of:

wherein R' is as defined above. 10

More preferably Tof : is selected from the group consisting

20 010493 wherein R' is as defined above.

More preferably T is selected from the group consistintçof :

wherein R' is as defined above.

Most preferably T is

.N y as methyl, ethyl, propylein R' is hydrogen,. In 10 wherein R' is H or C,., alkyl suchor tutyl and most preferably wheranother embodiment, T is a 1,2 thiazole optionallysubstituted with R' and\or is attached to J at the 2, 3, 4or Ξ position of the ring. 15

In p-articular embodiments, compounds of the invention arerepresented by formulas II, III, IV and V, wherein X, Y, B, F. to R4 and R8 are as previously defined.

’VAV Ï*-Jk 21 010433

In a particularly preferred embodiment, compounds of theinvention are represented by one of formulas VIIf VIII, IXand X: (VII) (IX)

10 15 20 25 wherein B is O, S, -CH2-, or -NH-; Y is selected front O, S, SO, SO2, N-Rs and CH-Re; 8, is an arginyl moiety or an analog or dérivative thereofoptionally substituted with an amino acid, a peptide ora heterocycle; ΐς is H or C,.t alkyl; R, is selected from H, NR^ and Cj.6 alkyl; and R4 and R, are independently selected from H; NR6R7; C6.u arylor C3_7 cycloalkyl optionally substituted with C,_6 alkyl;C,.u alkyl optionally interrupted by one or moreheteroatom or carbonyl group and optionally substitutedwith OH, SH, NR6Rj or a C6.u aryl, heterocycle or C3_,cycloalkyl group optionally substituted with halogen,hydroxyl, Cj_6 alkyl; an amino acid side Chain; and ahydrophobie group; R, is hydrogen, C,_6 alkyl optionally interupted with 1 or 2heteroatoms; Cé.16 aryl, C3.7 cycloalkyl or heterocyclicring or a hydrophobie group; and n is 1 or 2.

Preferred compounds according to formula VII include: 22 0005 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a] pyridine-3R- carboxamido (propyl ketoarginine) 0010 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-carboxamido (butyl ketoarginine) 10 15 0015 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-carboxamido(propylcarbmethoxyketoarginine) 0020 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(benzylketoarginine) 20 0025 6S-cyclohexyl methylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine -3R-carboxamido(carbmethoxypropyl cyclodithioketalarginine) 0030 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((S)-Arg-(R) -pipecolilic acid) 010493

25 23 0035 6S-benzylhexa hydro-5-oxo-5H- thiazolo [3,2-a]pyridine-3R- carboxamido (carboxamidopropyl cyclodithioketal arginine) 004 0 6 S- cyclohexylme thylhexahydro- 5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido ( ( S ) -Argnipecotamide) 10 0045 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-carboxamido ( ( S ) Argisonipecotamide) 15 0050 6S-benzylhexahydro-5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido ( carboxamidopentylcyclodithioketal arginine) 20 0055 6S-benzylhexahydro-5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido ( carbmethoxyprcpyl cyclodithioketalarginine) 25 0060 6 S - cyc lohexy lme thy lhexahydr o- 5-oxo-5H-thiazolo [3,2 —a]pyridine-3R-carboxamido(1-carboxy-3-thiobutylketoarginine)

ό

NH HN^NH,

H)' Ü 1 049 3 0055 6 S-cyc1ohexylme thylhexahydro- 5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(1-carboxy-3-thiobutylketoarginine) 007 0 6S-cyclohexylmethylhexahyd.ro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(1-cajsboxy-2-me thyl-3 - thiobutylketoarginine) 0075 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido{(3-thiobutyl sulfonic acid)ketoarginine) 0080 6S-cyclohexylmethylhexahydro- 5-oxo-5H-thiazolo[3,2-a ] pyr idine-3 R-carboxamido(iso-quinoliniummethyl ketoarginine)

NHhpAnh,

et 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R- carboxamido(propylcarbmethoxyketoarginine) 0090 cS-cyclohexylmethylhexahydro-5-cxo-5H-thiazolo[3,2-a ] pyr idine-3 R- carboxamido((prcpylketo)Arg-Fhe-Arg-NH2 )

O 25 0095 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R- carboxamido((propanoic acid) ketoarginine) 5 0100 6S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R-carboxamido (propyl carbmethoxy ketoarginine) 0105 6S-cyclohexylmethylhexahydro-10 5-oxo-5H-thiazolo [3,2- a]pyridine-3R-carboxamido (a-benzothiazolo keto arginine);and 0110 6S-cyclohexylpropylhexahydro-15 5-oxo-5H-thiazolo [3,2- a]pyridine-3R- carboxamido(propylcarbmethoxyketoarginine) 2& 010493 10 15 20 25 0205 6-Benzyl-5-oxo-hexahydro-tbiazolo[3,2-a]pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-bu.tyl]-amide 0210 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1-{benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide C215 6-3enzyl-5-oxo-hexahydro-tiiiazolo[3,2-a]pyridine-3-cazsboxylic acid [1-(bsnzothiazole-2-ca^sbonyl) -4-guanidino-butyl]-amide 022C 6-Senzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylicacid [1-(benzothiazole-2-carbonyl)-4-guanidino-bubyl]-amide

010493 0225 8a-Methyl-5-oxo-6- phene thy 1-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-bu ty l]-amide

0230 8a-Methyl-5-oxo-6- phene thy1-hexahydro- 10 thiazolo[3,2-a]pyridine-3 carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide

15 20 25 0240 8a-Methyl-5-oxo-6-(2- trifluoro methyl-quinolin-6-ylmethyl)-hexahydro-thiazolo[3,2-a] pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0245 6-Eenzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3- ' carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl ) butyl]-amide

o 1

HN o

II nh2 28 010403 0250 6-Benzyl-5-oxo-hexahydro-tbiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2- 5 c arbony 1 ) bu ty 1] - ami de

0255 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-{1-methyl-lH 10 iraidazole-2- carbonyl ) butyl]-amide 0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2-a]pyr idine - 3 - carboxy 1 i c 15 acid [4-guanidino-l- (tbiazole-2-carbonyl)-bu ty 1] - amide 0255 5-Oxo-6-(3-cyclohexyl- propyl)-hexahydro- 20 tfc.iazolo[3,2 -a]pyridine-3 - carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl )butyl]-amide 0275 3a-Mefchyl-5-oxo-6-(3- 25 phenyl-propyl)-hexahydro- thiazolo[3,2-a]pyridine-3 -carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl ) -butylj-amide

H

29 010493 0230 8a-Methyl-5-oxo-6-(3- phenyl-propyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 10

0235 8a-Methyl-5-oxo-6-(2- trifluoromethyl-guinolin-6-ylmethyl)-hexahydro-thiazolo[3,2 -a]pyridine-3 -carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

15 20 25 0295 6—(1,3-Dioxo-l,3-dihydro- isoindol-2-yl)-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 -carboxylicacid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0305 5-Oxo-6-(3-phenyl- propionyl amino)-hexahydrothiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

010493 0315 5-Oxo-6-(3-ph.enyl- propionyl amino)-hexahydrothiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

H

NH NH.

More preferred compounds according to formula (VII)inciude: 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a ] pyridine - 3 R- c arboxamido ( propy1 c arbomethoxyketoarginine); 0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-tliiazolo [3,2-a] pyr idine-3 R-c arboxamido ( (propylketo)Arg-Phe-Arg-NHJ ; 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 ketoarginine); 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-l-(thiazole-2- ·carbonyl)butyl]-amide; 31 010493 0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiu.zolo[3,2- a]pyridine-3-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl ) -butyl] -amide ; 0265 5-Oxo-6- (3-cyclohexyl-propyl) -hexahydro-thiazolo[3,2-5 a]pyridine-3-carboxylic acid [4-guanidino-l-(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-l-(thiazole-2-carbonyl)- 10 butyl]-amide; and 0315 5-0XO-6- (3-phenyl-propionylamino ) -hexahydro thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino1— ( thiazole-2-carbonyl ) -butylj-amide. 15 Most preferred compounds according to formula VII include:0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2- a]pyridine-3R-carboxamido (propylcarbo methoxyketoarginine); and 0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-20 a]pyridine-3R-carboxamido (α-benzothiozolo keto arginine). 32 010493

Preferred compounds according to formula VIII include: 0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo[l,2-a]pyridine-6-carboxylic acid[1-(benzothiazole-2-carbonyl)-4 - guanidino -bu ty 1] - ami de 10 15 20 0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0335 2-Benzoyl-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0340 4-0XO-2-(3-phenyl-propionyl)- octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid[4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide

010493 0345 4-Οχο-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid[4-guanidino-l- (5-methyl- 5 thiazole-2-carbonyl)-butyl]- amide 0350 2-{3-Cyclohexyl-propionyl)-4-oxo- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid[4-guanidino-l-(2-thiazole-carbonyl) -butyl]-amide 0355 5-Oxo-7- (3-phenyl-propionyl) - octahydro-2-thia-4a,Ί-diaza-naphthalene-4-carboxylic acid £5 [4-guanidino-l-(thiazole-2- carbonyl) -butylj-axnide 0365 4-Oxo-2-(4-phenyl-butyryl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid 20 [4-guanidino-l-( thiazole-2- carbonyl ) -butylj-axnide 0370 4-Oxo-2-phenylacetyl-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid 2.5 [4-guanidino-l-(thiazole-2- carbonyl ) -butylj-axnide

NH 010493 34 10 15 20 25 0375 2-(2-Amino-3-phenyl-· propionyl)-4-oxo-octahydro-pyrrolofl, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0380 2-[2-Amino-3-(4-hydroxy-phenyl)-propionyl]-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 0385 2-[2-Amino-3-(4-fluoro- phenyl) -propionyl]-4-oxo-o c t ahydr o -py r r ο 1 ο [ 1,2 - a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 0390 4-Oxo-2-(3-phenyl-propyl)- octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 0395 2-[2-Amino-3-(lH-indol-3-yl)-propionyl]-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide

♦n»· 35 010493 0400 4-Oxo-2-(3-thiophen-3-yl- propionyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]-amide 0405 4-Oxo-2-(S-thiophen^-yl- propionyl) -octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino- 1-(thiazole-2-carbonyl)-butyl]-amide 0410 2-(3-1 H-Imidazol-4-yl- propionyl)-4-oxo-octahydro-pyrrolofl, 2-a] pyrazine-6-carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-butyl]-amide 0415 2- (2-Amino-3-thiophen-3-yl-propionyl) -4-oxo-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1—(thiazole-2-carbonyl)-butyl]-amide 0420 4-Oxo-2-(1,2,3, 4-tetrahydro- isoquinoline-3-carbonyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

NH

y ‘r f * 36 010493 10 0425 2-(Hydroxy-phenyl-acetyl)-4- oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 0430 2-(2-Hydroxy-3-phenyl- propionyl)-4-oxo-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0435 4-Oxo-2-phenoxyacetyl-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

15 010493 37 0440 5 0445 10 0450 15 0455 20 0460 25 4-Oxo-2-(3-phenoxy-propionyl)-ocatahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -bu tyl]-amide4-Oxo-2-(2-phenyl-ethanesulfonyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-l-(thiazle-2-carbonyl)-butylj-amide 2-(Naphthalene-2-sulfonyl)-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide4- ( 6-(4-Guanidino-l- ( thiazole-2-carbonyl ) -butylcarbamoyl]-4-oxo-hexahydro-pyrrolo[l, 2-a]pyrazin-2yl)-4-oxo-3-(2 propyl-pentanoylamino)-butyric acidmethyl ester 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1) -bu tyl]-ami de 0465 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [3- 30 guanidino-propyl) -amide

-· . · ·* * · K » Z ,5* 38 010493 10 13 20 0470 4- (6-[4-Guanidino-l-(thiazole-2- carbonyl) -butylcarbamoyl]-4-oxo-hexahydro-pyrrolofl, 2-a]pyrazin 2-yl)-4-oxo-butyric acid 0475 4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(5-ethyl-thiazole-2-carbonyl)-4-guanidino-butyl]-amide 0480 4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-methyl-thiazole- 2-carbonyl) -butyl]-amide 0485 4-0XO-2- (3-phenyl-propionyl) - octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(4-methyl-thiazole- 2-carbonyl) -butyl]-amide 0490 4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid[l-(4-ethyl-thiazole-2-carbonyl) -4 -guanidino -bu ty 1] - ami de 0495 4-0ΧΟ-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (4-carbamimidoyl-pheny) -amide

NH

O 25 il i ji iicn jub iiimim ιιι i 39 010493 0500 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-phenyl-thiazole-2-carbonyl) -butyl]-axnide 0505 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2-carbonyl)-4-guanidino-butyl]-amide 0510 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [1-(4-carbarnimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0515 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0520 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethylj-amide 0525 4-0XO-2-(3-phenyl-propionyl)- octahydro-pyrrolo(l, 2-alpyrazine-6-carboxylic acid [1-( l-carbamiip.idoyl-piperidin-3 -ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl|-<unide

40 0530 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-2- 5 ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide 0535 [6-[4-Guanidino-l-( thiazole-2- carbonyl) -butylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)- 10 octahydro-pyrrolo[l,2- a]pyrazine-3-yl]-acetic acid 0540 3-[6-[4-Guanidino-l- ( thiazole-2-carbonyl) -butylcarbamoyl]-4-oxo-2 - (3-phenyl-propionyl)- 15 octahydro-pyrrolo[l, 2-a]pyrazin- 3- yl]-propionic acid 0545 [6-[l- ( l-Carbamimidoyl-piperin- 4- ylmethyl)-2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2- (3 — 20 phenyl-propionyl)-octahydro- pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 0550 3-[6-[l-( 1-Carbamimidoyl- piperidin-4-ylmethyl)-2-oxo-2- 25 thiazol-2-yl-ethylcarbamoyl]-4- oxo-2- ( 3 -phenyl-propionyl ) -octahydro-pyrrolo[l, 2-a]pyrazin- 3-yl)-acetic acid

010493

Al 0555 [G—[1— ( 1-Carbamimidoyl- p iperidin-3-y lmethy 1) - 2 -oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)- 5 octahydro-pyrrolo[l, 2-a]pyrazin- 3-yl)-acetic acid 0560 [6-(3-Guanidino- propylcarbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro- 10 pyrrolo[l, 2-a]pyrazin-3-yl ) - acetic acid 0565 3-[6-(3-Guanidino- propylcarbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro- 15 pyrrolo[l, 2-a]pyrazin-3-yl) - propionic acid 0570 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4- 20 guanidino-1-(thiazolc-2- carbonyl) -butyl]-methyl-amide 0575 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-alpyrazine-Ô-carboxylic acid [1- 25 (l-carbamimidoyl-piperidin-4- ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-me thyl-amide

42 010493 0580 [6- ([1-Carbamimidoyl-piperidin- 4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-methyl-carbamoyl) -4-oxo-2- (3-phenyl-propionyl) - 5 octahydro-pyrrolo[l, 2-a]pyrazin- 3 -yl]-acetic-acid 0585 4-Oxo-2- (3-phenyl-propionyl) -’ octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [Ι- ΙΟ ( l-carbainimidoyl-piperidin-3 - ylmethyl) -2-oxo-2-thiazol-2-yl-e thyl] -me thyl - ami de 0520 4-0xo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2- 15 a]pyrazine-6-carboxylic acid (3- guanidino-propyl) -methyl-amide 0525 2-(Naphthalene-2-carbonyl)-4- oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4- 2o guanidino-1-(thiazole-2- carbonyl) -butyl]-amide 0600 2-(Naphthalene-1-carbonyl)-4- oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4- 25 guanidino-1-(thiazole-2- carbonyl) -butyl]-amide

Ml, 010493 43 0605 2-(3-Naphthalen-l-yl- proplonyl)4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-{thiazole-2-carbonyl)-butyl]-amide 0610 2-(4-tert-Butyl-benzoyl)-4-oxo- octabydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butylj-amide 0615 2-(Benzo[l, 3]dioxole-5- carbonyl)-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0620 2-(3-Benzo[l, 3]dioxol-5-yl- propionyl)-4-oxo-octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0-25 2-[2- (2-Methyl-benzylidene) -but- 3 - enoy 1] - 4 - oxo - oc t ahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(1-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethylj-amide

NU,

0630 0635 2- [2- (2-Methyl-benzylidene) -but- 3- enoyl]-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 2- (2-Benzylidene-pent-3-enoyl) - 4- oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-amide 0 J Ο493

o

10 010493 10 15 20 0640 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid 4- c arbamimidoy 1 -benzy lamide 0645 4-0xo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[lz 2-aJpyrazine-6-carboxylic acid [4-imidazol-l-yl-1-(thiazole-2-carbonyl)-butyl]-amide 0650 0655

4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-(2-amino-imidazol-l-yl)-1-(thiazole-2-carbonyl) -butyl]-amide

4-Oxo-2-(3-phenyl-propionyl)-cctahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [3-(2-amino-6-methyl-pyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide

0570 4-Oxo-2-(3-phenyl-propionyl)-oc tahydro-pyrrο 1 ο[ 1,2 -a]pyr az ine-6-carboxylic acid [3-(2-amino-6-chloro-pyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide

25 010493 0675 5 0680 10 0685 15 0690 20 0695 25 4-Oxo-2- (3-phenyl-propionyl)-oc tahydro-pyrrolo[l, 2 -a]pyrazine-6-carboxylic acid [3-(6-amino- 'pyridin-2-yl)-1-(thiazole-2-carbonyl) -propyl]-amide 4-0XO-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [3-(2-amino-pyridin-4-yl)-1-(thiazole-2-carbonyl) -propyl]-axnide

4-Oxo-2~(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [2-(2-amino-pyridin-4-yl)-1-(thiazole-2-carbonyl) -ethyl]-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [2-( 6-amino-pyridin-2-yl)-1-(thiazole-2-carbonyl) -ethyl]-amide

