CA2208772A1 - Low molecular weight bicyclic thrombin inhibitors - Google Patents

Abstract

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

Description

W 0~6/19483 PCT/CA95/00708 LOW M~T~FrUI~R ~nEIGHT BICYCLIC
THROMBIN INHIBITORS

FIE~D OF THE lNv~:NlION
This invention relates to compounds useful for the treatment of thrombotic disorders, and more particularly to novel heterocyclic inhibitors of the enZyme thrombin.

BACKGROUND

Inordinate thrombus formation on blood vessel walls precipitates acute cardiovascular disease states that are the chief cause of death in economically developed societies. Plasma proteins such as fibrinogen, proteases and cellular receptors participating in hemostasis have emerged as important factors that play a role in acute and chronic coronary disease as well as cerebral artery disease by contributing to the formation of thrombus or blood clots that effectively ~;m;n;sh normal blood flow and supply. Vascular aberrations stemming from primary pathologic states such as hypertension, rupture of atherosclerotic pla~ues or denuded endothelium, activate kiochemical 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 ~ 35 exerts direct effects on diverse cell types that support and amplify pathogenesis of arterial thrombus disease.
The enzyme is the strongest activator of platelets causing them to aggregate and release substances (eg. ADP TXA NE) that further propagate the thrombotic cycle. Platelets in a fibrin mesh comprise the principal framework of a white r thrombus. Thrombin also exerts direct effects on 5 endothelial cells causing release of vasoconstrictor substances and translocation of adhesion molecules that become sites for attachment of immune cells. In addit-ion, the enzyme causes mitogenesis of smooth muscle cells 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 natural anticoagulant protein inhibitors of 30 thrombin activity (Markwardt F. Cardiovascular Drug Peviews, 10, 211, 1992). It is a biopharmaceutical that has demonstrated efficacy in experimental and clinical thrombosis. A potential drawback to the use of Hirudin as a therapeutic agent is likely antigenicity and lack of an 35 effective method of neutralization, especially in view of its extremely tight binding characteristics toward thrombin. The exceedingl~- high affinity for thrombin is unique and is attributed to a simultaneous interaction with the catalytic site as well as a distal "anion binding exosite" on the enzyme.

Thrombin activity can also be abrogated by Hirudin-like molecules such as hirulog (Maraganore, J.M. et al., Biochemistry, 29, 7095, 1990) or hirutonin peptides ~DiMaio, J. et al., J. Med. Chem., 35, 3331, 1992).

Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for thrombin's catalytic site, thereby inhibiting proteolysis of that protein or other protein substrates such as the thrombin receptor. A common strategy for designing enzyme inhibitory compounds relies on mimicking the specificity inherent in the primary and secondary structure of the enzyme's natural substrate. Thus, Blomback et al. first designed a thrombin inhibitor that was modeled upon the partial sequence of the fibrinogen A(Lsl)~ chain comprising its proteolytically susceptible region (Blomback, et al., J. Clin. Lab. Invest., 24, 59, 1969). This region of fibrinogen minimally includes the residues commencing with phenylalanine:

-la-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly -Gly-Gly-Val-Arg-Gly-Pro-Arg ~ scissile bond Systematic replacement of amino acids within this region has led to optimization of the tripeptidyl inhibitory sequence exemplified by the peptide (D)-Phe-Pro-Arg which corresponds to interactions within the P-P-P local binding sites on thrombin (Bajusz S. et al. in Peptides: Chemistry Structure and Biology: Proceedings of the Fourth American W 096/19~83 PCT/CA95/00708 Peptide Symposium, Walter R., Meienhofer J. Eds. Ann Arbor Science Publishers Inc., Ann Arbor MI, 1975, pp 603).

Bajusz et al. have also reported related compounds such as 5 (D)Phe-Pro-Arg-(CO)H (GYKI-14166) and (D)MePhe-Pro-Arg-(CO)H (GYKI-14766) (Peptides-Synthesis, Structure and- t Function: Proceedings of the Seventh American Peptid~
Symposium, Rich, D.H. & Gross, E. eds., Pierce Chemical Company , 1981, pp. 417). These tripeptidyl aldehydes are l0 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! residue, generating a l5 hemiacetal intermediate.

Related work in the area of thrombin inhibitory activity has exploited the basic recognition binding motif engendered by the tripeptide (D)Phe-Pro-Arg while 20 incorporating various functional or reactive groups in the locus corresponding to the putative scissile bond (i.e.
Pl-Pl' ) .

In U.S Patent 4,318,904, Shaw reports chloromethyl-25 ~-.etones (PPACK) that are reactive towards Ser and His .
These two residues comprise part of thrombinls catalytic triad (Bode, l~l. et al., EMBO Journal 8, 3467, 1989).

Other examples of thrombin inhibitors bearing the (D)Phe-30 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)-35 Still other congeners of this motif are those bearing phosphonates (Wang, C-L J., Tetrahedron Letters, 33, 7667, W O 96/19483 PCTICA9~/00708 1992) and ~-Keto esters (Iwanowicz, E.J. et al.,Bioorganic and Medicinal Chemistry Letters, 12, 1607, 1992).

Neises, B. et al. have described a trichloromethyl ketone c 5 thrombin inhibitor (MDL-73756) and Attenburger, J.M. et al. have revealed a related difluoro alkyl amide ketone (Tetrahedron Letters, 32, 7255, 1991).

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

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

Still others have focused their attention on peptides, peptide derivatives, peptidic alcohols, or cyclic peptides as anti-thrombotic agents (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 agents. The chemical structures described in these documents represent variations of side groups on the arginin~mide compound (U.S. 4,173,630; U.S.

W 096/19483 PCT/CA9~/00708 4,097,591; CA 1,131,621; U.S. 4,096,255; U.S. 4,046,876;
U.S. 4,097,472; CA 2,114,153).

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

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

One object of the present invention is to provide thrombin inhibitors that display inhibitory activity towards the target enzyme, thrombin.

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.

Still a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and ~ormulations thereof as anticoagulant and thrombin inhibitory agents Yet a further object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and formulations thereof for therapeutic treatment of various thrombotic maladies.

W 096/19483 PCT/CA9~/00708 A ~urther object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The enzyme inhibitors o~ the present invention are encompassed by the structure of general Formula I.

W 096/19~83 . PCT/CA95/00708 SU~ RY OF THE lNv~N~lIoN

The present invention provides for novel compounds that .
display thrombin inhibitory activity as re~lected in formula I:
,Y~_,A~

R~f ~R
z (I) wherein:
A is selected from (CH-RL) l~-! t S, SO, SO, O and NR wherein R8 is hydrogen, C alkyl optionally interupted with 1 or

2 heteroatomsi C,i; aryl, C cycloalkyl or heterocyclic ring or a hydrophobic group;
B is selected from S, SO, O, -M=, NH, -CH= and CR.R
wherein R6 and R7 are independently selected from hydrogen and C alkyl provided that when A is S, SO, SO, O, or NR~, then B is CR.R.. ;
D is selected from (CH-RL)I wherein R~ is hydrogen, C .
alkyl or -C(O)R; and CH with a double bond to B when B
is -N= or - CH=;
E is selected from CH and CH substituted with the -C(O)R, provided that only one of D and E is substituted with with -C(O) Rl;
X is selected from O, N-R~, or CH~Rr;
Y is selected ~rom O, S, SO, SO, N-R, and CH-RL provided that when X is N-R then Y is CH-R~. or O, and when X is O
then Y is CH-Rc i Z is selected from O, S and H;
Rl is a polar amino acid residuearginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;

R2 is selected from H and C alkyl optionally substituted with C aryl, a 6 member heterocycle or a C ~ cycloalkyl ring;
R3 is selected from H, NR~R and C1c alkyl; and ~ 5 R~ and Rs are independently selected from H; NR,.R; C~1~ aryl or C cycloalkyl optionally substituted with C,,. alkyl;
Cj1 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR R or a C , aryl, heterocycle or C, cycloalkyl group optionally substituted with halogen, hydroxyl, C!.. alkyl; an amino acid side chain; and a hydrophobic group.

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

DE~ATT~n DESCRIPTION OF THE lNv~:N~l~IoN

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

X--Y~A'B
R4~ ~ N '~

(I) wherein X, Y, Z, A, B, D, E and R to R; are as previously defined.

W 096/19483 PCT/CAg5/00708 The term "hydrophobic group" (HG) as used hereinafter, refers to any group which lacks affinity for, or displaces water. Hydrophobic groups include but are not limited to C alkyl, C ,alkenyl (e.g. vinyl, allyl) or C ,alkynyl (e.g. propargyl) optionally interrupted by a carbonyl group, (e.g. forming an acyl group); C i aryl, C
cycloalkyl, C aralkyl, C cycloalkyl substituted C, alkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group (e.g. ~orming 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 o~ the natural residue may incorporate a longer or shorter methylene chain from the alpha carbon (i.e. ethylene or butylene chain);
replacement of the guanidino group with a hydrogen bond donating or accepting group (i.e. amino, amidino or methox.y); replacement of the methylene chain with a ccnstrained group (i.e. an aryl, cycloalkyl or heterocyclic ring); elimination of the terminal carboxyl (i.e. des-carboxy) or hydroxyl (i.e. an aldehyde); or a combination thereof.

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

The term "aromatic" or "aryl" represents an unsaturated carbocyclic ring(s) of 6 to 16 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NR R) halogen or C alkyl. Aromatic rings include benzene, napththalene, phenanthrene and anthracene.
Pre~erred aromatic rings are benzene and naphthalene.
'~.
The term "cycloalkyl" represents a saturated carbocyclic ring o~ 3 to 7 carbon atoms which is optionally mono- or di-substituted with OH, SH, amino (i.e. NR~R.) halogen or C alkyl. Cycloalkyl groups include cyclo- propyl, butyl, pentyl, hexyl and heptyl. A preferred cycloalkyl group is cyclohexyl.
The term "aralkyl" represents a substituent comprising an aryl moiety attached via an alkyl chain (e.g. benzyl, phenethyl) wherein the sum total of carbon atoms for the aryl moiety and the alkyl chain is as specified. The aryl or chain portion of the group is optionally mono- or di-substituted with OH, SH, amino (i.e. NRR) halogen or C
alkyl The term "heteroatom/' as used herein represents oxygen, nitrogen or sulfur (O, N or S) as well as sulfoxyl or sul~onyl (SO or SO) unless otherwise indicated. It is understood that alkyl chains interrupted by one or more heteroatoms means that a carbon atom of the chain is replaced with a heteroatom having the appropriate valency.
P~ererrabl~, an alkyl chain is interrupted by 0 to 4 heteroatoms and that two adjacent carbon atoms are not both replaced.

The term "heterocycle" represents a saturated or unsaturated mono- or polycyclic (i.e. bicyclic) ring incorporating 1 or more (i.e. 1-4) heteroatoms selected from N, O and S. It is understood that a heterocycle is optionally mono- or di-substituted with OH, SH, amino (i e. NR R), halogen, CF, oxo or C alkyl. Examples of suitable monocyclic heterocycles include but are not limited to pyridine, piperidine, pyrazine, pipera-~ine, pyrimidine, imida~ole, thiazole, oxazole, furan, pyran and thiophene. Examples of sultable bicyclic heterocycles include but are not limited to indole, ~uinoline, isoquinoline, purine, and carbazole.

The term "hydrophobic amino acid" represents an amino acid residue that bears an alkyl or aryl group attached to the a-carbon atom. Thus glycine, which ~as no such group attached to the a-carbon atom is not a hydrophobic amino acid. The alkyl or aryl group can be substituted, provided that the substituent or substituents do not detract from the overall hydrophobic character of the amino acid.
Examples of hydrophobic amino acids include natural amino acid residues such as alanine; isoleucine; leucine;
phenylalanine; and non-naturally ocurring amino acids such as those described in "The Peptides", vol. 5, 1983, Academic Press, Chapter 6 by D.C. Roberts and F.
Vellaccio. Suitable non-naturally ocurring amino acids include cyclohexylalanine and l-aminocyclohexane-carboxylic By ~amino acid side chain" is meant the substituent attached to the carbon which is a to the amino group. ~or example, the side chain of the amino acid alanine is a methyl gro~p and while benzyl is the side chain for phenylalanine.

Preferably R2 is H or C alkyl. More preferably R is ~i, methyl or ethyl and most preferably R is H.

Preferably, R3 is H or Cj alkyl. More preferably, R is ~ methyl or ethyl, and most preferably R is H.

Preferably, one of R4 or Rs is a hydrophobic group such as a saturated or unsaturated carbocycle of 5 or 6 members optionally fused to another carbocyclic group while the other is H, C, alkyl optionally substituted by NRR or carboxy. The hydrophobic moiety may be linked via a spacer such as a Cl,~ alkyl chain optionally interrupted with 1 or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO) groups. More preferably, one of R~ and Rs is phenyl, cyclohexyl, indole, thienyl, ~uinoline, tetrahydroisoquinoline, naphthyl or benzodioxolane linked via C!; alkyl optionally interupted with a heteroatom or a carbonyl while the other is H, carboxymethyl or carboxyethyl.
Preferably, A is absent or CH .
Preferably, B is S or CH .
Preferably, D is CH .
Preferably, E is CH substituted with -C(O)R wherein R is as previously defined.
Preferably, X is CH-R, or N-R~.
Preferably, Y is CH-R or S.
Preferably, Z is O.

In a preferred embodiment, Rl is represented by one of formula VIa to VId:

R"'N~p~ ,T ~ R,lN
Vla K~J )0 7 Vlb G

U~ ~i wherein:
Rll is hydrogen or C alkyl;
K is a bond or -NH-;
G is C alkoxyi cyano; -NH; -CH -NH; -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)-NH or -CH -NH-C(NH)-NH; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH, -CH -NH, -C(NH)-NH., -NH-C(NH)-NH or -CH -NH-C(NH)-NH;
U is cyano, -NH, -C(NH)-NH. or -MH-C(NH)-NH.;
P is a bond, -C(O)- or a bivalent group:
OH
CH ~ CH
S~ ~S
CH , or -J is C alkylene optionally substituted with OH, NH and C alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and T is H, OH, amino, a peptide chain, Cjj alkyl, C , alkoxy, C aralkyl, or heterocycle optionally substituted.

Preferably Rll is H or methyl and most preferably H.
Preferably K is a bond.
Preferably G is -NH-C(NH)-NH attached via a methylene chain of 3-7 carbons or phenyl substituted with -C(NH)-NH
attached via a methylene chain of 0 to 3 carbons. More preferably G -NH-C(NH)-NH attached via a methylene chain of 3 atoms.
Pre~erably P is -C(O)-.
Preferably J is selected from: -CH -S-CH-CH-; -CH-O-CH -CH-; -CH -NH-CH -CH -; and a bond when n is 0. More pre~erably, 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 Bioory. Med.
Chem., 1995, 3:1145 :

l4 . CA 02208772 1997-06-20 ,,N~

(~NH (~NH ~NH

NH~ N~rMe ~ H2 N~JI~ " N~V~ ~ ~N ~, (q~NH (~NH (~NH
Nd~ NH2 N/~N N~\O
S--N
H2N o H2N
H~J~ H~ , ~.1T

( ~n~3H ~'2NH2 (~NH2 N

~T ~ N ~Jl~ N ~Jl~

N~N (~f~NH

~ T "~N ~

HN~NH2 T~zO T~O

N "~ --., NJ~", T~

HN~ NH H2N NH ~

o o o N ~,11~ ~ ~ N ~Jl~ ~ ~ N ~JI~T
(~ (~ (~
XJ ~NH2 H2N ~

~
(~ (~ (~
~ ~NH2 H2N

N~JI~ ,,N~JI~ _ 1~ q H~

~N~l T , ' ~1T
(~ (~ (~

NJ~NH2 HN ~NH

,~T ' .~T

H ~1 ~N~NH2 HN

T N~ll~ N~JI~

H2N~ ~NH2 "N~T N~l~ N,JI~

H~O ~N~NH

o o H~J ~T

HN~N~G
H2N~l'NH
NH

N ~J~ T

NH~ ~NH _~ NH, NH NH

wherein n=l-6, nl=1-2, n2=0-7 and T is as previously defined.

In a preferred embodiment, T is a peptide of 1 to 4 amino acid residues in length and preferably fibrinogen~s A or B
chain or fragment or derivative thereof. In another preferred embodiment, T is a heterocycle selected from the group consisting of:

--R' T~ ~ R

~ < X~ J X

wherein Xsl X10, Xll and Xl2 are each independently selected from the group consisting of N, or C-X where X7 is hydrogen, C
alkyl, or C aryli X and X13 are each independently selected from the group consisting of C, O, N, S, M-X,, or CH-X.;
r R' is hydrogen, C,i alkyl optionally carboxyl substituted, carboxyl, -CG_;C alkyl-CO -C alkyl, C~ . aralkyl, C
5 cycloalkyl, aryl or an aromatic heterocycle.

Preferably T is selected from the group consisting o~:
~N\[ ~j ~N~ ~N~,~

~1 ~ "~ s~ ~'~' ~3 N~3 N ~ N --~N~ ~o~ R'~ R

~N N N--N ~ S~

wherein R~ is as defined above.
More pre~erably T is selected ~rom the group consisting of:

/ 1 ~ 1' N --~

R R' W O 96/19483 PCT/CA9~/00708 wherein R' is as defined above.

More preferably T is selected from the group consisting of:
~ R' ~, wherein R' is as defined above.

Most preferably T is ~ ~ or R~ ~ ~

wherein R' is H or C , alkyl such as methyl, ethyl, propyl or butyl and most preferably wherein R' is hydrogen,. In another embodiment, T is a 1,2 thiazole optionally substituted with R' and\or is attached to J at the 2, 3, 4 or 5 position of the ring.
In particular embodiments, compounds of the invention are represented by formulas II, III, IV and V, wherein X, Y, B, R to R andR are as previously defined.

(II) R~R, R5~R

(IV) R ~ ~ R (V) N

3 PCT/CA95/00708 In a particularly preferred embodiment, compounds of the invention are represented by one of formulas VII, VIII, IX
and X:

~ R, ~ R, R5~--Y'Ih,R8 R5 ~ ~ B
R4?~¢ N ~CH2)n ~ $ R

wherein B is O, S, -CH.-, or -NH-;
Y is selected from O, S, SO, SO~, N-R5 and CH-R~3;
R1 is an arginyl moiety or an analog or derivative thereo~
optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or Cic alkyli R3 is selected from H, NR,R7 and C,~ alkyl; and R, and Rs are independently selected from H; NRtR; Cc, aryl or C . cycloalkyl optionally substituted with Cl alkyl;
C l alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NRR7 or a C,i~ aryl, heterocycle or Cj~
cycloalkyl group optionally substituted with halogen, hydroxyl, Cl, alkyli an amino acid side chaini and a hydrophobic group;
R8 is hydrogen, Cit alkyl optionally interupted with l or 2 heteroatoms; Ct. aryl, C3. cycloalkyl or heterocyclic ring or a hydrophobic group; and n is l or 2.
Preferred compounds according to formula VII include:

s~J~ r~ ~5 IIEET

0005 6S-benzylhexahydro-5-oxo-5H- ~ s thiazolo[3,2-a] pyridine-3R-Ph~ ~.~N~ ~
carboxamido (propyl ~
ketoarginine) NH

0010 6S-benzylhexahydro-5-oxo-5H- ~ s thiazolo[3,2-a] pyridine-3R- ~ ~ ~N~
carboxamido (butyl ~ J
ketoarginine) NH

0015 6s-benzylhexahydro-5-oxo-5H- ~ s thiazolo[3,2-a] pyridine-3R- Ph~ ~.~N~ ~ OCH3 carboxamido(propylcarbmethoxy ~ J
ketoarginine) NH

0020 6S-cyclohexylmethyl ~ ~ s hexahydro-5-oxo-5H- ~. N~
thiazolo[3,2-a]pyridine-3R- ~ J
carboxamido(benzylketo NH
arginine) HNlNH

0025 6S-cyclohexyl methyl ~ ~ s hexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine -3R- ~ J O
carboxamido(carbmethoxy ~NH
propyl HN NH2 cyclodithioketalarginine) 0030 6S-cyclohexylmethyl ~ ~ s hexahydro-5-oxo-5H- ~"~N~ H ~
thiazolo[3,2-a]pyridine-3R- O o~-N
carboxamido((S)-Arg-(R)-pipecolilic acid) lH

W O 96/19483 . PCT/CA95/00708 0035 6S-benzylhexa hydro-5-oxo-5H- ~ ~ s thiazolo [3,2-a]pyridine-3R- ~ O ~ h ~ NH2 carboxamido(carboxamidopropyl ~ O
cyclodithioketal arginine) ~NH

0040 6S-cyclohexylmethylhexahydro- ~ ~rS
5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((S)-Arg NH
HN NH.
nlpecotamlde ) 0045 6S-cyclohexylmethylhexahydro- ~ ~ s 5-oxo-5H-thiazolo[3,2- ~ ~N ~ X o a]pyridine-3R- ~ ~ O
carboxamido((S)Arg NH NH, isonipecotamide) HN NH2 0050 6S-benzylhexahydro-5-oxo-5H- ~ , rS
thiazolo[3,2-a]pyridine-3R- "~N~H ~ NH
carboxamido(carboxamidopentyl ~ ~
cyclodithioketal arginine) lH
HN NH

