LV12019B

LV12019B - BULK CYCLIC FIBERS INHIBITORS

Info

Publication number: LV12019B
Application number: LVP-97-141A
Authority: LV
Inventors: John GILLARD; John Dimaio; M.Arshad SIDDIQUI; Patrice Preville; Yves St-Denis; Micheline Tarazi; Sophie Levesque; Benoit Bachand; Jeremy John Edmunds; Annette Marian Doherty
Original assignee: Biochem Pharma Inc.
Priority date: 1994-12-22
Filing date: 1997-07-15
Publication date: 1998-07-20
Also published as: AP9701004A0; FI972466A0; JPH11508535A; CZ189997A3; OA10493A; EP0802916A1; LV12019A; CA2208772A1; HUT77651A; AU715378B2; NZ297360A; AU4062895A; SK83897A3; PL320965A1; EE9700113A; NO972892L; NO972892D0; BG101647A; CN1175259A; FI972466A

Description

LV 12019

LOW MOLECULAR WEIGHT BICYCLIC THROMBIN INHIBITORS

FIELD OF THE INVENTION

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

BACKGROUND

Inordinate thrombus formation on blood vessel walls precipitates acute cardiovascular disease States that are the chief cause of death in economically developed societies. Plasma proteins such as fibrinogen, proteases and cellular receptors participating in hemostāsis have emerged as important factors that play a role in acute and chronic coronary disease as well aš cerebral artery disease by contributing to the formation of thrombus or blood clots that effectively diminish normai blood flow and supply. Vascular aberrations stemming from primary pathologic States such as hypertension, rupture of atherosclerotic plaques or denuded endothelium, activate biochemical cascades that serve to respond and repair the injury site. Thrombin is a key regulatory enzyme in the coagulation Cascade; it serves a pluralistic role as both a positive and negative ’feedback regulator. However, in pathologic conditions the former is amplified through catalytic activation of cofactors required for thrombin generation as well as activation of factor XIII necessary for fibrin cross-linking and stabilization.

In addition to its direct effect on hemostasis, thrombin exerts direct effects on diverse celi types thaf'support and amplify pathogenesis of arterial thrombus disease.

The enzyme is the strongest activator of platelets causing l them to aggregate and release substances (eg. ADP TXA2NE) that further propagate the thrombotic cycle. Platelets in a fibrin mesh comprise the principal framework of a white thrombus. Thrombin also exerts direct effects on 5 endothelial celis causing release of vasoconstrictor substances and translocation of adhesion molecules that become sites for attachment of immune celis. In addition, the enzyme causes mitogenesis of smooth muscle celis and proliferation of fibroblasts. From this analysis, it is 10 apparent that inhibition of thrombin activity constitutes a viable therapeutic approach towards the attenuation of proliferative events associated with thrombosis.

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

Hirudin, derived from the glandular secretions of the leech hirido medicinalis is one of the high molecular weight natūrai anticoagulant protein inhibitors of 30 thrombin activity (Markwardt F. Cardiovascular Drug

Reviews, 10., 211, 1992) . It is a biopharmaceutical that has demonstrated efficacy in experimental and clinical thrombosis. A potential drawback to the use of Hirudin as a therapeutic aģent is likely antigenicity and lack of an 35 effective method of neutralization, especially in' view of its extremely tight binding characteristics toward thrombin. The exceedingly high affinity for thrombin is 2 LV 12019 unique and is attributed to a simultaneous interaction with the catalytic site as well as a distal "anion binding exosite" on the enzyme. 5 Thrombin activity can also be abrogated by Hirudin-like molecules such as hirulog· (Maraganore, J.M. et al., Biochemistry, 29., 7095, 1990) or hirutonin peptides (DiMaio, J. et al., J. Med. Chem., 3.5, 3331, 1992). 10 Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for thrombin's catalytic site, thereby inhibiting proteolysis of that protein or other protein substrates such as the thrombin receptor. A common strategy for designing enzyme 15 inhibitory compounds relies on mimicking the specificity inherent in the primary and secondary structure of the enzyme's natūrai substrāte. Thus, Blomback et al. first designed a thrombin inhibitor that was modeled upon the partial seguence of the fibrinogen A(LBi)a Chain comprising 20 its proteolytically susceptible region (Blomback, et al., J. Clin. Lab. Invest., 24., 59, 1969). This region of fibrinogen minimally includes the residues commencing with phenylalanine: 25 Ala-Asp-Ser-Glv-Glu-Glv-Asp-Phe-Leu-Ala-Glu-Glv -Gly-Gly-Val-Arg-Gly-Pro-Arg T scissile bond

Systematic replacement of amino acids within this region 30 has led to optimization of the tripeptidyl inhibitory sequence exemplified by the peptide (D)-Phe-Pro-Arg which corresponds to interactions within the Ρ,,-Ρ,-Ρ, local binding sites on thrombin (Bajusz S. et al. in Peptides: Chemistry 35 Structure and Biology: Proceedings cf tlie Fourth American 3

Peptide Symposium, Walter R. , Meienhofer J. Eds. Ann Arbor Science Publishers Inc., Ann Arbor Ml, 1975, pp 603).

Bajusz et al. have also reported related compounds such as (D)Phe-Pro-Arg-(CO)H (GYKI-14166) and (D)MePhe-Pro-Arg-(CO)H (GYKI-147 66) (Peptides-Synthesis, Structure and

Function: Proceedings of the Seventh American Peptide

Symposium, Rich, D.H. & Gross, E. eds., Pierce Chemical Company , 1981, pp. 417). These tripeptidyl aldehydes are effective thrombin inhibitors both in vitro and in vivo.

In the case of both GYKI-14166 and GYKI-14766, the aldehyde group is presumed to contribute strongly to inhibitory activity in view of its Chemical reactivity toward thrombin's catalytic Ser)S5 residue, generating a hemiacetal intermediate.

Related work in the area of thrombin inhibitory activity has exploited the basie recognition binding motif engendered by the tripeptide (D)Phe-Pro-Arg while incorporating various functional or reactive groups in the ločus corresponding to the putative scissile bond (i.e. Pl-Pl').

In U.S. Patent 4,318,904, Shaw reports chloromethyl-ketones (PPACK) that are reactive towards Ser155 and His57. These two residues comprise part of thrombin’s catalytic triad (Bode, W. et al., EMBO Journal £,. 3467, 1989) .

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

Stili other congeners of this motif are those bearing phosphonates (Wang, C-L J., Tetrahedron Letters, 33_, 7667, 4 LV 12019 1992) and α-Keto esters (Iwanowicz, E.J. et ai.,Bioorganic and Medicīnai Chemistry Letters, 12.. 1607, 1992).

Neises, B. et al. have described a trichloromethyl ketone thrombin inhibitor (MDL-73756) and Attenburger, J.M. et al. have revealed a related difluoro alkyl amide ketone (Tetrahedron Letters, 22., 7255, 1991) .

Maraganore et al. (European 0,333,356; W0 91/02750; U.S. 5,196,404) d'isclose a series of thrombin inhibitors that incorporate the D-Phe-Pro- moiety and hypothesize that this preferred structure fits well within the groove adjacent to the active site of thrombin. Variations on these inhibitors are essentially linear or cyclic peptides built upon the D-Phe-Pro moiety.

Another series of patents and patent applications have described attempts to develop effective inhibitors against thrombosis by using alpha-ketoamides and peptide aldehyde analogs (EP 0333356;WO 93/15756; WO 93/22344; WO 94/08941; WO 94/17817).

Stili others have focused their attention on peptides, peptide derivatives, peptidic alcohols, or cyclic peptides as anti-thrombotic aģents (WO 93/22344, EP 0276014; EP 0341607; EP 0291982). Others have examined amidine sulfonic acid moieties to achieve this same end (U.S. 4,781,866), while yet others have examined para or meta substituted phenlyalanine derivatives (WO 92/08709; WO 92/6549) . A series of Mitsubishi patents and patent applications have disclosed apparently effective argininamide compounds for use as antithrombotic aģents. The Chemical structures described in these documents represent variations* of side groups on the argininamide compound (U.S. 4,173,630; U.S. 5 4,097,591; CA 1,131,621; U.S. 4,096,255; U.S. 4,046,876; U.S. 4,097,472; CA 2,114,1535.

Canadian patent applications 2,076,311 and 2,055,850 disclose cyclic imino derivazives that exhibit inhibitory effects on cellular aggregation.

Many o£ the examples cited ab-ove are convergent by maintaining at least a linear acyclic tripeptidyl motif consisting of an arginyl unic whose basie side chain is required for interaction with a carboxylate group located at the base of the P, specificity cleft in thrombin. Two adjacent hydrophobic groups provide additional binding through favourable Van der h’aals interactions within a contiguous hydrophobic cleft on the enzyme surface designated the P.,-P, site.

One object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme, thrombin. A further object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme thrombin and are provided for in a pharmacologically acceptable state.

Stili a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof as anticoagulant and thrombin inhibitory aģents.

Yet a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof for therapeutic treatment of various thrombotic maladies. 6 LV 12019 A further object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The enzyme inhibitors of the present invention are encompassed by the structure of general Formula I. 7

SUMMARY OF THE INVENTION

The present invention provides for novel compounds that display thrombin inhibitory activity as reflected in formula I: R 1Ί

z (I) wherein: A is selected from (CH-Ra)0., , S, SO, SO,, 0 and NRa wherein R, is hydrogen, C,.4 alkyl optionally interupted with 1 or 2 heteroatoms; C4.u aryl, C3.7 cycloalkyl or heterocyclic ring or a hydrophobic group; B is selected from S, S02, Ο, -N=, NH, -CH= and CR6R, wherein Rs and R, are independently selected from hydrogen and Ct.6 alkyl provided that when A is S, SO, SO;, 0, or NR„, then B is CR^R,; D is selected from (CH-R,)0., wherein R, is hydrogen, Ct_6 alkyl or -C(0)R,; and CH with a double bond to B when B is -N= or -CH=; E is selected from CH, and CH substituted with the -C(0)R,, provided that only one of D and E is substituted with with -C(O)Rt; X is selected from 0, N-Rs, or CH-RS; Y is selected from 0, S, S0, S02, N-Rs and CH-Rg provided that when X is N-Rs then Y is CH-R„ or O, and when X is O then Y is CH-Re; Z is selected from O, S and H,;

Rx is a polar amino acid residuearginyl moiety or an analog or derivative thereof optionally substituted with~an amino acid, a peptide or a heterocycle; 8 LV 12019

Rj is selected from H and C,.f alkyl optionally substituted with C£ aryl, a 6 member heterocycle or a C3_7 cycloalkvl ring; R3 is selected from H, NR..R7 and C.., alkyl; and 5 R, and Rs are independently selected from H; NR6R7; C£.u aryl or C3.7 cycloalkyl optionally substituted with C,_6 alkyl; alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR£R7 or a C6..( aryl, heterocycle or C3_7 10 cycloalkyl group optionallv substituted with halogen, hydroxyl, C,.e alkyl; an amino acid side chain; and a hydrophobic group.

As will be appreciated from the disclosure to follow, the 15 molecules, compositions and methods of this invention are useful as anti-coagulants, or in the treatment and prevention of various diseases attributed to the undesirable effects of thrombin, as well as for diagnostic purposes. 20

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to molecules which inhibit 25 the enzyme, thrombin. These molecules are characterized by a heterobicyclic moiety as illustrated in Formula I:

wherein X, Y, Z, A, 3, D, E and R, to R„ are as previouslv defined. 9

The term "hydrophobic group" (HG) as used hereinafter, refers to any group which lacks affinity for, or displaces water. Hydrophobic groups include but are not limited to C,..,0 alkyl, C2.20 alkenyl (e.g. vinyl, allyl) or C;.20 aikvnyl (e.g. propargyl) optionally interrupted bv a carbonyl group, (e.g. forming an acyl group); 0(..,.. aryl, C,.7 cycloalkyl, Cc_2ū aralkyl, C6.20 cycloalkyl substituted C,..0 alkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group (e.g. forming an acyl group) and the ring portion is optionally substituted with C,.( alkyl such as methyl ethyl or t-butyl; or a hydrophobic amino acid side chain. Preferred hydrophobic groups include cyclohexyl, benzyl, benzoyl, phenylmethyl, phenethyl and para-t-butyl-phenylmethyl.

The term "arginyl moiety" represents an arginine amino acid residue or an analogue or derivative thereof. For example, an analogue or derivative of the natūrai residue may incorporate a longer or shorter methylene chain from the alpha carbon (i.e. ethylene or butylene chain); replacement of the guanidino group with a hydrogen bond donating or accepting group (i.e. amino, amidino or methoxy); replacement of the methylene chain with a constrained group (i.e. an aryl, cycloalkyl or heterocyclic ring); elimination of the termiņai carboxyl (i.e. des-carboxy) or hydroxyl (i.e. an aldehyde) ; or a combination thereof.

The term "alkyl" represents a straight or branched, saturated or unsaturated chain having a specified total number of carbon atoms.

The term "aromatic" or "aryl" represents an unsaturated carbocyclic ring(s) of 6 to 16 carbon atoms which' is optionally mono- or di-substituted with OH, SH, amino (i.e. NRtR7) halogen or C,.{. alkyl. Aromatic rings include 10 LV 12019 benzene, napththalene, phenanthrene and anthracene. Preferred aromatic rings are benzene and naphthalene.

The term "cycloalkyl" represents a saturated carbocyclic ring of 3 to 7 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NRSR7) halogen or C... alkyl. Cycloalkyl groups include cyclo- propyl, butvl, pentyl, hexyl and heptyl. A preferred cycloalkyl group is cyclohexyl.

The term " aralkyl'' represents a substituent comprising an aryl moiety attached via an alkyl chain (e.g. benzyl, phenethyl) wherein the sum total of carbon atoms for the aryl moiety and the alkyl chain is as specified. The aryl or chain portion of the group is optionally mono- or di-substituted with OH, SH, amino (i.e. NRjR,) halogen or C..t alkyl

The term "heteroatom" as used herein represents oxygen, nitrogen or sulfur (Ο, N or S) as well as sulfoxyl or sulfonyl (SO or S02) unless otherwise indicated. It is understood that alkyl chains interrupted by one or more heteroatoms means that a carbon atom of the chain is replaced with a heteroatom having the appropriate valency. Preferrably, an alkyl chain is interrupted by 0 to 4 heteroatoms and that two adjacent carbon atoms are not both replaced.

The term "heterocycle" represents a saturated or unsaturated mono- or polycyclic (i.e. bicyclic) ring incorporating 1 or more (i.e. 1-4) heteroatoms selected from N, 0 and S. It is understood that a heterocycle is optionally mono- or di-substituted with OH, SH, amino (i.e. NRCR7) , halogen, CF3, oxo or C,_6 alkyl. Examples of suitable monocyclic heterocycles include but are not limited to pyridine, piperidine, pvrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran and 11 thiophene. Examples of suitable bicyclic heterocycles include but are not limited to indole, guinoline, isoquinoline, purine, and carbazole. 5 The term "hydrophobic amino acid" represents an amino acid residue that bears an alkyl or aryl group attached to the α-carbon atom. Thus glycine, which has no such group attached to the α-carbon atom is not a hydrophobic amino acid. The alkyl or aryl group can be substituted, provided 10 that the substituent or substituents do not detract from the overall hydrophobic character of the amino acid. Examples of hydrophobic amino acids include natūrai amino acid residues such as alanine; isoleucine; leucine; phenylalanine; and non-naturally ocurring amino acids such 15 as those described in “The Peptides", vol. 5, 1983, Academic Press, Chapter 6 by D.C. Roberts and F.

Vellaccio. Suitable non-naturally ocurring amino acids include cyclohexylalanine and l-aminocyclohexane-carboxylic. 20

By "amino acid side chain" is meant the substituent attached to the carbon which is α to the amino group. For example, the side chain of the amino acid alanine is a methyl group and while benzyl is the side chain for 25 phenylalanine.

Preferably R3 is H or Cj.j alkyl. More preferably R2 is H, methyl or ethyl and most preferably R2 is H. 30 Preferably/ R, is H or C,.6 alkyl. More preferably, R., is H, methyl or ethyl, and most preferably R3 is H.

Preferably, one of R4 or Rs is a hydrophobic group such as a saturated or unsaturated carbocycle of 5 or 6 -members 35 optionally fused to another carbocyclic group while the other is H, alkyl optionally substituted by NRCR7 or 12 LV 12019 carboxy. The hydrophobic moiety may be linked via a spacer such as a C,.1S alkyl chain optionally interrupted with 1 or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO,) groups. More preferably, one of R4 and R; is phenyl, 5 cyclohexyl, indole, thienyl, quinoline, tetrahydroisoquinoline/ naphthyl or benzodioxolane linked via C,_u alkyl optionally interupted with a heteroatom or a carbonyl while the other is H, carboxymethyl or carboxyethyl. 10

Preferably, A is absent or CH_.

Preferably, B is S or CH,.

Preferably, D is CH2.

Preferably, E is CH substituted with -C(0)R, wherein R, is 15 as previously defined.

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

Preferably, Y is CH-Re or S.

Preferably, Z is 0. 20 In a preferred embodiment, is represented by one of formula Via to Vīd:

wherein:

Rxl is hydrogen or Cj.e alkyl; 25 K is a bond or -NH-; G is C,.4 alkoxy; cyano; -NH2; -ΟΗ2-ΝΗ3; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C (NH)-NH:; a Ct cycloalkyl or aryl substituted with cyano, -NH.,, -CH,-NH;, -C(NH)-NH.,, -NH- 13 C(NH)-NH. or -CH,-NH-C (NH) -NH,; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH2-NH,, -C(NH)-NH2, -NH-C (NH) -NH, or -CH,-NH-C (NH) -NH, ; U is cyano, -NH,, -C(NH)-NH, or -NH-C (NH) -NH, ; P is a bond, —C(0)— or a bivalent group:

OH

J is C[.c alkylene optionally substituted with OH, NH, and C.., alkyl and optionally interrupted by a heteroatom selected from 0, S and N; n is 0 or 1; and T is Η, OH, amino, a peptide chain, C,_u alkyl, C,.u alkoxy, C,.,0 aralkyl, or heterocycle· optionally substituted.

Preferably R21 is H or methyl and most preferably H. Preferably K is a bond.

Preferably G is -NH-C(NH)-NH2 attached via' a methylene Chain of 3-7 carbons or phenyl substituted with -C(NH)-NH, attached via a methylene chain of 0 to 3 carbons. More preferably G -NH-C (NH)-NH2 attached via a methylene chain of 3 atoms.

Preferably P is -C(O)-.

Preferably J is selected from: -CH2-S-CH,-CH2-; -CH,-0-CH,-CH,-; -CH,-NH-CH,-CH2-; and a bond when n is 0. More preferably, J is a bond while n is 0.

In particular embodiments of the invention, R. is selected from the following amino acid derivatives prepared according to the procedures described in Bioorg. Med.

Chem., 1995, 3:1145 : 14 LV 12019

15

16 LV 12019

17 ο ο

ΝΗ Ο Ο

ΝΗ ΝΗ 5 wherein η=1-6, nl=l-2, η2=0-7 and Τ is as previously defined.