2-[4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carbonyl]-3- ( thiazole-2-,carbonyl)-1,2,3,4-tetrahydro-isoquinoline-6--carboxamidine

010493 iti 0700 2-[4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carbonyl]-3- ( thiazole-2-carbonyl)-1,2,3,4-tetrahydro- 5 isoquinoline-7-carboxamidine 0705 n-[1-[4-Oxo-2-(3-phenyl- propionyl )-octahydro-pyrrolo[l, 2-a]pyrazine-6- 10 carbonyl]-5- ( thiazole-2- carbonyl)-pyrrolidin-3-yl]-guanidine

15 r* 0710 0715 20 0720 25 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(4-amino-cyclohexyl)-2-oxo-2-thiazol-2-yl-e thylj-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-ajpyrazine-6-carboxylic acid [l-(4-amino-cyclohexylmethyl)-2-oxo-2-thiazol-2 -yl-e thylj-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolofl, 2-ajpyrazine-6-carboxylic acid [1-(4-amino-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide

0725 0730 10 4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(4- aminome thyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide

4-Oxo-2-(3-phenyl-propionyl)-oc tahydr o—pyrr o lo[l, 2 - a]pyr az ine-6-carboxylic acid [l—(3— aminome thyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 15 20

0735 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine6-carboxylic acid (2-oxo-l-piperidin-4-ylmethyl-2-thiazol-2-yl-ethyl)-amide 0740 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine6-carboxylic acid (2-oxo-l-piperidin-3-yl-2-thiazol-2-yl-efchyl)-amide 0745

4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine- 6-carboxylic acid [1—(3—guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide

o 25 010493 0750

0755 10 0750 15 0765 20 4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine- 6-carboxylic acid [l-(4- guanidino-cyclohexylmethyl) -2- oxo-2-thiazol-2-yl-ethyl]-amide 4 -0xO“2 - ( 3 -phenyl-propionyl ) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(2-guanidino-cyclohexylmethyl) -2-oxo-2 - thi a z ο 1 - 2-y 1 - e thy 1] - amide4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2-carbonyl)-4-guanidino -bu ty 1] - amide 4-ÛXO-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1- ( 5-phenyl-thiazole-2-carbonyl) -butyl]-amide

25 077 0 4-0xo-2- (3-phenyl-propionyl) - octahydro-pyrido[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide

0775 S-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid[4-guanidinc-l-(thiazole-2- 30 carbonyl )-butyl]-amide

»»»1" ?» jr. «μ »i 010493 07 80 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

0735 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

o 0750 5-0XO-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

NH, 0755 S-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid[1 - ( 1-carbairtiiru.doy 1 -piperidin-4 -ylmethyl)-2-ox;-2-thiazol-2-yl-ethylj-amide

080 0 [4-[4-Guanidino-l- ( thiazole-2- carbonyl) -butylcarbamoyl]-5-oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a,7-diaza-naphthalen-6-ylj-acetic acid

5ί 010493 0805 S-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a/7-diaza-naphthalene-4-carboxylic acid[4-guanidino-l-(thiazole-2- 5 c arbony1 ) -bu ty 1 ] - ami de

0810 3-[4-[4-Guanidino-l- ( thiazole-2-c arbony 1 -bu ty 1 c arbamoy 1] - 5 - oxo -7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza- 10 naphthalen-6-yl]-propionic acid o

0815 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid[3-guanidino-propyl]-amide

15 0320 20 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid[1-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide o

Q325 4-Oxo-2-(3-pheny1-propiony1)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(hydroxy-thiazol-2-yl-methyl)-butyl]-amide

23 52 010493 4-Oxo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine~6~ carboxylic acid (4-guanidino-l- thiazol-2-ylmethyl-butyl) -amide 0830

Mil, 0835 0840 10 0845 15 0850 20 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-thiazol-2-yl-butyl)-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [4-methoxy-l-(thiazole-2-carbonyl)-butyl)-aïnide [6-[4-Methoxy-l- (thiazole-2-carbonyl) -butylcarbamoyl]-4-oxo- 2- (3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazin- 3- ylJ-acetic acid [2- (5-Methoxy-2- ([4-oxo-2- (3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carbonylj-amino) -pentanoyl) -thiazol-5-yl]-acetic acid

0855 25 4-Oxo-2-(3-phenyl-propionyl)-oc tahydro-pyrrolo[ 1,2- ajpyr az ine-β-carboxylic acid [4-amino-l-(thiazole-2-carbonyl)-butylj-amide

NIL 010493 0860 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [5-amino-l-(thiazole-2-carbonyl) -pentyl]-amide 10

0865 4-0xo-2- (3-phenyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [5-guanidino-1- ( thiazole-2-carbonyl) -pentyl]-amide 15

0870 2- (3-Naphthalen-2-yl-propionyl) 4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl ) -butyl]-amide

0875 20 0880 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1- ( l-methyl-lH-ixnidazole-2-carbonyl) -butyl]-amide

4-Oxo-2-(3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide

25 54 010493 0S35 8,8-Dimethyl-4-oxo-2-(3-phenyl- propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide

NU, 5 010493

Preferred compounds according to formula (VIII) inclucle:0325 3-Ainiiiomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo|l, 2- a]pyridine-6-carboxylic acid [1-(bonzothiazole-2-carbonyl) -4-guanidino-butyl]-amide 0330 3-Aminomefchyl-4-oxo-2-phcnylacetyl-octahydro-pyrrololl,2-a]pyrazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl) -4-guanidino-butyll-amicie 0515 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pvrazine-6-carboxylic acid [l-(3-carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 053C 4-QXO-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(l-carbamimidoyl-piperidin-2-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0545 [6-[1— (l-Carbamimidoyl-piperin-4-ylmethyl) -2-oxo-2-t hiaz ο 1- 2 -y 1 - e t hy le ar bamoy 1 ] -4-oxo-2-(3-pheny1-propionyl ) -octahydro-pyrrolofl, 2-a]pyrazin-3-yl ) -acePic acid 0550 3-[6-[l-(l-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-tb.iazol-2-yl-e thylcarbamoylJ-4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acePic acid 0555 [6-[l- (l-Carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4 -oxo-2 - ( 3 -pheny1-propionyl ) -octahydro-pyrrolo[l, 2-a]pyrazin-3-ÿl ) -acePic acid 56 010403 0560 [6- (3-Guanidino-propylcarbamoyl)-4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 0565 3-[6-(3-Guanidino-propylcarbamoyl ) -4-oxo~2~ (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyragin-3-yl) -propionic acid 0575 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl, 2-a]pyrazine-6-carboxylic acid [1-(1-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2.-yl-ethyl}-rnethyl-amide 0580 [6- ([l-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-methyl~carbamoyl ) -4-oxo-2- (3-phenyl-propionyl ) -octahydro-pyrrolo|lz 2-a}pyrazin-3-yl]-acetic-acid 0585 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carbcxylic acid [l-(l-carbamimidoyl-piperidin-3-ylmethyl ) -2-oxo-2-thiazol-2-yl-ethyl]-methyl-amide 0590 4-0XO-2- (3-phenyl-propionyl) -octahydro-pyrrololl, 2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-methyl-amide 0595 2- (Naphthalene-2-carbonyl) -4-oxo-oct:ahydro- pyrrolofl, 2-a]pyracine-6-carboxylic acid [4-guanidino-1- {thiazole-2-carbonyl ) -butyl]-amide 53 010493 0670 4-Oxo-2- (3-phenyl-propionyl)-octahydro-pyrrolo[l, 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[l,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[l, 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-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [2-(2-amino-pyridin-4-yl) -1- ( thiazole-2-carbonyl) -ethylj-amide 0690 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [2-(6-amino-pyridin-2-yl) -1- ( thiazole-2-carbonyl ) -ethylj-ainide 0695 2-[4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolofl, 1-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[l, 2-a]pyrazine-6-carbonyl]-3- ( thiazole-2-carbonyl)- 1.2.3.4- tetrahydro-isoguinoline-7-carboxamidine 0705 H-[l-[4-Oxo-2- (3-phenyl-propionyl) -octahydro- pyrrolo[l, 2-a]pyrazine-6-carbonyl]-5- ( thiazole-2-carbonyl) -pyrrolidin-3-yl]-guanidine 60 010 4 9 0815 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4u, 7-diaza-naphthalene-4-carboxylic acid |3~guanidino-propyl]-amide 5 0820 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7- diaza-naphthalene-4-carboxylic acid |1-(1-carbamim.idoyl-piperidin-3-ylmethyl ) -2-oxo-2-thiazol-2-yl-ethyl]-amide 10 0830 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid (4-guanidino-l-thiazol-2ylmethyl-butyl)-amide 0835 4-ÛXO-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-15 a]pyrazine-6-carboxylic acid [4-guanidino-l-thiazol-2 yl-butyl)-amide

More preferred compounds according to formula VIII 20 include: 0335 2-Benzoyl-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6- carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 25 0 550 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl, 2-a]pyrazine-6-carboxylic acid [4-(2-amino-imidazol-l-yl ) -I- ( thiazole-2-carbonyl ) -butyl]-amide 20 0555 4-0xc—2-(3-phenyl-propionyl)-octahydro-pyrrololl, 2- =]pyrazine-6-carboxylic acid [3-(2-amino-6-methvl-pyrimidin-4-yl )-1-( thiazole-2-carbonyl ) -propylJ-ami de ·* * î- ôi 010493 0715 4-Oxo-2- (3-phenyl-propionyl) -octahydrc>-pyrrolo| 1, ?.-a]pyrazine-6-carboxylic acid [l-(4-amino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 5 0720 4-QXO-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [l-(4-amino-benzyl)-2-oxo - 2 - thi a z ο 1 - 2-y 1 - e thy 1] - ami de 0725 4-Cxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-10 aJpyrazine-6-carboxylic acid [l-(4-aminomethyl- benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0735 4-Oxo-2-( 3-phenyl-propionyl)-octahydro-pyrrololl, 2-a]pyrazine-6-carboxylic acid (2-oxo-l-piperidin-4- 15 ylmethyl-2-thiazol-2-yl-ethyl)-amide 074C 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl,2- ajpyrazine—6—carboxylic acid (2-oxo-l-piperidin-3-yl-2-thiazol-2-yl-ethyl)-amide 20 075C 4-0XO-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(4-guanidino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 25 0760 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2-carhonyl) -4-guanidino-butyl]-amide 0765 4-0XO-2-(3-phenyl-propionyl)-octahydro-pyrrololl, 2-30 a]pyrazine-6-carboxylic acid [4-guanidino-l-(5-pheny1- thiazole-2-carbonyl) -butylj-amide ·«*irvnii·’'·» i 01 (1403 0770 4-Oxo-2- (3-phenyl-propionyl) -octahydro~pyrido| 1,2- a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butylj-amide 5 0775 5-Cxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a, 7- diaza-naphthalene-4-carboxylic acid [4-guanidino-l-( thiazole-2-carbonyl ) -butyl]-amide 0780 5-Cxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-10 diaza-naphthalene-4-carboxylic acid [1-(4- carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-arnide 0785 S-Qxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-15 diaza-naphthalene-4-carboxylic acid [l-(3- carbamimidoy 1 -benzy 1 ) -2 -oxo-2 - thiazo1 -2 -y 1 -ethy 1] -ami de 079C 5-Oo:o-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-20 diaza-naphthalene-4-carboxylic acid [1-(1- carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yi-ethyl]-amide 0805 5-0XO-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-25 diaza-naphthalene-4-carboxylic acid [4-guanidino-l- ( chiazole-2-carbonyl ) -butylj-amide 082: 30 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[ 1,2-a;pyrazine-6-carboxylic acid [4-guanidino-l-(hydroxy-thia.zol-2-yl-methyl) -butyl]-amide 63 010493 0840 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo| 1,2- a]pyrazine-6-carboxylic acid [4-methoxy-l-(thiacole-2-carbonyl) -butyl]-amide 0845 [6-[4-Methoxy-l- ( 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[l, 2-a]pyrazine-6-carbonyl]-amino) -pentanoyl) -thiazol-5-yl]-acetic acid 0855 4-Oxo-2-(3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-amino-l-(thiazole-2-carbonyl) -butyl]-amide 0860 4-ÛXO-2- (3-phenyl-propionyl) -octahydro-pyrrolofl, 2-a]pyrazine-6-carboxylic acid [5-amino-l-(thiazole-2-carbonyl) -pentylj-amide 0865 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolofl, 2- a]pyrazine-6-carboxylic acid [5-guanidino-l-(thiazole-2-carbonyl) -pentyl]-amide

Most preferred compounds according to formula VIII inc Lu de : 0345 4-Cxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[ 1,2- a]pyrazine-6-carboxylic acid [4-guanidino-l-( 5-metliyl-thiazole-2-carbonyl)-butyl]-amide ; and0340 4-Oxo-2- (3-phenyl-propionyl ) -octahydro-pyrrolo|.1., 2-a]pyrazine- 010493 6-carboxylic acid [4-guanidino-l-( t hia:-,ole-2-carbonyl ) -butyl]-amide.

Prefarred compounds according to formula IX include: 10 0890 3-àmino-4-oxo-2-phenyl-hexahydro-pyrrolo[2,1-b][l, 3]thiazine-6-carboxylicacid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl]-amide 15 20 089Ξ 3-Amino-2-benzyl-4-oxo—hexah.ydro-pyrrolo[2,1-b][lz 3]thiazine-6-carboxylicacid [1-(benzothiazole-2-carbonyl ) -4-guanidino-butyl]-amide 0900 3-Amino-2-cyclohaxyl-4-oxo-haxahydro-pyrrolo[2,1-b][l, 3]thiazine-6-carboxylicacid [1-(benzothiazole-2-carbonyl ) -4-guanidino-butyl]-amide

10 15 0 î I) l, 8 3

Prererred contpounds according to formula X include: 0905 7-Benzyl-6-oxo-octahydro- pyrido[2, l~c][l, 4]thiazine-4-carboxylic acid [1- (benzothiazole-2-carbonyl)-4-guanidino-bu ty 1] - ami de 0910 7-(4-tert-Butyl-benzyl)-6-oxo- octahydro-pyrido[2z1-c][l, 4]thiazine-4-carboxylicacid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0915 6-Oxo-octahydro-pyrido[2,1-c][l, 4]thiazine-4-carboxylicacid [4-guanidino-l-(thiazole-2-carbonyl) -bubylj-amide 0925 7-Benzyl-6-oxo:-octahydro- pyrido[2, l-c][l, 4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)butyl]-amide 0935 7-Benzyl-6-oxo-octahydro- pyrido[2, l-c][l, 4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)butyl |-amide

O o'

S

010493 10 0940 6-Oxo-7-phenet hy 1-oc tahydro- pyrido[2, l-cj[l, 4]thiazine-4-carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-butyl]-amide 0950 7-Benzyl-2,2,6-trioxo- octahydro-21>6__pyrido[2,1-cj[l, 4jthiazine-4-carboxylicacid [4-guanidino-l-(thiazole- 2- carbonyl) -butylj-amide

More preferred compounds according to formula X include:925 7-Benzyl-6-oxo-octahydro-pyrido[2, l-c]|l, 4jthiazine-4- carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) butylj-amide; and940 6-Oxo-7-phenethyl-octahydro-pyrido[2, l-c][l, 4]thiazine-4-carboxylic acid[4-guanidino-l- ( thiazole-2-carbonyl ) -butylj-amide.

Preferred compounds according to formula III include: 20 25 096C 4-oxo-l-(3-phenyl-propionyl)-octahydropyrrolo[1,2]pyrimidine-6-carboxylic acid[4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0965 4-oxo-l-(phenethy1suifonyl)- octahydropyrrolo[1,2]pyrimidine-6-carboxylic acid[4-cuanidino-l-(thiazole-2-carbonyl)-butyl]-amide

•-r» ·4**γ»“ϊι' 010493 07

For préparation of the compounds of formula (VII) variousméthode can be employed depending upon the particularstarting materials and/or intermediates involved. The 5 following scheme is one particular method of y J -tfi. — 63 010493 SCHEME 1

5 010493

Step 1:

The alkylation of a is done with appropriât»: bases acccrding to the procedures described in Evans et a.1 (J.Am. Chem. Soc., 1981, 103, 2127; ibid, 1982, 104, 1737;Aldrichimica Acta, 1982, '15, 23) to give 1_>.

Step 2:

Compound b upon hydroboration and oxidation followinçjconditions available in the literature (Synthesis, 1980,151' results in the aldéhyde c.

Step 3 :

The formation of adduct e from aldéhyde p with d is donebv soirring the reactant in aromatic solvents e.g. benzeneor toluene- in presence of catalytic amount of suitableacid e.g, p-toluenesulfonic acid.

Step 3':

The dnter conversion of aldéhyde c to aldéhyde a isreaddly achieved by appropriate protection deprotectionprotocals found in T. Greene, Protective Groups In OrganicSyntbesis, (John Wiley & Sons, 1981) .

Step 4:

The cylization of adduct ê to f may readily be achievedby appropriate Lewis acids e.g, trimethyl aluminum insuitable solvents e.g. dichloromethane, the methodologyfound in T. Greene, supra.

Aloematively, the compound f can be derived from theureatment of aldéhyde c with d in presence of suitablearcmatic solvents e.g, benzene.

The aster function (-C(O)O-R,„) of the bicyclic dnoermediste of formula X is then subjected to 70 010493 hydrolysation usina an app'ropriate agent ;;uch as HCl in anappropriate solvent such as ethyl ether to yield to thefree carboxylic acid. The resulting compound is thencoupied to RJî with a peptide coupling agent such as HOP in 5 an appropriate solvent such as DMF to yield to a bicycliccoupied compound of formula (VIII) . Suitable> conditionsfor peptide bond formation are well known in th art ofpeptide chemistry. For example see Princjples of PeptideSvntbesis, Bodanszky M., Springer-Verlag, Berlin, 10 Heidelberg, New York, Tokyo 1984; and The Peptides,

Analysis, Svnthesis, Bioloav, Vol, l.edited by Gross E.,and Keienhofer J., Academie Press , New York, SanFrancisco, London, 1979. 15 For préparation of the compounds of formula (VIII) variousmetheds can be employed depending upon the particularstarting materials and/or intermediates involved. Thefollcwing scheme is one particular method of préparation. 71 010493 SCHEME 2

OH

*;*·» .fr- ·;ν»»-ί**ί * r-A — 010493 wherein;

Pg is a nitrogen protecting group; each of RJ0? and Rai is independently a C,.,. alkyl; and X, RlZRjZ R< and R, are as previously defined.