0055 6S-benzylhexahydro-5-oxo-5H- ~ ~ rS
thiazolo[3,2-a]pyridine-3R- O ~ N ~ c~
carboxamido(carbmethoxy ~ G
propyl cyclodithioketal ~NH
arginine) HNlNH

0060 6S-cyclohexylmethylhexahydro- ~ ~ _s 5-oxo-5H-thiazolo[3,2- ~ ~H
t a]pyridine-3R-carboxamido(1- ~ ~.
carboxy-3-thiobutyl ~ 5 NH ,~
~ ketoarginine) HN~ O~

S ~ T

0065 6S-cyclohexylmethylhexahydro- ~ ~~rs 5-oxo-5H-thiazolo[3,2- ~ ~ ~ H
a]pyridine-3R-carboxamido(l- ~
carboxy-3-thiobutyl ~ s '=
ketoarginine) ~NH
NH2 OH .~
0070 6S-cyclohexylmethylhexahydro- ~ / s 5-oxo-5H-thiazolo[3,2- ~ '~N ~ H
a]pyridine-3R-carboxamido(l- ~
carboxy-2-methyl-3-thiobutyl ~ s ketoarginine) ~NH

0075 6S-cyclohexylmethylhexahydro- ~ / ~s 5-oxo-5H-thiazolo[3,2- .. ~N~
a]pyridine-3R-carboxamido((3- ~
thiobutyl sulfonic acid) NH S
ketoarginine) HN~
NH~ "~ --O
OOH
0080 6S-cyclohexylmethylhexahydro- ~ ~ s 5--oxo--5H--thiazolo[3,2-- ~ ~ N ~ H o N~
a]pyridine-3R- ~ c carboxamido(iso-quinolinium NH
methyl ketoarginine) HNlNH

0085 6S-cyclohexylmethylhexahydro- ~ ~ s 5-oxo-5H-thiazolo[3,2- .~,N ~
a]pyridine-3R- ~ ~ ~
carboxamido(propylcarbmethoxy ~ ~
ketoarginine) ~NH ~ - O
NH, 0090 6S-cyclohexylmethylhexahydro- -~ ~ s . ''~, N _~>
5-oxo-5H-thiazolo[3,2- ~ o~N ~ Nh a]pyridine-3R- ~ H
carboxamido((propylketo)Arg- H~J~ NH~
Phe-Arg-NH)NH ~ IH ~

SU~ST~ r~ 5.4~

O095 6s-benzylhexahydro-5-oxo-5H- ~ /~ S
thiazolo[3,2-a]pyridine-3R- ~.~'~N~
carboxamido((propanoic acid) ~ ~
ketoarginine) HN ~ o~~ OH
~ NH2 O100 6s-benzylhexahydro-5-oxo-5H~ s thiazolo[3,2-a]pyridine-3R- ~~~' ~N~
carboxamido(propyl ~ ~
carbmethoxy ketoarginine) 0105 6S-cyclohexylmethylhexahydro- ~~~ ~ s 5-oxo-5H-thiazolo [3,2- ~ l~ ~ ~ H
a]pyridine-3R-carboxamido (a- ~ s benzothiazolo keto arginine); ~NH ~N~
and HN

O110 6S-cyclohexylpropylhexahydro~ s 5-oxo-5H-thiazolo [3,2- ~~~~~~ '~ ~H
a]pyridine-3R- ~ ~
carboxamido(propylcarbmethoxy NH
ketoarginine) NH~ ~

SF~T~ i F~ ~T

0205 6-Benzyl-5-oxo-hexahydro- H
thiazolo[3,2-a]pyridine-3- ~ ~ ,N~
carboxylic acid [1- O . ~ s (benzothiazole-2-HN NH
carbonyl)-4-guanidino- 2 butyl]-amide 0210 6-Benzyl-5-oxo-hexahydro- H
thiazolo[3,2-a]pyridine-3- ~ ~ ~ ~N~
carboxylic acid [1- 0 ~ s (benzothiazole-2- l HN NH
carbonyl)-4-guanidino- 2 butyl]-amide 0215 6-Benzyl-5-oxo-hexahydro- H
thiazolo[3,2-a]pyridine-3- ~1! N~, carboxylic acid [1- O ~ s O NH
(benzothiazole-2-HN NH
carbonyl)-4-guanidino-butyl]-amide 0220 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2- ~-~ N ~ ~ ~N~
a]pyridine-3-carboxylic 0~ NH -acid [1-(benzothiazole-2- NH NH, carbonyl)-4-guanidino-butyl]-amide S~ T~T S~rT

0225 8a-Methyl-5-oxo-6-phenethyl-hexahydro-~,~N ~N~
thiazolo[3,2-a]pyridine-3- o ~ NH S
carboxylic acid [1- NH NH2 (benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0230 8a-Methyl-5-oxo-6-phenethyl-hexahydro- ~N~ HN~N~
thiazolol3,2-a]pyridine-3-~ ~ ~ NH S
carboxylic acid Ll- NH NH2 (benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0240 8a-Methyl-5-oxo-6-(2- CF3 N ~~~S
trifluoro methyl-quinolin- ~ N > ~ ~N~
6-ylmethyl)-hexahydro- O ~
thiazolo[3,2-a] pyridine-3- NH-~NH2 carboxylic acid [1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0245 6-Benzyl-5-oxo-hexahydro- H
thiazolo[3,2-a]pyridine-3- ~ N
carboxylic acid [4- O Y ~ s~
guanidino-1-(thiazole-2-carbonyl)butyl]-amide ~;UE~5T~l~U~E S~EET

0250 6-Benzyl-5-oxo-hexahydro- ,H
thiazolo[3,2-a]pyridine-3- ~ ;,N~ I <N~
carboxylic acid [4- O ~ S
- guanidino-l-(thiazole-2-carbonyl)butyl]-amide 0255 6-Benzyl-S-oxo-hexahydro- - H
thiazolo[3,2-a]pyridine-3- ~ <N
carboxylic acid [4- O 11 ~ N
guanidino-l-(l-methyl-lH NH Me imidazole-2-carbonyl)butyl]-amide 0260 6-Benzyl-8a-methyl-5-oxo- Me hexahydro-thiazolo[3,2- ¢~ "~ < 3 a]pyridine-3-carboxylic O ~NH
acid [4--guanidino--1-- HNlNH2 (thiazole-2-carbonyl)-butyl]-amide 0265 5-Oxo-6-(3-cyclohexyl- H
propyl)-hexahydro- Q~ , 3 thiazolo[3,2-a]pyridine-3- o ' C s ~ NH
carboxylic acid [4-- HNlNH2 guanidino-l-(thiazole-2-carbonyl)butyl]-amide 027S 8a-Methyl-5-oxo-6-(3- ~_ phenyl-propyl)-hexahydro- ~ A N ~ N
thiazolol3,2-a]pyridine-3- o " ~ s O NH
carboxylic acid [4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide ~ lJ E~T~ S iY ~ ET

W 096119483 . PCT/CA95/00708 0280 8a-Methyl-5-oxo-6-(3- ~
phenyl-propyl)-hexahydro- ~N~ N ~N
thiazolo[3,2-a]pyridine-3- " ~ ' s O NH
carboxylic acid [4-HN ~--NH2 guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0285 8a-Methyl-5-oxo-6-(2- CF~ N ~~~S
trifluoromethyl-~uinolin- ~I N~ HN ~N~
6-ylmethyl)-hexahydro- O ~ ~ NH
thiazolo[3,2-a]pyridine-3- ~NH - ~NH
carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0295 6-(1,3-Dioxo-1,3-dihydro- 0 ,~~ s isoindol-2-yl)-5-oxo- ~ ~ ~N3 hexahydro-thiazolo¦3, 2- 0 0 ~ ~H

a]pyridine-3-carboxylic acid L4-guanidino-1-(thiazole-2-carbonyl)-butyll-amide 0305 5-Oxo-6-(3-phenyl- , ~ s propionyl amino)-hexahydro ~ N~ N~'N3 thiazolo[3,2-a]pyridine-3- ~ o ~ _~H

carboxylic acid ~4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide T!I E L~ S~ ET
-W 096/19483 PCTlCAgS/00708 0315 5-Oxo-6-(3-phenyl- H S
propionyl amino)-hexahydro ~ ~ ~N ~ ~ ~ N3 thiazolo[3,2-a]pyridine-3- ~ ~ ~ ~H
carboxylic acid [4-NH NH2 guanidino-l-(thiazole-2-carbonyl)-butyl]-amide - More preferred compounds according to ~ormula (VII) include:
0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido(propylcarbo - methoxyketoarginine)i 0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo[3,2-a]pyridine-3R-carboxamido((propylketo)Arg-Phe-Arg-NH );
l0 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 (~-benzothiozolo keto arginine);
l5 0210 6-Benzyl-5-oxo-hexahydro-thiazolol3,2-a]pyridine-3-carboxylic acid [l-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amidei O ,0 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolol3,2-a]pyridine-3-carboxylic acid ~ benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide;0240 8a-Methyl-5-oxo-6-(2-tri~luoromethyl-~uinolin-6-ylmethyl)-hexahydro-thiazolo[3,2-a3pyridine-3-carboxylic acid ~1-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide;
0245 6-Benzyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-- 25 carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)butyl]-amide;

T ~ E~

-0260 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide;
0265 5-Oxo-6-(3-cyclohex~l-propyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid L4-guanidino-1-(thiazole-2-carbonyl)butyll-amide;
0285 8a-Methyl-5-oxo-6-(2-tri~luoromethyl-quinolin-6-ylmethyl)-hexahydro-thiazolo[3,2-a]pyridine-3-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide; and 0315 5-Oxo-6-(3-phenyl-propionylamino)-hexahydro thiazolo[3,2-a]pyridine-3-carboxylic acid ~4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide.

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-a]pyridine-3R-carboxamido (a-benzothiozolo keto arginine).

SUeSTll~U~E ~ T

Preferred compounds according to ~ormula VIII include:

- 0325 3-~mlnomethyl-2-benzoyl-4-oxo- ~
octahydro-pyrrolo[1,2-o~N~ Ho a]pyridine-6-carboxylic acid ~ N ~ , [l-(benzothiazole-2-carbonyl)-HN

- 4-guanidino-butyl]-amideHN~NH2 0330 3 -Arninomethyl-4-oxo-2- ~3~
phenylacetyl-octahydro-o~N~ Ho pyrrolo[l,2-a]pyrazine-6-carboxylic acid [1-lbenzothiazole-2-carbonyl)-4-: guanidino-butyl]-amide 0335 2-Benzoyl-4-oxo-octahydro- . ~ H
pyrrolo[l,2-a]pyrazine-6- ~ ~, ~ ~ ~
carboxylic acid ¦4-guanidino- ~ ~ ~ ~L s l-(thiazole-2-carbonyl)- NH NH2 butylJ-amide 0340 4-Oxo-2-(3-phenyl-propionyl)- ~ H
octahydro-pyrrolo~1,2- ~N~
a]pyrazlne-6-carboxyllc acid ~ o r~ NH S
¦4-guanidino-1-(thiazole-2- NHlLNH2 carbonyl)-butyl]-amide S'~51r5~Ul~E SiYEET

0345 4-Oxo-2-(3-phenyl-propionyl)- ~ H
octahydro-pyrrolo[1,2- ~N~ ~, a~pyrazine-6-carboxylic acid~ ~ NH S

[4-guanidino-1-(5-methyl-thiazole-2-carbonyl)-butyl]-amide 0350 2-(3-Cyclohexyl-propionyl)-4- ~ H
oxo- octahydro-pyrrolo[1,2- ~ N~ ~ ~ ~ N
a]pyrazine-6-carboxylic acid ~ ~ N~ s3 [4-guanidino-1-(2-thiazole- NH NH2 carbonyl)-butyl]-amide 0355 5-Oxo-7-(3-phenyl-propionyl)- ~ H
octahydro-2-thia-4a,7-diaza- ~ ~ ~ ~ O
naphthalene-4-carboxylic acid ~ ~~ ~ N~H s3 [4-guanidino-1-(thiazole-2- NH NH2 carbonyl)-butyl]-amide 0365 4-Oxo-2-(4-phenyl-butyryl)-octahydro-pyrrolol1,2-a]pyrazine-6-carboxylic acid ~ ~HN 5 [4-guanidino-1-(thiazole-2- H N~
carbonyl)-butyl]-amide NH
0370 4-Oxo-2-phenylacetyl-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2- ~
carbonyl)-butyl]-amide NH

JlrE ~ T

~375 2-(2-Amino-3-phenyl- ~
propionyl)-4-oxo-octahydro- ~ N~"
pyrrolo[l,2-a] pyrazine-6- ~ o~HN
carboxylic acid [4-guanidino- I~N~
~ l-(thiazole-2-carbonyl)- NH
butyl]-amide 0380 2-[2-Amino-3-(4-hydroxy- ~
phenyl)-propionyl]-4-oxo- HO ~ N '~-~
octahydro-pyrrolo[1,2-a] ~N
NH O
pyrazine-6-carboxylic acid [4- H~N~NH
~ guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0385 2-12-Amino-3-(4-fluoro- ~N~"
phenyl)-propionyl]-4-oxo- 0 ~HN
octahydro-pyrrolo[1,2-a] ~-N
pyrazine-6-carboxylic acid [4- NH
guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 0390 4-Oxo-2-(3-phenyl-propyl)- ~N~
octahydro-pyrrolo[1,2-a] ~ ~ s~
O o H \ ,~.= N
pyrazine-6-carboxylic acid 14- NH~, guanidino-l-~thiazole-2- H'N~NH
carbonyl)-butyl]-amide 0395 2-12-Amino-3-(lH-indol-3-yl)-propionyl]-4-oxo-octahydro-NH NH ~N~
pyrrolo[l,2-a]pyrazine-6-carboxylic acid [4-guanidino-NH
l-(thiazole-2-carbonyl)-butyl]-amide ~, .

w~ S ~! ~ E~

.

0400 4-Oxo-2-(3-thiophen-3-yl- ~
propionyl)-octahydro- ~ N~"
pyrrolo[l,2-a] pyrazine-6- o ~HN
carboxylic=acid [4-guanidino- NH~o l-(thiazole-2-carbonyl)- NH
butyl]-amide 0405 4-Oxo-2-(3-thiophen-2-yl- ~
propionyl)-octahydro- ~ N~"
pyrroloLl,2-a] pyrazine-6- o ~HN
carboxylic acid [4-guanidino-"N~NH ~ O
l-(thiazole-2-carbonyl)- NH
butyl]-amide 0410 2-(3-1 H-Imidazol-4-yl- ~
propionyl)-4-oxo-octahydro- N~l ~ N '~
pyrrolo[l,2-a] pyrazine-6- ~ o~HN~
carboxylic acid [4-guanidino- ff l-(thiazole-2-carbonyl)- Nll butyl]-amide 0415 ~
2-(2-Amino-3-thiophen-3-yl- ~ N~.
propionyl)-4-oxo octahydro ~ ~HN
pyrrolo[l,2-a] pyrazine-6- HN~H ~ ' carboxylic acid [4-guanidino- N~l l-(thiazole-2-carbonyl)-butyl]-amide 0420 4-Oxo-2-(1,2,3,4-tetrahydro- ~
iso~uinoline-3-carbonyl)- ~ N~"
octahydro-pyrroloLl,2-a] ,. C~HN
pyrazine-6-carboxylic acid [4- H~Nff guanidino-l-(thiazole-2- Nll carbonyl)-butyl]-amide .

5~J~-'5~ JTE ~ Fr W 096119483 . PCT/CA95/00708 0425 2-(Hydroxy-phenyl-acetyl)-4- ~ O
oxo-octahydro-pyrrolo[1,2-a]
- pyrazine-6-carboxylic acid [4- O ~HN S~
O \ )--N
guanidino-l-(thiazole-2- H2N~ H~O
~ carbonyl)-butyl]-amide NH
0430 2-(2-Hydroxy-3-phenyl- ~
propionyl)-4-oxo-octah~dro- ~N
~ pyrrolo[l,2-a] pyrazine-6- o ~HN
carboxylic acid [4-guanidino- H2N~ ~ ~
l-(thiazole-2-carbonyl)- N~
butyl]-amide 0435 4-Oxo-2-phenoxyacetyl- O~N~
octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [4- " HN~ N
guanidino-l-(thiazole-2- NH~ O
carbonyl)-butyl]-amide NH

F~ E -~

.

0440 4-Oxo-2-(3-phenoxy-propionyl)-ocatahydro-pyrrolo[1,2-~ a~pyrazine-6-carboxylic acid [4- 0 0 HN~_ ~N
guanidino-1-(thiazole-2- NH~ o carbonyl)-butyl]-amide NH
0445 4-Oxo-2-(2-phenyl- o~~, ethanesul~onyl)-octahydro- - ~ S~N '~
pyrrolo[1,2-a] pyrazine-6- ~ o~HN
carboxylic acid [4-guanidino-1- ff (thiazle-2-carbonyl)-butyl]- NH
amide 0450 2-(Naphthalene-2-sulfonyl)-4- o~~, oxo-octahydro-pyrrolo[1,2-a] ~S~N ~
pyrazine-6-carboxylic acid [4- O O HN ~N
guanidino-1-(thiazole-2- _ff carbonyl)-butyl]-amide NH
0455 4-(6-[4-Guanidino-l-(thiazole-2-carbonyl)-butylcarbamoyl]-4-oxo- ~N~
hexahydro-pyrrolo[1,2-a]
pyrazin-2yl)-4-oxo-3-(2 propyl -pentanoylamino)-butyric acid H.Nff methyl ester N~
0460 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2-a] ~ N~"
pyrazine-6-carboxylic acid [4- ~
guanidino-1)-butyl]-amide ~ H

0465 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2-a]
pyrazine-6-carboxylic acid [3- ~
guanidino-propyl) -amide ~HN

51~ T~ S~ T

0470 4-(6-[4-Guanidino-l-(thiazole-2- HO~N~' carbonyl)-butylcarbamoyl]-4-oxo-~ O ~HN
- hexahydro-pyrrolo[1,2-a]pyrazinNH~N
- 2-yl)-4-oxo-butyric acid HN~NH
0475 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolol1,2- ~ N~"N~ s a]pyrazine-6-carboxylic acid 11-~ ~N
(5-ethyl-thiazole-2-carbonyl)- ~
NH
4-guanidino-butyl]-amide 0480 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2- ~ N~"N~ s~
a]pyrazine-6-carboxylic acid [4-NH~N
guanidino-1-(5-methyl-thiazole- H~N~NH
2-carbonyl)-butyl]-amide 0485 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- ~N~
a]pyrazine-6-carboxylic acid [4-o ~-N
guanidino-l-(4-methyl-thiazole- H.N~
NH
2-carbonyl)-butyl]-amide 049~ 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2- ~ ~;~
a]pyrazine-6-carboxylic acid~l- Nt~
(4-ethyl-thiazole-2-carbonyl)- HN~
Nll 4-guanidino-butyl]-amide 0495 4-Oxo-2-(3-phenyl-propionyl)- '~
octahydro-pyrrolo[1,2- ~ ~;~
a]pyrazine-6-carboxylic acid (4- ~ H~
carbamimidoyl-pheny)-amide ~ N~

~;TIlFU~E S!~EET

W o 96/19483 PCT/CA95/00708 0500 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- ~ ~N~
a]pyrazine-6-carboxylic acid [4-guanidino-l-(5-phenyl-thiazole- NH
2-carbonyl)-butyl~-amide 0505 4-Oxo-2-(3-phenyl-propionyl)- o octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- ~ O H~N
(5-benzyl-thiazole-2-carbonyl)- H~N~
4-guanidino-butyl]-amide NH
0510 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrroloLl,2- ~ ~,~
a]pyrazine-6-carboxylic acid [1- ~ ' (4-carbarnimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0515 4-Oxo-Z-(3-phenyl-propionyl)- "
octahydro-pyrrolol1,2- ~N~'~
a]pyrazine-6-carboxylic acid ll- O ~H~
(3-carbamimidoyl-benzyl)-2-oxo- ~N~
2-thiazol-2-yl-ethyl]-amide Nll 0520 4-Oxo-2-(3-phenyl-propionyl)- '~
octahydro-pyrrolo[1,2- ~ ~;~
a]pyrazine-6-carboxylic acid (l-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl- ~,~
ethyl]-amide H~J~
0525 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2- ~ ~,~
a]pyrazine-6-carboxylic acid ~
(l-carbamimidoyl-piperidin-3- ~ '' ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide ~U~ TE SHEET