In a preferred embodiment, T is a peptide-of 1 to 4 amino 10 acid residues in length and preferably fibrinogen's A or B Chain or fragment or derivative thereof. In another preferred embodiment, T is a heterocycle selected from the group consisting of:

wherein X5, X10, Xu and X13 are each independently selected from the group consisting of N, or C-X. where X7 is hydrogen, Ct.4 alkyl, or Cj..^ aryl ; 18 15 LV 12019 X5 and X13 are each independently selected from the group consisting of C, 0, N, S, Ν-Χ-, or CH-X7; R' is hydrogen, C..16 alkyl optionally carboxyl substituted, carboxyl, -C0.IS alkyl-CO,-CI.1{ alkyl, C6.;. 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 preferablv T is selected from the group consisting of:

19 v/herein R' is as defined above.

More preferably T is selected from the group consisting of :

wherein R' is as defined above. Most preferably T is r·

10 wherein R' is H or C,.4 alkyl such as methyl, ethyl, propyl or butyl and most preferably wherein R' is hydrogen,. In another embodiment, T is a 1,2 thiazole optionally substituted with R' and\or is attached to J at the 2, 3, 4 or 5 position of the ring. 15

In particular eirbodiments, compounds of the invention are represented by formulas II, III, IV and V, wherein X, Y, B, R, to R4 and R, are as previously defined.

20 LV 12019

In a particularly preferred embodiment, compounds of the invention are represented by one of formulas VII, VIII, IX and X: (VII) (IX)

wherein B is 0, S, -CHj-, or -NH- ; ī is selected from 0, S, SO, S02, N-R; and CH-Re;

Rj, is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;

Rj is H or alkyl; R3 is selected from H, NRSR, and Cj_s alkyl; and R4 and R, are independently selected from H; NRjR,; Cs.16 aryl or C3_7 cycloalkyl optionally substituted with C,.s alkvl; Cj.ls alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally siibstituted with OH, SH, NR6Rj or a Ce.ls aryl, heterocycle or C3.7 cycloalkyl group optionally substituted with halogen, hydroxyl, alkyl; an amino acid side chain; and a hydrophobic group; R, is hydrogen, Cj.^ alkyl optionally interupted with 1 or 2 heteroatoms; Ce.I6 aryl, C..7 cycloalkyl or heterocyclic ring or a hydrophobic group; and n is 1 or 2.

Preferred compounds according to formula VII include: 21 0005 6 S-benzylhexahydrο-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (propyl ketoarginine) 0010 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-c arboxami do (buty1 ketoarginine) 0015 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a] pyridine-3R-carboxamido(propylcarbmethoxy ketoarginine) 0020 6S-cyclohexylmethyl hexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(benzylketo arginine) 0025 6S-cyclohexyl methyl hexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine -3R-carboxamido (,carbmethoxy propyl cyclodithioketalarginine) 0030 6S-cyclohexylmethyl hexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((S)-Arg-(R)-pipecolilic acid)

22 LV 12019 0035 6S-benzylhexa hydro-5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido(carboxamidopropyl cyclodithioketal arginine)

0040 6 S-cyc1ohexylme thy1hexahydro- 5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((S)-Arg nipecotamide)

0045 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3, 2-a]pyridine-3R-carboxamido((S)Arg isonipecotamide)

0050 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(carboxamidopentyl cyclodithioketai arginine)

0055 6 S-benzylhexahydro-5-oxo-5H- thiazolo[3,2-a]pyridine-3R-carboxamido(carbmethoxy propyl cyclodithioketal arginine)

0060 6 S-cyclohexylme thylhexahydro- 5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(Ιο arboxy-3 -thiobu ty1 ketoarginine)

23 0065 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(1-carboxy-3-thiobutyl ke toarginine) 0070 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridifte-3R-carboxamido(1-carboxy-2-methyl-3-thiobutyl ketoarginine) 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]pyridine-3R-carboxamido{iso-quinolinium methyl ketoarginine) 0085 6S-cycloh.exylmethylhexah.ydro-5-oxo-5H-thiazolo[3,2— a]pyridine-3R- carboxamido(propylcarbmethoxy ke toarginine) 0090 6 S-cyc1ohexylme thylhexahydro- 5-oxo-5H-thiazolo[3,2— a]pyridine-3R- carboxamido((propylketo)Arg-Phe-Arg-NH;,)

24 LV 12019 0095 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((propanoic acid) ketoarginine) 0100 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(propyl carbmethoxy ketoarginine) 0105 6 S-cyc1ohexylme thylhexahydro- 5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido (oc-benzothiazolo keto arginine); and 0110 6 S-cyc1ohexylpropylhexahydro- 5-oxo-5H-thiazolo [3,2-a-]pyridine-3R- carboxamido(propylcarbmethoxy ketoarginine)

25 0205 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide

H ο

o

HN NH, 0210 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 -carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-bu ty 1] - ami de 0215 6-Benzy1-5-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-bu ty 1] - ami de

H

Μθ

26 LV 12019 0225 8a-Methyl-5-oxo-6- phene thy1-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0230 8a-Methyl-5-oxo-6- phene thy1-hexahydro-thiazolo[3,2-a]pyridine-3- carboxylic acid [1-(benzothiazole-2-c arbony1)-4-guanidino-butyl]-amide 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-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 - carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) butyl] -amide

Me

H

27 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 -carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) butyl]-amide 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(l-methyl-lH imidazole-2-carbonyl) butyl]-amide 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 -carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-bu ty 1] - amide 5-Oxo-6-(3-cyclohexyl-propyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) butyl]- amide 8a-Methyl-5-oxo-6-(3-phenyl-propyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

H

ΜΘ

Me

HN==—WH 28 3 LV 12019 0280 8a-Methyl-5-oxo-6-(3 - phenyl-propyl) -hexahyd.ro-thiazolo[3,2 -a]pyridine-3 -carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 0285 8a-Methyl-5-oxo-6-(2- trifluoromethyl-quinolin-6-ylmethyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0295 6-(1,3-Dioxo-l,3-dihydro- isoindol-2-yl)-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3 -carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0305 5-Oxo-6-(3-phenyl- propionyl amino)-hexahydro thiazolo[3,2-a]pyridine-3 -carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

Me

29 0315 5-Οχο-6-(3-phenyl- propionyl amino)-hexahydro thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

More preferred compounds according to formula (VII) include: 5 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(propylcarbo methoxyketoarginine); 0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2— a]pyridine-3R-carboxamido ((propylketo)Arg-Phe-Arg-NH2) ; 10 0095 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((propanoic acid) ketoarginine); 0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2— a]pyridine-3R-carboxamido (α-benzothiozolo keto arginine); 15 0210 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guani dino -bu ty 1] - ami de; 20 Η

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-trif luoromethyl-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 ; 30 25 5 LV 12019 0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- a]pyridine-3-carboxylic acid [4-guanidino-l-(thiazole-2 -carbonyl) -butyl]-amide; 0265 5-Oxo-6- (3-cyclohexyl-propyl) -hexahydro-th.iazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-l-(thiazole-2 -carbonyl) bu tyl] -amide ; 10 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)-bu ty 1] -ami de; and 0315 5-Oxo-6- (3-phenyl-propionylamino) -hexahydro thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl]-amide. 15 Most preferred compounds according to formula VII include: 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido (propylcarbo methoxy ketoarginine); and 0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-20 a]pyridine-3R-carboxamido (α-benzothiozolo keto arginine). 31

Preferred compounds according to formula VIII include:

0325 3 - Aminome thy 1 - 2 -benzoy 1 - 4 - oxo - octahydro-pyrrolo[l, 2-a-]pyridine-6-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0330 3-Aminome thy 1-4-oxo-2 - phenylacetyl-octahydro-pyrrolo[l, 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)-bu ty 1] -amide 0340 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo[l, 2-ā]py^čizine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 32 LV 12019 0345 4-Oxo-2-(3-phenyl-propionyl) octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l- (5-methyl-thiazole-2-carbonyl)-butyl]-amide 0350 2-(3-Cyclohexy1-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,7-diaza-naphthalene-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 0365 4-Oxo-2-(4-phenyl-butyryl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 0370 4-Oxo-2-phenylacetyl-octahydro-pyrrolo[l/ 2-a-]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide

33 2-(2-Amino-3-phenyl-propionyl) -4-oxo-octahyd.ro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-butyl]-amide 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 2-[2-Amino-3- (4-fluoro-phenyl) -propionyl]-4-oxo-octahydro-pyrrolo[l/ 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 4-Oxo-2-(3-phenyl-propyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 2-[2-Amino-3-(lH-indol-3-yl)-pr op iony 1] - 4 - oxo -oc t ahydr o -pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide o

o

34 LV 12019 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 4-Oxo-2-(3-thiophen-2-yl-propionyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6- carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-butyl]-amide 2- (3-1 H-Imidazol-4-yl-propionyl)-4-oxo-octahydro-pyrrolo[l, 2-a] pyrazine-6- carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-butyl]-amide 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 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

O

o

35 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 043 5 4-Oxo-2-phenoxyac e ty 1 - octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide

o

36 LV 12019 4-Oxo-2-(3-phenoxy-propionyl)-ocatahydro-pyrrolo[l, 2-a-]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 4-Οχο-2-(2-phenyl-ethanesulfonyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-l-{thiazle-2-carbonyl)-butyl]-amide 2-(Naphthalene-2-sulfonyl)-4-oxo-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 4- (6-[4-Guanidino-l- (thiazole-2-carbonyl) -buty!carbamoyl]-4-oxo-hexahydro-pyrrolo[l, 2-a] pyrazin-2yl)-4-ΟΧΟ-3-(2 propyl -pentanoylamino)-butyric acid methyl ester 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [4-guanidino-1) -butyl]-amide 4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a] pyrazine-6-carboxylic acid [3-guanidino-propyl) -amide

o

37 0470 4- ( 6-[4-Guanidino-l- (thiazole-2-carbonyl) -butylcarbamoyl]-4-oxo-hexahydro-pyrrolo[l, 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 -bu tyl] -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-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(4-methyl-thiazole-2-carbonyl) -butyl]-amide 0490 4-0xo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l,2-a]py^a-zine-6-carboxylic acid[l-(4-ethyl-thiazole-2-carbonyl)-4 -guanidino -bu ty 1] -amide 0495 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-' a]pyrazine-6-carboxylic acid (4-carbamimidoyl-pheny)-amide

O

o

38 LV 12019 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]-amide 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-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [1-(4-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0515 4-Oxo-2- (3-ph.enyl-propionyl) - octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(3-carbaitiimidoyl-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-carbēunimidoyl^piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0525 4-Oxo-2-(3-phenyl-propionyl)- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-e thy 1] - amide

O

o

39 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-2-ylmethyl)-2-oxo-2-thiazol-2-yl- ethyl]-amide [6-[4-Guanidino-l-(thiazole-2-carbonyl) -butylcarbamoyl]-4-oxo- 2- (3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-3-yl]-acetic acid 3- [6-[4-Guanidino-l- (thiazole-2-carbonyl) -butylcarbamoyl]-4-oxo- 2- (3-phenyl-propionyl)- oc tahydr o -pyrrolo[l, 2 -a]pyraz in- 3- yl]-propionic acid [6—[1— (l-Carbamimidoyl-piperin- 4- ylmethyl)-2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2- (3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazin-3-yl) -acetic acid 3—[6 —[1— (l-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)-oc tahydro-pyrrolo[l, 2-a]pyrazin-3-yl)-acetic acid o

o

NH, o

o

40 LV 12019 [6-[l- (l-Carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l< 2-a]pyrazin-3-yl)-acetic acid [6- (3 -Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 3- [6- (3-Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -propionic acid 4- Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazolc-2-carbonyl) -butyl]-methyl-amide 4-0xo-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-ethyl]-methyl-amide o

o

41 [6- ([l-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-methyl-carbamoyl) -4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazin- 3- yl]-acetic-acid 4- Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine-6-carboxylic acid [1-{l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-e thy 1] -me thy 1 - amide 4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-methyl-amide 2-(Naphthalene-2-carbonyl)-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide 2-(Naphthalene-l-carbonyl)-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-amide o

o

NH, 42 LV 12019 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- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl) -butyl]-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-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l- (thiazole-2-carbonyl)-butyl]-amide 0625 2-[2-(2-Methyl-benzylidene)-but- 3-enoyl]-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(1-carbanvimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide

o

43 0630 2-(2-(2-Methyl-benzylidene)-but- 3- enoy 1] - 4 - οχο - oc tahydr ο-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(1-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 063 5 2- (2-Benzylidene-pent-3-enoyl) - 4- οχο-oc tahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-amide ο

o

44 LV 12019 4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid 4-c arbamixnidoyl-benzy lamide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine- 6-carboxylic acid [4-imidazol-l-yl-l-(thiazole-2-carbonyl)-butyl]-amide o

O

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 o

4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [3-(2-amino-6-methyl-pyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide

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 o

45 4-Οχο-2- (3-ph.enyl-propi.onyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [3 -(6-amino-pyridin-2-yl)-1-(thiazole-2-carbonyl) -propyl]-amide 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 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 4-0xo-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)-oc tahydro-pyrrolo[l, 2 -a]pyrazine-6-carbonyl]-3- (thiazole-2-carbonyl)-1,2,3,4-tetrahydro-isoQ[uinoline-6-carboxamidine

O

o

O

46 0700 0705 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-7-carboxamidine M-[l-[4-Oxo-2- (3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carbonyl]-5- (thiazole-2-carbonyl) -pyrrolidin-3-yl]-guanidine 0710 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine- 6-carboxylic acid [1-{4-amino-cyclohexyl)-2-oxo-2-thiazol-2-y 1 - e thy 1] - amide 0715 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(4-amino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0720 4-0xo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(4-amino-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide LV 12019 o

o

O

47 0725 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine-6-carboxylic acid [l-(4-aminomethyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine- 6-carboxylic acid [l-(3-aminomethyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 4-Oxo-2-(3-phenyl-propionyl)-o c t ahy dr o -pyr r ο 1 ο [ 1,2 - a]pyr az ine -6-carboxylic acid (2-oxo-l-piperidin-4-ylmethyl-2-thiazol-2-yl-ethyl)-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (2-oxo-l-piperidin-3-yl-2-thiazol-2-yl-ethyl)-amide 4-0xo-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

0730 o

0735 o

0740 o

0745 o

48 LV 12019 0750 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1—(4 — guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide

O

0755 0760 0765 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(2-guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 4-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-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-phenyl-thiazole-2-carbonyl) -butyl]-amide o

0770 4-0xo-2-(3-phenyl-propionyl)- octahydro-pyrido[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0775 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [4-guanidino-l- (th.iazole-2-carbonyl) -butyl]-amide o

NH

S N N 49 0780 0780 0785 0790 0795 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 5-0xo-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 5-0xo-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 5-0xo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1-(l-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0800 [4-[4-Guanidino-l-(thiazole-2- carbonyl) -butylcarbamoyl]-5-oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalen-6-yl]-acetic acid

o

50 LV 12019 0805 5-Oxo-7-(3-phenyl-propionyl)- octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide o

0810 3-[4-[4-Guanidino-l-(thiazole-2-carbonyl-butylcarbamoyl] - 5 -oxo -7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-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 -pr opy 1] -amide

0820 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-ethylļ-amide o

0825 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(hydroxy-thiazol-2-yl-methyl)-butyl]- amide o

NH? 51 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (4-guanidino-l-thiazol-2-ylmethyl-butyl ] -amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-carboxylic acid [4-guanidino-l-thiazol-2-yl-butyl]-amide 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-methoxy-l-(thiazole-2-carbonyl)-butyl]-amide [6-[4-Methoxy-l- (th.iazole-2-carbonyl) -butylcarbamoyl]-4-oxo- 2- (3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazin- 3- yl]-acetic acid [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 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-amino-l-(thiazole-2-carbonyl)-butyl]-amide

O

o

NH, 52 0860 0865 0870 0875 0880 4-Oxo-2-(3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [5-amino-l-(thiazole-2-carbonyl)-pentyl]-amide 4-Oxo-2-(3-phenyl-propionyl)-o c t ahy dr o -py r r o 1 ο [ 1,2 - a]pyr a z ine -6-carboxylic acid [5-guanidino-1-(thiazole-2-carbonyl)-pentyl]-amide 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 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(l-methyl-lH-imidazole-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 LV 12019

O

o

NH,

O

53 0885 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

NH, 54 LV 12019

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-c arbony 1) - 4 - guanidino -bu ty 1] - ami de 0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydro-pyrrolo[l/2-a]pyrazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl) -4-guanidino-butyl]-amide 0515 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxy 1 ic acid [1-(3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0530 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-{l-carbamimidoyl-piperidin-2 -ylmethyl) -2 -oxo-2 - thiazol-2 -yl-ethyl]-amide 0545 [6—[1— (l-Carbamimidoyl-piperin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 0550 3 —[6—[1— (l-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2 - thiazol-2-yl-ethylcarbamoyl]-4-ΟΧΟ-2- (3-phenyl- propionyl) -octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 0555 [6 — [ 1 — (l-Carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazin-3-yl) -acetic acid 55 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]pyrazin-3-yl) -propionic acid 0575 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-p iperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl] -methyl-aicri.de 0580 [6- ([l-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-methyl-carbamoyl) -4-oxo-2- (3-phenyl-propionyl) -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 [1-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-methyl-amide 0590 4-0xo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-methyl-amide 0595 2-(Naphthalene-2-carbonyl)-4-oxo-octahydro- pyrrolo(l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl]-amide 56 LV 12019 0625 2—[2 — (2-Meth.yl-benzylid.ene) -but-3-enoyl]-4-oxo- octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 5 0630 2-[2- (2-Methyl-benzylidene) -but-3-enoyl]-4-oxo- octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 10 0635 2- (2-Benzylidene-pent-3-enoyl) -4-oxo-octahydro- pyrrolo[l/2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-amide 15 0645 4-0xo-2- (3-phenyl-prbpionyl) -octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [4-imidazol-l-yl-l-{thiazole-2-carl5ohyl) -butyl]-amide 57 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-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [2-(2-amino-pyridin-4-yl) -1- (thiazole-2-carbonyl) -ethyl]-amide 0690 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [2-(6-amino-pyridin-2-yl)-1-(thiazole-2-carbonyl)-ethyl]-amide 0695 2-[4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carbonyl]-3- (thiazole-2-carbonyl) - 1.2.3.4- tetrahydro-isoguinoline-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-isoquinoline-7-carboxamidine 0705 N-[l-[4-Oxo-2-(3-phenyl-propionyl)-octahydro- pyrrolo[l, 2-a]pyrazine-6-carbonyl]-5- (thiazole-2-carbonyl) -pyrrolidin-3-yl]-guanidine 58 LV 12019 0710 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(4-amino-cyclohexyl)-2-0X0-2- thiazol-2-yl-ethyl]-amide 073 0 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(3-aminomethyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-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 0755 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-{2-guanidino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0795 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1-(1-c arbamimidoy 1 -p iper idin- 4 -y lme thyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0800 [4-[4-Guanidino-l-(thiazole-2-carbonyD- butylcarbamoyl]-5-oxo-7- (3-phenyl-propionyl) - octahydro-2-thia-4a·, 7-diaza-naphthalen-6-yl]-acetic acid 0810 3-[4-[4-Guanidino-l- (thiazole-2-carbonyl- butylcarbamoyl]-5-oxo-7- (3-phenyl-propionyl) - oc tahydro-2- thia-4a, 7-diaza-naphthalen-6-yl]-propionic acid 59 0815 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid (3-guanidino-pr opyl] - amide 0820 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1-(1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0830 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l,2- a]pyrazine-6-carboxylic acid (4-guanidino-l-thiazol-2-ylmethyl-butyl)-amide 0835 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [4-guanidino-l-thiazol-2-yl-butyl)-amide