The process in scheme 2 is briefly described as follows STEP 1 :

The amino and carboxylic functions of the unsaturatedcompound of formula (a) are protected with appropriaisprotecting groups. A variety of protecting groups knownfor reactive functional groups and suitable protection anddeprotection protocols may be found in T. Greene,Protective Groups In Organic Svnthesis, (John Wiley &

Sons, 19S1). The appropriate protecting group to use i®. a.particular synthetic scheme will dépend on ntany factors,,inclnding the presence of other reactive functional groupsand the reaction conditions desired for removal. Theunsaturated compound of formula is easily obtained bymethcds and protocols known to chemist skill on the art.The protected unsaturated compound of formula (a) issubjected to appropriate conditions to allow cyclisationusing an appropriate reagent such as mercuric acetate iraan inert solvent such as tetrahydrofuran (THF) to yield toa protected amino alcohol of formula (b) . STEP 2

The protected amino alcohol of formula (b) is oxidizedusine an appropriate oxidizing agent such as sulfurtrio’cice pyridine complex in an appropriate solvent suchas dicihcromethane or dimethylformamide to yield to aprotected amino aldéhyde of formula (c). Alternatively,inoemediate (C) can be made by the ozonolysj.s of aoor.pcurd of formula (a') prepared according to Collado etal, T. Oro. Chem,,1995, 60:5011. 010493 73 STEP 3

The protected amino aldéhyde of formula (c) is coupledwith an amino acid alkyl ester of formula (d) by firsfc 5 forming the imine followed by contacting the obtainedimine with an appropriate reagent such as sodiumtriacetoxy borohydride NaBH(OAc)3 to yield to a cyclicintermediate of formula (e). 10 STEF 4

The cyclic intermediate of formula (e) is functionalizedat tbe amino position to yield to the amino substitutedcyclic intermediate of formula (f) . Conditions appropriatefor such reactions are well known in the art and will 15 dépend on the nature of the Rs substituent. STEP 5

The amino protecting group of the cyclic intermediate offormula (f) is removed under appropriate conditions and 20 the resulting compound is then subjected to appropriate condition for internai ring closure such as low heat in aninert solvent or as a raw compound to yield to a bicyclicintermediate of formula (g) . The bicyclic intermediate offormula (g) can also be obtained by hydrolysing the ester 25 f une tien Î-CtOO-RjJ of the cyclic intermediate of formulaig) to the free carboxylic acid followed by standardpeptide coupling using an appropriate coupling reagentsuch as benzotriazole-l-yloxy-tris- i dimethylamino)phosphonium hexafluorophosphate (BOP) in an 30 inert solvent such as dimethyl formamide (DMF). STE? ΐ

The ester function i-C(0)0-R21) of the bicyclicintermediate of formula (g) is then subjected to 35 hydrelysatien using an appropriate agent such as HCl in anappropriate solvent such as ethyl ether to yield to the «10 4 9 3 free carboxylic acid. The rësulting compound is then coupled to RJI 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 5 for peptide bond formation are well known in th art ofpeptide chemistry. For example see Princjples of PeptideSvnthesis. Bodanszky M. , Springer-Verlag, Berlin,Heidelberg, New York, Tokyo 1984; and The Peptides,Analysis, Svnthesis, Biolocrv. Vol, l.edited by Gross E., 10 and Meienhofer J., Academie Press , New York, SanFrancisco, London, 1979.

For préparation of the compounds of formula (IX) variousmetnods can be employed depending upon the particular 15 starting materials and/or intermediates involved. The following scheme is one particular method of préparation. 75 010493

Rc Λ

OH V=S-Pg NH-Pg SCHEME 3

b

e

wherein:

Pg is a sulfur or amino protecting group; 5 L is a leaving group; each of R^î and Rai is independently a C^,, alkyl; and Ra, Ra,R4 and Rs are as previously defined.

The process depicted in scheme 3 is briefly described asSollows: 10 STEP 1 :

The carboxylic acid compound (a) is coupled to the cyclicamine compound (b) with a peptide coupling agent such asbenzccriasol-l-yloxy-tris- (dimethylamino ) phosphonium 15 hexarluorophosphate (BOP reagent) in the présence of a base such as n-methylmorpholine in an appropriât^. solventsuch as dimethylformamide (DMF) or dichloromethane (DCM)to yi-eld to an amido compound of formula (c) . Suitable 010493 conditions for peptide bond formation are well known in thart of peptide chemistry. For example see Princioles ofPentide Svnthesis. Bodanszky M., Springer-Verlag, Berlin,Heidelberg, New York, Tokyo 1984; and The Peptides,Analysis, Svnthesis, Biolocrv, Vol, l.edited by Gross E.,and Meienhofer J., Academie Press , New York, SanFrancisco, London, 1979. STE? 2

The compound of formula (c) is subjected to appropriateconditions to allow internai cyclisation to yield to abicyclic intermediate of formula (d). For exemple, acidmediated cyclisation using p-toluenesulfonic acid or TFAin an appropriate solvent such as dichloroethane. STE? 3

The ester function (-C (O) O-RjJ of the bicyclic intermediate of formula (d) is subjected to hydrolysisusing an appropriate agent such as lithium hydroxide(LiCH) in an appropriate solvent such as tetrahydrofuran(THE) to yield to the free carboxylic acid. The resultingcompound is then coupled to R,H with a peptide couplingagent such as BOP in an appropriate solvent such as DMF togive compound (e). Suitable conditions for peptide bondformation are well known in the art of peptide chemistry.For example see Princioles of Peptide Svnthesis. BodanszkyM., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo1984 ; and The Peptides, Analysis, Svnthesis, Biology, Vol.1.edâted by Gross E., and Meienhofer J., Academie Press ,New York, San Francisco, London, 1979.

For préparation of the compounds of formula (X) variousnethrds can be employed depending upon the particularstarsing materials and/or intermediates invoived.· Thefollcwinc scheme 4 is one particular method of --préparation. 010493 SCHEME 4

wherein: 5 each of R20 and R21 is independently a C,.c alkyl; and B, Rrj( R4, and R5 are as previously defined.

The process depicted in scheme 4 is briefjy described asfcllows: STBF 1:

The halogenated compound of formula (a) is converted to ahalcmethyl ketone of formula (b) using an appropriatereagent,such as diazomethane in an inert solvent such asdiefchyl ether at a température of about -25°C to about 0°C.The resulting mixture is then treated under acidicrend s ti nn.q to yield to the halomethyl ketone of formula(b) . ST5F 2

The halomethyl ketone of formula (b) is coupled with anamine acid alkyl ester of formula (c) with an appropriatebase such as sodium cyanoborohydride in an organic solventsuch as methanol (MeOH) to yield to a cyclic intermediateof formula (d)- 5ΤΞΡ 3

The cyclic intermediate of formula (d) is treated underacidic conditions using an appropriate acid such ascarpborsulfonic acid in an appropriate solvent such ascoluene to yield to a bicyclic intermediate of formula

The ester function (-C (O) O-R30) of the bicyclic intermediate of formula (e) is subjected to hydrolysation’asing an appropriate reagent such as LiOIl to yield to thecréé carboxylic acid. The resulting compound is thencoupled to RjH with a peptide coupling agent such as BOPen an appropriate solvent such as dimethylformamide toyield to a coupled bicyclic compound of formula (X) .üurcaiie conditions for peptide bond formation are wellkec.-.r in ch art of peptide chemistry. For example see 79 010493

Princinles of Peptide Svnthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and ThePeutides, Analysis, Svnthesis, Biology, Vol, l.edi t e d byGrcss E., and Meienhofer J., Academie Press , New York, 5 San Francisco, London, 1979.

Compounds of the présent invention are furthercharracterized by their ability to inhibit the catalyticactivity of thrombin, which is demonstrated in the assay 10 as follows. Compounds of the présent invention may beprepared for assay by dissolving them in buffer to givesolutions ranging in concentrations from 1 to ΙΟΟμΜ. Inan assay to détermine the inhibitory dissociationconstant, K,, for a given compound, a chromogenic or 15 flucrogenic substrate of thrombin would be added to asolution containing a test compound and thrombin; theresulting catalytic activity of the enzyme would bespectrophotometrically determined. This type of assay iswell known to those skilled in the art. 20

The compounds of the présent invention may be used asanti-coagulants in vitro or ex vivo as in thé case ofcontact activation with foreign thrombogenic surfaces suchas is found in tubing used in extracorporeal shunts. The 25 compounds of the invention may also be used to coat thesurface of such thrombogenic conduits. To this end, thecompounds of the invention are obtained as lyophilizedpowd—rs, redissolved in isotonie saline and added in anamourat sufficient to maintain blood in an anticoagulated 30 State.

The therapeutic agents of the présent invention may beadmirai s ter ed alone or in combination with phi^rmaceuticallyacceptable carriers. The proportion of each carrier is 35 determined by the solubility and Chemical nature of the compcund, the route of administration, and standard phanrnaceutical practice. For example, the compounds may 80 Oî 0493 be ûnjected parenterally; this being intiamuscularly,intravenously, or subcutaneously. For parentéraladministration, the compound may be used in the form ofstérile solutions containing other solutés, for example,sufricient saline or glucose to make the solutionisotonie. The compounds may be administered orally in theform of tablets, capsules, or granules containing suitableexcipients such as starch, lactose, white sugar and thelike. The compounds may also be administere^d sublinguallyin the form of troches or lozenges in which each activeingrédient is mixed with sugar or corn syrups, flavouringagents and dyes, and then dehydrated sufficiently to makethe mixture suitable for pressing into solid form. Thecompounds may be administered orally in the form ofsolutions which may contain colouring and/or flavouringagents.

Phys;cians will détermine the dosage of the présenttherapeutic agents which will be most suitable. Dosagesmay vary with the mode of administration and thepartrcular compound chosen. In addition, the dosage mayvary with the particular patient under treatment.

Khen the composition is administered orally, a largerçuantity of the active agent will typically be reguired toproduce the same effect as caused with a smaller quantitycives parenterally.

To further assist in understanding the présent invention,the following non-limiting examples of such thrombininhihitory compounds are provided. The followingexemples, of course, should not be construed asspecifically limiting the présent invention, variationspresently known or later developed, which would be withânthe purview of one skilled in the art and considered tofall withrn the scope of the présent invention asdescrrbed herein. The preferred compounds as of the 01υ 49 3 présent invention are synthesized using conventional préparative steps and recovery methods known to those skiiled in the art of organic and bio-organic synthesis, whiie providing a new a unique combination for the overall 5 synthesis of each compourid. Preferred synthetic routes for intermediates involved in the synthesis as well as theresulting anti-thrombotic compounds of the présentinvention follow. EXA2-IFLE 1

Boc

N

I ch3 A solution of fcert-butyloxycarbonyl-iodo-alanine-N,0-dimethyiamide (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-purgedround bcttom flask charged with zinc-copper couple (0.90 g) .The resuJLting mixture was sonicated under nitrogen until nostartinç material remained (as judged by TLC). Bis(tri-o-tolylphcsphine)palladium dichloride (0.35 g, 0.40 mmol) wasadded fcllowed by 4-iodobenzonitrile (1.72 g, 7.5 mmol) . Theresulting mixture was stirred under a nitrogen atmosphère withheating, allowed to cool, ethyl acetate (100 mL) was added,and the mixture filtered into a separatory funnel. Sequentialwasbing with aqueous HCl (50 mL; 0.1N), distilled H2O (3 x 50mL) , drying over Na2SO4, filtration, and concentration underreduced pressure yielded the crude product. Flash chrcmatography over silica gel (light petroleum-ethyl acetategradient) afforded the purified compound.

A solution or ter't-butyloxycarbonyl-iodo-alanine-N, O- «10493 dinethylamide (2.68g, 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) . 5

The resulting mixture was sonicated under nitrogen until nostarting material remained (as judged by TLC) . Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 ramol) wasadded followed by 3-iodobenzonitrile (1.72 g, 7.5 mmol). Theresulting mixture was stirred under a nitrogen îitmosphere with 2θ heating, allowed to cool, ethyl acetate (100 mL·) was added, and the mixture filtered into a separatory tunnel. Sequentialwasning with aqueous HCl (50 mL·; 0.1N), distilled H2O (3 x 50mL·) , drying over Na2SO4, filtration, and concentration underreduced pressure yielded the crude product. Flash m chromatography over silica gel (light petroleum-ethyl acetategradient) afforded the purified compound.

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethyiamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57,3397-34(34} in dry benzene (30 mL) , and dry N,N- 2θ dimethyiacetamide (2.0 mL) was added to a dry nitrogen-purgedround bottom flask charged with zinc-copper couple (0.90 g) .The resulting mixture was sonicated under nitrogen until nostanting material remained (as judged by TLC). Bis(tri-o-tolylphcsphine)palladium dichloride (0.35 g, 0.40 mmol) was 25 added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol) . Theresulting mixture was stirred under a nitrogen atmosphère withheating, allowed to cool, ethyl acetate (100 mL) was added,and the mixture filtered into a separatory funnel. Sequential ' *· · Vf (110 493 washinc with agueous HCl (50 mL; 0.1N), distilled H2O (3 x 50 mL) , drying over Na2SO4, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatograpby over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.

,OCHq ch3

To a solution of tert-butyloxycarbonyl-para-cyano- phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dryéthanol (20 mL) was added hydroxlyamine hydrochloride (0.416g, 6.0 nmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).The mixture was refluxed and then cooled. The precipitate wasfiltered, washed with cold éthanol, diisopropylether, driedwith MgSO4, concentrated under reduced pressure, and useddirectly in the next step. The semi-solid was suspended in amixture of acetic acid (20 mL), and dry éthanol (40 mL) withwarming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) wasadded, and hydrogen was bubbled through the mixture withwarming. The hydrogénation was continued until no startingmaterial could be detected as judged by TLC. The catalyst wasrentoved by filtration, the solution was concentrated underreduced pressure (50 mL), HCl (50 mL, 1 N) was added, and themixcure was concentrated once again to 50 mL. Tlie solutionwas chilled ovemight yielding the title compound.

Το a solution of tert-butyloxycarbonyl-meta-cyano- phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dryéthanol (20 mL) was added hydroxlyamine hydrochloride (0.416g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).The mixture was refluxed and then cooled. The precipitate wasfiltered, washed with cold éthanol, diisopropylether, driedwith MgSO<, concentrated under reduced pressure, and useddirectly in the next step. The semi-solid was suspended in amixture of acetic acid (20 mL) , and dry éthanol (40 mL) withwarming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) wasadded, and hydrogen was bubbled through the mixture withwarming. The hydrogénation was continued until no startingmaterial could be detected as judged by TLC. The catalyst wasremoved by filtration, the solution was concentrated underreduced pressure (50 mL) , HCl (50 mL, 1 N) was added, and themixture was concentrated once again to 50 mL. The solutionwas chilled ovemight yielding the title compound.

Ü i (i 4 !> 3

To a solution of fcerfc-butyloxycarbonyl-ortho-cyuno- phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dryéthanol (20 mL) was added hydroxlyamine hydrochloride (0.416g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).The mixture was refluxed and then cooled. The precipitate ®assfiltered, washed with cold éthanol, diisopropylether, driedwith MgSO4, concentrated under reduced pressure, and useddirectly in the next step. The semi-solid was suspended in amixture of acetic acid (20 mL), and dry éthanol (40 mL) withwarming. Subseguently, Pd/C catalyst (0.30 g, 10% Pd) wasadded, and hydrogen was bubbled through the mixture withwarming. The hydrogénation was continued until no startingmaterial could be detected as judged by TLC. The catalyst wasremoved by filtration, the solution was concentrated underreduced pressure (50 mL), HCl (50 mL, 1 N) was added, and themixture was concentrated once again to 50 mL. The solutionwas chilled ovemight yielding the title compound.

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-Butyioxycarbonyl-para-amidino-phenylalanine-N, O-dimethylamide(1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, andthe resulting mixture stirred. The reaction was quenched withsaturated agueous ammonium chloride. The mixture was dilutedwith ethyl acetate (150 mL) , and the organic layer washed withsaturated agueous ammonium chloride (2 x 50 mL), brine. (50mL ) , dried with MgSO4, filtered, and concentrated under reducedpressure. The crude material was purified on silica gel ü10493 (ethyl acetate/hexane) , and concentrâtes under reducodpressure.

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-Bu ty 1 oxy c arbony 1 - me ta - amidino -pheny 1a1anine-N,0- dime t hy 1 ami de(1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise andthe resulting mixture stirred. The reaction was quenched withsaturated aqueous ammonium chloride. The mixture was dilutedwith ethyl acetate (150 mL), and the organic layer washed withsaturated aqueous ammonium chloride (2 x 50 mL), brine (50mL)t dried with MgSO4, filtered, and concentrated under reducedpressure- The crude material was purified on silica gel(ethyl acetate/hexane), and concentrated under reducedpressure.