-. W O 96119483 PCT/CA95100708 - 0530 4-Oxo-2-(3-phenyl-propionyl)- "
-- octahydro-pyrrolo[1,2- ~ N~,N~ s a]pyrazine-6-carboxylic acid [1- c, ~ H~
. (1-carbamimidoyl-piperidin-2- H,N ~ O
; ylmethyl)-2-oxo-2-thiazol-2-yl- N~
ethyl]-amide 0535 [6-[4-Guanidino-1-(thiazole-2- "
carbonyl)-butylcarbamoyl]-4-oxo- ~N~ S~
2-(3-phenyl-propionyl)- HC (~ O ~, H~N
octahydro-pyrrolo[1,2-a]pyrazine-3-yl~-acetic acid ~NH
.
0540 3-[6-[4-Guanidino-1-(thiazole-2- '~' .- carbonyl)-butylcarbamoyl]-4-oxo- ~N~
2-(3-phenyl-propionyl)- HO~ ~ ~ H~
- octahydro-pyrrolo[1,2-a]pyrazin-3-yl]-propionic acid NH
0545 [6-[1-(1-Carbamimidoyl-piperin- "
4-ylmethyl)-2-oxo-2-thiazol-2- ~N~
yl-ethylcarbamoyl]-4-oxo-2-(3- HC) ~ ~t.
phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)-acetic acid 0550 3-[6-[1-(1-Carbamimidoyl- "
piperidin-4-ylmethyl)-2-oxo-2- ~ ~ ;~ ,~
~ thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)-- octahydro-pyrrolo[1,2-a]pyrazin-3-yl)-acetic acid '~

ITlJT~ S.~EET
-0555 [6-[1-(1-Carbamimidoyl- ~N~
piperidin-3-ylmethyl)-2-oxo-2- ~ ~N~
thiazol-2-yl-ethylcarb.amoyl]-4- HO o ~ o H~N
oxo-2-(3-phenyl-propionyl)- ~ NH
N _~
octahydro-pyrrolo[1,2-a]pyrazin- '~ ~NH2 3-yl)-acetic acid 0560 [6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3- ~ ~N~
phenyl-propionyl)-octahydro- HO <~ ~ <) Hr~
pyrrolo[1,2-a3pyrazin-3-yl)- ~HN
acetic acid H~N
0565 3-[6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3- ~ ~N~
phenyl-propionyl)-octahydro- ~K)~ o ~
pyrrolo[l,2-a]pyrazin-3-yl)- HN
propionic acid H.N~NH
0570 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2- ~ ~N~
a~pyrazine-6-carboxylic acid [4- " <~ ~-N
guanidino-1-(thiazolc-2-carbonyl)-butyl]-methyl-amide HN~NH
0575 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2- ~ ~,~
a]pyrazine-6-carboxylic acid (1-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl- r, ~, ethyl]-methyl-amide Sl~ T~

0580 [6-([1-Carbamimidoyl-piperidin- ~N~
4-ylmethyl)-2-oxo-2-thiazol-2- ~ 1 N~
yl-ethyl]-methyl-carbamoyl)-4- ~ ~ ~N~
. oxo-2-(3-phenyl-propionyl)- ~ "
-: octahydro-pyrrolo[1,2-a]pyrazin- N
HN~--NH~
3-yl]-acetic-acid 0585 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1- ~ o ~N
(1-carbamimidoyl-piperidin-3- ~ ~NH
ylmethyl)-2-oxo-2-thiazol-2-yl- NH
ethyl]-methyl-amide 0590 4-Oxo-2-(3-phenyl-propionyl)- '~
octahydro-pyrrolo[1,2- ~N~
a~pyrazine-6-carboxylic acid (3- o ~
guanidino-propyl)-methyl-amide ~H~NH

0595 2-(Naphthalene-2-carbonyl)-4- "
oxo-octahydro-pyrrolo[1,2 - ~N~N~ N~
a]pyrazine-6-carboxylic acid [4-guanidino-l-(thiazole-2- ~ "
carbonyl)-butyl]-amide ~LN~N, 0600 2-(Naphthalene-1-carbonyl)-4- ~ O
oxo-octahydro-pyrrolo[1,2- ~N~
- a]pyrazine-6-carboxylic acid [4- " (,' guanidino-1-(thiazole-2 carbonyl)-butyl]-amide NH

S~~~ l Z~JT~ S'! EE~T

0605 2-t3-Naphthalen-1-yl-proplonyl)4-oxo-octahydro-pyrrolo[1,2-a]pyrazine-6-0 0 HN ~N
carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-HN~NH

amide 0610 2-(4-tert-Butyl-benzoyl)-4-oxo- ~
octahydro-pyrrolo~1,2- ~N~
a]pyrazine-6-carboxylic acid [4- ~ ~N
guanidino-1-(thiazole-2-carbonyl)-butyl]-amide NH~NH.
061~ 2-(BenzoL1,3]dioxole-5- ~N~' carbonyl)-4-oxo-octahydro- O ~
pyrrolo[1,2-a]pyrazine-6-o O H ~ -N
carboxylic acid [4-guanidino-1- ~H~NH
(thiazole-2-carbonyl)-butyl]-NH.
amide 0620 2-(3-genzo~l~3]dioxol-5-yl- ~N~' propionyl)-4-oxo-octahydro-pyrrolo[1,2-a]pyrazine-6- O O H~N
carboxylic acid [4-guanidino-1-NH~
(thiazole-2-carbonyl)-butyl amide 0625 2-~2-(2-Methyl-benzylidene)-but- ~N~' 3-enoyl]-4-oxo-octahydro- ~N~
pyrrolo[1,2-a]pyrazine-6-carboxylic acid ¦1-(1- CN~
carbamimidoyl-piperidin-3-- ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide s ~ ~~ E riT

=

.
- 0630 2-~2-(2-Methyl-benzylidene)-but- ~N~' 3-enoyl]-4-oxo-octahydro- O ~HN
pyrrolo[l,2-a~pyrazine-6-- carboxylic acid [1-(1- ~
-. carbamimidoyl-piperidin-4- H~N~NH
~ ylmethyl)-2-oxo-2-thiazol-2--yl-ethyl]-amide 0635 2-(2-Benzylidene-pent-3-enoyl)- ~
4-oxo-octahydro-pyrrolo[1,2- ~ N
a]pyrazine-6-carboxylic acid (3- " ~
guanidino-propyl)-amide HN
H N

$~-J~iT~ ET

0640 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2-a]pyrazine- ~ ~N~

6-carboxylic acid 4- ~ o '~
carbamimidoyl-benzylamide H~N
0645 4-Oxo-2-(3-phenyl-propionyl)- o octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid ~4-imidazol-1- ~
yl-l-(thiazole-2-carbonyl)-butyl]-amide 0650 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid [4-(2-amino- ~
imidazol-l-yl)-l-(thiazole-2-carbonyl)-butyl]-amide NH

0655 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2-a]pyrazine- ~N~N~ N~
6-carboxylic acid [3-(2-amino-6-methyl-pyrimidin-4-yl)-1- ,~
(thiazole-2-carbonyl)-propyl]- N~
amide 0670 4-Oxo-2-(3-phenyl-propionyl)- '' octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [3-(2-amino-6- ~ O H~
chloro-pyrimidin-4-yl)-1- a~
(thiazole-2-carbonyll-propyl]- N~NH
~ amide STI ~ U~ ffE~T

W 096/19483 PCT/CA9~/00708 0675 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid [3-(6-amino-pyridin-2-yl)-1-(thiazole-2- ~ ~
carbonyl)-propyl]-amide ~N

0680 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine- ~N~.N~ N~
6-carboxylic acid [3-(2-amino- ~, O H~
pyridin-4-yl)-1-(thiazole-2- ~ "
carbonyl)-propyl]-amide Nii 0685 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~N~N~ N~
6-carboxylic acid [2-(2-amino- c, O H~
pyridin-4-yl)-1-(thiazole-2- ~ ~
carbonyl)-ethyl]-amide ~NH

0690 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrroloL1,2-a]pyrazine- ~N~ N~
6-carboxylic acid [2-(6-amino- ~. ~ H~
pyridin-2-yl)-1-(thiazole-2- ~N
~Nll carbonyl)-ethyl]-amide 0695 2-L4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~ ~'~
- 6-carbonyl]-3- ( thiazole-2- ~
carbonyl ) --1, 2, 3, 4--tetrahydro-- HAN~ N
s oc~ulno 1 ine - 6 - c arboxamldlne 5~ JTE ~

0700 2-[4-Oxo-2-(3-phenyl-propionyl)-~N~
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carbonyl]-3-(thiazole-2- H~
carbonyl)-1,2,3,4-tetrahydro- 5 N
.
isoquinoline-7-carboxamidine 0705 N-[1-[4-Oxo-2-(3-phenyl- ~N~
propionyl)-octahydro- ~N?
pyrrolo[l,2-a]pyrazine-6- ~H2O ~ O
carbonyl]-5-(thiazole-2- 5 N
carbonyl)-pyrrolidin-3-yl]-guanidine 0710 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid [1-(4-amino- ~ H~
cyclohexyl)-2-oxo-2-thiazol-2- ~ "
yl-ethyl]-amide H~N
0715 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid ll-(4-amino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0720 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a3pyrazine- ~N~
6-carboxylic acid [1-(4-amino- "
benzyl)-2-oxo-2-thiazol-2-yl- ~ ' ethyl]-amide ;T~ T~ E~

:' ~ ~

W O 96tl9483 PCT/CA95/00708 0725 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[l,2-a]pyrazine- ~N~
6--carboxylicacid [1--(4.-- 0o~H~ s - aminomethyl-benzyl)-2-oxo-2- ~ ~
thiazol-2-yl-ethyl3-amide H~N

0730 4-Oxo-2-(3-phenyl-propionyl)- '~
octahydro-pyrrolo[1,2-a]pyrazine- I~N~N~ N~;~
6-carboxylic acid [1-(3- O O H~S
aminomethyl-benzyl)-2-oxo-2- _~~
thiazol-2-yl-ethyl]-amide HN

0735 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a~pyrazine- ~N~ N~>
6-carboxylic acid (2-oxo-1- " o H~a piperidin-4-ylmethyl-2-thiazol- ,~"
2-yl-ethyl)-amide ~Hll>
0740 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl,2-a]pyrazine- ~N~.r~ N~>
- 6-carboxylic acid (2-oxo-1- " t~s piperidin-3-yl-2-thiazol-2-yl- H~ <
- ethyl)-amide 0745 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- (~N~O'~
6-carboxylic acid 11-(3-guanidino-cyclohexylmethyl)-2- ~--~
HN
oxo--2--thiazol--2--yl--ethyl]--amide H ~H~

SlJ~S I Ir~r S~Y~ET

WO96119483 ~CT/CA9~/00708 0750 4-Oxo-2-(3-phenyl-propionyl)- "
octahydro-pyrrolo[1,2-a]pyrazine- ~N~
6-carboxylic acid [1-(4- ~ ~ S
guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide HNyNH
NH, 0755 4-Oxo-2-(3-phenyl-propionyl'- ~
octahydro-pyrrolo[1,2-alpyrazine- ~N~'~
6-carboxylic acid [1-(2- o guanidino-cyclohexylmethyl)-2-H~N~ ~_~
oxo-2-thiazol-2-yl-ethyl]-amide NH
0760 4-Oxo-2-(3-phenyl-propionyl)- ~ , octahydro-pyrrolo[1,2-a]pyrazine- ~H N~
6-carboxylic acid [1-(5-benzyl-thiazole 2 carbonyl) 4 H~N~NH
guanidino-butyl]-amide 0765 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine- ~ ~Hr. "
6-carboxylic acid [4-guanidino-1 (5 phenyl thiazole 2 H,r.~NH
carbonyl)-butyl]-amide 0770 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrido[1,2-a]pyrazine- ~t~
6-carboxylic acid [4-guanidino-H~N~t~H ~ O
1-(thiazole-2-carbonyl)-butyl]- NH s~
amide 0775 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza- ~ N~N~
naphthalene-4-carboxylic acid~o NH
[4-~uanidino-1-(thiazole-2- NH ~N
carbonyl)-butyl]-amide ~ ~J E~lr!lrUTE S~ T

W 096/19483 . PCT/CA95/00708 0780 5-Oxo-7-(3-phenyl-propionyl)- ~
octahydro-2-thia-4a,7-diaza- (~ ~ 3 naphthalene-4-carboxylic acid ~o NH~ S
[1-(4-carbamimidoyl-benzyl)-2-~ oxo--2--thiazol--2--yl--ethyl]--amide H,N NH

- 0785 5-Oxo-7-(3-phenyl-propionyl)- "
~ octahydro-2-thia-4a,7-diaza- 0~~N~ S
naphthalene-4-carboxylic acid "~ NH~<S
~1-(3-carbamimidoyl-benzyl)-2- [~NH
oxo-2-thiazol-2-yl-ethyl]-amide ' NH, - 0790 5-Oxo-7-(3-phenyl-propionyl?- "
octahydro-2-thia-4a,7-diaza- ~'~'"'(I,~~
naphthalene-4-carboxylic acid ~c NH~<S
Il-(l-carbamimidoyl-piperidin-3- CIN~NH
ylmethyl)--2--oxo--2--thiazol--2--yl-- NH
ethyl]-amide ;' 0795 5-Oxo-7-(3-phenyl-propionyl)- '' octahydro-2-thia-4a,7-diaza- (~ ~o N3 naphthalene-4-carboxylic acid o NH~<' ll-(l-carbamimidoyl-piperidin-4--. ylmethyl)--2--oxo--2--thiazol--2--yl-- H.tJ~NH
ethyl]-amide 0800 L4-[4-Guanidino-l-(thiazole-2-- - carbonyl)-butylcarbamoyl]-5-oxo- ,~ J

7--(3--phenyl--propionyl)-- HO ~ NH
octahydro-2-thia-4a,7-diaza- ~rH
naphthalen-6-yl]-acetic acid STilr~ T
. = .

0805 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-~N~
naphthalene-4-carboxylic acid ~ O~NH
[4-guanidino-1-(thiazole-2-HN~NH~O
~ carbonyl)-butyl]-amideNH S~N

0810 3-[4-[4-Guanidino-l-(thiazole-2-carbonyl-butylcarbamoyl]-5-oxo- ~ ~'~
7-(3-phenyl-propionyl)-i"~' NH__~J~
octahydro-2-thia-4a,7-diaza- ~" S~N
naphthalen-6-yl]-propionic acid 0815 5-Oxo-7-(3-phenyl-propionyl)- "
octahydro-2-thia-4a,7-diaza- ~N~
naphthalene-4-carboxylic acid ~NH~
L3-guanidino-propyl]-amide N~l I IN NH

0820 5-Oxo-7-(3-phenyl-propionyl)- "
octahydro-2-thia-4a,7-diaza- ~ ~,~ " N3 naphthalene-4-carboxylic acid "(,~N, l-(l-carbamimidoyl-piperidin-3-~ J~
ylmethyl)-2-oxo-2-thiazol-2-yl- I,, ethyl]-amide 0825 4-Oxo-2-(3-phenyl-propionyl)- '' octahydro-pyrrolo~1,2-a]pyrazine- ~N~N~ N~
6-carboxylic acid [4-guanidino- ~ c l-(hydroxy-thiazol-2-yl-methyl)- ~ ~~
butyl]-amide NH

U ~ 5~1 T~ E ~

0830 4-Oxo-2-(3-phenyl-propionyl)-: octahydro-pyrrolo~1,2-a]pyrazine-6- ~ ~ N~
; carboxylic acid (4-guanidino-1- ~ ' thiazol 2 ylmethyl butyl) amide hrl~Hh 0835 4-Oxo-2-(3-phenyl-propionyi)- r octahydro-pyrrolo[1,2-a]pyrazine-6- ~N~
carboxylic acid f4-guanidino-1- ~ ,'~
thiazol-2-yl-butyl)-amide 0840 4-Oxo-2-(3-phenyl-propionyl)- ~
octahydro-pyrrolol1,2-a]pyrazine- ~ ~h~
6-carboxylic acid ¦4-methoxy-1- ~
(thiazole-2-carbonyl)-butyl]-amide 0845 l6-[4-Methoxy-l-(thiazole-2- ~ N~"
carbonyl)-butylcarbamoyll-4-oxo- ~ ~ N~
2-(3-phenyl-propionyl)- 110 0 ~rl ~', octahydro-pyrrolo[1,2-a]pyrazin-3-yl]-acetic acid ~~
0850 l2-(5-Methoxy-2-(l4-oxo-2-(3 phenyl-propionyl)-octahydro- ~N~
pyrrolo[l,2-a~pyrazine-6-carbonyl]-amino)-pentanoyl)-thiazol-5-yl]-acetic acid 0855 4-Oxo-2-(3-phenyl-propionyl)- ~ H
octahydro-pyrrolo~1,2-a]pyrazine-6-carboxylic acid [4-amino-1- o r~H~
(thiazole-2-carbonyl)-butyl]- ~ ~
amide NH .

S~iTiTlJT~ S~IIEET

. CA 02208772 1997-06-20 W O 96/19~83 PCT/CA95/00708 0860 4-Oxo-2-(3-phenyl-propionyl)- ~ NJ"
octahydro-pyrrolo[1,2-a]pyrazine- ~ ~ ~
6-carboxylic acid [5-amino-1- ~ O HN~5 ~thiazole-2-carbonyl)-pentyl]-amide ~-NH2 0865 4-Oxo-2-(3-phenyl-propionyl)- ~ ~ H
octahydro-pyrrolo[1,2-a]pyrazine- ~H~i 6-carboxylic acid ¦5-guanidino-l-(thiazole-2-carbonyl)-pentyl]- ~NH
amide NH~-NH

0870 2-(3-Naphthalen-2-yl-propionyl)-4-oxo-octahydro-pyrrolo[1,2- ~ ~"~ r~
a]pyrazine-6-carboxylic acid ~4-guanidino 1 (thiazole-2- HN~NH
rlH.
carbonyl)-butyl]-amide 0875 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo~1,2-a]pyrazine~ 1r~ N~
6-carboxylic acid [4-guanidino- ~ 1!
1-(1-methyl-lH-imidazole-2- HN~N,N,' carbonyl)-butyl]-amide 0880 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid 14-guanidino-1-(thiazole-2-carbonyl)-butyl]- H~
NH
amide 5 ~ ~ E l~T~ T

; l:

W 096119483 . PCT/CA95100708 0885 8,8-Dimethyl-4-oxo-2-(3-phenyl-propion~1)-octahydro- ~ ~N~
pyrrolo[1,2-a]pyrazine-6- ~
carboxylic acid [4-guanidino-1- HN~NH
(thiazole-2-carbonyl)-butyl]- NH
amide 54 .

Pre~erred compounds according to formula (VIII) include:
0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo[l~2 a]pyridine-6-carboxylic a.cid [l-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydro-pyrrolo~l,2-a]pyrazine-6-carboxylic acid [l-(benzothiazole-2-carbonyl)-4-guanidino-butyl]-amide 0515 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [l-(3-carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyll-amide 0530 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [l-(l-carbamimidoyl-piperidin-2-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0545 16-ll-(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 3-16-[l-(l-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxc,-2-thiazol-2-yl-ethylcarbamoyl]-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazin-3-yl)-acetic acid 0555 L6-[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 S ~ ~ ~T

.