More preferred compounds according to formula VIII include: 0335 2-Benzoyl-4-oxo-octahydro-pyrrolo[l, 2-a]pyrazine-6- carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0650 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 0655 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [3-(2-amino-6-methyl-pyrimidin-4-yl) -1- (thiazole-2-carbonyl) -propyl]-amide 60 LV 12019 0715 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(4-amino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0720 4-Οχο-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(4-amino-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0725 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [l-(4-aminomethyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 073 5 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-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]pyrazine-6-carboxylic acid (2-oxo-l-piperidin-3-yl-2-thiazol-2-yl-ethyl)-amide 0750 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 2-a]pyrazine-6-carboxylic acid [1-(4-guanidino-cyclohexylmethyl)-2-oxo-2-thiaz ο1-2-y1-e thy1] - amide 0760 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [1-(5-benzyl-thiazole-2-carbonyl) -4-guanidino-butyl]-aunide 07 65 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [4-guanidino-l-(5-phenyl-thiazole-2-carbonyl) -butyl]-amide 61 0770 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrido[l, 2- a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 0775 5-Οχο-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [4-guanicLino-l-(thiazole-2-carbonyl) -butyl]-amide 0780 5-0xo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1— (4— carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0785 5-0xo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1— (3 — carbamimidoyl-benžyl) -2-oxo-2-thiazol-2-yl-ethyl]-amide 0790 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [1-(1-carbamimidoy 1-piperidin-3-ylītiethy 1) -2-oxo-2 - thiazol-2-yl-ethyl]-amide 0805 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide 0825 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine-6-carboxylic acid [4-guanidino-l-(hydroxy-thiazol-2-yl-methyl) -butyl]-amide 62 LV 12019 0840 4-0xo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid' [4-methoxy-l-(thiazole-2-carbonyl) -butyl]-amide 0845 [6-[4-Methoxy-l-(thiazole-2-carbonyl)-butylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l/ 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-propioņyl)-octahydro-pyrrolo[l/ 2-a]|>yrazine-6-carboxylic acid [4-amino-l-(thiazole-2-carbonyl) -butyl]-amide 0860 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[li 2-a]pyrazine-6-carboxylic acid [5-amino-l-(thiazole-2-carbonyl) -pentyl]-amide 0865 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [5-guanidino-l-(thiazole-2-carbonyl) -pentyl]-amide

Most preferred compounds according to formula VIII include: 0345 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l, 2- a]pyrazine-6-carboxylic acid [4-guanidino-l-(5-methvl-thiazole-2-carbonyl) -butylļ-amide; and 0340 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo[l, 2-a]pyrazine- 63 5 6-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide.

Preferred compounds according to formula IX include:

HjN

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 0900 3-Amino-2-cyclohexyl-4-oxo- hexahydro-pyrrolo[2,1-b][l, 3]thiazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 64 LV 12019

Preferred compounds 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-butyl]-amide

0910 7-(4-tert-Butyl-benzyl)-6-oxo- octahydro-pyrido[2,1-c][l, 4]thiazine-4-carboxylic acid [1-(benzothiazole-2-carbonyl) -4-guanidino-butyl]-amide

0915 6-Oxo-octahydro-pyrido[2,1-c][l, 4]thiazine-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butyl]-amide

H

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

H

65 0940 6-Oxo-7-phenethyl-octahydro- pyrido[2 , l-c][l ( 4] thiazine-4-carboxylic acid [4-guanidino- 1- (thiazole-2-carbonyl)-buCv l]-amide 0950 7-Benzyl-2,2,6-trioxo- oc tahydro-21>6_pyrido[2 , 1-c][l, 4]thiazine-4-carboxylic acid [4-guanidino-l-(thiazole- 2- carbonyl) -butyl]-amide

H i ,

\ - _-N f; O i; O 9, ;io o r—ļ NH- d

NH

More preferred compounds according to formula X include: 92 5 7-Benzvl-6-oxo-octahydro-pyrido[2 , l-c][l, 4] thiazine-4 - carboxylic acid [4-guanidino-1-(thiazole-2- 5 carbonyl) butvl]-amide ; and 940 6-Oxo-7 -phenethyl- oc tahydro-pyrido[2 , l-c][l, 4j thiazine-4 - carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl) -butvl] -amide.

Preferred compounds according to formula III include: 0

0960 4-οχο-Ι-(3-phenyl-propionyl)-oc tahydropyrrolo[1,2]pyrimidin e-6-carboxylic acid[4-guanidino-1-(thiazole-2-carbonyl)-butvl]-amide 0965 4-οχθ-l-(phenethylsulfony1)- octahydropyrrolo[1,2]pvrimidin e-6-carboxylic acid[4-guanidino-1-(thiazole-2-carbonvl)-butvl]-amide 66 LV 12019

For preparation of the compounds of formula (VII) various methods can be employed depending upon the particular starting materiāls and/or intermediates involved. The 5 following scheme is one particular method of 67 SCHEME 1

Ο

68 5 LV 12019 step 1:

The alkylation of g is done with appropriate bases according to the procedures described in Evans et al (J. Am. Chem. Soc. , 1981, 103, 2127; ibid, 1982, 104, 1737; Aldrichimica Acta, 1982, 15, 23) to give £>.

Step 2:

Compound h upon hydroboration and oxidation following conditions available in the literature (Synthesis, 1980, 151) results in the aldehyde g.

Step 3:

The formation of adduct g from aldehyde g with d is done by stirring the reactant in aromatic solvents e.g. benzene or toluene in presence of catalytic amount of suitable acid e.g, p-toluenesulfonic acid.

Step 3':

The inter conversion of aldehyde g to aldehyde g is readily achieved by appropriate protection deprotection protocals found in T. Greene, Protective Groups In Organic Synthesis, (John Wiley & Sons, 1981).

Step 4:

The cylization of adduct £ to | may readily be achieved by appropriate Lewis acids e.g, trimethyl aluminum in suitable solvents e.g. dichloromethane, the methodology found in T. Greene, supra.

Step 4':

Alternatively, the compound f can be derived from the treatment of aldehyde g with d in presence of suitable aromatic solvents e.g, benzene.

Step 5:

The ester function (-C(O)O-RJ0) of the bicyclic intermediate of formula is then subjected to 69 hydrolysation using an appropriate aģent such as HC1 in an appropriate solvent such as ethyl ether to yield to the free carboxylic acid. The resulting compound is then coupled to I^H with a peptide coupling aģent such as BOP in 5 an appropriate solvent such as DMF to yield to a bicyclic coupled compound of formula (VIII). Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Princioles of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, 10 Heidelberg, New York, Tokyo 1984; and The Peotides,

Analvsis. Svnthesis, Biolocrv. Vol. l.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979. 15 For preparation of the compounds of formula (VIII) various methods can be employed depending upon the particular starting materiāls and/or intermediates involved. The following scheme is one particular method of preparation. 70 LV 12019

71 wherein;

Pg is a nitrogen protecting group; each of R„; and R21 is independently a C,_6 alkyl; and X, R1, Rj, R4 and Rs are as previously defined.

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

The amino and carboxylic functions of Che unsaturated compound of formula (a) are protected with appropriate protecting groups. A variety of protecting groups known for reactive functional groups and suitable protection and deprotection protocols may be found in T. Greene, Protective Groups In Orcranic Svnthesis. (John Wiley &

Sons, 1981). The appropriate protecting group to use in a' particular synthetic scheme will depend on many factors, including the presence of other reactive functional groups and the reaction conditions desired for removal. The unsaturated compound of formula is easily obtained by methods and protocols known to chemist skill on the art. The protected unsaturated compound of formula (a) is subjected to appropriate conditions to allow cyclisation using an appropriate reaģent such as mercuric acetate in an inert solvent such as tetrahydrofuran (THF) to yield to a protected amino alcohol of formula (b). STEP 2

The protected amino alcohol of formula (b) is oxidized using an appropriate oxidizing aģent such as sulfur trioxide pyridine complex in an appropriate solvent such as diclhoromethane or dimethylformamide to yield to a protected amino aldehyde of formula (c). Alternatively, intermediate (C) can be made by the ozonolysis of a compound of formula (a') prepared according to Collado et al, J. Org. Chem.,1995, 60:5011. 72 LV 12019 STEP 3

The protected amino aldehyde of formula (c) is coupled with an amino acid alkyl ester of formula (d) by first forming the imine followed by contacting the obtained imine with an appropriate reaģent such as sodium triacetoxy borohydride NaBH(0Ac)3 to yield to a cyclic intermediate of formula (e). STEP 4

The cyclic 'intermediate of formula (e) is functionalized at the amino position to yield to the amino substituted cyclic intermediate of formula (f). Conditions appropriate for such reactions are well known in the art and will depend on the nature of the R,substituent. STEP 5

The amino protecting group of the cyclic intermediate of formula (f) is removed under appropriate conditions and the resulting compound is th$n subjected to appropriate condition for internai ring closure such as low heat in an inert solvent or as a raw compound to yield to a bicyclic intermediate of formula (g). The bicyclic intermediate of formula (g) can also be obtained by hydrolysing the ester function (-C(O)O-Rj0) of the cyclic intermediate of formula (g) to the free carboxylic acid followed by Standard peptide coupling using an appropriate coupling reaģent such as benzotriazole-l-ylojcy-tris- (dimethylamino)phosphonium hexafluorophosphate (BOP) in an inert solvent such as dimethyl formamide {DMF). STEP 6

The ester function (-C (0) 0-R2l) of the bicyclic intermediate of formula (g) is then subjected to hydrolysation using an appropriate aģent such as ĪTCl in an appropriate solvent such as ethyl ether to yield to the 73 free carboxylic acid. The resulting compound is then coupled to RjH with a peptide coupling aģent 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 of peptide chemistry. For example see Princioles of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peotides, Analvsis. Svnthesis. Biolocrv. Vol. l.edited by Gross E., 10 and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.

For preparation of the compounds of formula (IX) various methods can be employed depending upon the particular 15 starting materiāls and/or intermediates involved. The following scheme is one particular method of preparation. 74 LV 12019 SCHEME 3

Ο α

MeO

STEP 1 -► b

MeO

c Υ= S-Pg NH-Pg STEP 2

wherein:

Pg is a sulfur or amino protecting group; 5 L is a leaving group; each of Rj0; and R^ is independently a C,.s alkyl; and R^ R3, R4 and R, are as previously defined.

The process depicted in scheme 3 is briefly described as £ollows: 10 STEP 1:

The carboxylic acid compound (a) is coupled to the cyclic amine compound (b) with a peptide coupling aģent such as benzotriazol-l-yloxy-tris-(dimethylamino)phosphonium 15 hexafluorophosphate (BOP reaģent) in the presence of a base such as n-methylmorpholine in an appropriate. solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to yield to an amido compound of formula (c). Suitable 75 conditions for peptide bond formation are well known in th art of peptide chemistry. For exaxnple see Princioles of Peotide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides. Analvsis. Svnthesis, Biolocrv. Vol. l.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979. STEP 2

The compound of formula (c) is subjected to appropriate conditions to allow internai cyclisation to yield to a bicyclic intermediate of formula (d). For example/ acid mediated cyclisation using p-toluenesulfonic acid or TFA in an appropriate solvent such as dichloroechane. STEP 3

The ester function (-0(0)0-11^) of the bicyclic intermediate of formula (d) is subjected to hydrolysis using an appropriate aģent such as lithium hydroxide (LiOH) in an appropriate solvent such as tetrahydrofuran (THF) to yield to the free carboxylic acid. The resulting compound is then coupled to RjH with a peptide coupling aģent such as BOP in an appropriate solvent such as DMF to give compound (e). Suitable conditions for peptide bond formation are well known in the art of peptide chemistry. For example see Principles of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides. Analvsis. Svnthesis, Biolocrv, Vol. 1.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.

For preparation of the compounds of formula (X) various methods can be employed depending upon the particular starting materiāls and/or intermediates involved. The following scheme 4 is one particular method of preparation. 76 LV 12019 SCHEME 4

O

B R. wherein: 5 each of R30 and R21 is independently a C,.6 alkyl; and B, R1( R3, R<( and Rs are as previously defined. 77

The process depicted in scheme 4 is briefly described as follows: STEP 1:

The halogenated compound of formula (a) is converted to a halomethyl ketone of formula (b) using an appropriate reaģent,such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25°C to about 0°C. The resulting mixture is then treated under acidic conditions to yield to the halomethyl ketone of formula (b) . STEP 2

The halomethyl ketone of formula (b) is coupled with an amino acid alkyl ester of formula (c) with an appropriate base such as sodium cyanoborohydride in an organic solvent such as methanol (MeOH) to yield to a cyclic intermediate of formula (d). STEP 3

The cyclic intermediate of formula (d) is treated under acidic conditions using an appropriate acid such as camphorsulfonic acid in an appropriate solvent such as toluene to yield to a bicyclic intermediate of formula (e) . STEP 4

The ester function (-C (0) 0-RJO) of the bicyclic intermediate of formula (e) is subjected to hydrolysation using an appropriate reaģent such as LiOH to yield to the free carboxylic acid. The resulting compound is then coupled to RjH with a peptide coupling aģent such as BOP in an appropriate solvent such as dimethylformamide to yield to a coupled bicyclic compound of formula (X). Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see 78 LV 12019

Principles of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analvsis, Svnthesis, Biolocrv, Vol. l.edited by Gross E., and Meienhofer J., Academic Press , New York, 5 San Francisco, London, 1979.

Compounds of the present invention are further characterized by their ability to inhibit the catalytic activity of thrombin, which is demonstrated in the assay 10 as follows. Compounds of the present invention may be prepared for assay by dissolving them in buffer to give Solutions ranging in concentrations from 1 to ΙΟΟμΜ. In an assay to determine the inhibitory dissociation constant, K,, for a given compound, a chromogenic or 15 fluorogenic substrate of thrombin would be added to a solution containing a tēst compound and thrombin; the resulting catalytic activity of the enzyme would be spectrophotometrically determined. This type of assay is well known to those skilled in the art. 20

The compounds of the present invention may be used as anti-coagulants in vitro or ex vivo as - in the case of contact activation with foreign thrombogenic surfaces such as is found in tubing used in extracorporeal shunts. The 25 compounds of the invention may also be used to coat the surface of such thrombogenic conduits. To this end, the compounds of the invention are obtained as lyophilized powders, redissolved in'isotonic saline and added in an amount sufficient to maintain blood in an anticoagulated 30 State.

The therapeutic aģents of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers. The proportion of each carrier is 35 determined by the solubility and Chemical nature oi the compound, the route of ādministration, and Standard pharmaceutical practice. For example, the compounds may 79 be injected parenterally; this being intramuscularly, intravenously, or subcutaneously. For parenteral ādministration, the compound may be used in the form of sterile Solutions containing other solutes, for example, sufficient saline or glucose to make the solution isotonic. The compounds may be administered orally in the form of tablets, capsules, or granules containing suitable excipients such as starch, lactose, white sugar and the like. The compounds may also be administered sublingually in the form of troches or lozenges in which each active ingredient is mixed with sugar or corn syrupsf flavouring aģents and dyes, and then dehydrated sufficiently to make the mixture suitable for pressing into solid form. The compounds may be administered orally in the form of Solutions which may contain colouring and/or flavouring aģents.

Physicians will determine the dosage of the present therapeutic aģents which will be most suitable. Dosages may vary with the mode of ādministration and the particular compound chosen. In addition, the dosage may vary with the particular patient under treatment.

When the composition is administered orally, a larger quantity of the active aģent will typically be required to producē the same effect as caused with a smaller quantity given parenterally.

To further assist in understanding the present invention, the following non-limiting examples of such thrombin inhibitory compounds are provided. The following examples, of course, should not be construed as specifically limiting the present invention, variations presently known or later developed, v-hich would be wxthin the purview of one skilled in the art and considered to fall within the scope of the present invention as described herein. The preferred compounds as of the 80 |_V 12019 present invention are synthesized using conventional preparative steps and recovery methods known to those skilled in the art of organic and bio-organic synthesis, while providing a new a unique combination for the overall 5 synthesis of each compound. Preferred synthetic routes for intermediates involved in the synthesis as well as the resulting anti-thrombotic compounds of the present invention follow. 81 EXAMPLE 1

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) , and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 4-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Seguential washing with aqueous HC1 (50 mL; 0.1N), distilled HjO (3 x 50 mL) , drying over Na2S04; filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light-petroleum-ethyl acetate gradient) afforded the purified compound.

A solution of tert-butyloxycarbonyl-iodo-alanine-N,O- 82 LV 12019 dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 3-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HC1 (50 mL; 0.1N), distilled Η,Ο (3 x 50 mL) , drying over Na2S04, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.

och3 N CH, A solution of fcert-butyloxycarbonyl-iodo-alanine-N, 0-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Seguential 83 washing with aqueous HC1 (50 mL; 0.1N), distilled H20 (3 x 50 mL) , drying over Na2S04, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.

^och3 n I ch3

To a solution of terfc-butyloxycarbonyl-para-cyano-phenylalanine-N,0-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgS04, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound. 84 LV 12019

Ν' CH3 OCH3

To a solution of tert-butyloxycarbonyl-meta-cyano-phenylalanine-N,0-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mntol) . The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgS04, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL) , HC1 (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound.

85

To a solution of terfc-butyloxycarbonyl-ort.ho-cyan.o-phenylalanine-N/0-dimēthylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgS04, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL), HC1 (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound. o

NH NH o

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 ty1oxycarbony1-para-amidino-phenylaianine-N,0-dime thy1amide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine. (50 mL), dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel 86 LV 12019 (ethyl acetate/hexane), and concentrated under reduced pressure.

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78° C, and the solution stirred. tert-Butyloxycarbonyl-me£a-amidino-phenylalanine-N,0-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

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) 87 dropwise at -78° C, and the solution stirred. fcert-Bu ty 1 oxy c arbony 1 - ortho-amidino -pheny 1 a lanine -N, 0 -dime thy lamide (1.15 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) and the organic layer washed with saturated agueous ammonium chloride (2 x 50 mL) , brine (50 mL), dried with MgS04> filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

„CH, OCH3 fcert-Butyloxycarbonyl-para-cyano-phenylalanine-N,0-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL) , and sponge Raney Ni (100 mg) added. The solution was shaken under H2 at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.

The residue was dissolved in ethyl acetate (250 mL) , and washed with 1 N NaOH (2 x 50 mL) , and brine (2 x 50 mL) . The solution was dried with MgS04, filtered, and concentrated under reduced pressure. 88 LV 12019

fcer t-Butyloxycarbonyl-me ta-cyano-pheny lalanine-N, O-dimethylamide (1.33 g,, 4.0 npiol) was.dissolved in ethanol saturated with ammoņia (3,0 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H, at room temperatūre (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.