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) 010493 t ert- dropwise at -78° C, and the solution stirred. terfc-Butyloxycarbonyl-orfcho-amidino-phenylalanine-N, O-dimethylamide 10 (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, andthe resulting mixture stirred. The reaction was quenched withsaturated aqueous ammonium chloride. The mixture was dilwtedwith ethyl acetate (150 mL) and the organic layer washed withsaturated aqueous ammonium chloride (2 x 50 mL) , brine (50mL), dried with MgSO„, filtered, and concentrated under reducedpressure. The crude material was purified on silica gel(ethyl acetate/hexane), and concentrated under reducedpressure.

t er z - Bu ty loxy c arbony 1 -para-cy ano -pheny 1 a 1anine-N,O- dimethylamide (1.33 g, 4.0 mmol) was dissolved in éthanolsaturated with ammonia (30 mL) , and sponge Raney Ni (100 mg)added. The solution was shaken under H2 at room température(40 psi) . The solution was filtered through celite, andconcentrated under reduced pressure to yield a clear residue.

The residue was dissolved in ethyl acetate (250 mL), andwashed with 1 N NaOH (2 x 50 mL) , and brine (2 x 50 mL) . Thesolution was dried with MgSO4, filtered, and concentrated underreduced pressure. «10493

t er t -Bu ty 1 oxy c arbony1 -meta- cyano -pheny1 alanine-Ν,Ο-dimethylamide (1.33 g, 4.0 mmol) was dissolved in éthanolsaturated with ammonia (30 mL), and sponge Raney Ni (100 mg)added. The solution was shaken under H2 at roont température 5 (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) , andwashed with 1 N NaOH (2 x 50 mL), and brine (2 x 50 mL). Thesolution was dried with MgSO4, filtered, and concentrated under 0 reduced pressure.

ter fc-Butyloxycarbonyl-or tho-cyano-phenylalanine-N, O- dimethylamide (1.33 g, 4.0 mmol) was dissolved in éthanolsaturated with ammonia (30 mL), and sponge Raney Ni (100 mg)added. The solution was shaken under H2 at room température(40 psi). The solution was filtered through ceJite, andconcentrated under reduced pressure to yield a clear residue.

The residue was dissolved in ethyl acetate (250 mL),"andwashed with 1 N NaOH (2 x 50 mL), and brine (2 x 50 mL). Thesolution was dried with MgSO„, filtered, and concentrated under 9'· reduced pressure. 010493

tert-Eutylox\*carbonyl-para-aminomethyl-phenylalcinine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10mL) under nitrogen with stirring. The solution was cooled,N,N‘-bis-(bemyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2mmol·), and Hçr21, (0.95 g, 3.5 mmol) added. The solution wasconcentrated under reduced pressure, the remaining residue wassuspendeâ in ethyl acetate (200 mL), and filtered throughcelite. The filtrate was concentrated under reduced pressure.

Flash cnromatography over silica gel (hexane/ethyl acetategradient' afforded the purified compound.

terC-EutydLcxy-carbonyl-.meta-aminomethyl-phenylalanine-N, O-dimerhylamide (1.00 g, 3.1 mmol) was dissolved in dry~THF ( i 0mL) under nitrogen with stirring. The solution was cooled,N,N'-bis-(benrylonycarbonyl)-S-methyl-isothiouren (1.14 g, 1.2 «10493 mmol) , and HgCl2 (0.95 g, 3.5 mmol) added. The solution w,is 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 acetategradient) afforded the purified compound.

t er c-Butyloxycarbonyl-or fcho-aminomethyl-phenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10mL) under nitrogen with stirring. The solution was cooled, N, N* -bis-(benzyloxycarbonyl) -S-methyl-isothiourea (1.14 g, 3.2mmol), and HgCl2 (0.95 g, 3.5 mmol) added, The solution wasconcentrated under reduced pressure, the remaining residue wassuspended in ethyl acetate (200 mL) , and filtered throughcelite. The filtrate was concentrated under reduced pressure.

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (Π 1)493 (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 ml*, 13.9 inrnoJ )dropwise at -78’ C, and the solution stirred. The protectedamiho acid (1.36 g, 3.3 mmol) in THF (15 mL) was then addeddropwise, and the resulting mixture stirred. The reaction wasquenched. with saturated aqueous ammonium chloride. Themixture was diluted with ethyl acetate (150 mL) , and theorganic layer washed with saturated aqueous ammonium chloride(2 x 50 mL) , brine (50 mL) , dried with MgSO<, filtered, andconcentrated under reduced pressure. The. crude material waspurifiée on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

15 !0

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. The protectedamino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then addeddropwise, and the resulting mixture stirred. The reaction wasquenched with saturated aqueous ammonium chloride. Themixture was diluted with ethyl acetate (150 mL) , and theorganic layer washed with saturated aqueous ammonium chloride(2 x 50 mL) , brine (50 mL) , dried with MgSO4, filtered, andconcentrated under reduced pressure. The crude material waspurified on silica gel (ethyl acetate/hexane) , andconcentrated under reduced pressure. 93 010493

Το 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. The protectedamino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then addeddropwise, and the resulting mixture stirred. The reaction wasquenched with saturated aqueous ammonium chloride. Themixture was diluted with ethyl acetate (150 mL) , and theorganic layer washed with saturated aqueous ammonium chloride(2 x 50 mL) , brine (50 mL) , dried with magnésium sulfate,filtered, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure.

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- dimerhylacetamide (2.0 mL) was added to a dry ni trogen-purgedround boctom flask charged with zinc-copper couple (0.90 g) .The resulting mixture was sonicated under nitrogen until no ν-τ « 1 U 4 9 3 starting material remained (as judged by TLC) . Bis{trj-c>-tolylphosphine)palladium dichloride (0.35 g, 0.10 mmol) wasadded fcllowed by 2-iodobenzonitrile (1.72 g, 7,5 mmol). The 5 resulting mixture was stirred under a nitrogen atmosphère withheating, allowed tocool, ethyl acetate (100 mL) was added,and the mixture filtered into a separatory tunnel. Sequentialwashing with aqueous HCl (50 mL; 0.1N), distilled H?0 (3 x 50mL) , drying over Na2SO4, filtration, and concentration under „ reduced uressure yielded the crude produc-t. Flash 0 chromatography over silica gel (light petroleum/ethyl acetategradient) afforded the purified compound.

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF(30 mL) vas added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78° C, and the solution stirred. The amino acid- 15 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 ammoniumchloride. The mixture was diluted with ethyl acetate (150 2Φ mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO4,filtered,, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure. 010493

,OCH, CH3 A solution oi terfc-butyloxycarbonyl - iodo-alanine-N, O- dimethvlamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57,3397-34C4) in dry benzene (30 mL) , and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purgedround boitom flask charged with zinc-copper couple (0.90 g) .The resuitinç mixture was sonicated under nitrogen until nostarting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) wasadded followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol) . Theresulting mixture was stirred under a nitrogen atmosphère withheating, allowed to cool, ethyl acetate (100 mL) was added,and Che mixture filtered into a separatory funnel. Sequentialwashing with agueous HCl (50 mL; 0.1N), distilled HZO (3 x 50mL) , drying cver Na2SO4, filtration, and concentration underreduced pressure yielded the crude product. Flashchromatoçrrapky over silica gel (light petroleum/ethyl acetategradient', afforded the purified compound.

,OCHq

N

I CH,

To a sciurion of terfc-butyloxycarbonyl-( 4-cyano ) 3 - pyridylalanine-N, O-dimethylamide (1.34 g, 4.0 mmol) in dry 010493 éthanol (20 mL) was added N,O-hydroxlyamine hydrochloride(0.416 ç, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0mmol) . The mixture was refluxed and then cooleçl. Theprecipitate was filtered, washed with cold éthanol, diisopropylether, dried with MgSO4, concentrated under reducedpressure, and used directly in the next step. The semi-solidwas suspsnded in a mixture of acetic acid (20 mb), and dryéthanol (40 mL) with warming. Subsequently, Pd/C catalyst(0.30 g, 10% Pd) was added, and hydrogen .was bubbled throughthe mixture with warming. The hydrogénation was. continueduntil no starting material could be detected as judged by TLC.

The catalyst was removed by filtration, and the solution wasconcentrated under reduced pressure (50 mL), HCl (50 mL, 1 N)was added, and the mixture was concentrated once again to 50mL. The solution was chilled overnight yielding the titlecompound.

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. The amino acid-N,0-dinethylamide (1.16 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 ammoniumchloride. The mixture was diluted with ethyl acétate (150mL) , and the organic layer washed with saturated aqueousammonium chloride (2 x 50 mL), brine (50 mL) , dried with MgSO4,filtered, and concentrated under reduced pressure. The crudematerial was purified en silica gel ethyl acetate/hexane), and 010493 concentrated under reduced pressure.

1. H2, PtO2/AcOII ->

tert-Butyloxycarbonyl-3-(4-pyridyl)alanine-N,O-dimethylamide(4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL) , and 5 PtO= (100 mg) added. The solution was shaken under H2 untilgas 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 10 with 1 N NaOH (2 x 50 mL) , brine (2 x 50 mL) , dried with MgSO4,filtered, and concentrated under reduced pressure to yield thetitle campound.

15 Pt O. (100 mg) added. gas uptake ceased. tert-Butyloxycarbonyl-3- (3-pyridyl) alanine-N, O-dimethylamide(4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), andThe solution was shaken under H2 untilThe solution was filtered through celite, and concentrated under reduced pressure yielding fcert-butyicxycarbonyi-3- (3-piperidyl) alanine-N, O-dimethylamide .

The residue vas dissolved in ethyl acetate (250 iuL) , washed « 1 Ü 4 9 3 with 1 N NaOH (2 x 50 mL) , brine (2 x 50 mL) , dried with MgSO4, filtered, and concentrated under reduced pressure to yield the title compound.

terfc-Butyloxycarbonyl-3- (2-pyridyl) alanine-N, O-dimethylamide(4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL) , andPtO2 (100 mg) added. The solution was shaken under H2 untilgas 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), washedwith 1 N NaOH (2 x 50 mL) , brine (2 x 50 mL) , dried with MgSO,,filtered, and concentrated under reduced pressure to yield thetitle compound.

terfc-Butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide(1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) undernitrogen with stirring. The solution was cooled, Ν,Ν'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), ·· 1 -1 010493 and HgClL (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue wassuspended in ethyl acetate (200 mL) , and filtered throughcelite. The filtrate was concentrated under reduced pressure.

Flash chromatography over silica gel (hexane/ethyl aceté^tegradient) afforded the title'compound.

ZhK^NHZ tert-Butyloxycarbonyl-3- (3-piperidyl)alanine-N, O-dimethylamide(1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) undernitrogen 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 wasconcentrated under reduced pressure, the remaining residue wassuspended in ethyl acetate (200 mL), and filtered throughcelice. The filtrate was concentrated under reduced pressure.

Flash chromatography over silica gel (hexane/ethyl acetategradient) afforded the title compound. 100

01 0403

HgCl2/THF

tert-Butyloxycarbonyl-3- (2-piperidyl) alanine-N, O-dimethylamide(l.OOg, 3.2 mmol) was dissolved in dry THF (10 mL) undernitrogen with stirring. The solution was cooled, Ν,Ν'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), 5 and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue wassuspended in ethyl acetate (200 mL) , and filtered throughcelite. The filtrate was concentrated under reduced pressure.

Flash chromatography over silica gel (hexane/ethyl acetate 0 gradient) afforded the title compound.

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 and the solution stirred. The guanidylated 4- piperidylalanine dérivative (2.00 g, 3.2 mmol) in anhydrousTHF (15 mL) was added dropwise, and the resuiting mixturestirred. The reaction was quenched with saturated aqueousammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO4, filtered, and concentrated under reduced pressure. 10

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 -78° C for 1h. The guanidylated 3-piperidylalanine dérivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resultingmixture stirred. The reaction was quenched with saturatedaçrueous ammonium chloride. The mixture was diluted with ethylacetate (150 mL), and the organic layer washed with saturatedaqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , driedwith MgSO4, filtered, and concentrated under reduced pressure.

To a solution ofwas added n-BuLi thiazole in anhydrous THF (1.23 g, 14.4 mmol)(1.6 M/hexane, 8.4 mL, 13.4 mmol) dropwise at .5 ο m ô 3 -78° C with stirring. The mixture was stirred at -78° C fc>r 1h. The guanidylated 2-piperidylalanine dérivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resultingmixture stirred. The reaction was quenched with saturatedagueous ammonium chloride. The mixture was diluted with ethylacetate (150 mL), and the organic layer washed with saturatedagueous ammonium chloride (2 x 50 mL) , brine (50 mL) , driedwith MgSO,, filtered, and concentrated under reduced pressure.

tert-Butyloxycarbonyl-para-nitro-phenylalanine-N, O- dimethylamide (13.88 g, 39.3 mmol) was dissolved in aceticacid (100 mL), and PtO2 (100 mg) added. The solution wasshaken under R, until gas uptake ceased. The solution wasfiltered through celite, concentrated under reduced pressure,taken up in H2O (150 mL) , and lyophilized. The semi-solid wasdissolved in ethyl acetate (350 mL) , washed with 1 N NaOH (3 x50 mL) , and brine (3 x 50 mL) . The solution was dried withMgSC\, filtered, and concentrated under reduced pressureyielding the title compound.

103 t) tf (;er*t-Butyloxycarbonyl-meta-nitro-phenylalanine-N, O- dimethylamide (13.88 g, 39.3 mmol) was dissolved in aceticacid (100 mL) , and PtO2 (100 mg) added. The solution wasshaken under H2 until gas uptake ceased. The solution wasfiltered through celite, concentrated under reduced pressure,taken up in H20 (150 mL) , and lyophilized. The semi-solid wasdissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 x50 mL) , and brine (3 x 50 mL). The solution was dried withMgSO<( filtered, and concentrated under r.educed pressureyielding the title compound.

terc-Butyloxycarbonyl-ortho-nitro-phenylalanine-N, O- dimethylamide (13.88 g, 39.3 mmol) was dissolved in aceticacid (ICO mL) , and PtO2 (100 mg) added. The solution wasshaZcen under IL, until gas uptake ceased. The solution wasfilrered through celite, concentrated under reduced pressure,taken up in HjO (150 mL) , and lyophilized. The semi-solid wasdissolved in ethyl acetate (350 mL) , washed with 1 N NaOH (3 x50 cL) , and brine (3 x 50 mL) . The solution was dried withMgSC4, filtered, and concentrated under reduced pressureyielding the title compound.

NHZ xVi· 010493 1. terc-Butyloxycarbonyl-3-(cis/trans-4- aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol)was dissolved in saturated aqueous sodium bicarbonate, and THF[60 mL, (1:1)3 with stirring. The solution was cooled and asolution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF(10 mL) was added dropwise. Excess solid sodium bicarbonatewas added, the THF was removed under reduced pressure, and theremaining aqueous phase was poured into ethyl acetate (250mL) , and mixed thoroughly. The aqueous phase was discardedand the remaining solution was washed with saturated aqueoussodium bicarbonate (2 x 50 mL) , 4 N aqueous sodium bisulfate(2 x 50 mL), and brine (2 x 50 mL) . The solution' was driedwith KgSO<, filtered, and concentrated under reduced pressure.

The seroi-solid was chromatographed on silica gel (ethylacetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)dropwise at -78° C and the solution stirred. The aboveprotected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL)was added dropwise, and the resulting mixture stirred. Thereaction was quenched with saturated aqueous ammoniumchloride. The mixture was diluted with ethyl acetate (150 mL). and the organic layer washed with saturated aqueousammonium chloride (2 x 50 mL), brine (50 mL), dried with MgSO4,filtered,, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure. 105 010493

1. fcert-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 asolution of benzyl chloroforma te (0.43 mL, 3.0 mmol) in THF(10 mL) was added dropwise. Excess solid sodium bicarbonatewas added, the THF was removed under reduced pressure, and theremaining aqueous phase was poured into ethyl acetate (250mL) , anc mixed thoroughly. The aqueous phase was discardedand the remaining solution was washed with saturated aqueoussodium bicarbonate (2 x 50 mL) , 4 N aqueous sodium bisulfate(2 x 50 mL) , and brine (2 x 50 mL) . The solution was driedwith Mg£04, filtered, and concentrated under reduced pressure.

The seni-solid was chromatographed on silica gel (ethylacetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)dropwise at -78° C and the solution stirred. The aboveprotected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL)was added dropwise, and the resulting mixture stirred. Thereaction was quenched with saturated aqueous ammoniumchloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueousammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO4,filtered, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure. 01049 106

1. Z-Cl, NaHCO/rHF:H2O ------->

2. Lithium thiazole/THF

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 asolution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF(10 mL) was added dropwise. Excess solid sodium bicarbonatewas added, the THF was removed under reduced pressure, and theremaining aqueous phase was poured into ethyl acetate (250mL) , and mixed thoroughly. The aqueous phase was discardedand the remaining solution was washed with saturated aqueoussodium bicarbonate (2 x 50 mL) , 4 N aqueous sodium bisulfate(2 x 50 mL) , and brine (2 x 50 mL) . The solution was driedwith MgSO«, filtered, and concentrated under reduced pressure.