W O 96/19~83 PCT/CA95/00708 0560 [6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl)-acetic acid . 0565 3-[6-(3-Guanidino-propylcarbamoyl)-4-oxo-2-(3-phenyl-- propionyl)-octahydro-pyrrolo[1;2-a]pyrazin-3-yl)-- propionic acid 0575 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid ll-(l-carbamimidoYl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-methyl-amide 0580 16-(11-Carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyll-methyl-carbamoyl)-4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazin-3-yl]-acetic-acid 0585 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [l~ carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-methyl-amide 0590 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-.- a~pyrazine-6-carboxylic acid (3-guanidino-propyl)-methyl-amide 0595 2-(Naphthalene-2-carbonyl)-4-oxo-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid ~4-guanidino-l-(thiazole-2-carbonyl)-butyl]-amide 6~ T

W O 96/19483 . PCT/CA95/00708 0625 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxO-octahydro-pyrroloLl,G-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0630 2-[2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1-(l-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0635 2-(2-Benzylidene-pent-3-enoyl)-4-oxo-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid t3-guanidino-propyl)-amide 0625 2-l2-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo-octahydro-pyrrololl,2-a]pyrazine-6-carboxylic acid 11-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0630 2-12-(2-Methyl-benzylidene)-but-3-enoyl]-4-oxo-octahydro-pyrrolo[1,2-a~pyrazine-6-carboxylic acid 11-(l-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0635 2-(2-Benzylidene-pent-3-enoyl)-4-oxo-octahydro-pyrroloLl,2-a]pyrazine-6-carboxylic acid (3-guanidino-propyl)-amide 0645 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl,2-alpyrazine-6-carboxylic acid l4-imidazol-1-yl-1-(thiazole-2-carbonyl)-butyl¦-amide ET

-W O96/19483 PCT/CAg5/00708 0670 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [3-(2-amino-6-chloro-pyrimidin-4-yl)-1-(thiazole-2-carbonyl)-propyl]-amide 5 0675 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid 13-(6-amino-PYridin-2-yl)-l-(thiazole-2-carbonyl)-propyl]-amide 0680 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [3-(2-amino-pyridin-4-yl)-l-(thiazole-2-carbonyl)-propyl]-amide :
0685 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrroloLl,2-a]pyrazine-6-carboxylic acid l2-(2-amino-pyridin-4-yl)-1-(thiazole-2-carbonyl)-ethyl]-amide -0690 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo~1,2-a]pyrazine-6-carboxylic acid ~2-(6-amino-pyridin-2-yl)-l-(thiazole-2-carbonyl)-ethyl]-amide 06~5 -14-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl,~-a~pyrazine-6-carbonyl]-3-(thiazole-2-carbonyl)- -1,2,3,4-tetrahydro-isoquinoline-6-carboxamidine 25 0700 2-~4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrr a]pyrazine-6-carbonyl]-3-(thiazole-2-carbonyl)-1,2,3,4-tetrahydro-isoquinoline-7-carboxamidine 0705 M-11-[4-Oxo-2-(3-phenyl-propionyl)-octahydro-.30 pyrrolo[l,2-a]pyrazine-6-carbonyl]-5-(thiazole-2-carbonyl)-pyrrolidin-3-yl]-guanidine . 9 ~ ~ ~ 5~ ~ E ~ '~ E ~
-0710 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid L1-(4-amino-cyclohexyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0730 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid il-(3-aminomethyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0745 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid 1l-(3-guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0755 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a~pyrazine-6-carboxylic acid [1-(2-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-carbamimidoyl-piperidin-4-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0800 l4-l4-Guanidino-1-(thiazole-2-carbonyl)-butylcarbamoyl]-5-oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalen-6-yl~-acetic acid 0810 3-14-14-Guanidino-1-(thiazole-2-carbonyl-butylcarbamoyl]-5-oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalen-6-yl~-propionic acid 5 W E~ T~T ~ ~ ~J E ~T

-0815 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid [3-guanidino-propyl]-amide 0820 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid ll-(l-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0830 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrololl,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 pre~erred compounds according to ~ormula VIII
20 include:

0335 "-Benzoyl-4-oxo-octahydro-pyrrololl,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 14-(2-amino-imidazol-l-yl)-l-(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)-l-(thiazole-2-carbonyl)-propyl~-amide .J~E $~:ET

0715 4-Oxo-2-~3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid ~1-(4-amino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0720 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [1-(4-amino-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0725 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrroloLl,2-a]pyrazine-6-carboxylic acid ll-(4-aminomethYl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0735 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid (2-oxo-1-piperidin-4-ylmethyl-2-thiazol-2-yl-ethyl)-amide 0740 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid (2-oxo-1-piperidin-3-yl-2-thiazol-2-yl-ethyl)-amide 0750 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolol1,2-a~pyrazine-6-carboxylic acid ll-(4-guanidino-cyclohexylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0760 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid ~1-(5-benzyl-thiazole-2-carbonyl)-4-guanidino-butyl]-amide 0765 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-phenyl-thiazole-2-carbonyl)-butyl]-amide 6l S~B~ E ~

0770 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrido[1,2-a]pyrazine-6-carboxylic acid L4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0775 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid L4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0780 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid ll-(4-: carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0785 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid 11-(3-carbamimidoyl-benzyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0790 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-~ 20 diaza-naphthalene-4-carboxylic acid 11-(1-carbamimidoyl-piperidin-3-ylmethyl)-2-oxo-2-thiazol-2-yl-ethyl]-amide 0805 5-Oxo-7-(3-phenyl-propionyl)-octahydro-2-thia-4a,7-diaza-naphthalene-4-carboxylic acid 14-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide 0825 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo¦1,2-a~pyrazine-6-carboxylic acid [4-guanidino-1-(hydroxy-thiazol-2-yl-methyl)-butyl]-amide SU5STITlJT~ T

0840 4-Oxo-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-(14-oxo-2-(3-phenyl-propionyl)-octahydro-pyrroloLl,2-a]pyrazine-6-carbonyl]-amino)-pentanoyl)-thiazol-5-yl]-acetic acid 0855 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [4-amino-l-(thiazole-2-carbonyl)-butyl]-amide 0860 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[l,2-a]pyrazine-6-carboxylic acid [5-amino-l-(thiazole-2-carbonyl)-pentyll-amide 0865 4-oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo1l,2-a1pyrazine-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-methyl-_ 30 thiazole-2-carbonyl)-butyl]-amidei andO340 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo1l,2-a]pyrazine-;TITU~E S~EET

W 096/19483 ~ PCT/CA95/00708 6-carboxylic acid [4-guanidino-1-(thiazole-2-~ carbonyl)-butyl]-amide.

Preferred compounds according to formula IX include: =
0890 3-Amino-4-oxo-2-phenyl- ~
hexahydro-pyrrolo[2,1- H~N~ o b][l,3]thiazine-6-carboxylic ~
acid [l-(benzothiazole-2- ~HNH
carbonyl)-4-guanidino-butyl]- H,N
amide 0895 3-Amino-2-benzyl-4-oxo-- 0 hexahydro-pyrrolo[2,1- ~ s b]ll, 31thiazine--6--carboxylic H~N~ --~ o acid [l-(benzothiazole-2-carbonyl)-4-guanidino-butyl¦- NHNH
amide H~N

0900 3-Amino-2-cyclohexyl-4-oxo-hexahydro-pyrrolo[2,1- ~ s b¦¦ 1, 31thiazine--6--carboxylic 0 ~_ ~N~3 acid ll-(benzothiazole-2- ~ ~H S
carbonyl)-4-guanidino-butyl]- HN~NH
amide !ilL~!~iil~3~lJTE ~.~EET

.

Pre~erred compounds according to formula X include:

- 0905 7-Benzyl-6-oxo-octahydro-pyrido[2,1-c][1,4]thiazine-4-carboxylic acid [1- NH~
(benzothiazole-2-carbonyl)-4- ~ s NH
guanidino-butyl]-amide H~N~NH

0910 7-(4-tert-Butyl-benzyl)-6-oxo- ~ H
octahydro-pyrido[2,1- ~ 5 c][l,4Jthiazine-4-carboxylic o~,~
acid Ll-(benzothiazole-2- ~ 5 carbonyl)-4-guanidino-butyl]- HN N~
amide 0915 6-Oxo-octahydro-pyridol2,1- . ~
c][l,4]thiazine-4-carboxylic ~ ~ ~ , 3 acid [4-guanidino-1-(thiazole- ~ ~ ~ NH S

2-carbonyl)-butyl]-amide 0925 7-Benzyl-6-oxo-octahydro- H
pyrido~2,1-c]¦1,41thiazine-4- ; ~ ~ , 3 carboxylic acid l4-guanidino- O ~ NH C
1-(thiazole-2-carbonyl)butyl~- NHl - NH~
amide 0935 7-senzyl-6-oxo-octahydro- H
pyrido[2,1-c][1,4]thiazine-4- ~ ~ ~ ~ ~N~
carboxylic acid [4-guanidino- 0 ~ NH ~
- 1-(thiazole-2-carbonyl)butyl]- NH~-NH2 amide S~B5TIT~JT~ 5!H~ET

. l . .

0940 6-Oxo-7-phenethyl-octahydro- H
pyridoL2,1-c~[1,4]thiazine-4- ~ ~ ~N ~s~ O
carboxylic acid l4-guanidino- O~ ~ NH s 1-(thiazole-2-carbonyl)- NH~LNH2 butyl]-amide 0950 7-Benzyl-2,2,6-trioxo- H
octahydro-21>6_pyridol2,1- ~ o c~ll,4]thiazine-4-carboxylic ~ ~ NH s acid l4-guanidino-1-(thiazole- NH NH2 2-carbonyl)-butyl]-amide More pre~erred compounds according to formula X include:
925 7-Benzyl-6-oxo-octahydro-pyridol2,1-C]¦1,4lthiazine-4-carboxylic acid l4-guanidino-1-(thiazole-2-carbonyl)butyl]-amidei and940 6-Oxo-7-phenethyl-octahydro-pyridol2,1-c]¦1,4lthiazine-4-carboxylic acid ¦4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide.

Pre~erred compounds according to ~ormula III include:
0960 4-oxo-1-(3-phenyl-propionyl)-octahydropyrrolo[1,2]pyrimidin ~ ~N~
e-6-carboxylic acid[4- ~ ~ c guanidino-1-(thiazole-2- ~ ~~~ ~N~3 carbonyl)-butyl]-amide N~HNH
H N
0965 4-oxo-1-(phenethylsul~onyl)- ~ sc,N
octahydropyrrolo[1,2]pyrimidin e-6-carboxylic acid[4- c ~N~
guanidino-1-(thiazole-2- ~ ~ S3 carbonyl) butyl]-amide LNH

TlJlrE 5~!~ET

For preparation o~ the compounds o~ ~ormula (VII) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The ~ollowing scheme is one particular method o~

STITUTE S~l~ET

R3~ ~ ~ STEP~

~OH STEP 3' Y~_ STEP-4' STEP-~ D ~ d O

R~ STEP-- ~ N e STEP-5 ~ R

~ "~

S~ ;TiTlJT~ S~iEET

W 096/19483 PCT/CA9~/00708 Step 1:
The alkylation of a is done with appropriate bases according to the procedures described in Evans et al (J.
Am. Chem. Soc., 1981, 103, 2127; ibid, 1982, 104, 1737;
Aldrichimica Acta, 1982, 15, 23) to give Step 2:
Compound b upon hydroboration and oxidation following conditions available in the literature (Synthesis, 1980, 151) results in the aldehyde c.

Step 3:
The formation of adduct e from aldehyde c with _ 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 o~ aldehyde c to aldehyde ~ is readily achieved by appropriate protection deprotection protocals ~ound in T. Greene, Protective Groups In Organic Synthesis, (John Wiley & Sons, 1981).

Step 4:
The cylization of adduct e to f may readily be achieved . 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 q with in presence of suitable ~ aromatic solvents e.g, benzene.

~ 35 Step 5:
The ester function (-C(O)O-R ) of the bicyclic intermediate of formula f is then subjected to ~W E~; ~ ITl ITE S~ET

-' CA 02208772 1997-06-20 .
~ hydrolysation using an appropriate agent such as HCl in an - appropriate solvent such as ethyl ether to yield to the free carboxylic acid. The resulting compound is then coupled to RlH with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to yield to a bicyclic coupled compound of formula (VIII). Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see Princi~les of Pe~tide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Pe~tides, Analysis. Synthesis, Bioloov, Vol. l.edited by Gross E., and Meienhofer J., Academic Press , Mew York, San Francisco, London, 1979.

For preparation of the compounds of ~ormula (VIII) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme is one particular method of preparation.

SUB~;TITU~E~ 5,L~

W O 96/19483 . PCTICA95/00708 HN STEP 1 ~ _ 2 1 N~
~ 21 a b O

ozonolysis ~ ~

~OR2~ STEP 3 R~

g Vlll S~ UTE SJLI E E1r . CA 02208772 1997-06-20 - wherein;
Pg is a nitrogen protecting group;
each of R20; and R2l is independently a C; alkyl; and X, Rl, R3, R, and Rs are as previously defined.
The process in scheme 2 is briefly described as follows:

~TEP 1:
The amino and carboxylic functions of the unsaturated compound of formula (a) are protected with appropriate protecting groups. A variety of protecting groups known for reactive functional groups and suitable protection and deprotection protocols may be found in T. Greene, Protective Groups In Orqanic 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 reagent such as mercuric acetate in an inert solvent such as tetrahydrofuran (THF) to yield to a protected amino alcohol of formula (b).

~TEP 2 The protected amino alcohol of formula (b) is oxidized using an appropriate oxidizing agent such as sul~ur 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 o~ ~ormula (a') prepared according to Collado et al, J. Org. Chem.,1995, 60:5011.

SU~3ST~T~T~ S!t~E~T
~ .

~ 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 con.tacting the obtained imine with an appropriate reagent such as sodium triacetoxy borohydride NaBH(OAc) to yield to a cyclic intermediate of formula (e).

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 Rssubstituent.

The amino protecting group of the cyclic intermediate of formula (f) is removed under appropriate conditions and the resuiting compound is then subjected to appropriate condition for internal ring closure such as low heat in an inert solvent or as a raw compound to yield to a bicyclic intermediate of formula (g). The bicyclic intermediate of formula (g) can also be obtained by hydrolysing the ester function (-C(O)O-R20) of the cyclic intermediate of formul.-(g) to the free carboxylic acid followed by standard peptide coupling using an appropriate coupling reagent such as benzotriazole-l-yloxy-tris-(dimethylamino)phosphonium hexafluorophosphate (BOP) in ar.
inert solvent such as dimethyl formamide (DMF).

The ester function (-C(O)O-R2l) of the bicyclic intermediate of formula (g) is then subjected to hydrolysation using an appropriate agent such as HCl in an appropriate solvent such as ethyl ether to yield to the SUBSTITlJTE ~ET

:
.CA 02208772 1997-06-20 .:
free carboxylic acid. The resulting compound is then coupled to R~H with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to yield to a bicyclic coupled compound o~ formula (VIII). Suitable conditions -5 for peptide bond formation are well known in th art of peptide chemistry. For example see Principles of Pe~tide Synthesis, Bodanszky M., Springer-Verlag, Berlin, -Heidelberg, New York, Tokyo 1984; and The Pe~tides, ~nalvsis, Svnthesis, Biolo~v, Vol. l.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 1979.

For preparation of the compounds of formula (IX) various methods can be employed depending upon the particular ; l5 starting materials and/or intermediates involved. The following scheme is one particular method of preparation.

T~JTE S i.Y ~ ~T

MeO

3~ MeO~ ~ ~ R~ HN (CH2)n R H~N~<(CH2)n . O ~_OR20 O ~ O R20 0 a O C
b Y= S-Pg NH-Pg R5 Y~_~R8 STEP 3 Rs~Y~I ~R~
O Rl H~ ~f N~ H2)n e d wherein:
Pg is a sul~ur or amino protecting group;
L is a leaving group;
each o~ R20; and R2l is independently a C alkyl; and Rl, R~

R, and Rs are as previously defined.
The process depicted in scheme 3 is briefly described as ~ollows:
STEP 1:
The carboxylic acid compound (a) is coupled to the cyclic amine compound (b) with a peptide coupling agent such as benzotriazol-1-yloxy-tris-(dimethylamino)phosphonium hexafluorophosphate (BOP reagent) in the presence of a - base such as n-methylmorpholine in an appropriate solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to yield to an amido compound of formula (c). Suitable S~ E3 STIT~JT~ S~

conditions ~or peptide bond formation are well known in th art of peptide chemistry. For example see Principles o~
Pe~tide Svnthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, Mew York, Tokyo 1984; and The Pe~tides, Analysis, Svnthesis, Bioloqv, Vol. l.edited by Gross E., and Meienhofer J., Academic Press , New York, San Francisco, London, 197g.

- l0 The compound of formula (c) is subjected to appropriate conditions to allow internal cyclisation to yield to a bicyclic intermediate of formula (d). For example, acid mediated cyclisation using p-toluenesulfonic acid or TFA
in an appropriate solvent such as dichloroethane.
1~

The ester function (-C(O)O-R ) of the bicyclic intermediate of formula (d) is subjected to hydrolysis using an appropriate agent such as lithium hydroxide (LioH) in an appropriate solvent such as tetrahydrofuran (THF) to yield to the ~ree carboxylic acid. The resulting compound is then coupled to RH with a peptide coupling agent such as BOP in an appropriate solvent such as DMF to give compound (e). Suitable conditions for peptide bond -25 formation are well known in the art of peptide chemistry.
For example see Princi~les of Peptide Svnthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo lg84; and The Peptides Analysis, Synthesis, Bioloav, Vol.
l edited by Gross E., and ~eienho~er J., Academic Press , New York, San Francisco, London, 1979.

For preparation o~ the compounds o~ formula (X) various methods can be employed depending upon the particular starting materials and/or intermediates involved. The following scheme 4 is one particular method of preparation.

T~T~ 1EE~

~¦~ STEP 1 ~L~,CI

RR4~ R ~ RR4~ R5 O OR2i ~ OR
a b O ~
H~B~OR2[i \~ STEP 2 OR~" ~

~ e ~ OR20 RlH ~

Rs~\ B
R 3~f ~

~ h--x o wherein:
each o~ R20 and R21 is independently a C alkyl; and 8, R1, R3, R" and Rs are as previously defined.

;TIT;~ITE S~Y~ Er ~, ? . .

W 096/19483 . PCT/CA95/00708 The process depicted in scheme 4 is briefly described as follows:
t STEP 1:
The halogenated compound of formula (a) is converted to a halomethyl ketone of formula (b) using an appropriate reagent,such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25~C to about 0~C.
The resulting mixture is then treated under acidic '~ 10 conditions to yield to the halomethyl ketone of formula (b).

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).

The cyclic intermediate o~ ~ormula (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).

The ester function (-C(O)O-R2C) of the bicyclic intermediate of formula (e) is subjected to hydrolysation using an appropriate reagent such as LioH to yield to the free carboxylic acid. The resulting compound is then coupled to R~H with a peptide coupling agent such as BOP
in an appropriate solvent such as dimethylformamide to yield to a coupled bicyclic compound of formula (X).
Suitable conditions for peptide bond formation are well known in th art of peptide chemistry. For example see S~J ! 3STllrUT~

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

Compounds of the present invention are further characterized by their ability to inhibit the catalytic activity of thrombin, which is demonstrated in the assay as follows. Compounds of the present invention may be prepared for assay by dissolving them in buffer to give solutions ranging in concentrations from 1 to lOO~M. In an assay to determine the inhibitory dissociation constant, K, for a given compound, a chromogenic or fluorogenic substrate of thrombin would be added to a solution containing a test compound and thrombin; the resulting catalytic activity of the enzyme would be spectrophotometrically determined. This type of assay is well known to those skilled in the art.
The compounds of the present invention may be used as anti-coagulants in vi tro or ex vivo as in the case of contact activation with foreign thrombogenic surfaces such as is found in tubing used in extracorporeal shunts. The compounds of the invention may also be used to coat the surface of such thrombogenic conduits. To this end, the compounds of the invention are obtained as lyophilized powders, redissolved in isotGnic saline and added in an amount sufficient to maintain blood in an anticoagulated state.

The therapeutic agents of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers. The proportion of each carrier is determined by the solubility and chemical nature of the compound, the route of administration, and standard pharmaceutical practice. For example, the compounds may S~J BSTIT~JT~

be injected parenterallyi this being intramuscularly, intravenously, or subcutaneously. For parenteral administration, the compound may be used in the ~orm 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 sranules containing suitable excipients such as starch, lactose, white sugar and the like. The compounds may also be administered sublingually in the form of troches or lozenges in which each active ingredient is mixed with sugar or corn syrups, flavouring agents and dyes, and then dehydrated sufficiently to make the mixture suitable for pressing into solid form. The compounds may be administered orally in the form of solutions which may contain colouring andior flavouring agents.

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

When the composition is administered orally, a larger guantit-~ of the active agent will typically be required tc produce the same e~fect as caused with a smaller quantity given parenterally.

To further assist in understanding the present invention, the following non-limiting examples o~ such thrombin inhibitory compounds are provided. The following examples, of course, should not be construed as speci~ically limiting the present invention, variations presently known or later developed, which would be within ~
the purview of one skilled in the art and considered to fall within the scope of the present invention as described herein. The preferred compounds as of the Sl )~3S ~ ITU~E SHEET

.

present invention are synthesized using conventional preparative steps and recovery methods known to those skilled in the art of organic and bio-organic synthesis, while providing a new a unique combination for the overall synthesis of each compound. Preferred synthetic routes for intermediates involved in the synthesis as well as the resulting anti-thrombotic compounds of the present invention follow.

TiTUTE S~IEET

W 096/19483 . PCT/CA95/00708 EX~PLE 1 O

Boc~ ~ ~OCH~1. Zn/Cu couple; ~Illra~u~ld Boc~ ~ ~OCH3 CH32. [o-CH3C6H4)3P]2PdCI2 ~ ~ CH3 4-iodobenzonitrile ~ 1' NC~

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 M,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 (0.3~ 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 ~unnel. Sequential washing with aqueous HCl (50 mL; 0.1N), distilled H O (3 x 50 mL), drying over Na~SO~, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.