The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 x 50 mL), and brine (2 x 50 mL) . The solution was dried with MgS04, filtered, and concentrated under reduced pressure.

tert-Butyloxycarbonyl-orfcho-cyano-phenylalanine-N,0-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H, at room temperatūre (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.

The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 x 50 mL) , and brine (2 x 50 mL) . The solution was dried with MgS04, filtered, and concentrated under 89 reduced pressure.

tert-Butyloxycarbonyl-pa.ra-aminomethyl-phenylalanine-N/ 0-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N‘-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL) , and filtered through celite. The filtrate was concentrated under reduced pressure.

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

tert-Butyloxycarbonyl-mefca-aminomethyl-phenylalanine-N, 0-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 90 LV 12019 mmol), and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.

tert-Butyloxycarbonyl-orfcho-aminomethyl-phenylalanine-N, 0-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N' -bis- (benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.

To a solution of thiazole (1.23 g, 15.0 mmol) in anhydrous THF 91 (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 protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated agueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

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 protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane) , and concentrated under reduced pressure. 92 LV 12019

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 protected amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was guenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. o o

A solution of fcerfc-butyloxycarbonyl-iodo-alanine-N,0-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetairu.de (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g) . The resulting mixture was sonicated under nitrogen until no 93 starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HC1 (50 mL; 0.1N), distilled H20 (3 x 50 mL) , drying over Na2S04, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum/ethyl acetate gradient) afforded the purified compound.

To a solution of thiazole (1.28 g, 15.0 romol) in anhydrous THF (30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -7 8° C, and the solution stirred. The amino acid-N,0-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 agueous ammonium 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 MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 94 LV 12019 ο Boc

\ CH3 ο 2. (o-CHaC^aOjPhPdCIj 2-cyano-5-bromopyridine 1. Zn/Cu couple; ultrasound

NC' N A solution of fcert-butyloxycarbonyl-iodo-alanine-N, 0-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HC1 (50 mL; 0.1N), distilled I^O (3 x 50 mL) , drying over Na2S04, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum/ethyl acetate gradient) afforded the purified compound.

NC o

N

o 1. NH2OH, DIEA/EtOH 2. H2, Pd/C/EtOH:HOAc H2N

NH

To a solution of tert-butyloxycarbonyl-(4-cyano)3-pyridylalanine-N,0-dimethylamide (1.34 g, 4.0 mmol) in dry ethanol (20 mL) was added N,0-hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol) . The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgS04, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10% Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC.

The catalyst was removed by filtration, and the solution was concentrated under reduced pressure (50 mL), HC1 (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound.

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-dimethylamide (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 ammonium 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 MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel ethyl acetate/hexane), and 96 LV 12019 concentrated under reduced pressure.

tert-Butyloxycarbonyl-3- (4-pyridyl)alanine-N, 0-dimethylaiiu.de (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and Pt02 (100 mg) added. The solution was shaken under H2 until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3- (4-piperidyl) alanine-N, 0-dimethylairu.de.

The residue was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgS04, filtered, and concentrated under reduced pressure to yield the title compound.

tert-Butyloxycarbonyl-3-(3-pyridyl)alanine-N,0-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO, (100 mg) added. The solution was shaken under H2 until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding fcert-butyioxycarbonyl-3- (3-piperidyl) alanine-N, 0-dimethylamide .

The residue was dissolved in ethyl acetate (250 mL), washed 97 with 1 N NaOH (2 x 50 inL) , brine (2 x 50 mL) , dried with MgS04, filtered, and concentrated under reduced pressure to yield the title compound. o

l.H2,Pt02/Ac0H 0

fcert-Butyloxycarbonyl-3-(2-pyridyl)alanine-N,0-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtOj (100 mg) added. The solution was shaken under Hj until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(2-piperidyl)alanine-N,0-dimethylamide.

The residue was dissolved in ethyl acetate (25.0 mL), washed with 1 N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgS04, filtered, and concentrated under reduced pressure to yield the title compound.

tert-Butyloxycarbonyl-3-(4-piperidyl)alanine-N,0-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N‘-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), 98 LV 12019 and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL) , and filtered through celite. The filtrate was concentrated under reduced pressure.

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

n NZ

fcert-Butyloxycarbonyl-3- (3-piperidyl) alanine-Nf 0-dimethylamide (1.00. g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N‘-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL) , and filtered through celite. The filtrate was concentrated under reduced pressure.

Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound. 99

terfc-Butyloxycarbonyl-3- (2-piperidyl) alanine-N, 0-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, Ν,Ν'-bis-(benzyloxycarbonyl) -S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl2 (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The filtrate was concentrated under reduced pressure.

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 derivative (2.00 g, 3.2 mmol) in anhydrous THF (15 mL) was added dropwise, and the resulting mixhure stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate 100 LV 12019 (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO,, filtered, and concentrated under reduced pressure.

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 1 h. The guanidylated 3-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, ahd the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgS04, filtered, and concentrated under reduced pressure.

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 101 -7 8° C with stirring. The mixture was stirred at -78° C for 1 h. The guanidylated 2-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was guenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgS04, filtered, and concentrated under reduced pressure.

tert-Butyloxycarbonyl-para-nitro-phenylalanine-N, O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL) , and Pt02 (100 mg) added. The solution was shaken under until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in Η,Ο (150 mL) , and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 x 50 mL), and brine (3 x 50 mL) . The solution was dried with MgS04, filtered, and concentrated under reduced pressure yielding the title compound.

102 LV 12019 tert-Butyloxycarbonyl-zneta-nitro-phenylalanine-N, 0-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL) , and Pt02 (100 mg) added. The solution was shaken under H2 until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in H20 (150 mL) , and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 x 50 mL), and brine (3 x 50 mL). The solution was dried with MgSO,, filtered, and concentrated under reduced pressure yielding the title compound. 0 o

tert-Butyloxycarbonyl-ortho-nitro-phenylalanine-N,0-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL) , and Pt02 (100 mg) added. The solution was shaken under H2 until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in HjO (150 mL) , and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL) , washed with 1 N NaOH (3 x 50 mL), and brine (3 x 50 mL). The solution was dried with MgSO,, filtered, and concentrated under reduced pressure yielding the title compound. o

2. LIthium tliiazole/THF o

NHZ' 103 1. terfc-Butyloxycarbonyl-3-(cisftrans-4- aminocyclohexyl)alanine-N,0-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL) , 4 N aqueous sodium bisulfate (2 x 50 mL) , and brine (2 x 50 mL) . The solution· was dried with MgS04, filtered, and concentrated under reduced pressure.

The semi-solid was chromatographed on silica gel (ethyl acetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol) dropwise at -78° C and the solution stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was guenched with saturated agueous ammonium 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 MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 104 LV 12019

1. tert-Butyloxycarbonyl-3- (cis/ trans-3- aminocyclohexyl) alanine-N, 0-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated agueous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate vas added, the THF was removed under reduced pressure, and the remaining agueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL) , 4 N aqueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgS04, filtered, and concentrated under reduced pressure.

The semi-solid was chromatographed on silica gel (ethyl acetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol) dropwise at -78° C and the solution stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 105

1. Cert-Butyloxycarbonyl-3-(cis/trans-2- aminocyclohexyl) alanine-N, 0-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF [60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The agueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4 N aqueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL) . The solution was dried with HgS04, filtered, and concentrated under reduced pressure.

The semi-solid was chromatographed on silica gel (ethyl acetate/ hexane). 2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 8.0 mL, 12.8 mmol) dropwise at -78° C and the solution stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO<, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 106 LV 12019

1. fcert-Butyloxycarbonyl-3-(cis/trans-4- aminocyclohexyl) alanine-N, 0-dimethylairu.de (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, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL) , and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) aiforded the purified product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -78° C and the solution stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) , and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL) , brine (50 mL) , dried with MgSO„, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 107 ο

ch3 Ν I och3 ο

ΝΖ 1. H3CS' χ ^ΤΊΗΖ

HgClj/THF

2. Lithium thiazole/THF 1. fcert-Butyloxycarbonyl-3-(cis/trans-3- aminocyclohexyl) alanine-N, 0-dimethylamide (2.0 g, 6.1 mmol) vas 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 irmol) , and HgCl2 (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL) , and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -78° C and the solution stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure. 108 LV 12019

1. tert-Butyloxycarbonyl-3-(cis/trans-2- aminocyclohexyl)alanine-N,0-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, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue'was suspended in ethyl acetate (300 mL) , and filtered. through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the purified product. 2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF was added n-BuLi (1.6 M/hexane, 15.9 mL, 25.4 mmol) dropwise at -7 8° C and the solution stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated agueous ammonium 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 MgS04, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexarie) , and concentrated under reduced pressure. 109

Examole 2

Synthesis of Intermediates

(4S,5R)-3-(l-oxo-3-phenylpropyl)-4-(phenyl)-5-(methyl)-2-oxazolidone (2). A solution of 10.0 g (1.0 equiv., 56.4 mmol) of (4S, 5R)-4-phenyl-5-methyl-2-oxazolidone (1) in 250 mL of dry THF, stirred at -7 8°C under argon, was treated dropwise with n-butyllithium (1.6 M in hexane, 1.1 eq., 38.8 mL). After stirring for 30 min., 8.4 mL (1.0 equiv., 56.4 mmol) of hydrocinnamoyl chloride was introduced dropwise over a 10 minūte period. The resulting mixture was warmed to 0°C, stirred for an additional hour, and quenched with saturated ammonium chloride. The solvent was removed in vacuo and the resulting white solid dissolved in ethyl acetate and ddH20. The aqueous phase was removed and extracted with two additional portions of ethyl acetate. The extracts were combined, washed with saturated sodium chloride, dried over sodium sulfate, and the solvent was removed in vacuo to afford a white .crystalline solid (2)(in 91% yield). m.p.95-96.5; [a]D-35.8 (c=l, CH2C12); [a]D-26.6 (c=1.018, CH2C12). 1-HNMR (CDC13) δ 0.89 (d, 3H, CH^, J=6.6 Hz) , 3.00-3.05 (m, 2H), 3.26-3.34 (m, 2H) , 4.73-4.78 (m, 1H), 5.64, d, 1H, J=7.4 Hz), 7.22-7.46 (m, 10H). 110 LV 12019

Oo 0 0

A solution of 5.0 g (1.00 equiv., 16.2 mmol) of (2) in 100 mL of dry THF, was cooled to -78°C. Enolization was 5 achieved with 17.8 mL (1.1 equiv:, 17.8 mmol) of lithium bis-trimethylsilylaiiu.de, which was added dropwise via syringe. The solution was stirred for 30 min. before 4.45 mL (3.0 equiv., 48.5 mmol) of allyl iodide was introduced and the reaction warmed to -15°C. After 1 h the reaction 10 was quenched with saturated ammonium chloride and extracted (3X) with ethyl acetate. The organic phase was washed with sodium metabisulfite, dried over sodium sulfate and the solvent removed in vacuo to afford an off-colour oil. Purification was achieved by flash 15 chromatography on silica gel using a stepwise gradient (15:1, 12:1, 10:1) to yield (3), a colourless oil (95%).

[CC]D 47.5 (c=3.12 , CH2C12). 20 1HNMR (CDC13) 5 0.82 (d, 3H, CH^, J=6.6), 2.31-2.40 (m, 1H, RCH=CHCH2), 2.49-2.57 (m, 1H, RCH=CHCH2), 2.84-3.00 (m, 2H, Ph-CH2), 4.32-4.37 (m, 1H, CH-(N)CO), 4.53-4.58 (m, 1H, CH3-CH-), 5.03-5.13 (m, 2H, ΑΒΧ, CH=CH2), 5.21 (d, 1H, Ph-CH, J=7.1 Hz), 5.81-5.89 (m, 1H, CH=CH2), 7.20-7.42 25 (m, 10H, ArH) ; 13c (CDCI3) δ 14.4, 36.2, 38.2, 43.9, 54.7, 78.4., 117.1, 125.4, 126.3, 128.2, 128.5, 129.0, 133.1, 134.8, 138.9, 152.4, 174.9. 111

A sairiple of the allvl compound, (3), (4.75 g, 13.6 mmol) in THF (100 mL) was treated with 13.6 mL (1.0 equiv., 13.6 mmol) of a 1.0 M solution of borane-tetrahydrofuran complex at 0°C and stirred for 2h. The solvent was evaporated and chloroform (100 mL) added via svringe. Oxidation of the organoborane was achieved by the additior. of 4.7 g (2.0 equiv., 27.2 mmol) of 3-chloroperoxybenzoic acid at 0°C, with warming to ambient temperature and stirring for an additional hour. The organic phase was washed with 5% Ν32003^, ddl^O, and dried over sodium sulfate. Due to the instability of the alcohol, a guick column was performed to remove the extreme polar and nonpolar material which originated from the 3-chloroperoxybenzoic acid. The alcohol (4) was obtained ir. a yield of 65%.

[a]D 39.3 (c = l.038, CH2C12). 1HNMR (CDCI3) δ 1 . 07 (d. 3H, ch3 -CH, J=6 .5 Kz), 1.31-1.?: (m. 3H, CH2-CH-H ) , 2 . 10-2 .19 (m, IK, ch2- CH- -K), 3.10-3.17 (m, 2H, Ph-CH2), 3 . 87 -3 .9 0 (m, 2H, CĶ2 OH) , 4 . .43-4.49 (tn. 1H, CH-CO), 4.70 -4 .75 (m, 1H, ch3 -CH) , 5 . 36 (d, ih, j_7 .1 Kz), 7.41- 7 . 63 (m, 10H, ArK ) ; 112 LV 12019

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 mixture changed from bright orange to a black colour. The reaction was monitored by TLC and after 30 min., if starting material remained, additional molecular sieves 10 were added. The solution was filtered through celite and the solvent was evaporated. The residue was dissolved in ethyl acetate and washed with saturated sodium chloride. If the orange colour persisted in the organic phase, additional filterings through celite pads were performed. 15 The aldehyde was obtained in a quantitative yield as a transparent, colourless oil (5). iHNMR (CDC13) § 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-CH0' J=13.3 & J=6.6), 2.92-2.99 (dd, 1H, CH2-CHO, J=13.2 & J=8.8), 4.19-4.22 (m, 1H, CH-CO), 4.46-4.51 (m, 1H, CH3-CH) , 5.13-^5.25 (m, 1H, Ph-CH) , 7.20-7.39 (m, 10H, Arfi) , 9.69 (s, 1H, CHO); 25 13C (CDCI3) δ 14.2, 23.8, 39.0, 41.2, 43.8, 54.9, 78.6, 125.3, 126.4, 128.2, 128.4, 128.5, 128.9, 132.8, 138.4, 152.4, 174.9, 201.1. 113 ο

5 ο

Ph

The aldehyde, (5)., (2.6 g, 7.10 mmol) was dissolved in benzene (70 mL) and a catalytic amount of p-5 toluenesulfonic acid was added, £ollowed by 1.58 g (1.2 eguiv., 8.52 mmol) of L-cysteine ethyl ester and 4 A molecular sieves. The reaction was allowed to stir overnight at ambient temperature followed be removal of solvent in vacuo. The residue was dissolved in 10 chloroform, washed with saturated sodium chloride, ddH20, and dried over sodium sulfate. The solvent was removed in vacuo to afford a gummy solid (6). o

C02Ēt 15 6 7 2.0M trimethylaluminum in hexane (2.4mL, 4.8mmol, 3 eguivalents) was added slowly to starting material (6) (800mg, 1.61mmol) stirring in anhydrous dichloromethane 20 under argon, using ovendried equipment. After stirring overnight, HPLC indicated that the reaction had gone to completion. 114 LV 12019

The mixture was quenched with excess methanol, then filtered on a short silica gel column (washing through with excess 10% methanol in ethylacetate. Evaporation gavē 784 mg of crude material that was purified using 2:1 hexane:EtOAc on a«silica gel column affording 258mg (0.81mmol being a 50% yield) of pure compound (7),a 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-ethyl ester' as a white/yellow solid. 'H NMR (CDCI3) d 1.28-1.31 (m, 3H) , 1.72-1.81 (m, 3H), 2.10-2.13 (m, 1H), 2.66 (dd, 1H, J= 11.5 and 6.0 Hz), 3.29-3.34 (m, 2H) , 4.19-4.29 (m, 2H), 4.88 (dd, 1H, J= 9.0 and 5.0 Hz), 5.22 (dd, 1H, J= 8.0 and 6.0 Hz), 7.18-7.23 (m, 3H), 7.28-7.31 (m, 2H).

7 8 LĪ0H.H20 (48mg, 1.12mmol) in lOmL of water was added to starting material (7) (240mg, 0.76mmol) dissolved in lOmL of dioxane. After 1 hour, TLC in 1:1 Hexane:EtOAc showed no starting material. The reaction was guenched with 10% citric acid, then extracted twice with dichloromethane. Drying and evaporating the combined organic layers gavē 354mg of crude product. This was redissolved in dichloromethane, then precipitated by adding excess hexane. The product was filtered to give 200mg (0.68mmol being a 90% yield)) of an ,off-white solid, (8), also known as 6S-benzylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxylic acid. Ή NMR (CD30D) d 1.71-1.82 (m, 3H), 2.12-2.17 (m, 1H), 2.67 (dd, 1H, J= 14 and 11 Hz), 2.77-2.81 (m, 1H), 3.30-3.40 Π5 (m, 3H) , 4.81 (dd, 1H, J= 8.5 and 4.9 Hz), 5.16 (t, 1H, J= 7.5 Hz), 7.18-7.31 (m, 5H).

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

11 10 12

The chiral auxiliary (11) (13.6g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78°C. Then n-BuLi (52.8 mL, 84.4 mmol, 1.2 equiv.) was added and left for 30 mins (dark orange solution). The acid chloride (10)(13.4 g, 76.6 mmol, 1 eq) was then added and left to stir overnight. Work-up was done by quenching with saturated NH4C1 extracting with ethyl acetate, washing the extracts with water and. brine, drying over sodium sulphate and concentration. A fast column, with dry loading, (6:1 hexane ethyl acetate) was run to purify the product. This afforded in a white solid (12) which was recrystallized 116 LV 12019 from ether and hexane to give the title compound in 78% yield.

[a]D=-20.1(c=l,EtOH); MP/BP mp=90.5-91.5oC 1H NMR (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, 2H) , 4.70-4.76 (m, 1H) , 5.65 (d, 1H, J= 7.2 Hz), 7.28-7.42 (m, 5H) .

The starting material (12)(9.13g, 29 mmol, leq) was dissolved in dry THF and cooled to -78°C, after which LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise over 40 mins. Then, 30 minūtes later, allyl bromide (7.5 mL, 86.9 mmol, 3 eq) was added slowly over 10 mins. The mixture was left to warm overnight. Work-up included quenching with sat. ammonium chloride, extraction with ethyl acetate, washing with 10% sodium thiosulphate, decolourising with charcoal, drying over sodium sulphate and concentration in vacuo. The product was obtained as a yellow oil (13) in 96% yield.