The semi-solid was chromatographed on silica gel (ethylacetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol)dropwise at -78° C and the solution stirred. The aboveprotected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL)was added dropwise, and the resuiting mixture stirred. Thereaction was quenched with saturated aqueous ammoniumchloride. The mixture was diluted with ethyl acetate (150 mL)„ and the organic layer washed with saturated aqueousammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO4,filtered, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetcite/hexane) ,and concentrated under reduced pressure. 010493

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 HgCl2 (1.81 g, 6.7mmol) aclded. The solution was concentrated under reducedpressure, the remaining residue was suspended in ethyl acetate(300 mL’ , and filtered through celite. The filtrate wasconcentrated under reduced pressure. Flash chromatographyover silica gel (hexane/ethyl acetate gradient) afforded thepurifiée product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol)dropwise at —78° C and the solution stirred. The aboveguanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) wasadded dnopwise, and the resuiting mixture stirred. Thereaction vas quenched with saturated aqueous ammoniumchloride. The mixture was diluted with ethyl acetate (150 mL) . and the organic layer washed with saturated aqueousammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO„,filtered, and concentrated under reduced pressure. The crudematerial vas purified on silica gel (ethyl acetate/hexane) ,and concenrrated under reduced pressure. 106 010493

Ο

H2N

HgCl2/THF

2. Lithium thiazole/THF ο

ZHN 1. tert-Butylo3sycarbonyl-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 HgCl2 (1.81 g, 6.7mmol) added. The solution was concentrated under reducedpressure, the remaining residue was suspended in ethyl acetate(30C mL), and filtered through celite. The filtrate wasconcentrated under reduced pressure. Flash chromatographyover silica gel (hexane/ethyl acetate gradient) afforded thepurified product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol)dropwise at -78° C and the solution stirred. The aboveguanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) wasadded dropwise, and the resulting mixture stirred. Thereaction was guenched with saturated agueous ammoniumchlcride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated agueousammonium chloride (2 x 50 mL), brine (50 mL), dried with MgSO4,filtered.,, and concentrated under reduced pressure. The crudematerial vas purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure. 108 01049

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) -5 S-methyl-isothiourea (2.18 g, 6.1 mmol) , and HgCl2 (1.81 g, 6.7 mmol) added. The solution was concentrated under reducedpressure» the remaining residue was suspended in ethyl acetate(300 mL)„ and filtered through celite. The filtrate wasconcentrated under reduced pressure. Flash chromatography 0 over silica gel (hexane/ethyl acetate gradient) afforded thepurified product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrousTHF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol)dropwise at —78° C and the solution stirred. The aboveguarridylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) wasadded dropwise, and the resulting mixture stirred. Thereaction was quenched with saturated aqueous ammoniumchlcride. The mixture was diluted with ethyl acetate (150mL) , and the organic layer washed with saturated aqueousammonium, chlcride (2 x 50 mL) , brine (50 mL) , dried with MgSO„,filtered,, and concentrated under reduced pressure. The crudematerial was purified on silica gel (ethyl acetate/hexane),and concentrated under reduced pressure. 110 010493

Exsmple 2

Synthesis of Intermediates0

Ph

THF/-7^C (4S, 5R) -3-(l-oxo-3-phenylpropyl)-4- (phenyl) -5- (methyl)-2-5 oxarolidone (2). A solution of 10.0 g (1.0 equiv., 56.4 mmol) of (4S, 5R) -4-phenyl-5-methyl-2-oxazolidone (1) in250 mL of dry THF, stirred at -78°C under argon, wastreated dropwise with n-butyllithium (1.6 M in hexane, 1.1eç., 38.8 mL·) . A.fter stirring for 30 min., 8.4 mL (1.0 10 equiv., 56.4 mmol) of hydrocinnamoyl chloride wasintroduced dropwise over a 10 minute period. Thexesulting mixture was warmed to 0°C, stirred for anadditional hour, and quenched with saturated ammoniumchloride. The solvent was removed in vacuo and the 15 resulting white solid dissolved in ethyl acetate and <ddH2O. The aqueous phase was removed and extracted withtwo additional portions of ethyl acetate. The extractswere combined, washed with saturated sodium chloride,edrieè over sodium sulfate, and the solvent was removed in 20 vacuc to afford a white crystalline solid (2) (in 91%yield). m.p.55-96.5; [cc]D-35.S (c=l, CH2C12); [a]D-26.6 (c=1.018, ŒvClç) . 25 ÀHKHF. (CDCI3) δ 0.89(d, 3H, CH3^, J=6.6 Hz), 3.00-3.05(n, 23), 3.26-3.34 (m, 2H) , 4.73-4.78 (m, 1H) , 5.64, d, LH, J=7.4 Hz), 7.22-7.46 (m, 10H). 11ί 01 0483

A solution of 5.0 g (1.00 eguiv., 16.2 mmol) of (2) in 100mL of dry THF, was cooled to -78°C. Enolization was 5 achieved with 17.8 mL (1.1 eguiv., 17.8 mmol) of lithiumbis-trimethylsilylamide, which was added dropwise viasyringe. The solution was stirred for 30 min. before 4.45mL (3.0 eguiv., 48.5 mmol) of allyl iodide was introducedand the reaction warmed to -15°C. After 1 h the reaction 10 was guenched with saturated ammonium chloride and extracted (3X) with ethyl acetate. The organic phase waswashed with sodium metabisulfite, dried over sodiumsulfate and the solvent removed in vacuo to afford an off-colcur oil. Purification was achieved by flash 15 chrcmatography on silica gel using a stepwise gradient (15:1, 12:1, 10:1) to yield (3), a colourless oil (95%).

[a]D 47.5 (c=3.12, CH2C12). 20 ^HNMP. (CDCI3) δ 0.82 (d, 3H, CH3, J=6.6), 2.31-2.40 (m, 1H, P.CH=CHCH2), 2.49-2.57 (m, 1H, RCH=CHCH2), 2.84-3.00(m, 2.H, Ph-CH2), 4.32-4.37 (m, 1H, CH-(N)CO), 4.53-4.58;m, LH, CH3-CH-), 5.03-5.13 (m, 2H, ABX, CH=CH2), 5.21 (d,LH, Ph-CH, J=7.1Hz), 5.81-5.89 (m, 1H, CH=CH2), 7.20-7.42 25 :m, L0H, ArH) ; -3C i CDCI3 ) δ 14.4, 36.2, 38.2, 43.9, 54.7, 78.4., 117.1, 12Ξ.4, 126.3, 128.2, 128.5, 129.0, 133.1, 134.8, 1-38.9,L52.4, 174.9. 010493 112

A sample or 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 5 mmol) of a 1.0 M solution of borane-tetrahydrofurancomplex at 0°C and stirred for 2h. The solvent wasevaporated and chloroform (100 mL) added via syringe.Oxidation of the organoborane was achieved by the additionof 4.7 g (2.0 equiv., 27.2.mmol) of 3-chloroperoxybenzoic 10 acid at 0°C, with warming to ambient température and stirring fer an additional hour. The organic phase waswashed with 5% Na2CO3,, ddH2O, and dried over sodium sulfate. Eue to the instability of the alcohol, a quickcoluzin was performed to remove the extreme polar and 15 nonpolar material which originated from the 3- chloroperoxybenzoic acid. The alcohol (4) was obtained ina yielc cf 65%.

[a]D 39.3 (c=1.038, CH2C12). 20 2ΗΝΜΕ (CECi3) δ 1.07 (d, 3H, CH3-CH, J=6.5 Hz), 1.81-1.93(m, 2H, CH2-CK-H), 2.10-2.19 (m, 1H, CH2-CH-H), 3.10-3.17im, 2H, Ρΰ-2Η2), 3.873.90 (m, 2H, CH2OH), 4.43-4.49 (m, 1H, CH-20), 4.70-4.75 (m, 1H, CH3-CH), 5.36 (d, 1H, Ph-CH, 25 2=7.1 Hz), ~.41-7.63 (m, 10H, ArH); 115

To a solution of the alcohol (4) (1.0 g, 2.7 mmol),dissolved in dichloromethane (27 mL) ; 876 mg (1.5 equiv., 5 4.1 mmol) of pyridinium chlorochromate and 1.0 g of 4 angstrom molecular sieves were introduced and the mixturechanged from bright orange to a black colour. Thereaction was monitored by TLC and after 30 min., ifstarting material remained, additional molecular sieves 10 were added. The solution was filtered through celite andthe solvent was evaporated. The residue was dissolved inethyl acetate and washed with saturated sodium chloride.If the orange colour persisted in the organic phase,additional filterings through celite pads were performed. 15 The aldéhyde was obtained in a quantitative yield as atransparent, colourless oil (5). 1HNK?. (CDC13) 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), 20 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 &0=8.3), 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); 25 -3C '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, 1-74.9, 201.1. ^aivîT'·7 177*7-4— - -, 010493 114

The aldéhyde, (5), (2.6 g, 7.10 mmol) was dissolved in benzene (70 mL) and a catalytic amount of p- 5 toluenesulfonic acid was added, followed by 1.58 g (1.2eguiv., 8.52 mmol) of L-cysteine ethyl ester and 4 Amolecular sieves. The reaction was allowed to stirovernight at ambient température followed be removal ofsolvent in vacuo. The residue was dissolved in 10 chloroform, washed with saturated sodium chloride, dd^O,and dried over sodium sulfate. The solvent was removedin vacuc to afford a gummy solid (6).

H CO2Et 15 6 7 2.0M trimethyialuminum in hexane (2.4mL, 4.8mmol, 3équivalents) was added slowly to starting material (6)(SOOmg, 1.61mrr.ol) stirring in anhydrous dichloromethane 20 under argon, using ovendried equipment. After stirringovernight, HPL-C indicated that the reaction had gone tocompletion. 010493

The mixture was quenched with excess methanol, thenfiltered on a short silica gel column (washing throughwith excess 10% methanol in ethylacetate. Evaporationgave 7 84 mg of crude matérial that was purified using 2:1hexane:EtOAc on a silica gel column afforcling 258mg(0.81mmol being a 50% yield) of pure compound (7),a 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-ethylester' as a white/yellow solid. ’ΉΝΜΕ (CDC13) 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= 9.0and 5.0 Ez), 5.22 (dd, 1H, J= 8.0 and 6.0 Hz), 7.18-7.23(m, 3H), 7.28-7.31 (m, 2H).

7 8 LÏ0E.H20 (48mg, 1.12mmol) in lOmL of water was added tostaminé matérial (7) (240mg, 0.76mmol) dissolved in lOmL of dioxane. After 1 hour, TLC in 1:1 Hexane:EtOAc showed nostaminé matérial. The reaction was quenched with 10% citricacid, then extracted twice with dichloromethane.. Drying andevaporating the combined organic layers gave 354mg of crudeproduct. This was redissolved in dichloromethane, thenprecipitated by adding excess hexane. The product wasfiltered to give 200mg (0.68mmol being a 90% yield)) of anoff-white solid, (8), also known as 6S-benzylhexahydro-5-oxo5E-thiazrlo[3,2-a]pyridine-3R-carboxylic acid. KMF. (ODSOD) d 1.71-1.82 (m, 3H) , 2.12-2.17 (m, 1H) , 2.67 ( dd , 1H, d= 14 and 11 Hz), 2.77-2.81 (m, 1H) , 3.30-3.40 116 0 î Ο 4 9 3 im, 3Η) , 4.81 (dd, 1Η, J= 8.5 and 4.9 Hz), 5.16 (t, 1H, J =7.5 Hz), 7.18-7.31 (m, 5H).

Oxalvl chloride (9) (25 g, 0.197 mol) was cooled to 0°C and cyclohexane propionic acid (20 ml, 0.14 mol) wasadded. This was left to stir overnight. The résultant 10 mixture was distilled to give an 84% yield of the colorless liguid (10), cyclohexyl propionic acid chloride.

1.1 10 12 15

The chiral auxiliary (11) (13.6g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78°C. Then n-BuLi(52.S irL>, 84.4 mmol, 1.2 equiv.) was added and left for 3 0mins (dark orange solution). The acid chloride (10)(13.4 20 g, 75.c nmol, 1 eq) was then added and left to stir overnight. Work-up was done by quenching with saturatedUE/Cl extracting with ethyl acetate, washing the extractswith water and brine, drying over sodium sulphate andconcentration. A fast column, with dry loading, (6:1 25 heyar-.e ethyl acetate) was run to purify the product. Thisaftorded in a white solid (12) which was recrystallized 117 0 ί 0 Λ 9 3 fron ether and hezane to give the title compound in 78%yield.

[a]D=-20.l(c=l,EtOH); MP/BP mp=90.5-91.5oC 1H MMR (CDC13, d 0.86-1.10 (m, 5H) , 1.18-1.30 (m, 4H),1.54-1.75 (m, 7H) , 2.86-2.97 (m, 2H0, 4.70-4.76 (m, 1H) ,5.65 (d, 1H, J= 7.2 Hz), 7.28-7.42 (m, 5H) .

The starcing matériel (12)(9.13g, 29 mmol, leq) wasdissolvec in dry THF and cooled to -78°C, after whichLiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwiseover 40 reins. Then, 30 minutes later, allyl bromide (7.5mL, 36.9 mmol, 3 eq) was added slowly over 10 mins. Themixture was left to warm overnight. Work-up includedquenchinc with sat. ammonium chloride, extraction withethyl acetate, washing with 10% sodium thiosulphate,decolourising with charcoal, drying over sodium sulphateand concentration in vacuo. The product was obtained as ayellcw oil (13) in 96% yield.

Ia]D=+S.5(c=1.0,EtOH) 'H HMF (CD213) 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) ,4.76-4.35 (m, 1H) , 5.00-5.07 (m, 2H0, 5.65 (d, 1H, J= 7Hz), 5.64-5.88 (m, 1H0, 7.27-7.46 (m, 5H) . 118 010493

2-methyl-2-butene was added dropwise to borane dimethylsulphide complex at -12°C. The reaction wasmaintained at this température for 15 minutes and then itwas warmed to 0°C, after which it was stirred for 2 hours.The disiamyl borane was then added to a mixture of thestarting material 13 in THF using a double-ended needle at0°C. The mixture was then stirred for 2 hours after whichthe solvents were removed and the residue dissolved indichloromethane. It was carefully added to a suspensionof pyridinium chlorochromate in dichloromethane containedin a flask equipped with a reflux condenser. After theinitial exothermic reaction had subsided, the mixture wasrefluxed at 50°C for 1 hour. The dark brown liquid wasdissolved in ethyl acetate and filtered through Florisil.The black residue of PCC was extracted with ethyl acetateand also filtered through the same Florisil pad.Concentration of the filtrâtes resulted in a 78% yield ofa yellow gummy product (14).

[a]I>=-17.8 (c=l.245,EtOH) ’HHMR (CDC13) 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, 1H0, 4.12-4.16 (m, 1H), 4.76-4.80 (m, 1H0,5.67 (d, 1H, J= 7Hz) , 7.27-7.46 (m, 5H), 9.77 (s, 1H). 119 01Ü493

^CO2Et 14 15

The resulting aldéhyde (14) (7.7g crude, 20.8 mmol 1 5 equiv.) was dissolved in 75 mL of toluene. To thesolution was added a catalytic amount of p-toluenesulphonic acid (50 mg), 10 g of 4Â molecularsieves, and L-cysteine ethyl ester (3.87 g, 20.8 mmol, 1equiv.). The mixture was stirred overnight, filtered and 10 concentrated. The residue was then purified by silica gelchrcmatography (6:1 hexane:ethyl acetate) afforded 6.36 gof the product (15) in 61% yield. 15 20 [a]D=-48.3(c=1.095,EtOH) ’H 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 (πύ 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).

"CO2Et 15 16 120 010493

The starting material (15) (1.97 g, 3.9 tnmol, 1 equiv.) was dissolved in 20 mL of dry dichloromethane and cooledto 0°C. Trimethylaluminum (5.9 mL, 11.8 mmol, 3 equiv.), 5 was added dropwise and the mixture was left stirring overnight. After complété reaction as evidenced by HPLC,methanol was added until a yellow solid mass was formed.Dichloromethane was added to dissolve the solid and thewhole mixture was stirred for 15-30 minutes and then 10 filtered. The residue after concentration in vacuo was run through a quick column (6:1 hexane:ethyl acetate), toremove auxilary and as many of the polar décompositionproducts as possible, affording in a 50% yield of a yellowoil (16) . 15 ’H NMR (CDC13) 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 and8.0 Hz), 4.11-4.25 (m, 2H) , 4.88 (dd, 1H, 11.0 and 5.o 20 Hz), 5.14 (dd, 1H, J= 10 and 6 Hz).

25 The starting material (16) (0.95 g, 2.9 mmol, 1 equiv.)was dissclved in 10 mL of dioxane. The solution wascooled te 10 C, and to it was added LiOH'H2O ( 0.123 g, 2.9mmol, 1 eq.) dissolved in 10 mL of water. The bath wasremovec and the mixture was stirred at room température 30 for 1 hour. . TLC showed complété reaction and the solventwas evaperated under vacuum. The remaining aqueous,layerwas washed with ether (2X), acidified with 10% citricacid, and extracted with dichloromethane (3X). The 121 010493 cotbined extracts were dried over sodium sulphate andconcentrated to give a white solid which wasrecrystallized from ether. Concentration of the filtrateand purification by silica gel column chromatography (2:1hexane: ethyl acetate) rësulted in more product (17) witha m.p. of 198.2-199°C. lH KMF. (DMSO-dô) 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) , 3.02 (dd, 1H, J= 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.83(dd, 1H, J= S and 4 Hz), 4.95 (dd, 1H, J= 8 and 5.5 Hz).

BOC-DiCbc Arg (18) (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0°C. Triethylamine (2.2mL) was added followed by 14.5 mmol of a IM toluenesolution of isopropyl chloroformate via a syringe. Thereaccicn -was allowed to s tir at 0°C for 3 0 minutes thenquicfcly filtered. The white solid was discarded. To thefiltxate was tubbled freshly prepared diazomethane untilthe coior of the solution turned yellow. The reactionmixture was allowed to stand overnight in a well ver.tilated fumehood which facilitated the discharge ofexcess diazomethane. Dry ether was added to precipitatethe ciazoketone. The product was filtered and dried undericupnr· to give light yellow flufcfy solid (4.6 g, 58%).

122 010493

Diazoketone (19) (lg, 1.77 mmol) was dissolved in THF (20mL) and. to this solution was added IM HCl in ether (20 mL)at 0°C. The reaction was allowed to stir at ambienttempérature overnight during which time a white 5 precipitate was formed. Further précipitation was achievedby adding ether. Filtration and drying the solid affordedthe product (20) (1.02 g , 100%). *H I2MR (DMSO-d6) d 1.65-1.77 (m, 3H) 2.06-2.50 (m, 1H) , 10 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) . UC-NMR (DMSO-d6) d 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 15 69.3, 128.6, 128. 7, 128.8, 128.9, 135. 2, 135 .9, 153.4, 157.4, 198.9.