O o Boc~ ~ ~OCH31. Zn/Cu couple; ultr~ound Boc~ ~ ~OCH, ~ CH32. Io-CH3C6H4)3P]~PdCl2 ~ e CH3 3-iodohenzonit~ile CN

A solution of tert-butyloxycarbonyl-iodo-alanine-M,O-~U~ST~T~JTE S~;EET

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

O O

Boc~ ~ ~OCH3 1. Zn/Cu couple; ullr~... oulld Boc~ ~ ~OCH3 C CH3 2. ~ CH3C,,H4)3P]~PdCI, ~ ~ CH3 2-iodohenzonitrile ~ ~ ~

solution of tert-butyloxycarbonyl-iodo-alanine-N,O-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL), and dry N,N-dimethylacetamide (2.0 mL) was added to a dry nitrogen-purged round bottom flask charged with zinc-copper couple (0.90 g).
The resulting mixture was sonicated under nitrogen until no starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential ~5~T~TUlrE SHEE~

. ?
~ r ' - W O96119483 PCTICA95100708 washing with a~ueous HCl (50 mL; O.lM), distilled H O (3 x 50 mL), drying over Ma~SO" filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum-ethyl acetate gradient) afforded the purified compound.
-o o H~l~ ~OCH3 1 NH20H. DlEA/EtO~l Boc~ ~ ~OCH3 NCJ3~ C~3 2 H~. Pd/C/E:tOH :HOA ~1 CH3 To a solution of ter~-butyloxycarbonyl-para-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).
The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL), and dry ethanol (40 mL) with warming. Subsequently, Pd/C catalyst (0.30 g, 10~ Pd) was added, and hydrogen was bubbled through the mixture with warming. The hydrogenation was continued until no starting material could be detected as judged by TLC. The catalyst was removed by ~iltration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 M) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound.

8~
SUE~STITUT~ S5~EET

Boc~ J~ ~OCH3 1 NNzOH. DlEA/EtOII Boc~N~ ~oCH3 CH3 2.H3,Pd/C~tOH:HOA~ ~ CH3 NC
H2N~ ~NH

To a solution o~ tert-butyloxycarbonyl-meta-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).
The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO;, 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 ~iltration, the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 M) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound.

3 I NH.OH.DlEA/EtOH ~ ~ I

CN z. U~.P~C~tOU:UOAc ~ NHZcH3 lJT~ S~ EE~

W 096/1948~ PCT/CA95100708 To a solution of tert-butyloxycarbonyl-ortho-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol ~20 mL) was added hydroxlyamine hydrochloride (O.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol).
The mixture was re~luxed and then cooled. The precipitate was ~iltered, washed with cold ethanol, diisopropylether, dried with MgSO4, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture o~ 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.

o o Boc~ ~ ~OCH3 Litbiumthi~zolc~HF ~ Boc~
~ CH3 H2N ~ ~ N

To a solution o~ thiazole (1.28 g, 15.0 mmol) in anhydrous THF
(30 mL) was added n-BuLi (1.6 M/hexane, 8.9 mL, 13.9 mmol) dropwise at -78 C, and the solution stirred. tert-Butyloxycarbonyl-para-amidino-phenylalanine-N,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 MgSO~, ~iltered, and concentrated under reduced pressure. The crude material was puri~ied on silica gel UT~ S~EET

(ethyl acetate/hexane), and concentrated under reduced pressure.

~N~ ~oCH3 LithiumthiazolrTHI

HN ~NH2 HN~NH2 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-meta-amidino-phenylalanine-N,O-dimethylamide (1 15 g, 3.3 mmoij in THF (i5 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 MgSO~, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

O O

HN~NI ~OCH3 Lithium thiazole/THF ~ Boc~ ~_~

7 ¢~ ,,NH2 N
HN NH

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) ~iU~ITU~E S~EE~

; CA 02208772 1997-06-20 -, W 096/19483 PCTICA95/00708 .
dropwise at -78~ C, and the solution stirred. tert-Butyloxycarbonyl-ortho-amidino-phenylalanine-N,O-dimethylamide (1.15 g, 3.3 mmol) in THF ~15 mL) was then added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL) and the organic layer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgSO~, filtered, and concentrated under reduced pressure. The crude material was puri~ied on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

O O

Boc ~ ~OCH3 1. H,. RaNUEtOH:NH 3 ~ Boc~ J~ I ~CH3 NCJ3~ CH3 H2N ,J3~ OCH3 tert-Butyloxycarbonyl-para-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H at room temperature ~40 psi). The solution was ~iltered 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 MgSO;, ~iltered, and concentrated under reduced pressure. 2 ~J BS~ITlJTE S~ EET

O O
H~ ~OCH3 1. H~,RaNVEtOH:NH3 N ~ l ~ CH3 CH3 ~-- OCH3 NC

tert-Butyloxycarbonyl-meta-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to yield a clear residue.
The residue was dissolved in ethyl acetate (250 mL), and washed with 1 N NaOH (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgSO4, filtered, and concentrated under reduced pressure.

~N ~ ~OCH3 l.H2,RaN~EtOH:NH3 Boc~ ~ N~ H3 -- CH ~NH2 OCH3 tert-Butyloxycarbonyl-ortho-cyano-phenylalanine-N,O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and sponge Raney Ni (100 mg) added. The solution was shaken under H at room temperature 7 (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 ~L). The solution was dried with MgSO;, filtered, and concentrated under ;TITUTE S!~EET

- .

: reduced pressure.

H~cs~NHz ~oC~N~ ~CH, ~3~ OCH3 H~CI2rrHF H J~l OCH3 H2N ZHNyN

NZ

tert-Butyloxycarbonyl-para-aminomethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the r~m~in;ng 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.

~N~JI~ ~CH H,CSJ~NHZ Eloc~ ~N~

¢~ OCH3 Hl~cl~/THl ~ ~ O~H3 - ZHN~NZ

tert-Butyloxycarbonyl-meta-aminOmethyl-phenylalanine-N,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 ~.
~!3ST~TUTE SHE~T

.

mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through ~ celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate - gradient) afforded the purified compound.

Boc~ ~N~ H3CSJ~NHZ Boc~ ~N~CH
~ OCH3 H~:CI2/THF ~ OCH3 I~NH2 ~N~NHZ

ZN

tert-Butyloxycarbonyl-ortho-aminomethyl-phenylalanine-M,O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.S mmol) added. The solution was concentrated under reduced pressure, the r~m~ining 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.

~N~ ~CH3 1. Lithi~m thiazole/THF ~N~
OCH3 ZHN~N~ J~ N

~, HN~NHZ NZ
NZ

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF

~B~@~ ITUT~ S! IEET

~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 (1S 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 a~ueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with M~SO~, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

I.Lithiomthi~ lc~UI ~ N

~H HN

ZN ~ NHZ ZHN ~ NZ

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 MgS0;, filtered, and concentrated under reduced pressure. The crude material was puri~ied on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

S~ TITUTE 5~!EET

:

O O

~~C ~ Nl ~ CH3 1, Li~iumthi~zolc~HF ~ N

~ N ~ NHZ N ~ NHZ

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 magnesium sulfate, ~iltered, and concentrated under reduced pressure The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

~N~ ~OCH3 1. Zn/Cu couple: ultra~ound ~NJI~ ~OCH3 CH3 2. Io-cH3c6H~)3p]2pdcl2 ~ CH3 2-amino-5-hromopyridin~

H2N~N~

A solution o~ 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 Slrlli ~I~E SH ~ ET

:.
~'~ W O 96/19483 PCT/CA95/00708 , . ~
starting material remained (as judged by TLC). Bis(tri-o-tolylphosphine)palladium dichloride (O.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HCl (50 mL; 0.lN), distilled H O (3 x 50 mL), drying over Na~SO, filtration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum/ethyl acetate gradient) af~orded the purified compound. -===

o o B~c~N ~ ~OCH~ I Lithi~mthi~Yulc~HF ~ ~c ~ N

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 ~, and the solution stirred. The amino acid-N,O-dimethylamide (1.07 g, 3.3 mmol) in anhydrous THF (15 mL) was then added dropwise and the resulting mixture stirred.
The rea~tion 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.

S~ Tllr~1 ~ E SHEET

o O

- Boc~ ~ ~OCH3 1. Zn/Cu couplc; ultrasound Boc~ ~ ~OCH3 CH3 2. [o-CH3C6HI)3P]2pdcl2 ~ CH3 2-cy~no~ romopyridin~ ll T
NC~N~

A solution of tert-butyloxycarbonyl-iodo-alanine-N,O-dimethylamide (~.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 7inc-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 ~ollowed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under a nitrogen atmosphere with heatin~, allowed to cool, ethyl acetate (100 mL) was added, and the mixture filtered into a separatory funnel. Sequential washing with aqueous HC (50 mL; 0.1N), distilled H O (3 x 50 mL), drying over MaSO, ~iltration, and concentration under reduced pressure yielded the crude product. Flash chromatography over silica gel (light petroleum/ethyl acetate gradient) afforded the purified compound.

o o HN ~ ~ OCH3 I.NH.OH.~lEA~tOH Boc~ ~ ~ OCH3 NC ~ CH3 '.H..P~C~tOH:HOA ~ CH3 NH

To a solution o~ tert-butyloxycarbonyl-(4-cyano)3-pyridylalanine-N,3-dimethylamide (1.34 g, 4.0 mmol) in dry 135T!TI! IT~ 5~1 EET

ethanol (20 mL) was added N,O-hydroxlyamine hydrochloride (0.416 g, 6.0 mmol), and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was ~iltered, washed with cold ethanol, diisopropylether, dried with MgSO~, concentrated under reduced pressure, and used directly in the next step. The semi-solid - was suspended in a mixture o~ 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 ~iltration, and the solution was concentrated under reduced pressure (50 mL), HCl (50 mL, 1 N) was added, and the mixture was concentrated once again to 50 mL. The solution was chilled overnight yielding the title compound.

o o Boc~ ~N~OCH3 1. Litbillm tlliazolc/THF Boc~
HZNb~ CH3 H2N~ N

- NH NH

To a solution o~ thiazole (1.28 g, 15.0 mmol) in anhydrous T~
(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 aci~-- N,O-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 MgSO;, ~ ~iltered, and concentrated under reduced pressure. The crude -~ material was puri~ied on silica gel ethyl acetate/hexane), and S~ T~TUTE SHEET

concentrated under reduced pressure.

~7 I. ll~ tO~/AcOi~ ~oc~ ~7~
N~ OCH3 HN~ OCH3 tert-Butyloxycarbonyl-3-(4-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-~4-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate ~250 mL), washed with 1 N NaOH ~2 x 50 mL), brine (2 x 50 mL), dried with MgSO4, ~iltered, and concentrated under reduced pressure to yield the title compound.

7 1. H2, PtO2/AcOH Boc~ ~7~CH3 ¢~ OCH3 H ~ OCH3 tert-Butyloxycarbonyl-3-~3-pyridyl)alanine-N,O-dimethylamide ~4.50 g, 14.4 mmol) was dissolved in acetic acid ~100 mL), and PtO ~100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl 3-~3-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate ~250 mL), washed ~ BSTiTU~E Sl iEET

with 1 N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgSO~, ç ~iltered, and concentrated under reduced pressure to ~ield the title compound.

~N~ ~CH, I.H~.P10~/AcOH ~oc ~1 OCH ~ ~ OCH3 -tert-Butyloxycarbonyl-3-(2-pyridyl)alanine-N,O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, and concentrated under reduced pressure yielding tert-butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide.
The residue was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgSO4, ~iltered, and concentrated under reduced pressure to yield the title compound.

O NZ O

~NJI~Nl ~CH3 H3CSJ~NHZ Boc~ ~ IN~CH3 ~ OCH3 H~C12/THF ~ OCH3 HN J ZHN~N J

tert-Butyloxycarbonyl-3-(4-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), SUE~S~ITUT~ SHEET

and HgCl~ (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and filtered through celite. The ~iltrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound.

Boc~ J~N~ H ~CS~NHZ I Boc~ ~N~
~ J OCH3 H~CI21THF ~ OCH3 ZN~NHZ

tert-Butyloxycarbonyl-3-(3-piperidyl)alanine-N,O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,M'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and ~iltered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) a~orded the title compound.

SUeSTlTUTE SHEET

-~N~ ~CII, H3CS ~IHZ l~oc~ J~N~

Cl' OCH3 H~CI /TEIF ~ OCtl3 NH N~NHZ

ZN

tert-Butyloxycarbonyl-3-(2-piperidyl)alanine-N,O-dimethylamide ~1.00 g, 3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (1.14 g, 3.2 mmol), and HgCl (0.95 g, 3.5 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (200 mL), and ~iltered through celite. The filtrate was concentrated under reduced pressure.
Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the title compound.

o o E;oc~ ~N~ 3 1. Ulhlumthl~u~lc/TUF

ZHN ~N~ ZHN ~N

NZ NZ

T ~ 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 mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate SUBSTITl~lTE ~HEET

(150 mL), and the organic Iayer washed with saturated aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgSO~, ~iltered, and concentrated under reduced pressure.

O ' o Boc~ ~No~H~H3 1. Lit~ mtlliazolclTHF E~oc~ ~N

ZN~NHZ ZN~NHZ

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

o . o ~CH3 1. Lithium thiazole/THF N~
OCH3 ~ N

~N~NHZ ~N~NHZ

ZN ZN

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 SU5BS~ITUTE ~iHE~T

~CA 02208772 1997-06-20 ;W 096/19483 PCT/CA95/00708 -78- C with stirring. The mixture was stirred at -78' C for 1 h. The guanidylated 2-piperidylalanine derivative 12.00 g, 3.2 mmol) in THF (15 mL) was added dropwise, and the resulting mixture stirred. The reaction was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer washed with saturated ~ aqueous ammonium chloride (2 x 50 mL), brine (50 mL), dried with MgSO~, filtered, and concentrated under reduced pressure.

o o Boc~ ~ IN~CH3 I. H2~ PtO2/AcOH ~N~D~ IN~cH3 O2N~ OCH3 H2N~ OCH3 tert-Butyloxycarbonyl-para-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in H O (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.

~N~ ~CH3 1. H2.PtOz/AcOH

OCH3 ~ OCH~
ozN H2 ~L~13STITUTE ~IEET

tert-Butyloxycarbonyl-meta-nitro-phenylalanine-M,o-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, ~ taken up in H O (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 o ~N~ ~CH3 1. H~. PtO2/ACOH BOC~ J~N~CH3 ~W~ OCH3 ~ - 1CH3 ~NO2 . ~NH2 tert-Butyloxycarbonyl-ortho-nitro-phenylalanine-N,O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) added. The solution was shaken under H until gas uptake ceased. The solution was filtered through celite, concentrated under reduced pressure, taken up in H O (150 mL), and lyophilized The semi-solid wa~
dissolved in ethyl acetate (350 mL), washed with 1 N NaOH (3 ::
50 mL), and brine (3 x 50 mL). The solution was dried with MgSOj, ~iltered, and concentrated under reduced pressure yielding the title compound.

~--i l.Z-Cl,N-HCO~/TNP:NiO ~ H~

H3N--~J OCH3 ~. Llthi~lmthi~ lc/TNp NHZ~ N

SUI~TITUTE SHEET

. CA 02208772 1997-06-20 '- W 096/19483 PCT/CA95/00708 1 . t er t -Bu tyloxycarbonyl-3-( ci s / trans - 4 -aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated a~leous 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 bisul~ate - (2 x 50 mL), and brine (2 x 50 mL). The solution was dried . with MgSO~, filtered, and concentrated under reduced pressure.
The semi-solid was chromatographed on silica gel (ethyl - acetate/ hexane).

2. To a solution o~ 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, ~iltered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

SlJBST~TlJTE SHI~ET

O O

~ Boc~ ~ ~CH3 I.Z-CI,NaHCO~HF:H2O ~oc~N ~ s~
ocH, 2.Litlliumtbi~zole/TllF ~ N _ 1. tert-Butyloxycarbonyl-3-( cis/ trans-3-aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF
[60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (O.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 r~m~i n; ng 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 MgSO~, 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 r 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.

SUBSTIT~5TE SHEET

~oc ~ ~ CH3 I.Z Cl~N4HCO~r~ HzO ~oc~
~ OC~3 l.Litlliumtli~zole~HF ~ N

.; NH2 NHZ
;

1. tert-Butyloxycarbonyl-3-( cis / trans-2 -aminocyclohexyl)alanine-N,O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF
[60 mL, (1:1)] with stirring. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF
(10 mL) was added dropwise. Excess solid sodium bicarbonate was added, the THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL), and mixed thoroughly. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4 N a~ueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgSO;, 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 a~ueous ammonium chloride (2 x 50 mLj, brine (50 mL), dried with MgSO, Y
filtered, and concentrated under reduced pressure. The crude material was puri~ied on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure.

~;IJ~ST~TU~E SIHEET

1. H,CSJ~NHZ ~N~
~ j OCH3HgCI2/TE~
H2N~ t. LithillmthiazolelTHF

1. tert-ButyloxycarbonYl-3-1 cis/ trans-4-aminocyclohexyl)alanine-N,O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry TH~ (20 mL) under nitrogen with stirring.
The solution was cooled to 0 C, M,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) a~orded 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.

SIJBSTITUTE SH!E~T

,.

N ~ ~ CH3 ~ H
OCH3H~CI2/THF _ N
. Lithium thilwle/THF [~

~NZ

ZHN

l tert-Butyloxycarbonyl-3-( cis/ trans-3-aminocyclohexyl)alanine-N,O-dimethylamide ~2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring.
The solution was cooled to 0' C, N,M'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The ~iltrate was concentrated under reduced pressure. Flash chromatography over silica gel (hexane/ethyl acetate gradient) afforded the puri~ied product.

2. To a solution o~ 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.

SUE~STITUT~ SHEE~

W O 96/19483 PCT/CA9~/00708 O ~Z O

E~oc~ J~ ~CH3 1. H3CSJ~NHZ Boc~ ~j_S
OCH3 H~CI7~H~ ~ N
2. Litllium tlli~ol~/THI; ~

~NH2 ~NH

ZN~NHZ

1. tert-Butyloxycarbonyl-3-( cis/ trans-2 -aminocyclohexyl)alanine-N,O-dimethylamide ( 2 .0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring.
The solution was cooled to 0 C, N,N'-bis-(benzyloxycarbonyl)-S-methyl-isothiourea (2.18 g, 6.1 mmol), and HgCl (1.81 g, 6.7 mmol) added. The solution was concentrated under reduced pressure, the r~m~in'ng 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 MgSO, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate/hexane), and concentrated under reduced pressure S ~ S ~ ~ ET

W 096/19~83 . PCT/CA95/00708 Exam~le 2 - Synthesis of Intermediates O O ' "~ 2 5 ~ N ~ O

THF/-7 ~ C 2 . . .
r ( 4S,5R)-3-(1-oxo-3-phenylpropyl)-4-(phenyl)-5-(methyl)-2-oxazolidone (2). A solution of 10.0 g (1.0 equiv., 56.4 mmol) of (4S, 5R)-4-phenyl-5-methyl-2-oxazolidone (1) in - 250 mL of dry THF, stirred at -78~C under argon, was treated dropwise with n-butyllithium (1.6 M in hexane, 1.1 ~- eq., 38.8 mL). After stirring for 30 min., 8.4 mL (1.0 - 10 equiv., 56.4 mmol) of hydroc;nn~moyl chloride was introduced dropwise over a 10 minute period. The resulting mixture was warmed to 0~C, stirred for an additional hour, and quenched with saturated ammonium ; chloride. The solvent was removed in vacuo and the resulting white solid dissolved in ethyl acetate and ddH2O. The aqueous phase was removed and extracted with two additional portions of ethyl acetate. The extracts -- were combined, washed with saturated sodium chloride, - dried over sodium sulfate, and the solvent was removed i.-20 vacuo to afford a white crystalline solid (2)(in 91~o yield).

m.p.95-96.5; [~]D-35.8 (c=1, CH2C12); [~]D-26.6 (c=1.01~
CH2C12 ) .

lHNMR (CDC13) ~ O.89(d, 3H, C_3 , J=6.6 Hz), 3.00-3.05 ~ (m, 2H), 3.26-3.34 (m, 2H), 4.73-4.78 (m, lH), 5.64, d, lH, J=7.4 Hz), 7.22-7.46 (m, lOH).

S~Slrll~UTE ~EET

CA 02208772 l997-06-20 O o o o "/ ~ ~ /Ph A solution o~ 5.0 g (1.00 equiv., 16.2 mmol) of (2) in 100 mL o~ dry THF, was cooled to -78~C. Enolization was achieved with 17.8 mL (1.1 e~uiv., 17.8 mmol) of lithium bis-trimethylsilylamide, which was added dropwise via~
syringe. The solution was stirred for 30 min. before 4.45 mL (3.0 equiv., 48. 5 r[unol) of allyl iodide was introduced and the reaction warmed to -15~C. After 1 h the reaction was ~uenched with saturated ammonium chloride and extracted (3X) with ethyl acetate. The organic phase was washed with sodium metabisulfite, dried over sodium sul~ate and the solvent removed in vacuo to a~ord an off-colour oil. Purification was achieved by ~lash chromatography on silica gel using a stepwise gradient (15:1, 12:1, 10:1) to yield (3), a colourless oil (95%).

[~]D 47-5 (c=3.12, CH2C12).