[<X]D=+9.5 (c = 1.0,EtOH) ‘H NMR (CDCI3) 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) , 4176-4.85 (m, 1H), 5.00-5.07 (m, 2H) , 5.65 (d, 1H, J- 7 Hz), 5.64-5.88 (m, 1H0, 7.27-7.46 (m, 5H) . 117

13 14 2-methyl-2-butene was added dropwise to borane dimethylsulphide complex at -12°C. The reaction was maintained at this temperature for 15 minūtes and then it was warmed to Q°C, after which it was stirred for 2 hours. The disiamyl borane was then added to a mixture of the starting material 13 ih THF using a double-ended needle at 0°C. The mixture was then stirred for 2 hours after which the solvents were removed and the residue dissolved in dichloromethane. It was carefully added to a suspension of pyridinium chlorochromate in dichloromethane contained in a flask equipped with a reflux condenser. After the initial exothermic reaction had subsided, the mixture was refluxed at 50°C for 1 hour. The dark brown liquid was dissolved in ethyl acetate and filtered through Florisil. The black residue of PCC was extracted with ethyl acetate and also filtered through the same Florisil pad. Concentration of the filtrates resulted in a 78% yield of a yellow gummy product (14).

[a]D=-17.8(c=l.245,EtOH) lH NMR (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, 1H) , 4.12-4.16 (m, 1H), 4.76-4.80 (m, 1H), 5.67 (d, 1H, J= 7Hz), 7.27-7.46 (m, 5H), 9.77 (s, 1H). 118 LV 12019

'όο2έ.\

W 14 15

The resulting aldehyde (14) (7.7g crude, 20.8 mmol 1 equiv.) was dissolved in 75 mL of toluene. To the solution was added a catalytic amount of p-toluenesulphonic acid (50 mg), 10 g of 4Ā molecular sieves, and L-cysteine ethyl ester (3.87 g, 20.8 mmol, 1 eguiv.). The mixture was stirred overnight, filtered and concentrated. The residue was then purified by silica gel chromatography (6:1 hexane:ethyl acetate) afforded 6.36 g of the product (15) in 61% yield.

[0C]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 (m, 1H), 3.24-3.34 (m, 1H), 3.77-4.29 (m, 4H), 4.46-4.81 (m, 2H), 5.66 (d, 1H, J= 7Hz), 7.27-7.46 (m, 5H).

119

The starting material (15) (1.97 g, 3.9 mmol, 1 equiv.) was dissolved in 20 mL of dry dichloromethane and cooled to 0°C. Trimethylaluminum (5.9 mL, 11.8 mmol, 3 equiv. ) , 5 was added dropwise and the mixture was left stirring overnight. After complete reaction as evidenced by HPLC, methanol was added until a yellow solid mass was formed. Dichloromethane was added to dissolve the solid and the whole mixture was stirred for 15-30 minūtes and then 10 filtered. The residue after concentration in vacuo was run through a quick column (6:1 hexane:ethyl acetate), to remove auxilary and as many of the polar decomposition Products as possible, affording in a 50% yield of a yellow oil (16). 15 lH 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 and 8.0 Hz), 4.11-4.25 (m, 2H), 4.88 (dd, 1H, J= 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 dissolved in 10 mL of dioxane. The solution was cooled to 10 C, and to it was added Li0H'H20 (0.123 g, 2.9 mmol, 1 eq.) dissolved in 10 mL of water. The bath was removed and the mixture was stirred at room temperature 30 for 1 hour. . TLC showed complete reaction and the solvent was evaporated under vacuum. The remaining aqueous laver was washed with ether (2X), acidified with 10% citric acid, and extracted with dichloromethane (3X). The 120 LV 12019 combined extracts were dried over sodium sulphate and concentrated to give a white solid which was recrystallized from ether. Concentration of the filtrate and purification by silica gel column chromatography (2:1 hexane: ethyl acetate) resulted in more product (17) with a m.p. of 198. 2-199°C. *H NMR (DMSO-d6) d 0.78-0,93 (m, 2H), 1.11-1.27 (m, 5H) , 1.34-1.36 (m, 1H), 1.51-1.56 (m, 1H), 1.60-1.75 (m, 1H) , 1.82-1.87 (m, 1H), 2.15-2.18 (m, 1H), 2.37-2.41 (m, 1H) , 3.03 (dd, 1H, J= 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.83 (dd, 1H, J= 9 and 4 Hz), 4.95 (dd, 1H, J= 8 and 5.5 Hz) .

BOC-DiCbz Arg (18) (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0°C. Triethylamine (2.2 mL) was added followed by 14.5 mmol of a 1M toluene solution of isopropyl chloroformate via a syringe. The reaction was allowed to .stir at 0°C for 30 minūtes then quickly filtered. The white solid was discarded. To the filtrate was bubbled freshly prepared diazomethane until the color of the solution turned yellow. The reaction mixture was allowed to stand overnight in a well ventilated fumehood which facilitated the discharge of excess diazomethane. Dry ether was added to precipitate the diazoketone. The product was filtered and dried under yacuum to give light yellow fluffy solid (4.6 g, 58%) . 121

Diazoketone (19) (lg, 1.77 mmol) was dissolved in THF (20 mL) and to this solution was added 1M HCl in ether (20 mL) at 0°C. The reaction was allowed to stir at ambient temperature overnight during which time a white precipitate was formed. Further precipitation was achieved by adding ether. Filtration and drying the solid afforded the product (20) (1.02 g , 100%). Ή NMR (DMSO-d6) d 1.65-1.77 (m, 3H) , 2.06-2.50 (m, 1H), 3.86-3.90 (m, 2H) , 4.29 (m, 1H) , 4.76 (d, 1H, J= 18Hz), 4.95 (d, 1H, J= 18 Hz), 7.35 (s, 2H) , 7.36 (s, 2H), 7.35-7.41 (m, 10H), 8.71 (br s, 3H), 10.1 (br s, 2H). •JC NMR (DMSO-d6) d 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3, 128.6, 128.7, 128.8, 128.9, 135.2, 135.9, 153.4, 157.4, 198.9.

To mimetic (17) (0.422 g, 1.42 mmol) in THF (50 mL) at 0°C and in presence of N-metbyl morpholine (0.19 mL), was slowly added 1M toluene solution of isopropylchloroformate (1.71 mL). The reaction was allowed to stir at 0°C for 30 minūtes then treated with aminochloromethylketone (20) in small portions. Once the addition was complete the reaction was further stirred for 15 minūtes followed by addition of N-methyl morpholine (0.19 mL) . The reaction was stirred at ambient temperature for 3,.hours, then extracted with ethyl acetate followed by washing with brine and 10% agueous citric acid. Removal of organic solvent gavē a white foam 122 LV 12019 (21) (1.03 g, 96%) which was used further without purification. 10 'h NMR (CDC13) d 0.07-0.97 (m, 1H), 1.15-1.41 (m, 7H), 1.62-1.91 (m, 10H), 2.10-2.16 (m, 1H) , 2.43-2.48 (m, 1H) 2.74-2.80 (m, 1H), 3.01-3.07 (m, 1H) , 3.87-3.94 (m, 1H) , 4.11-4.19 (m, 2H), 4.60-4.66 (m, 1H) , 4.74-4.86 (m, 2H) , 5.09-5.24 (m, 4H) , 7.30-7.39 (m, 10H) , 7.95 (d, 1H, J= 8 Hz), 9.4 (br s, 1H) , 9.56 (br s, 1H) . 123 EXAMPLE 3

(N-t-BOC-N-tosyl)butyrylketoarginine (240mg, 0.515 mmol), was deprotected using 30% TFA in dichloromethane. The deprotected arginine derivative, was coupled with the mimetic (8) (100 mg, 0.343 mmol) in DMF under basie conditions (Et3N, pH = 8-9), using BOP reaģent (228 mg, 0.52 mmol) as the dehydrating aģent. The reaction was typically complete within 2-4 hours. Extraction with ethyl acetate followed by successive washing with brine and 10% aqueous citric acid yielded the crude produet. The crude product was purified by column chromatography affording 180 mg (76%) of pure product. This product was then treated with HF to remove the tosyl group. Purification of the isolated deprotected product by HPLC afforded BCH-2737. EXĀMPLE 4

124 LV 12019

Chloromethylketone (21) (0.188 g, 0.245 mmol) was dissolved in THF (10 mL) treated with NMM (0.036 mL) followed by mercapto acetic acid (0.02 mL, 0.299 mmol).

The reaction was stirred at ambient temperature overnight. Extraction of the reaction mixture with ethyl acetate followed by successive washing with brine and 10 % aqueous citric acid and evaporation of organic solvent gavē the crude product which was purified by column chromatography to give foamy solid as the product (0.125 g, 62%).

This protected precursor (0.125 g, 0.154 mmol) was dissolved in DCM (5 mL) and cooled to -78°C. A 1M DCM solution of BBr3 (1.54 mL, 1.54 mmol) was slowly added. The reaction was stirred at ambient temperature for 5 hours, then cooled to -78°C again and treated with anhydrous methanol (2mL). The reaction was brought to room temperature and stirred for 2 additional hours. The solvents were removed under reduced pressure aiid the residue was partitioned between ether and water. The water layer was collected, lyophilized and the final product (23) obtained as a powder after HPLC purification and lyophilization.

The products of the reactions described above can be isolated in the free form or in the form of salts. In addition, the products can be obtained as pharmaceutically acceptable acid addition salts by reacting one of the free bases with an acid. In a similar manner, the product can be obtained as pharmaceutically acceptable salts by reacting one of the free carboxylic acids with a base. Likewise, treatment of the salts with a base or acid results in a regeneration of the free amide. 125 5 EXAMPLE 5 A general method of synth.esizing compound of formula II or III:

NH,

LiOH OOCHļCHjCOjM·

126 10 LV 12019

(16) EXAMPLE 6 Synth.esis of 5 STEP 1

Synthesis of 2-Benzyloxycarbonylamino-4-hydroxybutyric acid tert-butyl ester

NHZ

(Cyclohexyi)2NH.HOOC C02t-Bu

υ NMM.THF > 2) NaBH4, MeOH

10 (1) (2)

To a solution of the protected aspartic acid (1)(Bachem, 2.50 g, 4.95 mmols) in 50 mL of dry tetrahydrofuran 15 (THF) , at -10°C, under N2, was added N-methylmorpholine (109 flL, 0.2 eq) and isopropyl chloroformate (1.0 M/toluene : 384 μΧι, 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°C for 30 min. The mixted anhydride solution was then added to the NaBH4 suspension dropwise v\ia canula, and the final solution was stirred at -78°C for 3 hr. Acetic acid (2.8 mL, 10 eq) was then added and the solution was 127 warmed to r.t. (30 min). The solvents were evaporated, the residue taken up in EtOAc and washed with sat.aq. NaHC03 (2x) and brine. The organic layer was dried over MgS04, the solids were filtered and the solvent evaporated to give 1.53 g (4.95 mmols, 100%) of the alcohol (2) as a clear oil. *H NMR (CDC13, 400 MHz) : δ 7.40-7.31 (m, 5H, ArH) , 5.63 (d, 1H, J=7.3, NH), 5.13 (AB system, 2H, J=12.2, CH2Ph) , 4.43 (m, 1H, H-2), 3.69 (m, 2H, H-4), 2.17 (m, 1H, H-3), 1.63 (m, 1H, H-3), 1.48 (s, 9H, t-Bu). STEP 2 2-Benzyloxycarbonylamino-4-iodobutyric acid tert-butyl ester

To a solution of the alcohol (2) (1.53 g, 4.95 mmols) in a 1:1 mixture of CH3CN/Et20 (50 mL) , at -10°C, under N2, were added successively imidazole (607 mg, 1.8 eq) and Ph3P (2.21 g, 1.7 eq). Iodine (2.14 g, 1.7 eq) was then added in small portions over a period of 15 min. After the addition was completed, a white precipitate formed and the solution was brown. It was stirred at -10°C for 45 min. It was then poured in Et20 and the organic phase was washed with sat.aq. Na2S03, sat.aq. CuS04, H20 and dried over MgS04. The solids were filtered and the solvent evaporated to give a yellow oil that was purified by flash chromatography (silica gel, 5% to 20% EtOAc/

Hex). The iodide (3) was obtained in 83% yield (1.71 g) as a clear oil. 128 LV 12019 1H NMR (CDClj, 400 MHz) : 5 7.41-7.31 (ra, 5H, ArH) , 5.35 (bd, 1H, J=7.3, NH), 5.13 (s, 2H, CH2Ph), 4.30 (m, 1H, H- 2), 3.22-3.12 (m, 2H, H-4), 2.42 (m, 1H, H-3), 2.20 (m, 1H, H-3), 1.48 (s, 9H, t-Bu). STEP 2

Synthesis of 2-Benzyloxycarbonylamino-4-hexenoic acid tert-butyl ester

NHZ

C02t-Bu (3)

^^MgBr , Cul THF, -78°C

NHZ 'C02t-Bu (4)

To a suspension of Cul (2.27 g, 5 eq) in dry THF (20 mL) , at -78°C, under N2, was added slowly a 1.0M solution in THF of vinyl magnesium bromide (23.4 mL, 9.8 eq) . The solution was then warmed up to -10°C for 30 min (it tumed then black) and cooled back to -78°C. A solution of the iodide (3) (l.OOg, 2.39 mmols) in dry THF (3.5 mL) was then added slowly to the cuprate solution. The reaction mixture was stirred at -78°C for 2.5 hr.

Sat.aq. NH4C1 (50 mL) was added and the mixture was brought back to room temp. with vigorous stirring. It was then poured in Et20 and stirred for 5 min. The dark suspension was filtered through a cintered funnel and the phases were separated. The aqueous phase was extracted with Et20 (2x) and the combined organic extracts were dried over HgS04. The solīds were filtered, the solvents evaporated and the crude oil purified by flash chromatography (silica gel, 5% AcOEt/Hex) to give- 0.51 g (67%) of the pure alkene (4). 129 XK NMR (CDC13, 400 ΜΗζ) : 5 7.37-7.31 (m, 5Η, ArH) , 5.80 (m, 1H, H-5), 5.33 (d, 1H, J=7.8, NH), 5.12 (s, 2H, CH2Ph), 5.05 (d, 1H, J=17.2, H-6), 5.01 (d, 1H, J=10.4, H-6), 4.30 (q, 1H, J=7.4, H-2), 2.16-2.08 (m, 2H, H-4), 1.92 (m, 1H, H-3) , 1.74 (m, 1H, H-3), 1.48 (s, 9H, t-Bu) STEP 4

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

HO

1) Hg(OAc)2, THF 2) NaHCQ3, KBr ^ 3) NaBH4, 02 (4)

To a solution of the alkene (4) (50 mg, 0.157 mmol) in dry THF (3.1 mL) , atr.t., under N2, was added mercuric acetate (75 mg, 1.5 eq) . The solution was stirred at r.t. for 18 hr after which it was cooled down to 0°C. Sat.aq. NaHC03 (2 mL) was then added and the mixture was stirred at 0°C for 30 min. KBr (O.llg, 6 eq) was added and the mixture was stirred at r.t. for 2 hr. It was then poured in H20/Et20 and the phases were separated.

The aqueous phase was extracted with Et20 (2x) and the combined organic extracts were dried over MgS04. The solids were filtered and the solvents evaporated. Oxygen (02) was bubbled into a suspension of NaBH4 (3.3 mg, 0.55 eq) in dry DMF (0.4 mL) for 1 hr, and to this was added dropwise (syringe pump, 3 mL/hr) a solution of the organomercurial bromide in DMF (3.1 mL) with confcinuous introduction of 02. The bubbling was continued for 1 hr and Et20 (5 mL) was added. The grey suspension was 130 LV 12019 filtered through Celite and the filtrate was evaporated. The residue was chromatographed (silica gel, 6:4 Hex/EtOAc) to give the pyrrolidinol (5) (30 mg, 57%) as a clear oil. XH NMR (CDC13, 400 MHz) : δ 7.37-7.28 (m, 5H, ArH) , 5.22-5.09 (m, 2H, CH2Ph) , 4.30 (dd, 1H, J=1.4, 8.3, H-2), 4.24 (m, 1H, H-5) , 3.70-3.57 (m, 3H, CH2-0H) , 2.25 (m, 1H) , 2.13 (m, 1H) , 1.92 (m, 1H) , 1.70 (m, 1H) , 1.34 (s, 9H, t-Bu) . STEP 5

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

OH O

To a solution of the alcohol (5) (50 mg, 0.149 mmol) and Et3N (62ļlL, 3 eq) in dry CH2C12 (0.8 mL) is added slowly, under N2, at 0°C, a solution of S03-Pyridine complex (71 mg, 3 eq) in dry DMSO. The solution was stirred at 0°C for 30 min and 10% citric acid (2 mL) is added. The pH is brought to 4 with 1M NaOH and the aqueous phase is extracted with Et20 (3x). The combined organic extracts v/ere dried over MgS04. The solids were filtered and the solvents evaporated to give a crude oil which was purified by flash chromatography (silica gel, 7:3 Hex/EtOAc). The pure aldehyde (6) was obtained as a clear oil (45 mg, 90%). 131 XH NMR (CDC13, 400 ΜΗζ) : δ 9.68 + 9.56 (ds, 1Η, CHO) , 7.36-7.29 (τα, 5Η, ArH) , 5.23-5.11 (m, 2Η, CH2Ph) , 4.57-4.39 (m, 2Η, Η-2, Η-5), 2.30-1.97 (m, 4Η, Η-3, Η-4), 1.47 + 1.36 (2s, 9Η, t-Bu). 5 STEP 6

10

The pyrrolidine-aldehyde (6) is coupled with the protected diamino-propionic acid (7) by first forming the imine (8) {MgSO<( CHjC12) . Isolation of the imine (8) is done by filtration of the MgS04 and evaporation of the solvent. 15 The crude imine is then treated with NaBH(OAc): and actic acid (AcOH) in THF for 15 hours to obtain the amine (8) after extrative work-up.

132 LV 12019

The CB7 (7) protecting group of the amine (8) is removed by hydrogenation with palladium on charcoal 10% as a catalyst in methanol (MeOH). The catalyst is filtered and the MeOH evaporated to give the crude diamine (9) that can 5 be used without any:;^>urification. STEP 8

The cyclisation is done by heating the crude oil (9) from step 7, neat slightly above the boiling point of methanol. The bicyclic lactam (10) is purified by flash 15 chromatography. 133 STSP 9

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 give the bicyclic lactam-amide (11). 10 STEP 10

The BOC and t-butyl ester protecting groups of bicyclic 15 lactam amide (11) are removed under acidic conditions (HC1 in ethyl ether (Et20)). The amine salt (12) precipitates out of solution and is collected by filtration. 134 LV 12019 STEP 11

COOH (12)

CgH5CH2OCOCl k2co3, CH3CN

OH (13)

The primary amine of compound (12) is protected with a CBZ group by reacting it with benzyl chloroformate in acetonitrile (CH3CN) with K^COj as a base. Extrative work-up gives fully protected carboxylic acid (13) which can be use for step 12 without further purification..

The carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF using BOP as the coupling aģent in the presence of diisopropylethylamine (EtNiPr2) . Extraction with ethyl acetate (EtOAC) gives compound (15) as a solid which is purified by chromatography. 135 STEP 13

H2, Pd/C 10% MeOH (15)

(16) 5 The two CBZ(Z) protecting groups of compound (15) are removed by catalytic hydrogenation with Pd/C 10% as a catalyst. The catalyst is filtered and the solvent is evaporated to give the amino-guanidine (16). 136 LV 12019 EXAMPLE 7 SYNTHESIS OF COMPOUND (10)

(2) 10 4-methylmorpholine (NMM) was added to a solution of the carboxylic acid (2) (1.7g, 4.9mmol, 1.0eq), 4-hydroxyproline (3) (5.39mmol, l.leq), and BOP reaģent 15 (2.17g, 4.9mmol, 1.0eq) in anhydrous DMF (lOmL) at room temperature. The reaction mixture was stirred at room tenperature over night, quenched with brine (50mL) and ethyl acetate (lOOmL). The organic layer washed with 137 aqueous citric acid (10%, 2x50mL), sodium bicarbonate (10%, 2x50mL) and brine (50mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered and the solvent evaporated. The crude residue was purified by flash chromatography (5:4:1, ethyl acetate-hexane-methanol). l.lg of pure product (4) was recovered 48% yield. STEP 2

To a solution of 4-hydroxyproline derivative (4 ) (115mg, 240umol, 1.0eq) in dichloromethane (10 mL, anhydrous) at 0°C is added triethylamine (72mg, 720umol, 3.0eq) and methanesulfonyl chloride (28mg, 240uniol, 1.0eq) and the reaction mixture is stirred at roorn temperature. The mixture is then quenched with an aqueous solution of ammonium chloride and extracted with ethyl acetate. The organic layer is washed with 10% citric acid and brine, dried, filtered and the solvent is evaporated to dryness yielding compound (5). STEP 3 138 LV 12019

s z

HN

o (5)

1)Hg(QAc)2 CF3COCH/CPC O \ 2)Nc6H4

S HN Z

The enamine (5) (l.Oeg) is treated with mercuric acetate 5 (1.1 eq) in THF. The solvent is evaporated to dryness and the residue dissolved in methanol. The resulting organo-mercurial is reductivly cleaved with sodium borohydride (1.3eq). The resulting crude lactam thioether is purified by flash chromatography on silica gel affording compound 10 (6). STEP 4

(7) 15

To a solution of the lactam thioether (6) (1.0eq) in dry dichloromethane N-chlorosuccinimide (1.0eq) is added at 0°C. The reaction mixture is warmed to room temperature. When the reaction shows no more starting material the 20 solid is filtered and the solvent evaporated to dryness. The crude material (7) is use without any further purification for step 5. 139 STEP 5

(7) (8)

To a solution of the alpha-chlorothioether (7) (1.0eq) in 5 THF (anhydrous) a solution of phenylcuprate (1.0eq) (prepared according to litterature procedure) is added at low temperature. When the reaction mixture shows no starting chlorothioether, brine and ethyl acetate is added. The organic layer is dried, filtered and evaporated 10 to dryness to afford the desired product (8) . STgPJ?

15 The isolated bicyclic lactam (8) is hydrolysed with one equivalent of lithium hydroxyde in a 1:1 mixture of THF and water. The mixture is stirred at room temperature for 1 hour. The crude mixture is extracted with ether and the resulting solution is poured into 10% citric acic aqueous 20 solution and extracted wit.h dichloromethane to yield the corresponding carboxylic acid (9). 140 LV 12019 STEP 6

The crude carboxylic acid (9) is coupled with benzythiazole keto arginine in DMF using BOP as the coupling reaģent in the presence of diisopropylethylamine. Extraction with EtOAc gives a solid that is purified on silica gel to give the protected amide. The CBZ protecting group is removed with BBr3 in dichloromethane at room temperature finally gives the bicyclic benzothiazole keto arginine inhibitors (10) .·* .

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

141 COMPOUND #12

(12) 5 EXAMPLE 8 STEP 1

10

Commercially available glutaric acid monomethyl ester chloride (1) (20 ml,0.144 mol) was disolved in 40 ml of dry tetrahydrofuran (THF) and cooled to -15°C. Excess diazomethane freshly prepared in 300 ml of Ether was 15 introduced via cannula at -15°C to the solution. The mixture was left to warm up to room temperature overnight. Excess diazomethane was evacuated from the flask with a current of argon. To bring the reaction to completion, 75 ml of 1 N HC1 in Ether was added at 0°C and left to warm 20 up to room temperature for 5 hours. The volume of the solvent was reduced and then washed with 2x 5% NaHC03 142 LV 12019 dried over Na2C03 and evaporated to give crude chloromethylketone (20.46 g , 79%) wich was used in the next step without further purification. 1h NMR (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

α

Ha.H-C/s-OET / CHgCOONa/ NaCNBH3 MeOH.R.T 0^v-'OCH!CH3 HN' cr^ccH,

Crude chloromethylketone (2)(10.04 g, 56.15 mmol) was disolved in 300 ml of dry MeOH. Sodium acetate (2 eq, 9.21 g, 112.3 mmol) was added followed by L-Cysteine ethyl ester hydrochloride salt (1.3 eq, 13.55g, 72.98 mmol) and sodium cyanoborohydride (1.4 eq, 4.9 g, 78.59 mmol). The heterogeneous mixture was left to stir at room temperature for 2h30 min. 200 ml of methanol (MeOH) vas then added to disolve ali the solid and the pH was brought to 2 with IN HC1 .The mixture was then basified with saturated NaHCŪ3 until pH= 8. MeOH was evaporated and the remaining aqueous solvent was washed with ethyl acetate and dichloromethane. Solvents were combined, dried over Na2S04 and evaporated. The crude resicLue was purified by silica gel flash column chromatography using a gradient.of eluents ethyl acetate / hexane in the 143 following ratios: (3:7, 5:5, 6:4, 7:3) giving cyclic compound (3). NMR (CDCI3 ' 400MHz) of compound (3) d 1.21-1.27 (t, 5 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) 10 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 15 STEP 4

Cydic compound (3) (913mg, 3.32 mmol) was disolved in 50 ml of dry Toluene. (1S).-( + )-10-Camphorsulfonic acid ( 92 20 mg, 0.39 mmol) was added and the mixture was left to reflux for 4 days. When ali starting material was shown to be consumed (by TLC) , the mixture was worked up by evaporation of solvent, dissolving residue in ethyl acetate and washing with 2 x 5 % NaHC03. The Ethyl 25 acetate layer was dried over Na2SŪ4 and evaporated. The crude residue was purified by silica gel flash co'lumn chromatography using 60 % ETOAC / 40 %Hexane followed by 144 LV 12019 70 % ETOAC / 30 % Hexane giving 62.5% of Bicyclic compound (4) . 3-H NMR (CDCI3, 400MHz) of compound (4) d 1.27-1.31 (t, 5 3H, J=7 Hz), 1.5-1.6 (m, 1H), 1.72-1.87 (m, 2H), 2.02- 2.1 (m, 1H), 2.33-2.46 (m, 2H) , 2.52-2.59 (m, 2H), 2.83-2.88 (dd, 1H,J=14 ,4 Hz), 3.14-3.18 (d, 1H), 3.78-3.85 (m, 1H) , 4.2-4.27 (q, 2H, J= 3.9 Hz), 5.9- 5.92(t, 1H, J=3.4 Hz) . 10 STEP 5

H H

Bicyclic (4) (366mg,1.5 mmol) was dissolved in 25 ml of

THF and 5 ml H2O Lihium hydroxide. monohydrate (1.1 eq, 15 7.05 mg, 1.68 mmol) was added in 2.3 ml of H2O, at 0°C and the mixture was left to stirr at 0°C for 1 hr and at room teroperature for 3 hrs. THF was then evaporated and the remaining aqueous mixture was acidified by addition of Citric acid until pH»2. Extraction of agueous mixture 20 with 2 x CH2CL2 and 2 x ETOAC, drying of combined organic layers with Na2S04 and evaporation gavē a crude residue wich was purified by silica gel flash column chromatography using 70 % ETOAC / 30 %Hexane followed by 4.7 % HOAC / Ethyl acetate giving the pure acid (5) in 54 25 % yield . 16 % of starting material(4) was recovered. !η NMR (MeOD, 400 MHz) of compound (5) d 1.57-1.69 (m, 1H), 1.70-1.80 (m, 1H), 1.81-1.89 (m, 1H), 2.05-2.12 (m, 1H), 2.35-2.5 (m, 2H), 2.51-2.66 (m, 2H), 2.86- 145 5 2.91 (dd, 1H, J=13.8, 4 Hz), 3.12-3.17 (d, 1H), 3.3- 3.32 (m, 1H), 3.78-3.84 (m, 1H) , 5.76-5.78(t, 1H, J=3.53 Hz) . 13C NMR (MeOD, 400 MHz) d 17.052, 27.07, 28.928, 31.382, 32.096, 51.016, 55.138, 170,088, 171.24 STEP 6 10

To a solution of lithium bis (trimethylsilyl) amide (5ml of 1M THF solution, 5mmol) in THF (10 ml) is added at -78°C a solution of the carboxylic acid (5) (500mg, 2.32 mmol) . 15 20 the resulting solution is stirred at -78°C for 1 hour. Benzyl bromide (0.26 ml, 2.22 mmol) is then added and the mixture is allowed to reach room tempera tūre and stirred for 15 hours. The mixture is then poured into 10% HC1 (50 ml) and extracted wiht diclhoromethane (4x 60ml) . The combined organic phases are dried over MgS04 and the solvent remove by evaporation to yield to the crude alkylated amide (6). 146 LV 12019 STEP 7

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

The following compound is produced accordingly with the 15 exeption that the appropriate substitution of products were made in order to obtain the final compounds. COMPOUND #8

147 EXAMPLE 9

Determination of Kt Values for Heterocyclics 5

The affinity of inhibitors for thrombin was measured according to the procedures described in (DiMaio et al, J. Bio. Chem. , 1990, 265:21698) Inh.ibi.tion of amidolytic activity of human thrombin was measured fluorometrically 10 using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50 mM Tris-HCl buffer (pH 7.52 at 37°C) containing 0.1 M NaCl and 0.1% poly(ethylene glycol) 8000 at room temperature, and (Szewczuk et al., Biochemistry, 1992 31:9132). 15 The hydrolysis of the substrate by thrombin was monitored on a Varian-Cary 2000™ spectrophotometer in the fluorescence mode (λβΧ = 3 83 nm, λβπι = 455 nm) or on a Hitachi F2000™ fluorescence spectrophotometer (λΛ = 383 nm, λω = 455 nm), and the fluorescent intensity was calibrated 20 using AMC. The reaction reached a stead.y-state within 3 minūtes after mixing thrombin with the substrate and an inhibitor. The steady-state velocity was then measured for a few minūtes. The compounds of this invention were also pre-incubated with thrombin for 20 minūtes at room 25 temperature before adding the substrate. The steady-state was achieved within 3 min and measured for a few min. The kinetic data (the steady-state velocity at various concentrations of the substrate and the inhibitors) of the competitive inhibition was analyzed using the methods 30 described by Segel (1975). A non-linear regression program, RNLIN in the IMSL library (IMSL, 1987), LMDER in MINPACK library (More et al., 1980) or Microsoft™ Excell™ , was used to estimate the kinetic parameters (Ka Vmax and Kt) . 35 dTT assav 148 LV 12019

The fibrin clotting assay was performed in 50 mM Tris HC1 buffer (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 NIH unit/mL) and 0.03 % (w/v) of the final concentrations of human thrombin and bovine fibrinogen, respectively, as reported elsewhere (Szewczuk et al.,supra). The clotting time was plotted against the inhibitor concentrations and the IC50 was estimated as the inhibitor concentration required to double the clotting time relative to the control. Results are summarized in Tables 1 and 2 below.

Fibrin Clot Assav

The fibrin clot assay was performed essentially as described by Krtenansky et al, FEBS, 1987, 211:10. A serial dilution of the inhibitor was prepared in 50 mM tris HC1 buffer (pH7.8 at 23 °C) containing 0.1M NaCl and 0.1% (w/v) polyethylene glycol 8000. Human plasma (60pL, collected in 3.8% sodium citrate, blood/anticoagulant 9:1) was added to microtiter wells (microtiter plate, Falcon) containing 100pL of various inhibitor dilutions. The solution was mixed after which 50pL of human thrombin (InM final conc.) was added and mixed for 15 seconds. The turbidity of the clot was immediately monitored by microplate autoreader (Dynateck MR 5000) at 405nm and recorded every 3 min. The maximal turbidity in the absence of inhibitors was reached within a 60 min. ICS0 values were calculated at 30 minūtes as the inhibitor concentration that gavē half the optical density of the control.

Platelet Aaareoation and Secretion

Rat blood was collected into ACD (6/1 v/v) by cardiac puncture. Suspensions of washed platelets were prepared as described by Ardlie et al, (Br. J. Haematol. 1970, 19:7 149 and Proc. Soc. Exp. Biol. Med., 1971, 136:1021). The final suspending medium was a modified Tyrode solution (NaCl 138niM, KC1 2.9mM, HEPES 20mM, Nal^PO, 0.42mM, NaHC03 12mM, CaCl2 lmM, MgCl2 2mM, 0.1% glucose, 0.35% albumin, 5 apyrase lļiL/mL pH 7.4). Platelet counts were adjusted to 5000, ΟΟΟ/μΙ*.

To permit measurement of the extent of release of the contents of the dense granules, the platelets were 10 labelled in the first washing solution with “c-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 (5μΜ final conc.) was added to present the reuptake of released serotonin. 15

Platelet aggregation was recorded at 37°C in an aggregometer (BioData PAP-4) at a stirring speed of 1,100 rpm by measuring the variations of light transmission. Percentage of aggregation was determined 3 min. after the 20 addition of the stimulating aģent (human thrombin 0.1IU/mL final conc.) . Inhibitors were preincubated 1 minūte at 37°C before addition of stimulating aģent. ICS0 values represent the concentration that was necessary to inhibit platelet aggregation or secretion to 50% of the control. 25

Arterial Thrombosis Modei

FeClj Induced Carotid Arterial In-iurv Modei

The FeClj 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; 2 69-280, 1990 and Schumacher, W.A. et al. J. Pharmacology and Experimental Therapeutics 267; 1237-1242, 1993. 35 Male, Sprague-Dawley rats ( 375-410 g) were anesthetized with urethane ( 1500 mg\kg ip). Animals were laid on a 150 LV 12019 37°C heating pad. The carotid artery was exposed through a midline cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid sheath. Using forceps, the artery was lifted to provide sufficient 5 clearance to insert two small pieces of polyethylene tubing (PE-205’) underneath it. A temperatūre probe (Physitemp* MT23/3) was placed between one of the pieces of tubing and the artery. Injury was induced by topical application on the carotid artery above the temperature 10 probe of a small disc (3 mm dia.) of Whatman* No.l filter paper previously dipped in a 35% solution of FeCl3. The incision area was covered with aluminum foil in order to protect the FeCl3 from degradation by light. The vessel temperature was monitored for 60 minūtes after application 15 of FeCl3 as an indication of blood flow. Vessel temperature changes were recorded on a thermister (Cole-Palmer’ Modei 08533-41).

The time between the FeClj application and the time at 20 which the vessel temperature decreased abruptly (>2.4°C) was recorded as the time to occlusion of the vessel. Inhibitor compounds were given as an iv bolus (mg/kg) followed immediately by an iv infusion (pg/kg/min. via femoral vein) . The dose of inhibitor needed to double the 25 time to occlusion in comparison to control animals in which injury was induced in the absence of inhibitor was determined. 151

Table 1

Antiplatelet activity μΜ

Ki μΜα Aggregation * 5-HT Secretion dTTICgj μΜ Plasma fibrin clot assay ICW μΜ 4 ND ND 47 >450 4.0 21 19 89.5 >450 Ίό >100 >100 162 >450 2.2 18 14.2 22 >450 53 >100 >100 >625 >450 8.6 >100 >100 67 320 34 >100 >100 319 >450 19 >100 >100 207.5 >450 74 ND ND 415 >450 62 ND ND >450 32.7 47.5 52 42 200 4.4 22 2.1 25 78 0.048 0.4 0.38 0.375 ND 0.031 ND ND 0.33 ND 152 LV 12019

0095 26 ND ND 0100 19 ND ND 165 ND *suspension of washed platelets from rats * Inhibitory dissociation constant for human a-thrombin 153

Table 2

Compound ki dTT Route M.O.T (nM) (nM) ivb+inf (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 0295b . 5000 520 0.75-50 19+/-2.7 0240 18 0.75-50 17+/-2.6 —- ģ 0/75-50 20.13+/-3.4 0255 500 0260 16 0.75-50 14.83+/-0.2 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 -- 138 '0775-50.......... ''''15.Ϊ+7-Ϊ4.2'...... 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 120 154 LV 12019

Table 2 (continued) Compound Ki (nM) dTT (nM) Route ivb+inf M.O.T (min)+/-sem 0925a+b 10 0.75-50 19.6+/-0.2 6925b.......... ..........30......... 0925a 7 0.75-50 20.3+/-3.5 0940a 16 0.75-50 15.2+/-0.82 0940b 160 0950a 150 0950b 1000 a = early eluting on RP HPLC singla isomer b = late eluting on RP HPLC single isomer a + b = mixture 155 LV 12019 WE CLAIM: 1. A compound of formula (I) :

wherein: A is selected from (CH-Re)0_, , S, SO, S03, 0 and NR„ wherein R, is hydrogen, alkyl optionally interupted with 1 or 2 heteroatoms; C6.1S aryl, C3.7 cycloalkyl or heterocyclic ring or a hydrophobic group; B is selected from S, SOs, Ο, -N=, NH, -CH= and CRSR, wherein R, and R, are independently selected from hydrogen and Cj_4 alkyl provided that when A is S, SO, S02, 0, or NRg/ then B is CR4R,; D is selected from (CH-R,)0_2 wherein R, is hydrogen, Cy t alkyl or -0(0)^; and CH with a double bond to B when B is -N= or -CH=; E is selected from CHj and CH substituted with the -CCOJRj, provided that only one of D and E is substituted with with -C(0)R,; X is selected from 0, N-Rj, or CH-RS; Y is selected from 0, S, SO, S02, N-Rj and CH-R, provided that when X is N-Rj then Y is CH-R, or 0, and when X is 0 then Y is CH-R8; Z is selected from 0, S and Hj,* R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle; R, is selected from H and C,_6 alkyl optionally substituted with C6 aryl, a 6 member hetērocycle or a C,_7 cycloalkyl ring; 156 5 5 10 2 . R3 is selected from H, NR^ and C,_4 alkyl; and R( and R, are independently selected from H; NR^; C4_l6 aryl or C3_7 cycloalkyl optionally substituted with Cj.e alkyl; Ct_l4 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R, or a C4_14 aryl, heterocycle or Cj_7 cycloalkyl group optionally substituted with halogen, hydroxyl, C^6 alkyl; an amino acid side Chain; and a hydrophobic group. A compound according to claim 1, wherein R3 is one of formula Via to Vīd:

wherein: 15 R21 is hydrogen or Ct_4 alkyl; K is a bond or -NH-; 20 G is Ct.4 alkoxy; cyano; -NHj,* -CHj-NHj,* -C(NH)-NH,; -NH-C(NH) -NHj; -CHj-NH-C (NH) -NH2; a C4 cycloalkyl or aryl substituted with cyano, -NHj, -CH2-NHj, -C(NH)-NH2, -NH-C(NH) -NHj or -CH2-NH-C (NH) -NH^; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CHj-NHj, -C (NH) -ΝΗ2, -NH-C(NH)-ΝΗ2 or -CHj-NH-C (NH) -NHj; U is cyano, -NH2, -C(NH) -NH2 or -NH-C(NH) -NHj; P is a bond, -C(O)- or a bivalent group: 157 25 LV 12019