To mimetic (17) (0.422 g, 1.42 mmol) in THF (50 mL) at 0°C and in presence of N-methyl morpholine (0.19 mL) , wasslowly added IM toluene solution of isopropylchloroformate(1.71 mL). The reaction was allowed to stir at 0°C for 30 25 minutes then treated with aminochloromethylketone (20) insmall portions. Once the addition was complété the reactionwas further stirred for 15 minutes followed by addition ofK-ir.ethyl morpholine (0.19 mL) . The reaction was stirred atambient température for 3 hours, then extracted with ethyl 30 ace-are followed by washing with brine and 10% agueous citric acid. P.emoval of organic solvent gave a white foam 123 010493 (21) (1.03 g, 96%) which was used further without purification. ’H NMR (CDC13) d 0. 07-0.97 (m, 1H ), 1.15-1.41 (si, , 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 £, 1H) .' , « ri Ί < < r ·ι « 124 010493

EXAMPLE 3

1. TFA/DCM -».

2. DMF/BOP

Ph

NHToi

Pti A''"

CO,H

NH, (N-t-BOC-N-tosyl)butyrylketoarginine (240mg, 0.515 mmol),was deprotected using 30% TFA in dichloromethane. Thedeprotected arginine dérivative, was coupled with the 10 mimetic (8) (100 mg, 0.343 mmol) in DMF under basic conditions (Et3N, pH = 8-9), using BOP reagent (228 mg, 0.52 mmol) as the dehydrating agent. The reaction wastypically complété within 2-4 hours. Extraction with ethylacetate followed by successive washing with brine and 10% 15 aqueous citric acid yielded the crude productk The crudeproduct was purified by column chromatography affording 180mg (76%) of pure product. This product was then treatedwith HF to remove the tosyl group. Purification of theisolated deprotected product by HPLC afforded BCH-2737. 20 EXAMPLE 4

123 010493

Chloroxnethylketone (21) (0.188 g, 0.245 mniol) wasdissolved in THF (10 mL) treated with NMM (0.036 mL)followed by mercapto acetic acid (0.02 mL·, 0.299 mmol)„ 5 The reaction was stirred at ambient température overnight.Extraction of the reaction mixture with ethyl acetatefollowed by successive washing with brine and 10 % aqueouscitric acid and évaporation of organic solvent gave thecrude product which was purified by'column chromatography 10 to give foamy solid as the product (0.125 g, 62%).

This protected precursor (0.125 g, 0.154 mmol) wasdissolved in DCM (5 mL) and cooled to -78°C. A IM DCMsolution of BBr3 (1.54 mL, 1.54 mmol) was slowly added. The 15 reaction was stirred at ambient température for 5 hours,then cooled to -78°C again and treated with anhydrousmethanol (2mL). The reaction was brought to roomtempérature and stirred for 2 additional hours. Thesolvents were removed under reduced pressure and the 20 residue was partitioned between ether and water. The waterlaver was collected, lyophilized and the final product (23)obtained as a powder after HPLC purification and lyophilisation. 25 The products of the reactions described above can beisolated in the free form or in the form of salts. Inaddition, the products can be obtained as pharmaceuticallyacceptable acid addition salts by reacting one of the freebases with an acid. In a similar manner, the product can 30 be obcained as pharmaceutically acceptable salts by reacting one of the free carboxylic acids with a base.Likewise, treatment of the salts with a base or acidresulrs in a régénération of the free amide. 010493 12υ EXAMPLE 5 A general method of synthesizing compouncl of formula II orHZ: 5

2. deprotection as- in example 3 COCH2CH2CO2Me

NH Λ, CBz NH NHL CBz

10 k * V ' - XZ ι 010493 EZ=MPLE_ÊSynthesis of

(16) 5 sthp i

Synthesis of 2-Benzyloxycarbonylamino-4-hydroxybutyricacid tert-butyl ester

(Cydohexyi),NH.HOOC 10

NHZ CO2t-Bu

NMM.THF2) NaBH., MeOH

NHZ

HO 'CO2t-Bu (1) (2)

Te a solution of the protected aspartic acid (1) (Bachem,2.50 g, 4.95 mois) in 50 mL of dry tetrahydrofuran 15 (THF) , at -10°C, under N2, was added N-methylmorpholine(109 pi, 0.2 eq) and isopropyl chloroformate (1.0M. toluene : 384 (IL, 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) , 20 at -78°C, under N2. This suspension was stirred at -78°Cfer 30 min. The mixted anhydride solution was then addedte the NaBH4 suspension dropwise via canula, and__thefinal solution was stirred at -78°C for 3 hr. Aceticacid (2.8 mL, 10 eq) was then added and the solution was 010493 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 overMgSO4, the solids were filtered and the solvent 5 evaporated to give 1.53 g (4.95 minois, 100¾) of thealcohol (2) as a clear oil. XH MMR (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), 10 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). S TEP 2 2-Eenzyloxycarbonylamino-4-iodobutyric15 ester

Imidazole, -10°CCHjCN/EtjO

NHZ HO'^X^CO2t-Bu (2) 20 To s solution of the alcohol (2) (1.53 a 1:1 mixture of CH3CN/Et2O (50 mL) , atwere added successively imidazole (607Ph,E (2.21 g, 1.7 eq). Iodine (2.14 g, acid tert-butyl

g, 4.95 mmols) in -10°C, under N2,mg, 1.8 eq) and 1.7 eq) was then added in small portions over a period of 15 min. After25 the addition was completed, a white precipitate formed and the solution was brown. It was stirred at -10°C for45 min. It was then poured in Et2O and the organic phasewas washed with sat.aq. Na2SO3, sat.aq. CuSO4, H2O anddried over MgSO4. The solids were filtered and the30 solvent evaporated to give a yellow oil that was purifiedbv flash chromatography (silica gel, 5% to 20% EtOAc/

Hex' . The iodide (3) was obtained in 83% yield (1.71 g)as a clear oil. 129 010433 1E NMR (CDC13, 400 MHz) : Ô 7.41-7.31 (m, 5H, ArH) , 5.35(od, 1H, J=7.3, MH), 5.13 (s, 2H, CH2Ph) , 4.30 (m, 1H, H-2), 3.22-3.12 (m, 2K, H-4), 2.42 (m, 1H, H-3), 2.20 (m,ΙΕ, H-3), 1.48 (s, 9H, t-Bu). STE? 3

Synthesis of 2-Benzyloxycarbonylamino-4-hexenoic acidtero-butyl ester

NHZ

CO2t-Bu

^'MgBr , CulTHF, -78°C

NHZ 'CO2t-Bu (3) (4)

To a suspension cf Cul (2.27 g, 5 eg) in dry THF (20 mL),at -78°C, under K2, was added slowly a 1.0M solution inTHE of vinyl magnésium bromide (23.4 mL, 9.8 eq).. Thesolution was then warmed up to -10°C for 30 min (itcumed then black) and cooled back to -78°C. A solutioncf the iodide (3) (1.00g, 2.39 mmols) in dry THF (3.5 mL) vas then added slowly to the cuprate solution. Theréaction mixture was stirred at -78°C for 2.5 hr.

Sat.aq. NH4C1 (50 mL) was added and the mixture wasbrcught back to room temp. with vigorous stirring. Itvas then poured in Et2O and stirred for 5 min. The dark suspension was filtered through a cintered funnel and thephe-es were separated. The aqueous phase was extractedvitra Et-O (2x) and the combined organic extracts werecried over McSO4. The solids were filtered, the solvents evaporated and the crude oil purified by flash chrcnnatcgraphv (silica gel, 5% AcOEt/Hex) to give- 0.51 g(£7%) of the pure alkene (4). ΐ3υ 010493 ΙΕ MMR (CDC13, 400 MHz) : δ 7.37-7.31 (ni, 5H, ArH) , 5.80(m, 1H, H-5), 5.33 (d, 1H, J=7.8, NII) , 5.12 (ε, 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), 5 1.92 (m, 1H, H-3), 1.74 (m, 1H, H-3) , 1.48 (s, 9H, t-Bu). STEP 4

Synthesis of l-Benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinecarboxylic acid tert-butyl ester 10 ΝΉΖ

(4)

1) Hg(OAc)2, THF 2) NaHCO3, KBr 3) NaBH4, O2

15 To a solution of the alkene (4) (50 mg, 0.157 rranol) in dryTHF (3.1 mL), at r.t., under N2, was added mercuricacetate (75 mg, 1.5 eq) . The solution was stirred atr.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 20 stirred at 0°C for 30 min. KBr (0.11g, 6 eq) was addedand the mixture was stirred at r.t. for 2 hr. It wasthen poured in H2O/Et2O and the phases were separated.

The aqueous phase was extracted with Et2O (2x) and thecombined organic extracts were dried over MgSO4. The 25 solids were filtered and the solvents evaporated. Oxygen(02) was bubbled into a suspension of NaBH4 (3.3 mg, 0.55eq) in dry DMF (0.4 mL) for 1 hr, and to this was addeddropwise (syringe pump, 3 mL/hr) a solution of theorganomercurial bromide in DMF (3.1 mL) with continuous 30 introduction of O2. The bubbling was continued for 1 hrand Et2O (5 mL) was added. The grey suspension was '•,’lTlVfrx.T·· 13- 010493 filtered through Celite and the filtrate was evaporated.

The residue was chromatographed (silica gel, 6:4Hex/EtOAc) to give the pyrrolidinol (5) (30 mg, 57%) as a clear oil. 5 iH KMR (CDC13, 400 MHz) : δ 7.37-7.28 (m, 5H, ArH), 5.22-5. OS (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). 10 STEP 5

Synthesis of l-Benzyloxycarbonyl-5-carboxy-2-pyrrolidlnecarboxylic acid tert-butyl ester

To a solution of the alcohol (5) (50 mg, 0.149 mmol) andEt3N (62)1L, 3 eq) in dry CH2C12 (0.8 mL) is added slowly,under N-, at 0°C, a solution of SO3-Pyridine complex (71 20 mg, 3 eq) in dry DMSO. The solution was stirred at 0°Cfor 30 min and 10% citric acid (2 mL) is added. The pHis brouçht to 4 with IM NaOH and the aqueous phase isextracted with Et2O (3x). The combined organic extractswere dried over MgS04. The solids were filtered and the 25 solvents evaporated to give a crude oil which waspurifiée, by flash chromatography (silica gel, 7:3Hex EtOAc) . The pure aldéhyde (6) was obtained as aclear oil (45 mg, 90%). lû - ο ί 0493 ΙΕ NMR (CDC13, 400 MHz)7.36-7.29 (m, 5Η, ArH),4.3 9 (m, 2H, H-2, H-5) ,+ 1.36 (2s, 9HZ t-Bu). δ 9.68 +5.23-5.112.30-1.97 9.56 (ds, 1H, CHO), (m, 2HZ CH2Ph), 4.57-(m, 4HZ H-3, H-4), 1.47 STEP 6

The pyrrolidine-aldehyde (6) is coupled with the protecteddiamino-propionic acid (7) by first forming the imine (8)(MgSO<z CHjClj) . Isolation of the imine (8) is done byfiltration of the MgSO4 and évaporation of the solvent.

The crude imine is then treated with NaBH(OAc)3 and acticacid (AcOH) in THF for 15 hours to obtain the amine (8)aftet extrative work-up. STEF 7

(8) (9) 133 010493

The CB7 (7) protecting group of the amine (8) is removedby hydrogénation with palladium on charcoal 10% as acatalyst in methanol (MeOH). The catalyst is filtered andthe MeOH évaporated to give the crude diamine (9) that can 5 be used without any purification. STEP 8

(10)

The cyclisation is done by heating the crude oil (9) fromstep 7, neat slightly above the boiling point of methanol.The hicyclic lactam (10) is purified by flash 15 chroma tography. nrv'viv'r p*’ .*·»« · 010493

5 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 givethe bicyclic lactam-amide (11) . 10 STEF 10

(11)

HCl/Et2O -î>

The EOC and t-butyl ester protecting groups of bicyclic15 lactam amide (11) are removed under acidic conditions (HCl in ethyl ether (Et20)). The amine sait (12) précipitâtesout c-f solution and is colléeted by filtration. '.•î'vv;;' 135 010493 STE? 11

(12) (13) 5 The primary amine of compound (12) is protected with a CBZgroup by reacting it with benzyl chloroformate inacetanitrile (CH3CN) with KjCO3 as a base. Extrative work-up cives fully protected carboxylic acid (13) which can beuse for step 12 without further purification.. 10 STEF 12

O

ZHN N- (13)

(15) 15 The carboxylic acid (13) is coupled with benzothiazoleketoarginine (14) in DMF using BOP as the coupling agentin the presence of diisopropylethylamine (EtNiPr2) .Extraction with ethyl acetate (EtOAC) gives compound (15)as a solia which is purified by chromatography. 20 136 010493 ST2P 13

(16) 5 The two CBZ(Z) protecting groups of compound (15) areremoved by catalytic hydrogénation with Pd/C 10% as acatalyst. The catalyst is filtered and the solvent isevaporated to give the amino-guanidine (16) . 137 010493 EXAMPLE 7 SYNTHESIS OF COMPOUND (10)

(2) 10 4-methylmorpholine (NMM) was added to a solution of thecarboxylic acid (2) (1.7g, 4.9mmol, l.Oeq), 4- hydroxyproline (3) (5.39mmol, 1. leq) , and BOP reagent 15 (2.17g, 4.9mmol, l.Oeq) in anhydrous DMF (lOmL) at room température. The reaction mixture was stirred at roomtenperature over night, quenched with brine (50mL) andethyl acetate (lOOmL). The organic layer washed with TT-r* S»»*’** 010493 aqueous citric acid (10%, 2x50mL) , sodium bicarbonate (10%, 2x50mL) and brine (50mL). The resulting organic layer was dried over anhydrous magnésium sulfate, filtered and the solvent évaporated. The crude residue was purified 5 by flash chromatography (5:4:1, ethyl acetate-hexane-methanol). 1.1g of pure product (4) was recovered 48%yield. STEP 2 10 13ί

To a solution of 4-hydroxyproline dérivative (4 ) (115mg, 15 240umol, l.Oeq) in dichloromethane (10 mL, anhydrous) at0°C ïs added triethylamine (72mg, 720umol, 3.0eq) andmethanesulfonyl chloride (28mg, 240umol, l.Oeq) and thereacdion mixture is stirred at room température. Themixture is then quenched with an aqueous solution of 20 ammonium chloride and extracted with ethyl acetate. Theorganic layer is washed with 10% citric acid and brine,«dried, filtered and the solvent is evaporated to drynessyielding compound (5) . 25 STEP 3 139 010493

l)Hg(Œc)2

CF3OOOH/CPC 2)NcëH4

The enamine (5) (l.Oeg) is treated with mercuric acetate5 (1.1 eg) in THF. The solvent is evaporated to dryness and the residue dissolved in methanol. The resulting organo-mercurial is reductivly cleaved with sodium borohydride(l.ûeg). The resulting crude lactam thioether is purifiedby flash chromatography on silica gel affording compound 10 (6). STEF 4

15 20

To a solution of the lactam thioether (6) (l.Oeg) in drydichloromethane N-chlorosuccinimide (l.Oeg) is added at0°C. The reaction mixture is warmed to room température.When the reaction shows no more starting material thesolic. is filtered and the solvent evaporated to dryness.The crude material (7) is use without any further_.purification for step 5. 3.4v STEP 5 010493

Το a solution of the alpha-chlorothioether (7) (l.Oeq) in5 THF (anhydrous) a solution of phenylcuprate (l.Oeq) (prepared according to littérature procedure) is added atlow température. When the reaction mixture shows nostarting chlorothioether, brine and ethyl acetate isadded. The organic layer is dried, filtered and évaporated 10 to dryness to afford the desired product (8) . STEP 6

(8) (9) 15 The isolated bicyclic lactam (8) is hydrolysed with oneéquivalent of lithium hydroxyde in a 1:1 mixture of THFand rater. The mixture is stirred at room température for1 hour. The crude mixture is extracted with ether and theresulting solution is poured into 10% citric acic'aqueous 20 solution and extracted with dichloromethane to yiéld thecorresponding carboxylic acid (9) . 141 010493 STEF 6 ο

5 The crude carboxylic acid (9) is coupled with benzythiazole keto arginine in DMF using BOP as thecoupling reagent in the presence of diisopropylethylamine.Extraction with EtOAc gives a solid that is purified onsilica gel to give the protected amide. The CBZ protecting 10 group is removed with BBr3 in dichloromethane at room température finally gives the bicyclic benzothiazole keto I arginine inhibitors (10)

The following compounds are produced accordingly with the15 execption that the appropriate substitution of products were made in order to obtain the final compounds. COMPODND #11

20 142 010493

COMPOUND #12

(12) 5 EXAMPLE 8

10

Commercially available glutaric acid monomethyl esterchloride (1) (20 ml, 0.144 mol) was disolved in 40 ml ofdry tetrahydrofuran (THF) and cooled to -15°C. Excessdiazomethane freshly prepared in 300 ml of Ether was 15 introduced via cannula at -15°C to the solution. The mixture was left to warm up to room température overnight.Excess diazomethane was evacuated from the flask with acurrent of argon. To bring the reaction to completion, 75ml of 1 N HCl in Ether was added at 0°C and left tn warm 20 up to room température for 5 hours. The volume of the solvent was reduced and then washed with 2x 5% NaHCC>3 ,'A"W î·. ï ” 010193 J-~r -· dried over Na2CO3 and evaporated to give crude chloromethylketone (20.46 g , 79%) wich was used in thenext step without further purification. 5 !η MMR (CDCL3, 400MHz) d 1.16-1.2 (t,lH), 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 10