20 lHNMR (CDCl3) ~ 0.82 (d, 3H, CH3, J=6.6), 2.31-2.40 (m, lH, RC_=CHCH2), 2.49-2.57 (m, lH, RC_=CHCH2), 2.84-3.00 (m, 2H, Ph-CH2), 4.32-4.37 (m, lH, C_-(N)CO), 4.53-4.58 (m, lH, CH3-CH-), 5.03-5.13 (m, 2H, ABX, CH=CH2), 5.21 (d lH, Ph-CH, J=7.1 Hz), 5.81-5.89 (m, lH, C_=CH2), 7.20-7.42 - 25 (m, lOH, Ar_);

3C (CDC13) ~ 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.

$a~ ~36~ S~~ ~ ~T
-CA 02208772 l997-06-20 O o - ~ ~NJ~O 1. BH3/THF/o~c ~NJ~O
" ~- MCPBA/CHC13/R~ Me /Ph A sample of the allyl compouna, (3), (4.75 g, 13.6 mmol) - in THF (100 mL) was treated with 13 . 6 rnL ( 1. O equiv., 13 . 6 ; 5 mmol) of a 1.0 M solution of borane-tetrahydro~uran complex at 0~C and stirred for 2h. The solvent was evaporated and chloroform (100 mL) added via syringe.
~ Oxidation of the organoborane was achieved by the addition o~ 4.7 g (2 . 0 equiv., 27 . 2 mmol) of 3-chloroperoxybenzoic - 10 acid at 0~C, with warming to ambient temperature and stirring for an additional hour. The organic phase was washed with 5% Na2CO3, ddH2O, and dried over sodium sulfate. Due to the instability o~ the alcohol, a quick column was performed to remove the extreme polar and nonpolar material which originated from the 3-chloroperoxybenzoic acid. The alcohol (4) was obtained in a yield o~ 65%.

[ ~ D 3 9 .3 (c = l . 0 3 &, CH~ C 12 ) .

HNMR (CDC13) ~ 1.07 (d, 3H, CH3-CH, J=6.5 Hz), 1.81-1.93 (m, 3H, CH2-C_-H), 2 . 10-2 . 19 (m, lH, CH2-C_-H), 3 . 10-3 . 17 (m, 2H, Ph-C_ 2 ), 3 . 873 . 90 (m, 2H, CH2OH), 4 . 43-4 . 49 (m, lH, C_-CO), 4.70-4.75 (m, lH, CH3-CH), 5.36 (d, lH, Ph-C_, 25 J=7 . 1 Hz), 7 . 41-7 . 63 (m, lOH, Ar_);

~i~e~TiT~IT~ IE~

O O O o " ~ N~ ~O 1. PCC/mol. sieves~ ' ~ 'NJ ~ O
I Me /Ph 2. CH2C12/RT ~ Me /Ph ~ ~ OH 4 H J~O

To a solution of the alcohol (4) (1.0 g, 2.7 mmol), dissolved in dichloromethane (27 mL), 876 mg (1.5 equiv., 4.1 mmol) o~ 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 a~ter 30 min , i~
starting material remained, additional molecular sieves were added The solution was filtered through celite and the solvent was evaporated. The residue was dissolved in ethyl acetate and washed with saturated sodium chloride.
I~ the orange colour persisted in the organic phase, additional filterings through celite pads were per~ormed.
The aldehyde was obtained in a quantitative yield as a transparent, colourless oil (5).

HMMR (CDCl3) ~ 0.84 (d, 3H, CH3-CH, J=6.6Hz), 1.87-1.94 (m, lH, CH2-C_(H)-CHO), 2.04-2.13 (m, lH, CH2-C_(H)-CHO), .45-2.50 (m, 2H, Ph-CH2), 2.79-2.85 (dd, lH, CH2-CHO, J=13.3 & J=6.6), 2.92-2.99 (dd, lH, C_2-CHO, J=13.2 &
J=8.8), 4.19-4.22 (m, lH, C_-CO), 4.46-4.51 (m, lH, CH3-CH), 5.13-5.25 (m, lH, Ph-C_), 7 .20-7.39 (m, lOH, Ar-?~

9.69 (s, lH, CHO);

3C (CDCl3) ~ 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.

S lL~ B~;Ti T ~ S H E ~T

oo o O
1. PTSA/IJ-CYS Et-ester/ 11 11 mol. sieves ~N O

Me Ph Ph~/RT ~ ~' ~e / Ph H O S NH
O
~ 5 ~ 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 e~uiv., 8.52 mmol) of L-cysteine ethyl ester and 4 A
molecular sieves. The reaction was allowed to stir ~; overnight at ambient temperature followed be removal of solvent in vacuo. The residue was dissolved in chloroform, washed with saturated sodium chloride, ddH20, and dried over sodium sulfate. The solvent was removed in Vacuo to afford a gummy solid (6).

NJI\o Me3Al l DcM~

roomtemp. s /~H Me Ph H
S NH

CO2Et 2.OM trimethylaluminum in hexane (2.4mL, 4.8mmol, 3 e~uivalents) was added slowly to starting material (6) (800mg, 1.61mmol) stirring in anhydrous dichloromethane under argon, using ovendried equipment. After stirring overnight, HPLC indicated that the reaction had gone to completion.

ll4 ~' IJ ~ T~T~
.

The mixture was ~uenched with excess methanol, then - ~iltered on a short silica gel column (washing through with excess 10% methanol in ethylacetate. Evaporation gave 7&4 mg o~ crude material that was puri~ied using 2:1 hexane:EtOAc on a silica gel column a~fording 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 (CDC13) d 1.28-1.31 (m, 3H), 1.72-1.81 (m, 3H), 2.10-2.13 (m, lH), 2.66 (dd, lH, J= 11.5 and 6.0 Hz), 3.29-3.34 (m, 2H), 4.19-4.29 (m, 2H), 4.88 (dd, lH, J= 9.0 and 5.0 Hz), 5.22 (dd, lH, J= 8.0 and 6.0 Hz), 7.18-7.23 (m, 3H), 7.28-7.31 (m, 2H).

Z~

H H

LioH~H2o (48mg, 1.12mmol) in 10mL o~ water was added to starting material (7) (240mg, 0.76mmol) dissolved in 10mL o~
dioxane. After 1 hour, TLC in 1:1 Hexane:EtO~c showed no starting material. The reaction was c~uenched with 10% citric acid, then extracted twice with dichloromethane. Drying and evaporating the combined organic layers gave 354mg of crude product. This was redissolved in dichloromethane, then precipitated by adding excess hexane. The product was ~iltered to give 200mg (0.68mmol being a 90% yield)) of an off-white solid, (8), also known as 6S-benzylhexahydro-5-oxc-5H-thiazolo[3,2-a]pyridine-3R-carboxylic acid ,. .
H NMR (CD30D) d 1.71-1.82 (m, 3H), 2.12-2.17 (m, lH), 2.67 (dd, lH, J= 14 and 11 Hz), 2.77-2.81 (m, lH), 3.30-3.40 ll5 S~ STll~JFE SHE. T

= ~ :

(m, 3H), 4.81 (dd, lH, J= 8.5 and 4.9 Hz), 5.16 (t, lH, J=
7.5 Hz), 7.18-7.31 (m, 5H).

O O
l! (cocl)2 il /\/\~ OH ~ ~ Cl 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 liquid (10), cyclohexyl propionic acid chloride.

1. n-BuLi / THF / -780C
HN O I /~'~~~ N O

CH 'Ph 2 '~ ~' CH Ph ~CI
I

Th- chiral auxiliary (11) (13.6g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78C. 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 NH~Cl 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 puri~y the product This afforded in a white solid (12) which was recrystallized S~E~S~5~ Fl~

.

from ether and hexane to give the title compound in 78%
yield.

[a]D=-20.1(c=l,EtOH)i MP/BP mp=90.5-91.5OC
lH NMR (CDCl,3) d 0.86-1.10 (m, 5H), 1.18-1.30 (m, 4H), 1.54-1.75 (m, 7H), 2.86-2.97 (m, 2H0, 4.70-4.76 (m, lH), 5.65 (d, lH, J= 7 2 Hz), 7.28-7.42 (m, 5H).

1. LiHMDS/THF/ -78~(: ~

C~l~ rh r ~ ~h The starting material (12)(9.13g, 29 mmol, leq) was dissolved in dry THF and cooled to -78C, after which LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise over 40 mins. Then, 30 minutes later, allyl bromide (7.5 mL, 86.9 mmol, 3 eq) was added slowly over 10 mins. The mixture was le~t 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 Cl yellow oil (13) in 96% yield.

[~]D=+9.5(c=1.0,EtOH) ~H NMR (CDC13) d 0.92-1.10 (m, 5H), 1.10-1.39 (m, 5H), 1.63-1.75 (m, 6H), 2.27-2.42 (m, 2H), 4.01-4 14 (m, lH), 4.76-4.85 (m, lH), 5.00-5.07 (m, 2H0, 5.65 (d, lH, J= 7 Hz), 5.64-5.88 (m, lH0, 7.27-7.46 (m, 5H).

ll7 SUE~Slrl~UTE ~HEET

1. BH(cH(cH3)cH(cH3)2)2 o O

~N O THF/O C/2h ~, ~N O
./ 2. PCC / DCM / reflux(lh) ~J ~ /
CH3 Ph J CH3 Ph O

2-methyl-2-butene was added dropwise to borane dimethylsulphide complex at -12~C. The reaction was maintained at this temperature for 15 minutes and then it was warmed to 0'C, after which it was stirred for 2 hours.
The disiamyl borane was then added to a mixture o~ the starting material 13 in THF using a double-ended needle at 0 C. The mixture was then stirred ~or 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.
oncentration o~ the ~iltrates resulted in a 78% yield o~
a yellow gummy product (14).

t~]D=-l7~8(c=l.245~EtoH) H NMR (CDCl3) d 0.89-1.18 (m, 5H), 1.20-1.47 (m, 8H), 1.60-1.74 (m, 6H), 1.83-2.00 (m, lH), 2.48-2.53 (m, 2H), 3.90-4.10 (m, lH0, 4.12-4.16 (m, lH), 4.76-4.80 (m, lH0, 5.67 (d, lH, J= 7Hz), 7.27-7.46 (m, 5H), 9.77 (s, lH).

S ~ E s ~ ~ ~r . .. .

Il Toluene / p-TSA / ~
1'/~/--~/\ N o mol. sieves "~ N O
CH3 Ph ~, ~ CH3 Ph ' C02Et S NH

- 'C02Et 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 o~ 4A molecular sieves, and L-cysteine ethyl ester (3.87 g, 20.8 mmol, 1 equiv.). The mixture was stirred overnight, ~iltered and concentrated. The residue was then purified by silica gel chromatography (6:1 hexane:ethyl acetate) afforded 6.36 g o~ the product (15) in 61% yield.

[~]D=-48.3(c=1.095,EtOH) H NMR (CDCl3) d 0.84-0.98 (m, 4H), 1.11-1.38 (m, 7H), 1.50-1 90 (m, 10H), 2.80-2.99 (m, lH), 3.24-3.34 (m, lH), . 77-4.29 (m, 4H), 4.46-4.81 (m, 2H), 5.66 (d, lH, J=
7Hz), 7.27-7.46 (m, 5H).

o 1~l N/r~O Me3AI / DCM ~ , S

" 0~C RT " ' ~ ~ ~ N
~ CH3 Ph O C~2Et S~ N H

' C02Et ll9 Sl~Wl~ S ~ E ET

- The starting material (15) (1.97 g, 3.9 mmol, 1 e~uiv.) 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 minutes and then filtered. The residue after concentration in vacuo was run through a quick column (6 1 hexane:ethyl acetate), to remove auxilary and as many of the polar decomposition products as possible, affording in a 50% yield of a yellow oil (16).
H MMR (CDC13) d 0.83-0.98 (m, 2H), 1.09-1.38 (m, 10H), 1.57-2.00 (m, llH), 2.12-2.18 (m, lH), 2.49-2.54 ( m, lH), 3.10 (dd, lH, J= 11 and 6 Hz), 3.27 (dd, lH, J= 11.5 and 8.0 Hz), 4.11-4.25 (m, 2H), 4.88 (dd, lH, J= 11.0 and 5.o Hz), 5.14 (dd, lH, J= 10 and 6 Hz).

/\~S LiOH / H20-Diox ~ ~', S
" N ~> 1 0~C RT ~ \ N ,/

O C~2Et CO2H

The starting material (16) (0.95 g, 2.9 mmol, 1 equiv.) was dissolved in 10 mL o~ dioxane. The solution was cooled to 10 C, and to it was added LiOHH0(0.123 g, 2.9 mmol, 1 eq.) dissolved in 10 mL of water. The bath was removed and the mixture was stirred at room temperature for 1 hour. TLC showed complete reaction and the solvent was evaporated under vacuum. The remaining aqueous layer was washed with ether (2X), acidified with 10% citric acid, and extracted with dichloromethane (3X). The W O 96/19483 PCT/CA95/nO708 combined extracts were dried over sodium sulphate and concentrated to give a white solid which was - recrystallized from ether. Concentration of the ~iltrate and purification by silica gel column chromatography (2:1 hexane: ethyl acetate) resulted in more product (17) with a m.p. of 198.2-l99 C.

H NMR (DMSO-d6) d 0.78-0,93 (m, 2Hj, 1.11-1.27 (m, 5H), 1.34-1.36 (m, lH), 1.51-1.56 (m, lH), 1.60-1.75 (m, lH), 1.82-1.87 (m, lH), 2.15-2.18 (m, lH), 2.37-2.41 (m, lH), 3.03 (dd, lH, J= 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.83 (dd, lH, J= 9 and 4 Hz), 4.95 (dd, lH, J= 8 and 5.5 Hz).

O H
OH ~, ~/ -- CHN2 Cl H3N CH2CI
O / O
Cbz ~ Cbz j N N Cl~
HN~NH HN~NH ~
HN NH
Cbz Cbz Cbz BOC-DiCbz Arg (18) (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to OC. Triethylamine (2.2 mL) was added followed by 14.5 mmol of a lM toluene solution of isopropyl chloroformate via a syringe. The reaction was allowed to stir at OC for 30 minutes then quickly filtered. The white solid was discarded. To the filtrate was bubbled freshly prepared diazomethane until the color of the solution turned yellow. The reaction mixture was allowed to stand overnight in a well ventilated fumehood which facilitated the discharge of excess diazomethane. Dry ether was added to precipitate the diazoketone. The product was filtered and dried under vacuum to give light yellow fluffy solid (4.6 g, 58%).

EE~

.

Diazoketone (19) (lg, 1.77 mmol) was dissolved in THF (20 mL) and to this solution was added lM 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 af~orded the product (20) (1.02 g , 100%).

H NMR (DMSO-d6) d 1.65-1.77 (m, 3H), 2.06-2.50 (m, lH), 3.86-3.90 (m, 2H), 4.29 (m, lH), 4.76 (d, lH, J= 18Hz), 4.95 ~d, lH, J= 18 Hz), 7.35 (s, 2H), 7.36 (s, 2H), 7.35-7.41 (m, 10H), ~.71 (br s, 3H), 10.1 (br s, 2H).

C NMR (DMSO-d6) d 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3, 128.6, 128.7, 128.8, 128.9, 135.2, 135.9, 153.4, 157.4, 198.9.

, S ~ H N ~ Cl N ~ + ~ ~ ~ N ~>
HO NC~z o ~ HN ~
HN ~ NCbZ =~ \ , NC~z ~ Cl To mimetic (17) (0.422 g, 1.42 mmol) in THF (50 mL) at 0C
and in presence of N-methyl morpholine t0.19 mL), was slowly added lM toluene solution of isopropylchloroformate (1.71 mL). The reaction was allowed to stir at 0 C for 30 2~ minutes then treated with aminochloromethylketone (20) in small portions. Once the addition was complete the reaction was further stirred for 15 minutes followed by addition of N-methyl morpholine (0.19 mL). The reaction was stirred at ambient temperature for 3 hours, then extracted with ethyl acetate followed by washing with brine and 10% ac~eous citric acid Removal of organic solvent gave a white foam S ~ T- 5 !~

~ . , W 096119483 PCT/CA9~/00708 (21) (1 03 g, 96%) which was used further without puri~ication.

H MMR (CDCl3) d 0.07-0.97 (m, lH), 1.15-1.41 (m, 7H), 1.62-1.91 (m, lOH), 2.10-2.16 (m, lH), 2.43-2.48 (m, lH), 2.74-2.80 (m, lH), 3.01-3.07 (m, lH), 3.87-3.94 (m, lH), 4.11-4.19 (m, 2H), 4.60-4.66 (m, lH), 4.74-4.86 (m, 2H), 5.09-5.24 (m, 4H), 7.30-7.39 (m, lOH), 7.95 (d, lH, J= 8 Hz), 9.4 (br s, lH), 9.56 (br s, lH).

.~ .

GOC--N ~ FA/DCM r~ ~ o ~ 2. DMF/ BOP

~N~IIH~- Çr ~O,H ~ :

(N-t-BOC-N-tosyl)butyrylketoarginine (240mg, 0.515 mmol), was deprotected using 30% TFA in dichloromethane. The deprotected arginine derivative, was coupled with the mimetic (8) (100 mg, 0.343 mmol) in DMF under basic conditions (Et~N, pH = 8-9), using BOP reagent (228 mg, O.52 mmol) as the dehydrating agent. The reaction was typically complete within 2-4 hours. Extraction with ethyl acetate followed by successive washing with brine and 10%
a~ueous citric acid yielded the crude product. The crude product was puri~ied by column chromatography affording lB0 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.

s ~ s ~-~ N ~> 1. HS-CH2-C02H ' ~ ~ N ~>
o ~ j~ NHCbz ~ o , HN =
O =~ ~NCbZ O =~ \ N'- -- ~ 2. BBR3/DCM
Cl HO ~ s l24 SU!~3STITUlrE SII~E~T

W O 96/19483 PCT/CA9~/00708 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 o~ the reaction mixture with ethyl acetate followed by successive washing with brine and 10 % aqueous citric acid and evaporation of organic solvent gave the crude product which was purified by column chromatography to give foamy solid as the product (0.125 g, 62%).

This protected precursor (0.125 g, 0.154 mmol~ was dissolved in DCM (5 mL) and cooled to -78C. A lM DCM
solution o~ BBr (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 and the residue was partitioned between ether and water. The water layer was collected, lyophilized and the final product (23) obtained as à 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.

slJ~ S~

~X~PLE 5 A general method of synthesizing compound o~ formula II or e ~COOMe ~5~

Hs/~C~~Me NaCNBH3 ~ NH COOMe RT 16 hrs NH2 COOMe $1 KHMDS/THF ~/i N~J TosYI N3 O
O COOMe NH ~ COCH CH.CO M.
LiOH _ ~N,1, NH
CE17 NH~ NH. C[lz [~ 1. same procedure [~
f~S as for2Q ~f S

3~ ~J NH~N~J
o Co-NH~cocH2cH2co2Me ~ CO-NH~cocH CH~CO,,Me ~J 2. deprotection as ~' NH in example 3 NH
CBz NH ~ NH2 CBz NH ~NH2 SU ~S i ITlJ~ S~ EET

.

EX~PLE 6 Synthesis of :
~ .

~--N--O ~N~

~--NH
(16) Synthesis of 2-Benzyloxycarbonylamino-4-hydroxybutyric acid tert-butyl ester 1) ~o~cl NHZ NHZ
I NMM.THF
(Cy~l~hexyl)2NH.HOOC ~ CO2t-Bu 2)NaBH~,MeOH HO - COlt-Bu (1) (2) To a solution of the protected aspartic acid (l)(Bachem, 2.50 g, 4.95 mmols) in 50 mL o~ dry tetrahydro~uran (THF), at -10~C, under N~, was added N-methylmorpholine (109 ~L, 0.2 eq) and isopropyl chloro~ormate (1.0 M/toluene : 384 ~L, 1.1 eq). The soluion was stirred at

-10~C for 60 min. In another ~lask, NaBH4 (375 mg, 2 eq) was suspended in a dry 5:1 mixture of THF/MeOH (50 mL), at -78~C, under N~. This suspension was stirred at -78~C
for 30 min. The mixted anhydride solution was then added to the NaBH4 suspension dropwise via canula, and the final solution was stirred at -78~C for 3 hr. Acetic acid (2.8 mL, 10 eq) was then added and the solution was Sl~Ç~STIT~ S~I~ET

-warmed to r.t. (30 min). The solvents were evaporated, the residue taken up in EtOAc and washed with sat.a~.
NaHCO3 (2x) and brine. The organic layer was dried over MgSO4, the solids were filtered and the solvent - 5 evaporated to give 1.53 g (4.95 mmols, 100%) of the alcohol (2) as a clear oil.

H MMR (CDCl3, 400 MHz) : â 7.40-7.31 (m, 5H, ArH), 5.63 (d, lH, J=7.3, ~IH), 5.13 (AB system, 2H, J=12.2, CH2Ph), 4.43 (m, lH, H-2), 3.69 (m, 2H, H-4), 2.17 (m, lH, H-33, 1.63 (m, lH, H-3), 1.48 (s, 9H, t-Bu).