OH

5 J is alkylene optionally substituted with OH, NHj and alkyl and optionally interrupted by a heteroatom selected from 0, S and N; n is 0 or 1; and T is H, OH, amino, a peptide Chain, Cj_ie alkyl, alkoxy, C6_20 aralkyl, or heterocycle optionally substituted. ΙΟ 3 A compound according to claim 2, wherein T is a heterocycle selected from the group consisting of:

wherein 15 X,, X10, Xu and X12 are each independently selected from the group consisting of N, or C-Χ, where X, is hydrogen, Cj_, alkyl, or C4.„aryl; 20

Xs and X13 are each independently selected from the group consisting of C, 0, N, S, Ν-Χ,, or CH-Χ^- and R' is hydrogen, C,.u alkyl optionally carboxyl substituted, carboxyl, -C0.u alkyl-C02-C1.ls alkyl, C6.20 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. 4. A compound according to claim 3, wherein T is selected from the group consisting of: 158 25

5 5 10 15 6 wherein R' is hydrogen, Ct.is alkyl optionally carboxyl substituted, carboxyl, —C0_lS- alkyl-C02-C,_16 alkyl, C5_20 aralkyl, C3_7 cycloalkyl, aryl or an aromatic heterocycle. * A compound according to claim 4, wherein T is selected from:

wherein R' is hydrogen, C,.,t alkyl optionally carboxyl substituted, carboxyl, -C0_u alkyl-C02-C1.u alkyl, C6.20 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. A compound according to claim 1, wherein one of R4 and Rs is a hydrophobic group selected from C,_20alkyl, C2.2Ū 159 LV 12019 alkenyl or C2_20 alkynyl optionally interrupted by a carbonyl group, C4.u aryl, C3_7 cycloalkyl, C6.20 aralkyl, C6.20 cycloalkyl substituted Cj.20 alkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group and the ring portion is optionally substituted with alkyl; and a hydrophobic artiino acid side chain. 7. A compound according to claim. 6, wherein R3 is H. 8. A compound according to claim 1, wherein Z is O. 9. A compound according to claim 1, wherein R, is H. 10. A compound of the formula (VII):

O wherein

Rt is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle; R, is H or Cl-6 alkyl;

Rj is selected from H, NR4R7 and Cj_4 alkyl; and R4 and Rj are independently selected from H; NR6R7; C615 aryl or C3_7 cycloalkyl optionally substituted with C] S alkyl; C,.I4 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRjR, or a Cs.u aryl, heterocycle or C3.7 cycloalkyl group optionally substituted with halogen, hydroxyl, C,.4 alkyl; an eimino acid side chain; and a hydrophobic group. 160 11. A compound according to claim 10, wherein Rļ is one of formula Via to Vīd:

wherein: IĻ is hydrogen or alkyl; K is a bond or -NH-; G is C1M alkoxy; cyano; -NH,; -ΟΗ,-ΝΗ,; -C (NH) -NH,; -NH-C(NH) -NH,; -CH,-NH-C (NH) -NH,; a C( cycloalkyl or aryl substituted with cyano, -NH,, -CH,-NH,, -C (NH) -NH,, -NH-C(NH) -NHj or -CH,-NH-C(NH) -NH,; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH,-NH,, -C (NH) -NH,, -NH-C (NH) -NH, or -CH,-NH-C (NH) -NH,; U is cyano, -NH,, -C(NH)-NH, or -NH-C(NH) -NH,; P is a bond, -C(O)- or a bivalent group:

OH

J is Ct.4 alkylene optionally substituted with OH, NH, and Cj.4 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, Ct.ie alkyl, alkoxy, Cs_,0 aralkyl, or heterocycle optionall·y substituted. 161 LV 12019 12. A compound according to claim 11, wherein T is a heterocycle selected from the group consisting of:

Χϋ— Χι, wherein

Xs, Χ^, Xxl and ^ are each independently selected from the group consisting of N, or C-Χ, where X, is hydrogen, alkyl, or Cs uaryl; 10 X4 and Xl3 are each independently selected from the group consisting of C, Ο, N, S, Ν-Χ,, or ΟΗ-Χ,; R' is hydrogen, Ct.jS alkyl optionally carboxyl substituted, carboxyl, -C0_14 allc/l-COj-Cj.j,. alkyl, C6.J0 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. 15 13 . A compound according to claim 12, wherein T is selected from the group consisting of: 162

and R' is hydrogen, Οχ.ιβ alkyl optionally carboxyl substituted, carbo:xyl, -Ce_M alkyl-COj-Cj_16 alkyl, Cs_20 aralkyl, C3_7 cycloalkyl/ aryl or an aromatic 5 heterocycle. 14. A compound according to claim 13, vherein T is selected from:

R·

10 and R' is hydrogen, C,.^ alkyl optionally carboxyl substituted, carboxyl, -C0_16 alkyl-C02-Cl_l6 alkyl, C6.20 aralkyl, C3_7 cycloalkyl, aryl or an aromatic heterocycle. 15 15. A compound according to claim 10, wherein R, and R3 are H. 163 LV 12019 16. A compound according to claim 10, wherein R4 is alkyl optionally interupted with a heteroatom or a carbonyl, and optionally substituted with a Cs.15 aromatic, C3_7 cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF3 or oxo. 17. A compound according to claim 10, wherein Rj is H. 18. A compound according to claim 12, wherein: R3 is H; R4 is Cj.u alkyl optionally interupted with a heteroatom or a carbonyl, and optionally substituted with a Ce.ls aromatic, C3_7 cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF3 or oxo; and Rj is H. 19. A compound according to claim 10, selected from: 0085 6S-cyclohexylmethylhexahydro-5-oxo-5H- thiazolo [3,2-a]pyridine-3R-carboxamido (propylcarbo methoxy ketoarginine); and 0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a]pyridine-3R-carboxamido (a-benzothiozolo keto arginine). 20. A compound according to claim 1, of formula (VIII):

164 R, is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle; R, is H or Cl-6 alkyl; R, is selected from H, NRjR, and alkyl; and R4 and Rs are independently selected from H; NR^R,; Ce.l6 aryl or C3_7 cycloalkyl optionally substituted with Cj.4 alkyl; alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRSR, or a Ce_14 aryl, heterocycle or C3_7 cycloalkyl group optionally substituted with halogen, hydroxyl, Cj.e alkyl; an amino acid side chain; and a hydrophobic group. 21. A coropound according to claim 20, wherein R, is one of formula Via to Vīd:

wherein:

Rtl is hydrogen or Cj_4 alkyl; K is a bond or -NH-; G is Cļ_4 alkoxy; cyano; -NH,; -CH,-NH,; -C (NH) -NH,; -NH-C (NH) -NH,; -CH,-NH-C (NH) -NH,; a Cs cycloalkyl or aryl substituted with cyano, -NH,, -CH,-NH,, -C(NH)-NH,, -NH-C (NH) -NH, or -CH,-NH-C (NH) -NH,; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH.-NH,, -C(NH)-NH,, -NH-C (NH) -NH, or -CH,-NH-C (NH) -NH,; ϋ is cyano, -NH,, -C(NH)-NH, or -NH-C (NH)-NH,; 165 LV 12019 P is a bond, -C(O)- or a bivalent group:

OH

J is C1.i alkylene optionally substituted with OH, NHj and Cļ_6 alkyl and optionally interrupted by a heteroatom selected from 0, S and N; n is 0 or 1; and T is H, OH, amino, a peptide Chain, alkyl, alkoxy, Cs_20 aralkyl, or heterocycle optionally substituted. 22. A compound according to claim 21, wherein T is a heterocycle selected from the group consisting of:

wherein

Xs, Xt0, X11 and X13 are each independently selected from the group consisting of N, or C-Χ, where X, is hydrogen, alkyl, or Ct.l6 aryl; X4 and X13 are each independently selected from the group consisting of C, Ο, N, S, Ν-Χ,, or CH-Χ,; R' is hydrogen, 0^,^ alkyl optionally carboxyl substituted, carboxyl, -C0.IS alkyl-C02-C1.u alkyl, C6.20 aralkyl, C2.7 cycloalkyl, aryl or an aromatic heterocycle. 23. A compound according to claim 22, wherein T i-s selected from the group consisting of: 166

and R' is hydrogen, ClMs alkyl 0ptionally carboxyl substituted, carboxyl, alkyl-COa-Ct.M alkyl, C4.20 aralkyl, C3.7 cycloalkyl/ aryl or an arOmatic 5 heterocycle. 24. A compound according to claim 23, wherein T is selected from:

10 and R' is hydrogen, alkyl optionally carboxyl substituted, carboxyl, -C0_16 alkyl-C02-C1.u alkyl, C6.J0 aralkyl, C3_, cycloalkyl, aryl or an aromatic heterocycle. 15 25. A compound according to claim 20, wherein Rj and R3 are both H. 167 LV 12019 26. A compound according to claim 20, wherein R4 is H or Cļ.4 alkyl substituted with COOH. 27. A compound according to claim 20, wherein R,, R3 and R4 are H and Rj is C,.ie alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R, or a Cs.16 aryl, heterocycle or C3.7 cycloallcyl group optionally substituted with halogen, hydroxyl or Ct_4 alkyl. 28. A compound according to claim 22, wherein:

Rj, R3 and R4 are H; and

Rs is C,_1S alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR^ or a C4_IS aryl, heterocycle or C3.7 cycloalkyl group optionally substituted with halogen, hydroxyl or Ct.4 alkyl. 29. A coirpotind according to claim 20, selected from: 0345 4-Oxo-2-(3-phenyl-propionyl)-octahydro- pyrrolo[l,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-methyl-thiazole-2-carbonyl)-bu ty 1] - amide; and 0340 4-Oxo-2- (3-phenyl- propionyl) -octahydro-pyrrolo [1,2-alpyrazine-6-carboxvlic acid [4-guanidino-l-(thiazole-2- carbonyl) -butyll-amide. 30. A compound according to claim 1, of formula (IX) : 168

wherein Y is selected from 0, S, SO, S02, N-Rs and CH-R,;

Rx is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;

Rj is H or Cl-6 alkyl; R, is selected from H, NRjR, and alkyl; and R4 and R, are independently selected from H; NRSR,; C6.16 aryl or C3_7 cycloalkyl optionally substituted with Cj.4 alkyl; Cj.lt alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRsRj or a CS.1S aryl, heterocycle or C3_7 cycloalkyl group optionally substituted with halogen, hydroxyl, Cj_6 alkyl; an amino acid side Chain; and a hydrophobic group; R, is hydrogen, Cj.4 alkyl optionally interupted with 1 or 2 heteroatoms; CS.U aryl, C3_7 cycloalkyl or heterocyclic ring or a hydrophobic group; and n is 1 or 2; 31. A compound according to claim 30, wherein Rt is one of formula Via to Vīd:

169 LV 12019 wherein: R1X is hydrogen or Cj_6 alkyl ; K is a bond or -NH-; G is alkoxy; cyano; -NH,; -CHj-NHj,* -CfNHj-NHj,· -NH-C(NH) -NHj; -CHj-NH-C (NH) -NHj; a C6 cycloalkyl or aryl substituted with cyano, -NHj, -CHj-NHj, -C(NH)-NHj, -NH-C (NH) -NHj or -CHj-NH-C (NH) -NH,; or a 5 or 6 mernber, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH2-NH2, -C (NH) -NHj, -NH-C(NH) -NH, or -CHj-NH-C (NH) -NHj,* U is cyano, -NK,, -C(NH) -NH, or -NH-C (NH)-NH,; P is a bond, -C(O)- or a bivalent group:

OH

J is Ct_6 alkylene optionally substituted with OH, NHj and Cļ_£ alkyl and optionally interrupted by a heteroatom selected from O, S and N; n is 0 or 1; and T is Η, OH, amino, a peptide Chain, C,_ie alkyl, C,.ls alkoxy, C6_20 aralkyl, or heterocycle optionally substituted. 32. A compound according to claim 31, wherein T is a heterocycle selected from the group consisting of:

wherein

Xs, X10, Xlx and X12 are each independently selec-ted from the group consisting of N, or 0-Χ7 where X, is hydrogen, C,., alkyl, or C(_u aryl; 170 X, and X13 are each independently selected from the group consisting of C, 0, N, S, Ν-Χ,, or CH-X,; R' is hydrogen, alkyl optionally carboxyl substituted, carboxyl, -C0_lt alJ^l-COj-Cļ.^ alkyl, Cs_20 5 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. 33. A compound according to claim 32, wherein T is selected from the group consisting of: 10

and R' is hydrogen, alkyl optionally carboxyl substituted, carboxyl, -Ce.u alkyl-C02-C1.16 alkyl, C5.20 aralkyl, C3.7 cycloalkyl, aryl or an aromatic heterocycle. 34. A compound according to claim 33, wherein T is selected from: 171 15 LV 12019

and R' is hydrogen, cļļ6 alkyl optionally carboxyl substituted, carboxyl, -C0_u alkyl-C02-C1_u alkyl( C6_20 aralkyl/ C3_7 cycloalkyl, aryl or an aromatic heterocycle. 35. A compound according to claim 30, wherein Rj and R3 are both H. 36. A compound according to claim 30, wherein R4 is H, NR-eR? ot alkyl substituted COOH. 37. A compound according to claim 30, wherein Rj is C6_u aryl, C4_20 aralkyl, or alkyl substituted with C3_7 cycloalkyl. 38. A compound according to claim 32, wherein n is 1;

Rj, Rj and R4 are H; and

Rs is C5.ie aryl, C6_20 aralkyl, or Cj.14 alkyl substituted with 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-hutyl]-amide; and 172 0900 3-Amino-2-cyclohexyl-4-oxo-hexahydro-pyrrolo[2,1-b][l,3]thiazine-6-carboxylic acid [1-(benzothiazole-2-carbonyl) -4-guanidino-butyl]-axtd.de. 40. A compound according to claim 1, of formula (X) :

O wherein B is Ο, S, -CHj-, or -NH-; R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle; R, is H or Cl-6 alkyl; R, is selected from H, NRjR, and alkyl; and R4 and Rj are independently selected from H; NRjR,; C4_14 aryl or C3_7 cycloalkyl optionally substituted with Cj.4 alkyl; Cj_14 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR4R, or a C4_l4 aryl, heterocycle or C3.7 cycloalkyl group optionally substituted with halogen, hydroxyl, Cj_4 alkyl; an amino acid side Chain; and a hydrophobic group. 41. A compound according to claim 40, wherein Rj is one of formula Via to Vīd: 173 LV 12019

wherein: R,, is hydrogen or alkyl; K is a bond or -NH-; 5 G is alkoxy; cyano; -NH,; -CH,-NH,;. -C (NH)-NH,; -NH- 10 C (NH) -NH,; -CH,-NH-C (NH) -NH,; a C6 cycloalkyl or aryl substituted with cyano, -NH,, -CH,-NH,, -C(NH)-NH,, -NH-C (NH) -NH, or -CH,-NH-C (NH) -NH,; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH,, -CH,-NH,, -C (NH) -NHj, -NH-C (NH)-NH, or -CH,-NH-C (NH) -NH,; U is cyano, -NH,, -C(NH)-NH, or -NH-C (NH)-NH,; P is a bond, -C(O)- or a bivalent group:

OH

15 J is C,^ alkylene optionally substituted with OH, NH, and C,.t alkyl and optionally interrupted by a heteroatom selected.from O, S and N; n is 0 or 1; and T is H, OH, amino, a peptide chain, C^ alkyl, c,.u alkoxy, Cs.,0 aralkyl, or heterocycle optionally substituted. 42. A compound according to claim 41, wherein T is.a heterocycle selected from the group consisting of: 174 20 5

R' wherein

Xs< Χ^, Χχί and Χ13 are each independently selected from the group consisting of N, or C-Χ, where X, is hydrogen, Ct.4 alkyl, or C4_ltaryl; X4 and X13 are each independently selected from the group consisting of C, Ο, N, S, Ν-Χ,, or CH-Χ,; 10 R' is hydrogen, C1.lt alkyl optionally carfaoxyl substituted, carboxyl, -Ce_16 alkyl-C03-C1_16 alkyl, C4.20 aralkyl, C2.7 cycloalkyl, aryl or an aromatic heterocycle. 15 43. A coirpound according to claim 42, wherein T is selected from the group consisting of: 175 LV 12019

and Rr is hydrogen, Cws alkyl optionally carboxyl substituted, carboxyl, -C,.^ alkyl-C02-C1.ls alkyl, C€_20 aralkyl, C3_7 cycloalkyl/ aryl.pr an aromatic 5 heterocycle. 44. A compound according to claim 43, wherein T is selected from:

and R' is hydrogen, Ct.u alkyl optionally carboxyl 10 substituted, carboxyl, -Ca_ls alkyl-C02-C1.ls alkyl, Cs.20 aralkyl, C3_7 cycloalkyl, aryl or an aromatic heterocycle. 45. A compound according to claim 40, wherein Rj .and R3 are both H. 176 15 46. A compound according to claim 40, wherein R4 is alkyl substituted with C4_JS aryl optionally substituted with Cj.14 alkyl. 5 47. A compound according to claim 40, wherein Rs is H. 48. A compound according to claim 42, wherein B is S;

Rj, R3 and Rs are H; and 10 R4 is Cj.ie alkyl substituted with Cs.u aryl optionally substituted with C,.IS alkyl. 49. A compound according to claim 40, selected from: 925 7-Benzyl-6-oxo-octahydro-pyrido[2,1- 15 20 25 c][l, 4]thiazine-4-carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)butyl]-amide; and 940 6-Οχο- 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 of thrombotic disorders in a mammai, comprising administering to said mammai an effective amount of a compound according to claim 1. 51. A method according to claim 50, wherein said thrombotic disorder is venous thrombosis. 52. A method according to claim 50, wherein said 30 thrombotic disorder is a pulmonary embolism. 53. A method according to claim 50, wherein said thrombotic disorder is arterial thrombosis. 177 LV 12019 54. A method according to claim 50, wherein said thrombotic disorder is myocardial infarction. 55. A method according to claim 50, wherein said 5 thrombotic disorder is cerebral infarction. 56. A process for producing a compound according to claim 1. 10 57. A process for producing a compound according to any one of claims 10, 20, 30 or 40. 178

Claims

EN 12019 DESCRIPTION OF THE INVENTION

A compound of formula (I) 0) wherein: A is selected from (CH-RgVi, S, SO, SO2.0 and NR8 wherein Rg is hydrogen, alkyl optionally substituted with 1 or 2 heteroatoms; Ce is aryl, C3) .7 dichloroalkyl or heterocyclic ring or hydrophobic group; B is selected from S, SO2, Ο, -N =, NH, -CH = and CR CRR7, wherein R «and R7 are independently selected from hydrogen and alkyl at that; if A is S, SO, SO2.0 or NRg then B is CReR ?; D is selected from (CH-R8) («where Rg is hydrogen, C1-6 alkyl or -C (O) R, and CH with a double bond with B when B is -N = or -CH =; E is selected from CH 2 and CH substituted with C (O) R 1, wherein only one of D and E is substituted with -C (O) R 1; X is selected from O, N-R5, or CH-Rj; Y is selected from O, S, SO, SO2, N-Rs and CH-Rg, wherein when X is N-R5, Y is CH- Rg or O, and when X is O, Y is CH-R8; Z is selected from O, S and H2; R1 is an arginyl moiety or an analogue or derivatives thereof optionally substituted with an amino acid; R 2 is selected from H and alkyl optionally substituted with C 6 aryl, 6 membered heterocycle or C 3-7 cycloalkyl ring; R3 is selected from H, NReR? and C 1-6 alkyl; and Rtun Rsir independently selected from H; NReR ?; Ceyl aryl or C3-7 cycloalkyl are optionally substituted with C1-4 alkyl; C1-6 alkyl optionally interrupted by one or more heteroatom carbonyl groups and optionally substituted by OH, SH NRJI7 or 0.16 aryl, heterocycle or C3.7 cycloalkyl optionally substituted by halogen, hydroxy, C1-6 alkyl; amino acid side chain; and hydrophobic group.