Crude chloromethylketone (2)(10.04 g, 56.15 mmol) wasdisolved in 3 00 ml of dry MeOH. Sodium acetate (2 eq, 15 9.21 g, 112.3 mmol) was added followed by L-Cysteine ethyl ester hydrochloride sait (1.3 eq, 13.55g, 72.98mmol) and sodium cyanoborohydride (1.4 eq, 4.9 g, 78.59mmol). The heterogeneous mixture was left to stir atroocn température for 2h30 min. 200 ml of methanol (MeOH) 20 was then added to disolve ail the solid and the pH wasbrought to 2 with IN HCl .The mixture was then basifiedwith saturated NaHCO3 until pH= 8. MeOH was evaporatedand the remaining aqueous solvent was washed with ethylacetate and dichloromethane. Solvents were combined, 25 dried over Na2SÛ4 and evaporated. The crude residue was purified by silica gel flash column chromatography usinga gradient, of eluents ethyl acetate / hexane in the 144 010493 following ratios: (3:7, 5:5, 6:4, 7:3) giving cycliccompound (3) . 1h NMR (CDCI3 , 400MHz) of compound (3) d 1.21-1.27 (tt 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), 2.72- 2.76 (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

Cyclic compound (3) (913mg, 3.32 mmol) was disolved in 50 ml of dry Toluene. (1S)-(+)-10-Camphorsulfonic acid ( 92mg, 0.39 mmol) was added and the mixture was left toreflux for 4 days. When ail starting material was shownte be consumed (by TLC), the mixture was worked up byévaporation of solvent, dissolving residue in ethylace ta te and washing with 2 x 5 % NaHCC>3 . The Ethylacetate layer was dried over Na2SÛ4 and evaporated. Thecrade residue was purified by silica gel flash cO'lumnchronnatography using 60 % ETOAC / 40 %Hexane followed by 010493 14ο 70 % ETOAC / 30 % Hexane giving 62.5% of Eicycliccompound (4). XH NMR (CDC13, 400MHz) of compound (4) d 1.27-1.31 it„3H, J=7Hz), 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.S3-2.88 (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). . 10

lîoh.h2o,thf/h2o-»

O°C->R.T

15 20 25

Eicyclic (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 Η£θ, at 0°C and the mixture was left to stirr at 0°C for 1 hr and atrooc température for 3 hrs, THF was then evaporated andthe remaining aqueous mixture was acidified by additionof Citric acid until pH=2. Extraction of aqueous mixturewitfc. 2 x CH2CL2 and 2 x ETOAC, drying of combinedorganic layers with Na2SÛ4 and évaporation gave a cruderesidue wich was purified by silica gel flash columnchrcmatography using 70 % ETOAC / 30 %Hexane followed by4.7 % HOAC / Ethyl acetate giving the pure acid (5) in 54% yield . 16 % of starting material(4) was recovered. 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) , 2.03-2.12 (m, 1Ή), 2.35-2.5 (m, 2H) , 2.51-2.66 (m, 2H), 2.86- ’.’TFWr ’ *: ' 14 ύ 010493 2.21 (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 MMR31.382, (MeOD, 400 MHz) d32.096, 51.016, 17.052, 55.138, 27.07, 170,088, 28.928, 171.24 STEF 6 10

15 20

To a solution of lithium bis (trimethylsilyl) amide (5mlof IM THF solution, 5mmol) in THF (10 ml) is added at -78°Ca solution of the carboxylic acid (5) (500mg, 2.32 mmol).the resulting solution is stirred at -78°C for 1 hour.Eenzyi bromide (0.26 ml, 2.22 mmol) is then added and themixture is allowed to reach room température and stirredfor 15 hours. The mixture is then poured into 10% HCl (50mi) and extracted wiht diclhoromethane (4x 60ml) . Thecombined organic phases are dried over MgSO4 and thesolvent remove by évaporation to yield to the crudealkylated amide (6 ) . "Tssry*·, i-il 010493 STEP 7

5 The crude aklylated araide (6) is coupled with benzythiazole keto arginine in DMF using BOP as thecoupling reagent in the presence of diisopropylethylamine.Extraction with EtOAc gives a solid that is purified onsilica gel to give the protected amide. The CBZ protecting 10 group is removed with BBr3 in dichloromethane at room température finally gives the bicyclic benzothiazole ketoarginine inhibitors (7). 15

The following compound is produced accordingly with theexeption that the appropriate substitution of productswere made in order to obtain the final compounds. COMPOUND #8

010493 14ü EXAMPLE 9 Détermination of Kt Values for Heterocyclics 5

The affinity of inhibitors for thrombin was measuredaccording to the procedures described in (DiMaio et al, J.Bio. Chem., 1990, 265:21698) Inhibition of amidolyticactivity of human thrombin was measured fluorometrically 10 using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50mM Tris-HCl buffer (pH 7.52 at 37°C) containing 0.1 M NaCland 0.1% poly(ethylene glycol) 8000 at room température, and(Szewczuk et al., Biochemistry, 1992 31:9132). 15 The hydrolysis of the substrate by thrombin was monitoredon a Varian-Cary 2000™ spectrophotometer in thefluorescence mode (ÀeX =383 nm, Àem = 455 nm) or on aHitachi F2000™ fluorescence spectrophotometer (λ^ = 383 nm,λο = 455 nm), and the fluorescent intensity was calibrated 20 using AMC. The reaction reached a steady-state within 3minutes after mixing thrombin with the substrate and aninhibitor. The steady-state velocity was then measured fora few minutes. The compounds of this invention were alsopre-incubated with thrombin for 20 minutes at room 25 tenperature before adding the substrate. The steady-statewas achieved within 3 min and measured for a few min. Thekinetic data (the steady-state velocity at variousconcentrations of the substrate and the inhibitors) of thecompétitive inhibition was analyzed using the methods 30 described by Segel (1975). A non-linear régression program,RNLIN in the IMSL library (IMSL, 1987), LMDER in MIMPACKlibrary (More et al., 1980) or Microsoft™ Excell™ , wasused to estimate the kinetic parameters (K* and K,) . 35 dTT assay 149 010493

The fibrin clotting assay was performed in 50 mM Tris HClbuffer (pH 7.52 at 37 °C) containing 0.1 M NaCl and 0.1%poly(ethylene glycol) 8000 with 9.0 x 10-10 M (0.1 NIHunit/mL) and 0.03 % (w/v) of the final concentrations of 5 human thrombin and bovine ' fibrinogen, respectively, asreported elsewhere (Szewczuk et al.,supra). The clottingtinte was plotted against the inhibitor concentrations andthe ICS0 was estimated as the inhibitor concentrationrequired to double the clotting time .relative to the 10 control. Results are summarized in Tables 1 and 2 below.

Fibrin Clôt Assav

The fibrin clôt assay was performed essentially as15 described by Krtenansky et al, FEBS, 1987, 211:10. A serial dilution of the inhibitor was prepared in 50 mMtris HCl buffer (pH7.8 at 23 °C) containing 0.1M NaCl and0.1% (w/v) polyethylene glycol 8000. Human plasma (60pL,collected in 3.8% sodium citrate, blood/anticoagulant 9:1) 20 was added to microtiter wells (microtiter plate, Falcon)containing 100pL of various inhibitor dilutions. Thesolution was mixed after which 50pL of human thrombin (lnMfinal conc.) was added and mixed for 15 seconds. Theturbidity of the clôt was immediately monitored by 25 microplate autoreader (Dynateck MR 5000) at 405nm andrecorded every 3 min. The maximal turbidity in theabsence of inhibitors was reached within a 60 min. IC„values were calculated at 30 minutes as the inhibitorconcentration that gave half the optical density of the 30 control.

Platelet Aooreqation and Sécrétion

Rat blood was collected into ACD (6/1 v/v) by cardiac 35 purcture. Suspensions of washed platelets were preparedas described by Ardlie et al, (Br. J. Haematol. 1970, 19:7 150 010493 and Proc. Soc. Exp. Biol. Med., 1971, 136:1021). Thefinal suspending medium was a modified lyrode solution(NaCl 138mM, KCl 2.9mM, HEPES 20mM, NaH2PO4 0.42mM, NaHCO312mM, CaCl2 ImM, MgCl2 2mM, 0.1% glucose, 0.35% albumin, 5 apyrase lpL/mL pH 7.4) . Platelet counts were adjusted fco5000,000/pL.

To permit measurement of theextent of release of thecontents of the dense granules, the platelets were 10 labelled in the first washing solution with uC-serotonin(5-HT) (lpCi/lOmL of washing fluid) and release of UC-serotonin was determined as described in Holmsen et al,(Enzymology, 1989, 169:206). Inipramine (5pM final conc.)was added to présent the reuptake of released serotonin. 15

Platelet aggregation was recorded at 37°C in anaggregometer (BioData PAP-4) at a stirring speed of 1,100rpm by measuring the variations of light transmission.Percentage of aggregation was determined 3 min. after the 20 addition of the stimulating agent (human thrombin O.lIU/mLfinal conc.). Inhibitors were preincubated 1 minute at37°C before addition of stimulating agent. I'CS0 valuesrepresent the concentration that was necessary to inhibitplatelet aggregation or sécrétion to 50% of the control. 25

Arterial Thrombosis Model

FeCb, Induced Carotid Arterial Injurv Model

The FeCl3 induced injury to the carotid artery in rats was 30 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 andExperimental Therapeutics 267; 1237-1242, 1993. 35 Male, Sprague-Dawley rats ( 375-410 g) were anesthetizedwith urethane ( 1500 mg\kg ip) . Animais were laid on a 010493 37°C heating pad. The carotid artery was exposed through ex midline cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid sheath.Usine forceps, the artery was lifted to provide sufficient 5 clearance to insert two small pièces of polyethylenetubing (PE-205) underneath it. A température probe(Physitemp* MT23/3) was placed befween one of the pièces oftubing and the artery. Injury was induced by topical - application on the carotid artery above the température 10 probe of a small dise (3 mm dia. } of Whatman’ No.l filterpaper previously dipped in a 35% solution of FeCl3 . The incision area was covered with aluminum foil in order toproteçt the FeCl3 from dégradation bylight. The vesseltempérature was monitored for 60 minutes after application 15 of FeCl3 as an indication of blood flow. Vessel température changes were recorded on a thermister .(Gole-Palmer’ Model 08533-41) .

The time between the FeCl, application^andthe..time at 20 which the vessel température 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 (pg/kg/min. viafémoral vein). The dose of inhibitor needed to double the 25 time to occlusion in comparison to control animais in which injury was induced in the absence of inhibitor wasdetermined. 152 010493

Table 1 cmpd Ki |iM° Antiplcrtelet activity μΜ dTT icæ μΜ Plasma fibrin clôt assay ICt>, μΜ Aggregation 5-HT Sécrétion 0005 4 ND ND ... 47 >450 0010 4.6 21 19 89.5 >450 0015 Ίό >100 >100 162 >450 0020 2.2 18 14.2 22 >450 0025 53 >100 ; >100 >625 >450.; 0030 8.6 >100 >100 67 320 0035 34 >100 >100 319 >450 0040 19 >100 >100. 207.5 >450 0045 74 ND ND 415 >450 0050 62 ND ND >450'· 0065 32.7 47.5 52 42 200 0070 4.4 22' 2.1 25 78 0080 0.048 0.4 0.38 0.375 ND 0090 0.031 ND ND 0.33 ND —

X

0095 26 ND

ND 0104S3

0100 19 ND

ND 165 ND *suspension of washed.platelets. from rats“ Inhibitory dissociation constant for human a-thrombin ï>-> orna:

Table 2

Compound ki (nM) dTT (nM) Route ivb+inf M.O.T (min)+/-sem 0220 18 0225 550 0.75-50 23+/-7 235 0245 5 0.5-30 27+/-3 8 0.75-50 22.6+/-2.6 0250 40 350 0.25-20 23+/-8 0.75-50 22+/-3 0295a 1500 0.75-50 20+/-1 ...... .....r-„............... ...........-..... .................. __________________________________ 0295b 5000 520 0.75-50 19+/-2.7 0240 18 0.75-50 17+/-2.6 0210 8 0.75-50 2043+/-3.4 0255 500 0260 16 0.75-50 14.83+Λ0.2 0305a 220 0305b 12000 .................................. ..... ............................... ....................... ................... 0265a 4 0.75-50 21.5+/-10 0265b 18 0.75-50 14.83+/-2.3 ................................. ..... ______... ............................... ..........................~~...... ............................... 0285 10 0.75-50 11.33+/-1.34 150 ................................ .. ....... ............„................. .................. ...... ............................................ 0315a+b 45 0315b 10 0.75-50 30.33+/-8.4 0335 25 138 0.75-50 45.8+/-14.2....... 0.5-30 41.5+/-7.27 0.25-20 27.5+/-11,3 0340 0.6 0.25-20 36+/-9,6 0.75-50 42.25+/-11,9 0345 2 0.75-50 50+/-5.86 0915 1600 0.75-50 15+/-1,3 0935 120 ίϋί» 010493

Table 2 (continued)

Compound Ki (nM) dTT (nM) Route ivb+inf M.O.T (min)+/-sem O925a+b 10 0.75-50 19.6+/-0.2 0925b 30 0925a 7 . 0775-50 ..... ......2Ô.3+/-3.5........ """" 094Ôa.......... —ïë”...... 0775-50 '”'"Îë2+ÿ-ÔÏ827·' 0940b ——·· 0950a 150 0950b 1000 a = early eluting on RP HPLC single isomerb = late eluting on RP HPLC single isomera + b = mixture

Claims

WE CLAIM: «iOt 010493 -.1 h compound of formula (I):

10 15 20 25 wherein: A is selected from (CH-R8)0., , S, SO, SO2, O and NRewherein Rg is hydrogen, C,_6 alkyl optionallyinterupted with 1 or 2 heteroatoms; C6_16 aryl, C3_7cycloalkyl or heterocyclic ring or a hydrophobie ..........group; B is selected from S, SO2, O, -N=, MH, -CH= and CR6R7wherein R„ and R, are independently selected fromhydrogen and Cj.e alkyl provided that when A is S,SO, SO2, O, or NR8, then B is CR6R7; D is selected from (CH-R9)„_2 wherein R, is hydrogen, C6 alkyl or -C(O)R1Z· and CH with a double bond to B .....when 3 is -N= or -CH-; Eis selected from CH2and CH substituted with the -C(O)Rt, provided that only one ofD and E issubstituted with with -C(O)Rj; X is selected from O, N-R5, or CH-RS; χ Y is selected from O, S, SO, SO2, N-Rs and CH-R8 provided that when X is N-Rs then Y is CH-Ra or O,and when X is O then Y is CH-R8; Z is selected from O, S and H2; R2 is an arginyl moiety or an analog or dérivative ' thereof optionally substituted with an amino acida peptide or a heterocycle; . · R2 is selected from H and C,_6 alkyl optionally. substituted with C6 aryl, a 6 member heterocycle ca C3.7 cycloalkyl ring; 30 R3 is selected from H, NR6R7 and C,.( alkyl; and R4 and Rs are independentlv selected from H; NR(R..; ct.,, aryl or C3„7 cycloalkyl optionally substituted with C,_6 alkyl; Cj.,6 alkyl optionally interrupted by one 5 or more heteroatom or carbonylgroup and optionally substituted with OH, SH, NRfR7 or a C6_1G aryl,heterocycle or C3.7 cycloalkyl group optionally . . -.substituted with .halogèn e thydroxyl,, : alkyl y anamino acid side chain; and a hydrophobie group. 10
2. A compound according to claim 1, wherein Rj is one offormula Via to VId:

15 20 25 wherein: Rlt is hydrogen or alkyl; K is a bond or -NH-; - G is Cw alkoxy; cyano; -NH2; -CH2-NH2; -Ç(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C (NH) -NH2; a C6 cycloalkyl oraryl substituted with cyano, -NH2, —CH2-NH2, -C(NH)-NH2, -NH-C (NH)-NH2 or -CH2-NH-C(NH)-NH2; or a 5 or 6member, saturated or unsaturated heterocycleoptionally substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C (NH)-NH2; U-is cyano, -NH., -C (NH) -NH.or --NH-C (NH) -NIL,; -P is a bond, -C(O)- or a bivalent group: 010493 10 3. OH

J is C,.,; alkylene optionally substituted with OH, NHL,and C,.t alkyl and optionally interrupted by aheteroatom selected from O, S and N; n is 0 or 1; and T is H, OH, amino, a peptide chain, C,_16 alkyl, C,.16alkoxy, C6_20 aralkyl, or heterocycle optionallysubstituted. A compound according to claim 2, wherein T is a -heterocycle selected from the group consisting of:

Τ5 20 wherein X5, X1Q, Xtl and X13 are each independently selected fromthe group consisting of N, or C-X7 where X7 ishydrogen, Ct.4 alkyl, or C6_8 aryl; X6 and X13 are each independently selected from thegroup consisting of C, O, N, S, N-X,, or CH-X7; andR' is hydrogen, Ο1-16 alkyl optionally carboxylsubstituted, carboxyl, -C0_16 alkyl-CO2-C1_16 alkyl, C6 20aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle.
4. A compound according to claim 3, wherein T isselected from the group consisting of: -- 25 010493

wherein R' is hydrogen, Cj.u alkyl optionally carboxylsubstituted, carboxyl, -C0_16 alkyl-CO2-Cj.16 alkyl, C6.20aralkyl, C3.7 cycloalkyl, aryl or an aromatic 5 heterocycle. 5. A coinpound according to claim 4selected from: wherein T is

or

s-' 10 wherein R' is hydrogen, C,_u alkyl optionally carboxylsubstituted, carboxyl, -C0.u alkyl-CO2-Cj_1(. alkyl, C6.20aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle. 15 6. A compound according to claim 1, wherein one of R„ and R5 is a hydrophobie group selected from alkyl, C2.2c 01049: alkenvl or C7.2O alkynyl optionally interrupted by et carbonyl group, Ct.1(. aryl, C,.7 cycloalkyl, C6.2(, .aralkyl, C6.2c cycloalkyl substituted C.,,,, alkyl, wherein the aliphatic portion is optionally interrupted by a 5 carbonyl group and the ring portion is optionally substituted with C,.c alkyl; and a hydrophobie aminoacid side chain. 7 . ' A compound according to claim 6, wherein is H. 8. A compound according to claim 1, wherein Z is 0. 9 . A compound according to claim 1, wherein R2 is H7 15 10. A compound of the formula (VII);

wherein Rt is an arginyl moiety or an analog or dérivative20 thereof optionally substituted with an amino acid, a peptide or a heterocycle; Rs is H or Cl-6 alkyl; R3 is selected from H, NR,R.; and C,_6 alkyl; andR4 and R, are independently selected from H; NRJt,; Cé.ie25 aryl or C3.7 cycloalkyl optionally substituted with C,.c alkyl; C,.16 alkyl optionally interrupted by oneor more heteroatom or carbonyl group and optionallysubstituted with OH, SH, NR6R7 or a C61c aryl,heterocycle or C3_7 cycloalkyl group optionallysubstituted with halogen, hydroxyl, C,.6 alkyl; anamino acid side chain; and a hydrophobie group. 30 Ibl 01U4SS
11. A compound according to clciim 10, wherein R. is ont offormula Via to VId:

wherein: 5 R11 is hydrogen or C,_6 alkyl; K is a bond or -NH-; G is C,.4 alkoxy; cyano; -NH2; -CH2-NH2; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C(NH)-NH2; a C6 cycloalkyl oraryl substituted with cyano, -NH2, -CE^-NHj, -C(NH)- 10 NH2, -NH-C(NH) -NH2 or -CH2-NH-C (NH) -NH2; or a 5 or 6 mernber, saturated or unsaturated heterocycleoptionally substituted with cyano, ~NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CHj-NH-ÇÎNHÎ-NI^; U is cyano, -NH2, -C(NH)-NH2 or -NH-C (NH)-NH2; ..... 15 P is a bond, -C(O)- or a bivalent group: on

σ is C,.6 alkylene optionally substituted with OH, NH2and C,_6 alkyl and optionally interrupted by aheteroatom selected from O, S and N; n is 0 or 1; and T is H, OH, amino, a peptide chain, C,.,6 alkyl, 0,.,,.alkoxy, C6.20 aralkyl, or heterocycle optionallysubstituted. 20 162 Oî 0493 A compound according to clair.; 11, wherein τ is aheterocycle seiected from Lhe group consisting of :

5 wherein X5, X10, X13l and X12 are each independently seiected fromthe group consisting of N, or C-X, where X7 ishydrogen, Ct.4 alkyl, or C5_8 aryl; Xe and X13 are each independently seiected from the10 group consisting of C, O, N, S, N-X7, or CH-X7; R' is hydrogen, Cw6 alkyl optionally carboxylsubstituted, carboxyl, ~C0_1(. alkyl-CO2-C1.li alkyl, Ού_20aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle. 15
13. A compound according to claim 12, wherein T isseiected from the group consisting of: X. 010493

and R' is hydrogen, Ct.16 alkyl optionally carboxylsubstituted, carboxyl, -C0.lc alkyl-CO2-Cj_16 alkyl, C6.20aralkyl, C3.7 cycloalkyl, aryl or an aromatic 5 heterocycle. 14. A compound according to claim 13, wherein T isselected from: P’

10 and R' is hydrogen, C,.lt alkyl optionally carboxyl substituted, carboxyl, -C0_16 alkyl-CO2-Cj_16 alkyl, C6.20aralkyl, C3.7 cycloalkyl, aryl or an aromatic :heterocycle. 15 15 A compound according to claim 10, wherein R. and R3are H. 164 010493 16. l·. coitpound according to claim 10, wherein R. is c.alkyl optionally interupted with a heteroatom or acarbonyl, and optionally substituted with a C(i.16aromatic, C3.7 cycloalkyl or heterocycle ring whereinthe ring is optionally substituted with CF3 or oxo.
17. A compound according to claim 10, wherein R5 is H.
18. A compound according to claim 12, wherein: R3 is H; R4 is C,_16 alkyl optionally interupted with aheteroatom or a carbonyl, and optionallysubstituted with a CÉ.,6 aromatic, C3_7 cycloalkyl orheterocycle ring wherein the ring is optionallysubstituted with CF3 or oxo; and R5 is H.
19. A compound according to claim 10, selected from: 0085 6S-cyclohexy lmethylhexahydro-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 (a-benzothiozoloketo arginine).
20. A compound according to claim 1, of formula (VIII): R, R. wherein Ο 1 (J49 R, is an arginyl moiety or an anaiog or dérivativethereof optionally substituted with an amino acid,a peptide or a heterocycle; R3 is H or Cl-6 alkyl; 5 Rj is selected from H, NRtR., and C,_6 alkyl; and R4 and Rs are independently selected from H; NRCR7; C6_J(aryl or C3.7 cycloalkyl optionally substituted withC,_6 alkyl; C,.^ alkyl optionally interrupted by oneor more heteroatom or carbonyl group and optionally 10 substituted with OH, SH, NRJt, or a aryl, heterocycle or C3.7 cycloalkyl group optionallysubstituted with halogen, hydroxyl, Ο3.6 alkyl; anamino acid side chain; and a hydrophobie group. 15 21. A compound according to claim 20, wherein Rt is one of formula Via to VId;

wherein: Ri:l is hydrogen or C3_6 alkyl; 20 K is a bond or -NH-; G is Cj_4 alkoxy; cyano; -NH2; -CH2-NH2; -C(NH)-NH2; -NH- C(NH)-NH2; -CI^-NH-CiNH) -NH2; a CG cycloalkyl oraryl substituted with cyano, -NR,, -CH,-NH2, -C(NH)- NH2, -NH-C (NH) -NH2 or -CH,-NH-C (NH) -NH2 ; or a 5 or 6 25 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CH-2-NH2, -C(NH)-NHZ, -NH-C (NH)-NH2 or -CH,-NH-C (NH)-NH2 ; U is cyano, -NH2, -C(NH)-NH2 or -NH-C (NH)-NH2; 01048 4. \>k. P is a bond, -C(O)- or a bivé-.lent group: OH ,OH Cri 'Cii, CH, J is C,.t, alkylene optionally substituted with OH, NH2and C,.f> alkyl and optionally intërrupted by aheteroatom selected from O, s and N; n is 0 or 1; and T ifs H, OH, amino, a peptide chain, Cj_I6 alkyl, C,.l6alkoxy, C6_20 aralkyl, or heterocycle optionallysubstituted.
22. A compound according to claim 21, wherein T is a heterocycle selected from the group consisting of :

wherein X5, Xi0, Xxl and X12 are each independently selected fromthe group consisting of N, or C-X, where X7 ishydrogen, C14 alkyl, or C5_8 aryl; " X6 and X13 are each independently selected from thegroup consisting of C, O, N, S, N-X7, or CH-X7; R' is hydrogen, alkyl optionally carboxyl substituted, carboxyl, -C0.16 alkyl-CO2-Cj_16 alkyl, C6_20aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle.
23. A compound according to claim 22, wherein T rsselected from the group consisting of: λ U t 010483

and R' is hydrogen, Cj.16 alkyl optionally carboxylsubstituted, carboxyl, ~CO.U alkyl-CO2-C,.U alkyl, C6 20aralkyl, C3.7 cycloalkyl, aryl or an arômatic 5 heterocycle.
24. A compound according toselected from:

10 and R' is hydrogen, C,.,6 substituted, carboxyl, aralkyl, C3_7 cycloalkyl,heterocycle. claim 23, wherein T is

alkyl optionally carboxylC0.u alkyl-CO2-C1.16 alkyl, Cc.2caryl or an arômatic 15 25. A compound according to claim 20, wherein R2 and R3 are both H. 16& 010493
26. A compound according to claim 20, wherein R4 is H or C,.t alkyl substituted with COOH.
27. A compound according to claim 20, wherein R2, R3 and R4 are II and R5 if. alkyl optionally interrupted by one or more heteroatora or carbonyl group andoptionally substituted with OH, SH, NR6R7 or a C6_uaryl, heterocycle or C3_., cycloalkyl group optionallysubstituted with halogen, hydroxyl or C,.c alkyl.
28. A compound according to claim 22, wherein: R2, R3 and R4 are H; and Rs is Cj_,6 alkyl optionally interrupted by one or moreheteroatom or carbonyl group and optionallysubstituted with OH, SH, NR6R; or a C6.16 aryl,heterocycle or C3.7 cycloalkyl group optionallysubstituted with halogen, hydroxyl or Cj.6 alkyl.
29. A compound according to claim 20, selected from: 0345 4-Oxo-2- (3-phenyl-propionyl) -octahydro- pyrrolofl, 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[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazolè-2-carbonyl ) -butyl]-amide.
30. A compound according to claim 1, of formula (IX) : 010493

wherein Y is selected from O, S, SO, SO2, N-Rs and CH-RB; Rt is an arginyl moiety or an analog or derivéïtive 5 thereof optionally substituted with an amino acid, a peptide or a heterocycle; Ra is H or Cl-6 alkyl; R3 is selected from H, NR^R, and C,.6 alkyl; and R4 and Rs are independently selected from H; NR6R7; C6_u 10 aryl or C3_, cycloalkyl optionally substituted with C,,.6 alkyl; Cj_16 alkyl optionally interrupted by oneor more heteroatom or carbonyl group and optionallysubstituted with OH, SH, NR6R7 or a C6.16 aryl,heterocycle or C3.7 cycloalkyl group optionally 15 substituted with halogen, hydroxyl, Cj_c alkyl; an amino acid side chain; and a hydrophobie group; R„ is hydrogen, Ct.6 alkyl optionally interupted with 1or 2 heteroatoms; Cê.u aryl, C3.7 cycloalkyl orheterocyclic ring or a hydrophobie group; and 20 n is 1 or 2 ;
31. A compound according to claim 30, wherein R3 vis one offormula Via to VId:

l/G 010493 wherein : RX1 à."· hydrogen or C,.< alkyl; K is a bond or -NH-; G is C,.4 alkoxy; cyano; -NH..; -CH-NH.,; -C(NH)-NI-I2; -NH- 5 C(NH)-NH., ; -CH2-NH-C (NH) -NH. ; a C6 .cycloalkyl or aryl substituted with cyano, -NH,, -CH,-NH2, -C(NII)-NH,, -NH-C (NH) -NHL, or -CH..-NH-C (NH)-ΝΊΊ, ; or a 5 or 6member, saturated or unsaturated heterocycleoptionally substituted with cyano, -NH,, -CH,-NH2, - 10 C(NH)-NH2, -NH-C(NH)-NII2 or -CH2-NH-C(NH)-NH2; ü is cyano, -NH2, -C(NH)-NH2 or -NH-C (NH)-NH2; P is a bond, -C(O)- or a bivalent group; OH

J is Cj_c alkylene optionally substituted with OH, NH215 and C,_6 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,.u alkyl, Cj.16alkoxy, C6_20 aralkyl, or heterocycle optionally 20 substituted.
32. A compound according to claim 31, wherein T is a heterocycle selected from the group consisting of;

25 wherein X XK, Xr and X12 are each independently selecrted fromthe group consisting of N, or C-X7 where X7 ishydrogen, C. , alkyl, or C aryl; 171 010493 X, and X., are each independently selected front the group consisting of C, O, N, S, I\f-X7, or CH-X,,· R' is hydrogen, C,.,6 alkyl optionally carboxyl· substituted, carboxyl, "Co_1(, alkyl-CCf-C..,,. alkyl, C, ,!C 5 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. 33 A compound according to claim 32, wherein T isgroup consisting of:

10

R‘

R'

and R' is hydrogen, Cj.u alkyl optionally carboxylsubstituted, carboxyl, -Co u alkyl-CO2-C1I6 alkyl, C^20aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle.
34. A compound according to claim 33, wherein T isselected front: 15 010493

or

e and R' is hydrogen, Cj.,fc alkyl optionally carboxylsubstituted, carboxyl, -C0.16 alkyl-CC^-C,.,, alkyl, C6 ,(, aralkyl, C3 Ί cycloalkyl, aryl or an aromaticheterocycle.
35. A conipound according to claim 30, wherein R2 and R3are both H.
36. A compound according to claim 30, wherein R4 is H, NR6R7 or Cj_6 alkyl substituted COOH.
37. A compound according to claim 30, wherein Rs is C6_uaryl, C6_20 aralkyl, or Cj.16 alkyl substituted with C3_7cycloalkyl.
38. A compound according to claim 32, whereinn is 1; R2, R3 and R4 are H; and R6 is C616 aryl, C6.20 aralkyl, or C,.16 alkyl substitutedwith C3_7 cycloalkyl.
39. A compound according to claim 30, selected from: 0890 3-Amino-4-oxo-2-phenyl-hexahydro-pyrrolo[2,1- b][l, 3]thiazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide; 0895 3-Amino-2-benzyl-4-oxo--hexahydro-pyrrolo[2,1-b][l, 3]thiazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl·]-amide; and 010493 •ï. / U 0900 3-Amino-2-cyclohexy1-4 -oxo-hexahvdro~pyrrolo| 2,1 -b][l, 3]thiazine-6-carboxylic acid |1-(benzothiazole-2-carbonyl) -4-guanidino-butyl |-amide.
40. A compound according to claim 1, of formula (X) :

wherein B is O, S, -CH2-, or -NH-; Rx is an arginyl moiety or an analog or dérivativethereof optionally substituted with an amino acid,a peptide or a heterocycle; R2 is H or Cl-6 alkyl; R3 is selected from H, NR6R7 and C^,. alkyl; and R4 and R5 are independently selected from H; NRéR7; C6_16aryl or C3.7 cycloalkyl optionally substituted withCj_6 alkyl; C,.,6 alkyl optionally interrupted by oneor more het.roatom or carbonyl group and optionallysubstituted with OH, SH, NR6R7 or a C6_1G aryl,heterocycle or C3_7 cycloalkyl group optionallysubstituted with halogen, hydroxyl, C^6 alkyl; anamino acid side chain; and a hydrophobie group. A compound according to claim 40, wherein Rj is one offormula Via to VId: 41 174 01Ü 49 3

wherein: Rlt is hydrogen or C,.6 alkyl ; K is a bond or -NH-; 5 G is C,_4 alkoxy; cyano; ~NH2; -CH2-NH2; . -C(NH)-NH2; -NH- C(NH)-NH2; -CH2-NH-C (NH) -NH2; a C6 cycloalkyl oraryl substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH) -NH2 or -CH2-NH-C (NH) -NH2; or a 5 or 6member, saturated or unsaturated heterocycle 10 optionally substituted with cyano, -NH2, -CE^-NH^ - C(NH)-NH2, -NH-C(NH)-NH2 or -CI^-NH-C (NH) -NH2; U is cyano, -NH2, -C(NH)-NH2 or -NH-C (NH) ~NH2; P is a bond, -C(O)- or a bivalent group: OH

15 J is C,_6 alkylene optionally substituted with OH, NK, and C,_6 alkyl and optionally interrupted by aheteroatom selected from O, S and N; n is 0 or 1; and T is H, OH, amino, a peptide chain, Ο1-16 alkyl, Cj.16 20 alkoxy, C6.20 aralkyl, or heterocycle optionally substituted.
42. A compound according to claim 41, wherein T is_aheterocycle selected from the group consisting of: 0 î Ο493

wherein Xs, Xxl and X13 are each indeipendently selected from 5 the group consisting of N, or C-X7 where X7 is hydrogen, C,.4 alkyl, or Cs_„ aryl; Xe and X13 are each independently selected from thegroup consisting of C, 0, N, S, N-X,, or CH-X7; R' is hydrogen, C,.u alkyl optionally carboxyl 10 substituted, carboxyl, -C0_16 alkyl-CO2-Cj.16 alkyl, C6_20 aralkyl, C3.7 cycloalkyl, aryl or an aromaticheterocycle.
43. A compound according to claim 42, wherein T is 15 selected from the group consisting of: i/υ 010493

and R' is hydrogen, C,.u alkyl optionally carboxylsubstituted, carboxyl, -C0_u alkyl-CO-p-C^ alkyl, C6.20aralkyl, C3.7 cycloalkyl, aryl or an aromatic5 heterocycle.
44. A compound according to claimselected from: 43, wherern T is

R'

and R' is hydrogen, alkyl optionally carboxyl 10 substituted, carboxyl, -Co_16 alkyl-CO2-C!_16 alkyl, C6_20 aralkyl, C3_7 cycloalkyl, aryl or an aromaticheterocycle.
45. A compound according to claim 4G, wherein R2 ^nd R3are both H. 15 01 0 4 9 3 vu
46. A compound according to daim 40, wherein R. is C,_1(alkyl substituted with C,..u aryl optionallysubstituted with C,.,6 alkyl.
47. A compound according to daim 40, wherein R, is H.
48. A compound according to daim 42, whereinB is S; R2, Ro and Rs are H; and R4 is Cj.H alkyl substituted with C^I6 aryl optionallysubstituted with Cj.^ alkyl.
49. A compound according to claim 40, selected fromk925 7-Benzyl-6~oxo-octahydro-pyrido[2,1- c][l, 4]thiazine-4-carboxylic acid j4-guanidino-l-( thiazole-2-carbonyl)butyl]-amide; and940 6-Oxo-7-phenethyl-octahydro-pyrido[2 , l-c][l, 4]thiazine-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl ) -butyl]-amide.
50. A method for the treatment or prophylaxis ofthrombotic disorders in a mammal, comprisingadministering to said mammal an effective amount of acompound according to claim 1.
51. A method according to claim 50, wherein saidthrombotic disorder is venous thrombosis.
52. A method according to claim 50, wherein saidthrombotic disorder is a pulmonary embolism.
53. A method according to claim 50, wherein saidthrombotic disorder is arterial thrombosis. ίΐί
54. A method according to daim 50, wherein saidthroiubot.ic disorder is myocardial infarction.
55. A method according to daim 50, wherein said 5 throinbotic disorder is cérébral infarction.
56. A process for producing a compound according to claim1. 10 57. A process for producing a compound according to any one o£ daims 10, 20, 3 0 or 40. 010493