2-Benzyloxycarbonylamino-4-iodobutyric acid tert-butyl ester NHZ Ph~P.I, ~
HO~CO~t-Bu Imid;~ -1()~C I CO2t-Bu CEI3CN/Et2O
(2) (3) To a solution of the alcohol (2) (1.53 g, 4.95 mmols) in a 1:1 mixture of CH3CM/Et2O (50 mL), at -10~C, under N-~, were added successively imidazole (607 mg, 1.8 eq) and Ph3P (2.21 g, 1.7 eql. Iodine (2.14 g, 1.7 eq) was then added in small portions over a period of 15 min. After 25 the addition was completed, a white precipitate formed and the solution was brown. It was stirred at -10~C for 45 min. It was then poured in Et2O and the organic phase was washed with sat.aq. Ma2SO3, sat.aq. CuSO4, H~O and dried over MgSO4. The solids were filtered and the 30 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.

i28 S~iTl~lJTE S~.~ET

lH NMR (CDCl3, 400 MHz) : ~ 7.41-7.31 (m, 5H, ArH), 5.35 (bd, lH, J=7.3, MH), 5.13 (s, 2H, CH2Ph), 4.30 (m, lH, H-2), 3.22-3.12 (m, 2H, H-4), 2.42 (m, lH, H-3), 2.20 (m, lH, H-3), 1.48 (s, 9H, t-Bu).

Synthesis of 2-Benzyloxycarbonylamino-4-hexenoic acid tert-butyl ester NHZ ~ M~Br,CuI NHZ
I ~ CO2t-Bu THF.-78~C CO,t-Bu (3) (4) To a suspension of CuI (2.27 g, 5 eq) in dry THF (20 mL), at -78~C, under N2, was added slowly a l.OM solution in THF of vinyl magnesium bromide (23.4 mL, 9.8 eq). The solution was then warmed up to -10~C for 30 min (it turned then black) and cooled back to -78~C. A solution of the iodide (3) (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. NH4Cl (50 mL) was added and the mixture was 25 brought back to room temp. with vigorous stirring. It was then poured in Et20 and stirred for 5 min. The dar~.
suspension was filtered through a cintered funnel and the phases were separated. The aqueous phase was extracted with Et~O (2x) and the combined organic extracts were dried over MgSO~. The solids were filtered, the solvents evaporated and the crude oil purified by flash f . chromatography (silica gel, 5% AcOEt/Hex) to give 0.51 g (67%) of the pure alkene (4).

5~ TIT~ E ET

: CA 02208772 1997-06-20 W 096119~83 PCT/CA95/00708 H NMR (CDCl3, 400 MHz) : ~ 7.37-7.31 (m, 5H, ArH), 5.80 (m, lH, H-5), 5.33 (d, lH, J=7.8, MH), 5.12 (s, 2H, CH2Ph), 5.05 (d, lH, J=17.2, H-6), 5.01 (d, lH, J=10.4, H-6), 4.30 (q, lH, J=7.4, H-2), 2.16-2.08 (m, 2H, H-4), 1.92 (m, lH, H-3), 1.74 (m, lH, H-3), 1.48 (s, 9H, t-Bu).

Synthesis of l-Benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinecarboxylic acid tert-butyl ester HO
NHZ 1) H~(OA~)7~
~) NaHCO~. KBr ~ ~CO2t-Bu 3) NaBH~- ~2 ~ ZN~_ Ot-Bu (4) (5) To a solution of the alkene (4)(50 mg, 0.157 mmol) in dry THF (3.1 mL), at r.t., under N2, was added mercuric acetate (75 mg, 1.5 eq). The solution was stirred at r.t. for 18 hr after which it was cooled down to 0~C.
Sat aq. NaHCO3 (2 mL) was then added and the mixture was stirred at 0~C for 30 min. KBr (0.llg, 6 eq) was added and the mixture was stirred at r.t. for 2 hr. It was then poured in H~O/Et~O and the phases were separated.
The aqueous phase was extracted with Et~O (2x) and the combined organic extracts were dried over MgSO4. The solids were filtered and the solvents evaporated. Oxygen (O~) was bubbled into a suspension of NaBH~ (3.3 mg, 0.55 eq) in dry DMF (0.4 mL) for 1 hr, and to this was added dropwise (syringe pump, 3 mL/hr) a solution of the organomercurial bromide in DMF (3.1 mL) with continuous introduction of O,. The bubbling was continued for 1 hr and Et~O (5 mL) was added. The grey suspension was r 3STlTlJT ~
.

~iltered through Celite and the ~iltrate was evaporated.
The residue was chromatographed (silica gel, 6:4 Hex/EtOAc) to give the pyrrolidinol (5) (30 mg, 57%) as a clear oil.

H MMR (CDC13, 400 MHz) : ~ 7.37-7.28 (m, 5H, ArH), 5.22-5.09 (m, 2H, CH2Ph), 4.30 (dd, lH, .~=1.4, 8.3, H-2), 4.24 (m, lH, H-5), 3.70-3.57 (m, 3H, CH2-OH), 2.25 (m, lH), 2.13 (m, lH), 1.92 (m, lH), 1.70 (m, lH), 1.34 (s, 9H, t-Bu).

Synthesis of 1-Benzyloxycarbonyl-5-carboxy-2-pyrrolidinecarboxylic acid tert-butyl ester OH U
SO~-Pyr,E~N ~ ~
ZN ~ D MSO/CH~C~ ()~C ZN ~
~ Ot-Bu ~ Ot-Bu O O
(5) (6) To a solution of the alcohol (5) (50 mg, 0.149 mmol) and Et,N (62~L, 3 eq) in dry CH2Cl2 (0.8 mL) is added slowly, under N , at 0~C, a solution o~ SO3-Pyridine complex (71 mg, 3 eq) in dry DMSO. The solution was stirred at 0~C
~or 30 min and 10% citric acid (2 mL) is added. The pH
is brought to 4 with lM NaOH and the aqueous phase is extracted with Et2O (3x). The combined organic extracts were dried over MgSO4. The solids were ~iltered and th~
solvents evaporated to give a crude oil which was puri~ied by ~lash chromatography (silica gel, 7:3 Hex/EtOAc). The pure aldehyde (6) was obtained as a clear oil (45 mg, 90%).

IT~E S ~ ~ ET

t ' WO 96/19~83 PCT/CA95/00708 .

H NMR (CDCl3, 400 MHz) : ~ 9.68 + 9.56 (ds, lH, CHO), 7.36-7.29 (m, 5H, ArH), 5.23-5.11 (m, 2H, CH2Ph), 4.57-. 4.39 (m, 2H, H-2, H-5), 2.30-1.97 (m, 4H, H-3, H-4), 1.47 - + 1.36 (2s, 9H, t-Bu).

(7) NH
\\ BOCHN~
H ~ MgSO4, CH2C12 HN ~ ~
z ~ OBut ~ CO Me ~ OBut - ,~ 2) NaBH(OAC)~
~ AcOH, THF ~ ~
BX - -(6) (8) The pyrrolidine-aldehyde (6) is coupled with the protected - diamino-propionic acid (7) by first forming the imine (8) (MgSO, CH Cl). Isolation o~ the imine (8) is done by filtration of the MgSO; and evaporation of the solvent.
The crude imine is then treated with NaBH(OAc) and actic acid (AcOH) in THF for 15 hours to Dbtain the amine (8) a~ter extrative work-up.
, ~

HII ~ 1) H2 Pd/C /MeOH D ~ H N
CO M~ --OBut ~--CO M~OBUt BoNH ~NH
(8) (9) SUBSTITUT~ Sl IEE~

The CB7 (7) protecting group o~ the amine (8) is removed by hydrogenation with palladium on charcoal 10% as a - catalyst in methanol (MeOH). The catalyst is ~iltered and the MeOH evaporated to give the crude diamine (9) that can be used without any purification.

/ CO M- ~ OBut Heat ~ ~, . OBut (9) (10) The cyclisation is done by heating the crude oil (9) ~rom step 7, neat slightly above the boiling point of methanol.
The bicyclic lactam (10) is purified by flash chromatography.

~i~B~T~T~TE ~ El!

. . =

=

HN/--~> ~cocl ~ H
~ ~OBut Pyridine D ~ ~ ~OBut s - ,NH ,NH
C~
(10) ~11) - 5 The secondary amine of the bicyclic lactam (10) is protected as an amide using benzoyl chloride in pyridine.
Evaporation o~ the pyridine and extractive work-up give the bicyclic lactam-amide (11).
-o H o = ~ HCl/Et20 OBUt ~ O OH

BCC Cl- NH
~11) (12) The BOC and t-butyl ester protecting groups of bicyclic lactam amide (11) are removed under acidic conditions (HCl in eth~l ether (Et 0)). The amine salt (12) precipitates- -out o~ solution and is collected by ~iltration.

-W 096/19483 . PCT/CA95/00708 ~ c6H5CH2OCOCl ~ N

~ COOH K2CO3, CH3cN ~ OH
a- N~; o O

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

(14) ~ ~' ~ J ' ~ ~
ZH~ ~ BOP, El~lPr2,DlVF H~
~ --NHZ
(l;!
(15) The carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF using BOP as the coupling agent in the presence of diisopropylethylamine (EtNiPr).
Extraction with ethyl acetate ~EtOAC) gives compound (15) as a solid which is purified by chromatography.

~3 ~

Z~ , F~/C ~0 MeGI-' Hl: HM~
NH
(1'1 (1~ !

The two CBZ(Z) protecting groups o~ 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).

T~ C~ T
.

W 096/19483 PCT/CA9~/00708 SYNTHESIS OF COMPOUND (10) ~S

2 ~O~HN~N~

NH
(1 o ) N H

H N~_ ~ OH
f~ + O 1) BOP/DMF ~ ~
o \ NMM HN~ ~_ S (3) Z O o O~

J~ OH (4 H N
Z O

(2) 4-methylmorpholine (NMM) was added to a solution of the carboxylic acid (2) (1.7g, 4.9mmol, l.Oeq), 4-hydroxyproline (3) (S.39mmol, l.leq), and BOP reagent (2.17g, 4.9mmol, l.Oeq) 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 l37 ~j ;2~ S ~

: W 096/19483 . PCT/CA9S/00708 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 ~lash chromatography (5:4:1, ethyl acetate-hexane-methanol). l.lg of pure product (4) was recovered 48%
yield.

OH (3~
l)triethylarnine ~ ~\
z J~ $o ~ a z ~ $o ~ o \ ooc o (4) (5) To a solution of 4-hydroxyproline derivative (4 ) (115mg, 240umol, l.Oeq) in dichloromethane (10 mL, anhydrous) at 0~C is added triethylamine (72mg, 720umol, 3.Oeq) and methanesulfonyl chloride (28mg, 240umol, l.Oeq) and the reaction mixt~re is stirred at room temperature. The mixture is then ~uenched with an agueous 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).

.
138 ~=

.

~ l)Hg(~AC)2 ~S~ ~
HN~ --~_ CF30C0H/0~C z ~ o o ~ 2)NaBH4 ( 6 ) ( 5 ) The ~n~mine (5) (l.Oeq) is treated with mercuric acetate (1.1 e~) 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 (6).

HN~_ NCS/Da~ HN
'O \ Z O
(6) (7) To a solution of the lactam thioether (6) (l.Oeq) in dr~
dichloromethane N-chlorosuccinimide ~l.Oeq) is added at 0~C. The reaction mixture is warmed to room temperature.
When the reaction shows no more starting material the solid is filtered and the solvent evaporated to dryness.
~ The crude material (7) is use without any ~urther puri~ication for step 5.

c ~ g ~ E~ ~

:: ' -:

- Cl~S~ ~S~

I ~ $ P~ ~ $
Z O O T H F Z O O

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

~~ H2O~ HF ~ OH

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

~ q ~ L ~ 5~E~Flj .e z o~_ 2 ) BBr3 / CH2C12 0 HN~N~
NH
(1 O) )--NH
H N

The crude carboxylic acid (9) is coupled with benzythiazole keto arginine in DMF using BOP as the coupling reagent in the presence o~ 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 BBr in dichloromethane at room temperature finally gives the bicyclic benzothiazole keto arginine inhibitors (10).

The ~ollowing compounds are produced accordingly with the execption that the appropriate substitution of products were made in order to obtain the ~inal compounds.

COMPOUND ~11 ~S~

O --~ ~,N~

NH

(11) J-NH
H N

T; l! ~ R ~ T

- . ~

COMPOUND $12 V\~s~

HzN~ ~_ ' NH
)--NH

(12) EX~iMPLE 8 O O
J~CI 1. CH2-N=N T H F /Elher -15~C I Cl 2. HG/Elher o~ cCH 3 O~ OCH, Commercially available glutaric acid monomethyl ester - chloride ~1) (20 ml,0.144 mol) was disolved in 40 ml of dry tetrahydrofuran (THF) and cooled to -15~C. Excess diazomethane freshly prepared in 300 ml of Ether was introduced via cannula at -15~C to the solution. The - mixture was left to warm up to room temperature overnight.
Excess diazomethane was evacuated from the flask with a current of argon. To bring the reaction to completion, 75 ml of 1 N HC1 in Ether was added at 0~C and left to warm up to room temperature for 5 hours. The volume of.the solvent was reduced and then washed with 2x 5% NaHCO3 l42 Sk ~ ~f S ~ ~T~ R

dried over Na2CO3 and evaporated to give crude chloromethylketone (20.46 g , 79%) wich was used in the next step without ~urther puri~ication.

1H NMR (CDCL3, 400 MHz) d 1.16-1.2 (t,lH), 1.83-1.9 (m,2H), 2.27-2.35 (m,2H), 2.6-2.64(t,lH), 3.6 (s,3H), 4.04 (s,2H).

o ,CI o~~CH2CH3 H a. H-C~s-OE T / CH 3COONa/ NaCNB H 3 HN /\
MeOH, R.T ~ S
o//~OCH
2 ~

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 e~, 4.9 g, 78.59 mmol). The heterogeneous mixture was left to stir at room temperature for 2h30 min. 200 ml of methanol (MeOH) was then added to disolve all the solid and the pH was brought to 2 with lN HCl .The mixture was then basified with saturated NaHCO3 until pH= 8. MeOH was evaporated and the remaining aqueous solvent was washed with ethyl - acetate and dichloromethane. Solvents were combined, dried over Na2SO4 and evaporated. The crude residue was ~ purified by silica gel flash column chromatography using a gradient.of eluents ethyl acetate / hexane in the l43 T~ w a ~ ~ ~! E t l~

c : W 096119483 PCT/CA95/00708 - following ratios: (3:7, 5:5, 6:4, 7:3) giving cyclic- compound (3).

lH MMR (CDCl3 , 400MHz) o~ 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, lH, J=10.9), 2.72-- 2.76 (dd, lH, J=10.7 Hz), 2.8-2 86 (m, lH), 3.6-3.64 (d, 4H, .J=2.55 Hz), 3.63 (s, 3H), 4.13-4.2 (m, 2H) 13C NMR (CDC13, 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 o ~ OCH2CH3 H
H N--T oluene / H + ,~ S
R efl K ~ N ~
0~ OCH 2CH 3 O~OCH~

Cyclic compound (3) (913mg, 3.32 mmol) was disolved in 50 ml of dry Toluene. (lS)-(+)-10-Camphorsulfonic acid ( 92 mg, 0.39 mmol) was added and the mixture was le~t to reflux for 4 days. When all 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 % NaHCO3. The Ethyl :25 acetate layer was dried over Ma2SO4 and evaporated. The crude residue was purified by silica gel ~lash column -chromatography using 60 % ETOAC / 40 %Hexane followed by = l44 ~;~!3STiTlJTE 5g~EET
~ . , 70 % ETOAC / 30 % Hexane giving 62.5% of Bicyclic compound (4).

H MMR (CDC13, 400MHz) of compound (4) d 1.27-1.31 (t, 3H, J=7 Hz), 1.5-1.6 (m, lH), 1.72-1.87 (m, 2H), 2.02-2.1 (m, lH), 2.33-2.46 (m, 2H), 2 52-2.59 (m, 2H), 2.83-2.88 (dd, lH,J=14 , 4 Hz), 3.14-3.18 (d, lH), 3.78-3.85 (m, lH), 4.2-4.27 (q, 2H, J= 3.9 Hz), 5.9-5.92(t, lH, J=3.4 Hz).

H H
,/~'~/\S LiOH.H20,THF/H20 ' . S
N ~/ > N
o~OCH2CH3 OOH

Bicyclic (4) (366mg,1.5 mmol) was dissolved in 25 ml of THF and 5 ml H2O Lihium hydroxide. monohydrate (1.1 eq, 7.05 mg, 1.68 mmol) was added in 2.3 ml of H2O, at 0~C
and the mixture was left to stirr at 0~C for 1 hr and at room temperature for 3 hrs. THF was then evaporated and the remaining aqueous mixture was acidified by addition of Citric acid until pH=2. Extraction of aqueous mixture with 2 x CH2CL2 and 2 x ETOAC, drying of combined organic layers with Ma2SO4 and evaporation gave a crude residue wich was purified by silica gel flash column chromatography using 70 % ETOAC / 30 %Hexane followed b~-4.7 % HOAC / Ethyl acetate giving the pure acid (5) in 5' % yield . 16 % of starting material(4) was recovered.

H MMR (MeOD, 400 MHz) of compound (5) d 1.57-1.69 (m, lH), 1.70-1.80 (m, lH), 1.81-1.89 (m, lH), 2.05-2.12 (m, lH), 2.35-2.5 (m, 2H), 2.51-2.66 (m, 2H), 2.86-l45 S ~ 5~ S ~Y ~ ET

- - =

. W 096/19483 PCT/CA95100708 - 2.91 (dd, lH, J=13.8, 4 Hz), 3.12-3.17 (d, lH), 3.3-- 3.32 (m, lH), 3.78-3.84 (m, lH), 5.76-5.78(t, lH, - 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 I~LiHMDS ~78~C

O OH 2)BrBr,-78~C~ RT o OH

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

W O 96/19~83 PCT/CA95/00708 1 ) H3N'~N~

N11CE~Z S

NH~
2 ) BBr3 / CH2C
NH
~NH

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

The ~ollowing compound is produced accordingly with the exeption that the appropriate substitution of products were made in order to obtain the ~inal compounds.
COMPOUMD #8 \I H
~ s ~~N~/

~ NH--\<,N~
~ S
NH
~'NH

TIT~ SHE~T

. W 096/19483 . PCT/CA9~/00708 . .

EX~MPLE 9 : Determination of K Values for Heterocyclics The affinity cf inhibitors for thrombin was measured according to the procedures described in tDiMaio et al, J.
Bio. Chem., 1990, 265:21698) Inhibition of amidolytic activity o~ human thrombin was measured fluorometrically -using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50 mM Tris-HCl buffer (pH 7.52 at 37~C) containing 0.1 M NaCl and 0.1% poly(ethylene glycol) 8000 at room temperature, and (Szewczuk et al., Biochemistry, 1992 31:9132).

. .
The hydrolysis of the substrate by thrombin was monitored on a Varian-Cary 2000TM spectrophotometer in the fluorescence mode (~eX = 383 nm, ~em = 455 nm) or on a Hitachi ~2000TM fluorescence spectrophotometer (~6:: = 383 nm, - ~ = 455 nm), and the fluorescent intensity was calibrated en using AMC. The reaction reached a steady-state within 3 minutes after mixing thrombin with the substrate and an inhibitor. The steady-state velocity was then measured for a few minutes. The compounds of this invention were also pre-incubated with thrombin for 20 minutes at room temperature before adding the substrate. The steady-state was achieved within 3 min and measured for a few min. The kinetic data (the steady-state velocity at various concentrations of the substrate and the inhibitors) of the competitive inhibition was analyzed using the methods - 30 described by Segel (1975). A non-linear regression program, RNLIN in the IMSL library (IMSL, 1987), LMDER in MIMPACK
library (More et al., 1980) or MicrosoftTM ExcellTM , was used to estimate the kinetic parameters (K, VL:~: and K).

dTT assa~

l48 $,.~ T~ E~T
.

The fibrin clotting assay was performed in 50 mM Tris HCl buffer (pH 7.52 at 37 ~C) cont~;nin~ 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 hllm~n thrombin and bovine fibrinogen, respectively, as reported elsewhere (Szewczuk et al.,supra). The clotting time was plotted against the inhibitor concentrations and the ICs, 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 HCl buffer (pH7.8 at 23 C) containing O.lM NaCl and 0.1% (w/v) polyethylene glycol 8000. Human plasma (60~L, collected in 3.8% sodium citrate, blood/anticoagulant 9:1) was added to microtiter wells (microtiter plate, Falcon) containing lOO~L of various inhibitor dilutions. The solution was mixed after which 50~L of human thrombin (lnM
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. IC
values were calculated at 30 minutes as the inhibitor concentration that gave half the optical density of the control.