2. A compound according to claim 1 wherein R (is one of the formulas Via and Vid:

Wherein: Ru is hydrogen or alkyl; K is a bond or -NH-; G is CM alkoxy; cyano; -NH2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2; -CH2-NH-C (NH) -NH2; C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; or 5 or 6 membered, saturated and unsaturated heterocycles optionally substituted with cyano, -NH 2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2 or CH 2 -NH-C (NH) -NH 2; U is cyano, -NH2; -C (NH) -NH 2 or -NH-C (NH) -NH 2; P is a bond, -C (O) - or a bivalent group:

J is Q 6 alkylene optionally substituted with OH, NH 2 and C 1-4 alkyl and optionally interrupted by a heteroatom selected from 0, S and N; nirO or 1; and T is H, OH, an amino group, a peptide chain, a C 1-6 alkyl group, a C 1-6 alkoxy group, a Cs-m with an alkyl group, or an optionally substituted heterocycle.

A compound according to claim 2, wherein T is a heterocycle selected from the group consisting of

wherein: Xs, Xw, Xn and X, 2 are each independently selected from the group consisting of N, or C-X7 wherein X7 is hydrogen, C1-4 alkyl, or Cs.g aryl; X 6 and X 13 are each independently selected from the group consisting of C, Ο, N, S, N-X 7, or CH-X 7; and R 'is hydrogen, C1-6 alkyl optionally substituted with carboxyl, carboxyl, -C0-6; alkyl-CO 2 -C 1-16 alkyl, C 1-6 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

A compound according to claim 3, wherein T is selected from the group consisting of

3 EN 12019

/, N

N-N N / O-N-N

/ wherein R 'is hydrogen, C 1-16 alkyl optionally substituted by carboxyl, carboxyl, -C 0-16 alkyl-CO 2 -C 1, alkyl 6, C 1-6 alkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

5. The compound of claim 4, wherein T is selected from:

wherein R 'is hydrogen, C1-16 alkyl optionally substituted with carboxyl, carboxyl, -C0-6alkyl-CO2-C, C1-6 alkyl, C1-6 aralkyl, C3-7cycloalkyl, aryl or aromatic heterocycle.

6. A compound according to claim 1, wherein one of R4 and R5 is a hydrophobic group selected from alkyl, C2-2o alkenyl or a dnyl optionally substituted with carbonyl, C5-16 aryl, C3-7 cycloalkyl, Ce-M aralkyl, cycloalkyl with C1-20 alkyl, wherein the aliphatic moiety is optionally interrupted by a carbonyl group and the ring portion is optionally substituted by a Cw alkyl group; and a hydrophobic amino acid side chain.

7. The compound of claim 6, wherein R3 is H.

The compound of claim 1, wherein Z is 0.

9. The compound of claim 1, wherein R2 is H.

10. A compound of formula (VII):

Wherein: R 1 consists of an arginyl group or an analogue or derivatives optionally substituted with an amino acid, peptide or heterocycle; R2 is H or C1-6 alkyl; R3 is selected from H, NReR7 and alkyl; and R 1 and R 8 are independently selected from H; NR ^; 0.16 aryl or C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; C 1-16 alkyl optionally interrupted by one or more heteroatoms or carbonyl groups and optionally substituted by OH, SH, NR 17 or C 1-4 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted with halogen, hydroxy, C 1-6 alkyl ; amino acid side chain; and hydrophobic group.

A compound according to claim 10, wherein R 1 is one of the formulas of formula V to Vid: Via

VIb

^ 0-8 G

(J) n wherein: R 11 is hydrogen or C 1-6 alkyl; K is a bond or -NH-; G is alkoxy; cyano; -NH2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2; -CH2-NH-C (NH) -NH2; C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; or 5 or 6 membered, saturated and unsaturated heterocycles optionally substituted with cyano, -NH 2; -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or CH 2 -NH-NH (NH) -NH 2; U is cyano, -NH2; -C (NH) -NH 2 or -NH-C (NH) -NH 2; P is a bond, -C (O) - or a bivalent group: .OH 'CH' CH,

, or

J is C 1-6 alkylene optionally substituted with OH, NH 2 and C 1-6 alkyl and optionally interrupted by a heteroatom selected from O, S and N; nirO or l; and T is H, OH, an amino group, a peptide chain, a C 1-6 alkyl group, a C 1-6 alkoxyl group, a C 6-2 aralkyl group, or an optionally substituted heterocycle.

The compound of claim 11, wherein T is a heterocycle selected from the group consisting of

Wherein: X5, Xi0, Xu and X12 are each independently selected from the group consisting of N, or C-X7 wherein X is hydrogen, CM alkyl, or C5.8 aryl; X6 and Χ13 are each independently selected from the group consisting of C, Ο, N, S, N-X7, or CH-X7; and R 'is hydrogen, C 1-16 alkyl optionally substituted with carboxyl, carboxyl, -C 0.16; alkyl-CO 2 -C 1-6 alkyl, OΜ aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

A compound according to claim 12, wherein T is selected from the group consisting of

wherein R 'is hydrogen, Q, 6 alkyl optionally substituted with carboxyl, carboxyl, -C 1-16 alkyl-CO 2 -C 1-6 alkyl, C 1-6 aralkyl, C 1-4 cycloalkyl, aryl or aromatic heterocycle.

A compound according to claim 13, wherein T is selected from

Wherein R 'is hydrogen, C 1-6 alkyl optionally substituted by carboxyl, carboxyl, -C 1-16 alkyl-CO 2 -C 1-6 alkyl, C 1-6 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 10, wherein R 2 and R 3 are H.

The compound of claim 10, wherein R 4 is C 1-6 alkyl optionally interrupted with a heteroatom or carbonyl group and optionally substituted with a C < s.

17. The compound of claim 10, wherein R5 is H.

The compound of claim 12, wherein R 3 is H; R4 is optionally substituted by a heteroatom or carbonyl group, and optionally substituted with a C6.16 aromatic, C3.7 cycloalkyl or heterocyclic ring, wherein the ring is optionally substituted by CF3 or oxo; and Rs is H.

A compound according to claim 10 selected from; 0085 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-alpyridine-3R-carboxamide (propylcarbomethoxy ketoarginine); and 0105 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-alpyridine-3R-carboxamide (α-benzothiazole ketoarginine).

20. The compound of claim 1 having the formula (ΥΠΙ):

O wherein R 1 is a part of an arginyl group or an analogue or derivatives optionally substituted with an amino acid, a peptide or a heterocycle; R2 is H or C1-6 alkyl; R 3 is selected from H, NR 'R 7 and C 1-6 alkyl; and R 1 and R 5 are independently selected from H; NR <sR7; 0.16 aryl or C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; CM6 alkyl is optionally interrupted by one or more heteroatoms or carbonyl groups and optionally substituted by OH, SH, NR1R7 or C 1-8 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted by halogen, hydroxy, C 1-6 alkyl; amino acid side chain; and hydrophobic group.

A compound according to claim 20, wherein R 1 is one of the formulas of formula V to V 1

T G VIb

K. G 7 EN 12019

wherein: R 1 is hydrogen or C 1-6 alkyl; K is a bond or -NH-;

(J) n T G is CM alkoxy; cyano; -NH2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2; -CH2-NH-C (NH) -NH2; C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; or a 5- or 6-membered saturated or unsaturated heterocycle optionally substituted with a cyano, -NH2; -CH 2 NH 2; C (NH) -NH 2 -NH-C (NH) -NH 2 or CH 2 -NH-NH (NH) -NH 2; U is cyano, -NH2; -C (NH) -NH 2 or -NH-C (NH) -NH 2; P is a bond, -C (O) - or a bivalent group: .OH CH CH,

, or

J is Cms alkylene optionally substituted with OH, NH2 and alkyl and optionally interrupted by a heteroatom selected from O, S and N; nirO or l; and T is H, OH, amino acid, peptide chain, 0Μ6 alkyl, CM6 alkoxy, C6 M aralkyl, or optionally substituted heterocycle.

A compound according to claim 21, wherein T is a heterocycle selected from the group consisting of

wherein: X5, Χω, Xu and Xn are each independently selected from the group consisting of N, or C-X7 wherein X7 is hydrogen, C1-4 alkyl, or C5- aryl; Xf and X13 are each independently selected from the group consisting of C, Ο, N, S, N-X7, or CH-X7; and R 'is hydrogen, C1-6 alkyl optionally substituted by carboxyl, carboxyl, -Cq.16; alkyl-CO 2 -C 1-6 alkyl, Ο. aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 22, wherein T is selected from the group consisting of 8

wherein R * is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, carboxyl, -C 1-6 alkyl-CO 2 -CM 6 alkyl, C 1-6 aralkyl, C 1-6 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 23, wherein T is selected from

wherein R 'is hydrogen, C 1-6 alkyl optionally substituted by carboxyl, carboxyl, -C 1-16 alkoxy-CO 2 -C 1-16 alkyl, C 1-6 aralkyl, C 1-6 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 20, wherein R2 and R3 are both H.

The compound of claim 20, wherein R * is H or C 1-4 alkyl substituted with COOH.

The compound of claim 20, wherein R 1 R 3 and R * are H and R 5 is C 1, 6 alkyl optionally interrupted by one or more heteroatoms or carbonyl groups and optionally substituted by OH, SH, NR * R 7 or C 1. 16 aryl, heterocycle or C 3-7 cycloalkyl are optionally substituted with halogen, hydroxy or Cw alkyl.

The compound of claim 22, wherein R 1, R 3 and R 1 are H; and R 5 is C 1-6 alkyl optionally interrupted with one or more heteroatoms or carbonyl and optionally substituted with OH, SH, NR 'R 7 or C 1-6 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted with halogen, hydroxy, C 1-6 alkyl.

29. A compound according to claim 20 selected from: 0345 4-Oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1,2-alpyrazine-6-carboxylic acid [4-guanidino-1- (5- methyl thiazole-2-carbonyl-butyl] -amide; and 0340 4-Oxo-2- (3-phenylpropionyl) octahydropyrrolo [1,2-alpyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide. 9 EN 12019

30. The compound of claim 1 having the formula (IX):

wherein Y is selected from O, S, SO, SO 2, N-R 5 and CH-R *; R! is arginyl or an analogue or derivatives thereof optionally substituted by an amino acid, a peptide or a heterocycle; R 2 is H or C 1-6 alkyl; R3 is selected from H, NR6R7 and Ct <alkyl; and R4 and R8 are independently selected from H; NR «R7; aryl or C 3-7 cycloalkyl optionally substituted with Cu alkyl; CM6 alkyl groups are optionally interrupted by one or more heteroatom carbonyl groups and optionally substituted by OH, SH1NRJF1 or Ceearyl, heterocycle or C3-7cycloalkyl optionally substituted by halogen, hydroxy, C1-6 alkyl; amino acid side chain; and hydrophobic group. R8 is hydrogen, C1-6 alkyl is optionally interrupted by 1 or 2 heteroatoms; C6i6 aryl, C3-7cycloalkyl or heterocyclic ring or hydrophobic group; and n is 1 or 2;

The compound of claim 30, wherein R 1 is one of the formulas of formula V to Vid: r 'Via P > ^ 0-7 x (J) n VIb RtN < / > 0-6

'(J) n VIc ^ (J) n > 1-3 wherein: Rn is hydrogen or alkyl; K is a bond or -NH-; G is Cw alkoxy; cyano; -NH2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2; -CH 2 NH-C (NH) -NH 2; C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; or 5 or 6 membered, saturated or unsaturated heterocycle optionally substituted with cyano, -NH 2; -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or CH 2 -NH-NH (NH) -NH 2; U is cyano, -NH 2, -C (NH) -NH 2, or -NH-C (NH) -NH 2; P is a bond, -C (O) - or a bivalent group: OH. OH CH ΓΊ CH, or Ύ J is C 1-6 alkylene optionally substituted with OH, NH 2 and C 1-6 alkyl and optionally interrupted with a heteroatom selected from O, S and N; nirO or 1; and 10 T is Η, ΟΗ, an amine, a peptide chain, a C 1-6 alkyl group, a C 1-6 alkoxy group, a C 1-4 aralkyl group, or an optionally substituted heterocycle.

The compound of claim 31, wherein T is a heterocycle selected from the group consisting of

wherein: X5, X10, Xu and X12 are each independently selected from the group consisting of N, or C-X7 wherein X7 is hydrogen, C1-4 alkyl, or C5.8 aryl; X 6 and X 13 are each independently selected from the group consisting of C, Ο, N, S, N-X 7, or CH-X 7; and R 'is hydrogen, C 1-16 alkyl optionally substituted with carboxyl, carboxyl, -C 0-16 alkyl-CO 2 -C 1, 16 alkyl, C 0- aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 32, wherein T is selected from the group consisting of

wherein R 'is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, carboxyl, -C 1-6 alkyl-CO 2 -CM 6 alkyl, C 1-6 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 33, wherein T is selected from

Where R 'is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, carboxyl, -QM 6 alkyl-CO 2 -C 1-16 alkyl, aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 30, wherein R2 and R3 are both H.

36. The compound of claim 30, wherein R 1 is H, NR e R 7 or C 1-6 alkyl substituted with COOH.

The compound of claim 30, wherein R 5 is C 1 -C 6 alkyl, aralkyl, or C 1-6 alkyl substituted with C 3-7 cycloalkyl.

The compound of claim 32, wherein n is 1; R2, R3 and R are H; and R 5 is C 6, aryl, C 1-6 aralkyl, or C 1-6 alkyl substituted with C 3-7 cycloalkyl.

A compound according to claim 30 selected from 0890 3-Amino-4-oxo-2-phenyl-hexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1 - (benzothiazole) 2-carbonyl) -4-guanidino-butyl] -amide; 0895 3-Amino-2-benzyl-4-oxo-hexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] - amide; and 0900 3-Amino-2-cyclohexyl-4-oxo-hexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino] butyl] -amide.

40. The compound of claim 1 having the formula (X):

wherein B is selected from O, S, CH 2 -, or -NH-; R 1 is arginine or an analogue or derivatives thereof optionally substituted with an amino acid, peptide or heterocycle; R2 is H or C1-6 alkyl; R 3 is selected from H, NR ' R < 7 > and R 1 and R 8 are independently selected from H; NR «R7; C < m6 also lgrapras or C3.7 cycloalkyl optionally substituted with C1-4 alkyl; C1-16 alkyl optionally interrupted by one or more heteroatom carbonyl groups and optionally substituted by OH, SH, NReR7 or C 1-4 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted by halogen, hydroxy, alkyl; amino acid side chain; and hydrophobic group.

41. The compound of claim 40, wherein R 1 is one of the formulas of formula V to Vid: Via

G VIb

K. G 12

wherein Ru is hydrogen or alkyl; K is a bond or -NH-; G is CM alkoxy; cyano; -NH2; -CH2-NH2; -C (NH) -NH 2; -NH-C (NH) -NH 2; -CH2-NH-C (NH) -NH2; C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; or 5 or 6 membered, saturated or unsaturated heterocycle optionally substituted with cyano, -NH 2; -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH 2, or -CH 2 -NH-NH (NH) -NH 2; U is cyano, -NH2; -C (NH) -NH 2 or -NH-C (NH) -NH 2; P is a bond, -C (O) - or a bivalent group: -OH 'CH

, or

S J is C 1-6 alkylene optionally substituted with OH, NH 2 and C 1-6 alkyl and optionally interrupted by a heteroatom selected from O, S and N; nirO or l; and T is Η, OH, an amino group, a peptide chain, a CM6 alkyl group, a C 1, 6 alkoxy group, a C 1-4 aralkyl group, or an optionally substituted heterocycle.

The compound of claim 41, wherein T is a heterocycle selected from the group consisting of

wherein X5, X10, Xu and Xi2 are each independently selected from the group consisting of N, or C-X7 wherein X7 is hydrogen, C1-4 alkyl, or C5.8 aryl; X6 and Χ13 are each independently selected from the group consisting of C, Ο, N, S, N-X7, or CH-X7; and R 'is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, carboxyl, -C 0-6; alkyl-CO 2 -C 1-16 alkyl, C 1-6 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 42, wherein T is selected from the group consisting of 13 EN 12019

wherein R 'is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, carboxyl, -C 1-16 alkyl- CO- C 1-6 alkyl, C 6-20 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

44. The compound of claim 43 wherein T is selected from the group consisting of r

wherein R 'is hydrogen, C16 alkyl optionally substituted with carboxyl, carboxyl, -C0.16 alkyl-CO 2 -C 1-6 alkyl, C 1-6 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle.

The compound of claim 40, wherein R2 and R3 are both H.

The compound of claim 40, wherein R * is C 1-6 alkyl substituted with C 6 6 aryl optionally substituted with allyl.

47. The compound of claim 40, wherein R8 is H.

48. The compound of claim 42 wherein BirS14 R2, R3 and R5 are H; and R 1 is C 1-6 alkyl substituted with C 1-6 aryl optionally substituted with C 1-6 alkyl.

A compound according to claim 40 selected from 925 7-Benzyl-6-oxo-octahydro-pyrido [2,1c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1 (thiazole-2-carbonyl) ) butyl] amide; and 940 6-Oxo-7-phenethyl-octahydro-pyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1 (thiazole-2-carbonyl) -butyl] -amide;

A method of treating and preventing thrombotic disorders in a mammal comprising administering an effective amount of the compound of claim 1.

51. The method of claim 50, wherein the thrombotic phenomenon is venous thrombosis.

52. The method of claim 50, wherein the thrombotic phenomenon is pulmonary embolism.

53. The method of claim 50, wherein the thrombotic event is arterial thrombosis.

54. The method of claim 50, wherein the thrombotic phenomenon is a myocardial infarction.

55. The method of claim 50, wherein the thrombotic phenomenon is a cerebral infarction.

56. The method of making a compound according to claim 1.

57. A method for producing a compound according to any one of claims 10, 20, 30 or 40.