Platelet A~qre~ation and Secretion ., Rat blood was collected into ACD (6/l v/v) by cardiac puncture. Suspensions of washed platelets were prepared as described by Ardlie et al, (Br. J. Haematol. 1970, 19:7 l49 ~5~ ;1riTE~ 6~E~

and Proc. Soc. Exp. Biol. Med., 1971, 136:1021). The ~inal suspending medium was a modi~ied Tyrode solution (MaCl 138mM, KCl 2.9mM, HEPES 20mM, NaH~PO4 0.42mM, NaHCO
12mM, CaCl lmM, MgCl. 2mM, 0.1% glucose, 0.35% albumin, ~ 5 apyrase l~L/mL pH 7.4). Platelet counts were adjusted to 5000,000/~L.

, .
.~ To permit measurement of the extent of release of the . .
-' contents of the dense granules, the platelets were -~ lO labelled in the first washing solution with I~C-serotonin (5-HT) (l~Ci/lOmL of washing fluid) and release of '~C-serotonin was determined as described in Holmsen et al, (Enzymology, 1989, 169:206). Inipramine (5~M ~inal conc.) was added to present the reuptake of released serotonin.
1~
Platelet aggregation was recorded at 37Cc in an aggregometer (BioData PAP-4) at a stirring speed o~ 1,100 rpm by measuring the variations of light transmission.
; Percentage of aggregation was determined 3 min. after the addition of the stimulating agent (human thrombin O.lIU/mL
~inal conc.). Inhibitors were preincubated 1 minute at 37 C before addition of stimulating agent. ICs values represent the concentration that was necessary to inhibit platelet aggregation or secretion to 50% of the control.
~rterial Thrombosis Model FeCl Induced Carotid Arterial Iniurv Mcdel The FeCl induced injury to the carotid artery in rats was induced according to the method described by Kurz, K.D., Main, R.W., Sandusky, G.E~, Thrombosis Research 60; 269-280, 1990 and Schumacher, W.A. et al. J. Pharmacology and Experimental Therapeutics 267; 1237-1242, 1993.

3~ ~ale, Sprague-Dawley rats ~ 375-410 g) were anesthetized with urethane ~ 1500 mg\kg ip). Animals were laid on a l50 LI ~ T~ T

37~C heating pad. The carotid artery was exposed through a midline cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid sheath. Using forceps, the artery was lifted to provide sufficient clearance to insert two small pieces of polyethylene tubing (PE-205) underneath it. A temperature probe (Physitemp MT23/3) was placed between one of the pieces of tubing and the artery. Injury was induced by topical application on the carotid artery above the temperature probe of a small disc (3 mm dia.) of Whatman No.l filter paper previously dipped in a 35% solution of FeC13. The incision area was covered with aluminum foil in order to protect the FeCl from degradation by light. The vessel temperature was monitored for 60 minutes after application of FeC13 as an indication of blood flow. Vessel temperature changes were recorded on a thermister (Cole-Palmer Model 08533-41).

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

l5l 5~;T~TUTE ~ T

' :, Table 1 ,.
- Antiplatelet activity !lM
5-HT dTTlCs~Plasma fibrin cmpd Ki ~lM~A~gregationSecretion !lMclot assay IC~
!lM
0005 4 ND ND 47 >450 . .
,.
0010 4.6 21 19 89.5 >450 , 0015 16 >100 >100 162 >450 - 0020 2.2 18 14.2 22 >450 0025 53 >100 >100 >625 >450 0030 8.6 >100 >100 67 320 0035 34 >100 >100 319 >450 0040 19 >100 >100 207.5 >450 0045 74 ND ND 415 ,450 0050 62 ND ND >450 0065 32.7 47.5 52 42 200 0070 4.4 22 2.1 25 78 0080 0.048 0.4 0.38 0.375 ND

. 0090 0.031 ND ND 0.33 ND

-EF~ S!~
' *suspension of washed platelets from rats a Inhibitory dissociation constant for human a-thrombin T~T~

W 096/19483 PCT/CAg51~0708 , ' Table 2 ~= Compound ki dTT Route M.O.T
(nM) (nM) ivb+inf(min)+/-sem ~- 0225 550 0.75-50 23+/-7 !~ 0245 5 0.5 30 27+/ 3 8 0.75-5022.6+/-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 0210 8 0.75-5020.13+/-3.4 0260 16 0.75-5014.83+/-0.2 0305a 220 0305b 12000 0265a 4 0.75-5021.5+/-10 0265b 18 0.75-5014.83+/-2.3 0285 10 0.75-5011.33+/-1.34 0315a+b 45 0315b 10 0.75-5030.33+/-8.4 0335 25 138 0.75-5045.8+/-14.2 0.5-3041.5+/-7.27 0.25-2027.5+/-11,3 0340 0.6 0.25-20 36+/-9,6 0.75-5042.25+/-11,9 0345 2 0.75-5050+/-5.86 0915 1600 0.75-50 15+/-1,3 ~54 S ~
~.., W o 96/19483 PCT/CA95/00708 Table 2 (continued) Compound Ki dTT Route M.O.T
(nM) (nM) ivb+inf (min)+/-sem 0925a+b 10 0.75-50 19.6+/-0.2 0925b 30 0925a 7 0.75-50 20.3+/-3.5 0940a 16 0.75-~0 15.2+/-0.82 0940b 1 60 0950a 1 50 0950b 1 000 a = early eluting on RP HPLC single lsomer b = late eluting on RP HPLC single isomer a + b = mixture C ~ E~

_

Claims (57)

WE CLAIM:

1. A compound of formula (I):

(I) wherein:
A is selected from (CH-R8)0-1 , S, SO, SO2, O and NR8 wherein R8 is hydrogen, C1-~ alkyl optionally interupted with 1 or 2 heteroatoms; C~-1~ aryl, C~-~
cycloalkyl or heterocyclic ring or a hydrophobic group;
B is selected from S, SO2, O, -N=, NH, -CH= and CR6R7 wherein R6 and R7 are independently selected from hydrogen and C1-6 alkyl provided that when A is S, SO, SO2, O, or NR9, then B is CR~R7;
D is selected from (CH-R9) 0-2, wherein R9 is hydrogen, C1-~ alkyl or -C(O)R1; and CH with a double bond to B
when B is -N= or -CH=;
E is selected from CH2 and CH substituted with the -C(O)R, provided that only one of D and E is substituted with with -C(O)R1;
X is selected from O, N-R5, or CH-R5;
Y is selected from O, S, SO, SO2, N-R5 and CH-R5 provided that when X is N-R5 then Y is CH-R6 or O, and when X is O then Y is CH-R9;
Z is selected from O, S and H2;
R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is selected from H and C1-~ alkyl optionally substituted with C~ aryl, a 6 member heterocycle or a C3-~ cycloalkyl ring;

R3 is selected from H, NR6R~ and C1-~ alkyl; and R4 and R5 are independently selected from H; NR6R7; C6-16 aryl or C3-7 cycloalkyl optionally substituted with C1-6 alkyl; C1-16 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C6-1~ aryl, heterocycle or C3-7 cycloalkyl group optionally substituted with halogen, hydroxyl, C1-6 alkyl; an amino acid side chain; and a hydrophobic group.

2. A compound according to claim 1, wherein R1 is one of formula VIa to VId:

VIa VIb VIc VId wherein:
R11 is hydrogen or C1-~ alkyl;
K is a bond or -NH-;
G is C1-4 alkoxy; cyano; -NH; -CH~-NH~; -C(NH)-NH;
-NH-C(NH)-NH2; -CH2-NH-C(NH)-NH~; a C~ cycloalkyl or aryl substituted with cyano, -NH, -CH2-NH~, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH~, -CH~-NH~, -C(NH)-NH3, -NH-C(NH)-NH~ or -CH~-NH-C(MH)-NH~;
U is cyano, -NH~, -C(NH)-NH2 or -NH-C(NH)-NH~;
P is a bond, -C(O)- or a bivalent group:

, , , or J is C1-6 alkylene optionally substituted with OH, NH2 and C1-6 alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and T is H, OH, amino, a peptide chain, C~-1~ alkyl, C1-1~
alkoxy, C~~ aralkyl, or heterocycle optionally substituted.

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

wherein X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X where X7 is hydrogen, C~-4 alkyl, or C9-~ aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X~, or CH-X~; and R' is hydrogen, C1-~~ alkyl optionally carboxyl substituted, carboxyl, -C~-~~ alkyl-CO-~C1-~~ alkyl, C~~~
aralkyl, C~~~ cycloalkyl, aryl or an aromatic heterocycle.

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

wherein R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C0-1~ alkyl-CO~-C1-~ alkyl, C~
aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

5. A compound according to claim 4, wherein T is selected from:

or wherein R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C0-~ alkyl-CO~-C1-~ alkyl, C~
aralkyl, C3-~ cycloalkyl, aryl or an aromatic heterocycle.

6. A compound according to claim 1, wherein one of R4 and R5 is a hydrophobic group selected from C1-~ alkyl, C1-2~

alkenyl or C~ alkynyl optionally interrupted by a carbonyl group, C~ aryl, C~ cycloalkyl, C~
aralkyl, C~ cycloalkyl substituted C1-20 alkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group and the ring portion is optionally substituted with C1-~ alkyl; and a hydrophobic amino acid side chain.

7. A compound according to claim 6, wherein R~ is H.

8. A compound according to claim 1, wherein Z is O.

9. A compound according to claim 1, wherein R~ is H.

l0. A compound of the formula (VII):

wherein R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or C1-6 alkyl;
R3 is selected from H, NR,R. and C1-~ alkyl; and R4 and R5 are independently selected from H; NR~R; C~
aryl or C~ cycloalkyl optionally substituted with C~ alkyl; C1-~ alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR,R. or a C~ aryl, heterocycle or C~ cycloalkyl group optionally substituted with halogen, hydroxyl, C~ alkyl; an amino acid side chain; and a hydrophobic group.

160

11. A compound according to claim 10, wherein R1 is one of formula VIa to VId:

VIa VIb VIc VId wherein:
R11 is hydrogen or C~ alkyl;
K is a bond or -NH-;
G is C1-4 alkoxy; cyano; -NH; -CH2-NH2; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C(NH)-NH2; a C6 cycloalkyl or aryl substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH~; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH~ or -CH2-NH-C(NH)-NH2;
U is cyano, -NH2, -C(NH)-NH2 or -NH-C(NH)-NH2;
P is a bond, -C(O)- or a bivalent group:

, , , or J is C1-~ alkylene optionally substituted with OH, NH, and C1-~ 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, C1-~ alkyl, C1-~
alkoxy, C~ aralkyl, or heterocycle optionally substituted.

12. A compound according to claim 11, wherein T is a heterocycle selected from the group consisting of:

wherein X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X, where X, is hydrogen, C1-4 alkyl, or C~ aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X7, or CH-X7;
R' is hydrogen, C1-~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO-C1-~ alkyl, C~
aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

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

and R' is hydrogen, C1-~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C~ alkyl, C~
aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

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

or and R' is hydrogen, C1-~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C~ alkyl, C~
aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

15. A compound according to claim 10, wherein R~ and R~
are H.

16. A compound according to claim 10, wherein R4 is C1-~
alkyl optionally interupted with a heteroatom or a carbonyl, and optionally substituted with a C~
aromatic, C~ cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF3 or oxo.

17. A compound according to claim 10, wherein R5 is H.

18. A compound according to claim 12, wherein:
R3 is H;
R4 is C1-~ alkyl optionally interupted with a heteroatom or a carbonyl, and optionally substituted with a C6-~ aromatic, C~ cycloalkyl or heterocycle ring wherein the ring is optionally substituted with CF3 or oxo; and R5 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 (.alpha.-benzothiozolo keto arginine).

20. A compound according to claim 1, of formula (VIII):

wherein R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or C1-6 alkyl;
R3 is selected from H, NR6R7 and C1-~ alkyl; and R4 and R5 are independently selected from H; NR6R7; C6-16 aryl or C3-7 cycloalkyl optionally substituted with C1-6 alkyl; C1-16 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C1-16 aryl, heterocycle or C~ cycloalkyl group optionally substituted with halogen, hydroxyl, C1-6 alkyl; an amino acid side chain; and a hydrophobic group.

21. A compound according to claim 20, wherein R1 is one of formula VIa to VId:

VIa VIb VIC VId wherein:
R11 is hydrogen or C1- alkyl;
K is a bond or -NH-;
G is C1-4 alkoxy; cyano; -NH2; -CH2-NH2; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C(NH)-NH2; a C cycloalkyl or aryl substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2;
U is cyano, -NH2, -C(NH)-NH or -NH-C(NH)-NH2;

P is a bond, -C(O)- or a bivalent group:

, , , or J is C1-~ alkylene optionally substituted with OH, NH~
and C1-6 alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and T is H, OH, amino, a peptide chain, C1-~ alkyl, C1-~
alkoxy, C~ aralkyl, or heterocycle optionally substituted.

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

wherein X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X- where X7 is hydrogen, C1-4 alkyl, or C5-~ aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X-, or CH-X-;
R' is hydrogen, C1-~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C~ alkyl, C
aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

23. A compound according to claim 22, wherein T is selected from the group consisting of:

and R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C1-~ alkyl, C~
aralkyl, C3-~ cycloalkyl, aryl or an aromatic heterocycle.

24. A compound according to claim 23, wherein T is selected from:

or and R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C1-~ alkyl, C~
aralkyl, C3-~ cycloalkyl, aryl or an aromatic heterocycle.

25. A compound according to claim 20, wherein R2 and R~
are both H.

26. A compound according to claim 20, wherein R4 is H or C1-6 alkyl substituted with COOH.

27. A compound according to claim 20, wherein R2, R3 and R4 are H and R5 is C1-~ alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C6-~
aryl, heterocycle or C3-7 cycloalkyl group optionally substituted with halogen, hydroxyl or C~ alkyl.

28. A compound according to claim 22, wherein:
R2, R3 and R4 are H; and R5 is C1-~ alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C6-~ aryl, heterocycle or C3-7 cycloalkyl group optionally substituted with halogen, hydroxyl or C1-6 alkyl.

29. A compound according to claim 20, selected from:
0345 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(5-methyl-thiazole-2-carbonyl)-butyl]-amide; and0340 4-Oxo-2-(3-phenyl-propionyl)-octahydro-pyrrolo[1,2-a]pyrazine-6-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide.

30. A compound according to claim 1, of formula (IX):

wherein Y is selected from O, S, SO, SO2, N-R5 and CH-R8;
R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or C1-6 alkyl;
R3 is selected from H, NR6R7 and C1-6 alkyl; and R4 and R5 are independently selected from H; NR6R7; C1-16 aryl or C~ cycloalkyl optionally substituted with C~ alkyl; C~ alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C~ aryl, heterocycle or C~ cycloalkyl group optionally substituted with halogen, hydroxyl, C~ alkyl; an amino acid side chain; and a hydrophobic group;
R8 is hydrogen, C~ alkyl optionally interupted with 1 or 2 heteroatoms; C6-16 aryl, C~ cycloalkyl or heterocyclic ring or a hydrophobic group; and n is 1 or 2;

31. A compound according to claim 30, wherein R is one of formula VIa to VId:

VIa VIb VIc VId wherein:
R11 is hydrogen or C~ alkyl;
K is a bond or -NH-;
G is C1-4 alkoxy; cyano; -NH2; -CH2-NH2; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C(NH)-NH2; a C6 cycloalkyl or aryl substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH7 or -CH2-NH-C(NH)-NH2; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2;
U is cyano, -NH2, -C(NH)-NH2 or -NH-C(NH)-NH2;
P is a bond, -C(O)- or a bivalent group:

, , , or J is C1-6 alkylene optionally substituted with OH, NH
and C1-6 alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and T is H, OH, amino, a peptide chain, C1-16 alkyl, C~
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 X5, X10, X,, and X12 are each independently selected from the group consisting of N, or C-X~ where X7 is hydrogen, C1-4 alkyl, or C5-~ aryl;

X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X7, or CH-X7;
R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C0-16 alkyl-CO2-C1-16 alkyl, C6-20 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:
and R' is hydrogen, C1-~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C~ alkyl, C~
aralkyl, C3-7 cycloalkyl, aryl or an aromatic heterocycle.

34. A compound according to claim 33, wherein T is selected from:

or and R' is hydrogen, C1-16 alkyl optionally carboxyl substituted, carboxyl, -C0-16 alkyl-CO2-C1-16 alkyl, C6-20 aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

35. A compound according to claim 30, wherein R2 and R3 are both H.

36. A compound according to claim 30, wherein R4 is H, NR6R7 or C~ alkyl substituted COOH.

37. A compound according to claim 30, wherein R6 is C6-16 aryl, C~ aralkyl, or C1-16 alkyl substituted with C3-7 cycloalkyl.

38. A compound according to claim 32, wherein n is 1;
R2, R3 and R4 are H; and R5 is C1-16 aryl, C6-20 aralkyl, or C1-16 alkyl substituted with C~ cycloalkyl.

39. 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. A compound according to claim 1, of formula (X):

wherein B is O, S, -CH2-, or -NH-;
R1 is an arginyl moiety or an analog or derivative thereof optionally substituted with an amino acid, a peptide or a heterocycle;
R2 is H or C1-6 alkyl;
R3 is selected from H, NR6R7 and C~ alkyl; and R4 and R5 are independently selected from H; NR6R7; C~
aryl or C3-7 cycloalkyl optionally substituted with C~ alkyl; C1-16 alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR6R7 or a C~ aryl, heterocycle or C3-7 cycloalkyl group optionally substituted with halogen, hydroxyl, C~ alkyl; an amino acid side chain; and a hydrophobic group.

41. A compound according to claim 40, wherein R1 is one of formula VIa to VId:

VIa VIb VIc VId wherein:
R11 is hydrogen or C~ alkyl;
K is a bond or -NH-;
G is C~ alkoxy; cyano; -NH; -CH2-NH2; -C(NH)-NH2; -NH-C(NH)-NH2; -CH2-NH-C(NH)-NH2; a C~ cycloalkyl or aryl substituted with cyano, -NH, -CH2-NH2, -C(NH)-NH2, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2; or a 5 or 6 member, saturated or unsaturated heterocycle optionally substituted with cyano, -NH2, -CH2-NH2, -C(NH)-NH, -NH-C(NH)-NH2 or -CH2-NH-C(NH)-NH2;

U is cyano, -NH, -C(NH)-NH2 or -NH-C(NH)-NH2;
P is a bond, -C(O)- or a bivalent group:

, , , or J is C~ alkylene optionally substituted with OH, NH
and C~ alkyl and optionally interrupted by a heteroatom selected from O, S and N;
n is 0 or 1; and T is H, OH, amino, a peptide chain, C~ alkyl, C~
alkoxy, C~ aralkyl, or heterocycle optionally substituted.

42. A compound according to claim 41, wherein T is a heterocycle selected from the group consisting of:

wherein X5, X10, X11 and X12 are each independently selected from the group consisting of N, or C-X, where X7 is hydrogen, C1-4 alkyl, or C5-2 aryl;
X6 and X13 are each independently selected from the group consisting of C, O, N, S, N-X7, or CH-X,;
R' is hydrogen, C~ alkyl optionally carboxyl substituted, carboxyl, -C0-16 alkyl-CO2-C1-16 alkyl, C6-20 aralkyl, C~ cycloalkyl, aryl or an aromatic heterocycle.

43. A compound according to claim 42, wherein T is selected from the group consisting of:

and R' is hydrogen, C~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C1-16 alkyl, C6-20 aralkyl, C3-7 cycloalkyl, aryl or an aromatic heterocycle.

44. A compound according to claim 43, wherein T is selected from:

or and R' is hydrogen, C~ alkyl optionally carboxyl substituted, carboxyl, -C~ alkyl-CO2-C~ alkyl, C~
aralkyl, C3-7 cycloalkyl, aryl or an aromatic heterocycle.

45. A compound according to claim 40, wherein R2 and R3 are both H.

46. A compound according to claim 40, wherein R4 is C1-16 alkyl substituted with C6-16 aryl optionally substituted with C1-16 alkyl.

47. A compound according to claim 40, wherein R5 is H.

48. A compound according to claim 42, wherein B is S;
R2, R3 and R5 are H; and R4 is C1-16 alkyl substituted with C~ aryl optionally substituted with C1-16 alkyl.

49. A compound according to claim 40, selected from:
925 7-Benzyl-6-oxo-octahydro-pyrido[2,1-c][1,4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)butyl]-amide; and940 6-Oxo-7-phenethyl-octahydro-pyrido[2,1-c][1,4]thiazine-4-carboxylic acid [4-guanidino-1-(thiazole-2-carbonyl)-butyl]-amide.

50. A method for the treatment or prophylaxis of thrombotic disorders in a mammal, comprising administering to said mammal an effective amount of a compound according to claim 1.

51. A method according to claim 50, wherein said thrombotic disorder is venous thrombosis.

52. A method according to claim 50, wherein said thrombotic disorder is a pulmonary embolism.

53. A method according to claim 50, wherein said thrombotic disorder is arterial thrombosis.

54. A method according to claim 50, wherein said thrombotic disorder is myocardial infarction.

55. A method according to claim 50, wherein said thrombotic disorder is cerebral infarction.

56. A process for producing a compound according to claim 1.

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