SK83897A3 - 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

(54) Title of the invention: Low molecular weight bicyclic thrombin inhibitors

(13) Document's type: A3 (51) Int. Cl. ⁶ : OJ C 07D 513/04, A 61K 31/435, OJ C 07D 487/04, A 61K 31/495 U (C 07D 513/04, C 07D 277: 00 C 07D 211: 00) (C 07D 513/04) C 07D 279: 00, C07D 221: 00) (C 07D 487/04, C07D 241: 00) C 07D 209: 00)

(57) Annotation:

Described are heterocyclic competitive inhibitors of the thrombin enzyme having the formula (I) in which A can be (CH-R 5) 0, S, SO, SO ₂ , O and NR ₈ , wherein R ₈ is hydrogen, C 1. C ₆ alkyl optionally interrupted with 1 or 2 heteroatoms; C 1-6 aryl, C 3-7 cycloalkyl or a heterocyclic ring or a hydrophobic group; B may be S, SO ₂ , O, -N =, NH-, -CH =, and CR 6 R 7, wherein R ₇ and R ₇ may independently be hydrogen or C 1. ₆ alkyl with the proviso that when A is S, SO, SO _2, O, _S or NR, then B is CR ₆ R _7; D can be (CH-R ₉₎ o_ _2, wherein R ₉ is hydrogen, alkyl or _d6-C (O) R |; and CH with a double bond to B when B is -N = or -CH =; E may be CH ₂ or CH substituted with -C (O) R _b with the proviso that only one D and E is substituted with C (O) R _b X may be O, NR ₅ , or CH-R ₅ ; Y may be O, S, SO, SO _2, NR ₅ or CH-R _8, p podminenkou that when X is _NR, Y is _CH-R or O, and when X is O, Y is CH -R ₈ ; Z may be O, S and H ₂ ; R 1 is a polar amino acid residue of arginyl or an analogue, or a derivative, or a derivative thereof, optionally substituted with an amino acid, a peptide or a heterocycle; R ₂ can be H or alkyl optionally substituted _d6 with C ₆ aryl, 6-membered hetero ring, or C _3. 7, cycloalkyl ring; R ₃ may be H, NR ₆ R _7, or C _b6 alkyl and R 4 and R ₅ can be independently H; NR ₆ R _7> C ₆ . ₁₆ aryl or C _{3. 7} cycloalkyl optionally substituted with C alkyl _{b6; BL6} C alkyl optionally interrupted by one or more heteroatom or carbonyl group and optionally substituted with OH, SH, NR ₆ R _7, or C _b | ₆ aryl, heterocycle or C _{3. 7} cycloalkyl of LITEL substituted with halogen, hydroxyl, C alkyl _b6; an amino acid side chain; and a hydrophobic group. These compounds can be used in vitro as anticoagulants and in vivo for the treatment and prophylaxis of thrombotic diseases such as vascular thrombosis, pulmonary embolism and arterial thrombosis inducing acute ischemic events, such as myocardial infarctions or cerebral infarctions.

Low molecular weight thrombin inhibitors

Technical field

The present invention relates to compounds useful in the treatment of thrombotic diseases, more particularly novel heterocyclic thrombin inhibitors.

BACKGROUND OF THE INVENTION

Irregular thrombus formation along the walls of blood vessels accelerates acute cardiovascular disease states, which are the leading cause of death in economically developed societies. Plasma proteins such as fibrinogen, proteases and cellular receptors involved in hemostasis seem to be important factors that play a role in acute and chronic coronary disease, as well as cerebral artery disease, by assisting in the formation of thrombus or blood clots which effectively reduce normal blood flow and intake. Vascular aberrations that stem from primary pathological conditions, such as hypertension, rupture of arterosclerotic plaques or exposed endothelium, activate biochemical cascades that serve as a response and repair of the damaged site. Thrombin is a key regulatory enzyme in the coagulation cascade; performs numerous tasks, both positive and negative feedback regulator. However, under pathological conditions, this regulatory role is multiplied through catalytic activation by the cofactors required for both thrombin formation and activation of the ΧΙΠ factor required for crosslinking and stabilization of fibrin.

In addition to a direct effect on hemostasis, thrombin has direct effects on various cell types that promote and enhance the pathogenesis of arterial thrombosis. The enzyme is the most potent platelet activator causing their aggregation and release of substances (such as ADP TXA, NE) that further promote a thrombotic cycle. Platelets in the fibrin network represent an essential framework for white thrombosis. Thrombin also has direct effects on endothelial cells, where it causes the release of vasoconstrictors and translocation of adhesion molecules, which become sites of attachment of immune cells. In addition, the enzyme causes mitogenesis of smooth muscle cells and fibroblast proliferation. From this analysis, it is clear that inhibition of thrombin activity is a viable therapeutic approach to slow down the proliferative phenomena accompanying thrombosis.

The basic endogenous neutralizing factor of thrombin activity in mammals is antithrombin III (ΑΊΊΠ), which is circulating plasma macroglobulin with a low affinity for the enzyme. Heparin exhibits clinical efficacy in vascular thrombosis by increasing ATIH / thrombin binding during catalysis. However, heparin also catalyzes the inhibition of other proteases in the coagulation cascade, and its efficacy in platelet-dependent thromboses is greatly reduced or abolished due to the inaccessibility of the thrombus-binding enzyme. After prolonged administration of heparin, adverse side effects such as thrombocytopenia, osteoporosis, and triglyceridemia have been observed.

Hirudin, obtained from the glandular secretions of the leech hiridio medicinalis, is one of the natural anticoagulant protein inhibitors of high molecular weight thrombin activity (Markwardt F. Cardiovascular Drug Reviews, 10, 211, 1922). It is a biopharmaceutical that demonstrates efficacy in experimental and clinical thromboses. The potential drawback of the use of hirudin as a therapeutic agent is the probable antigenicity and lack of effective methods of neutralization, especially in view of its extremely strong thrombin binding characteristics. The excessively high affinity for thrombin is unique and is attributed to both the simultaneous interaction with the catalytic site and the presence of a remote "anion-binding exo-site" in the enzyme itself.

Thrombin activity can also be abolished by hirudin-like molecules such as hirulog (Maraganore, JM 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 the catalytic site of thrombin and thereby inhibit proteolysis of this protein or other protein substrates such as the thrombin receptor. A common strategy in designing inhibitory compounds relies on mimicking specific properties in the primary and secondary structure of natural enzyme substrates. Thus, Blomback et al. they first proposed a thrombin inhibitor that was modeled according to a partial sequence of fibrinogen A (LB1) and a chain containing a segment that undergoes proteolysis (Blomback, et al., J. Clin. Lab. Invest., 24, 59, 1969). This segment of fibrinogen contains minimal residues starting from phenylalanine:

-3Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg-Gly-Pro-Arg

T cleaved bond

The systematic substitution of amino acids within this segment led to the optimization of tripeptide inhibitory sequences, as in the case of the (D) -Phe-Pro-Arg peptide, which is analogous to the interactions within the P-P-P local binding site of the thrombin. (Bajusz S. et al., In Peptides: Chemistry Structure and Biology: Proceedings of the Fourth American Peptide Symposium, Walter R., Meienhofer J. Eds. Ann Arbor Science Publishers Inc., Ann Arbor MI, 1975, pp. 603) .

Bajusz et al. also disclosed similar compounds such as (D) Phe-Pro-Arg- (CO) H (GYKJ14166) and (D) MePhe-Pro-Arg- (CO) H (GYKI 14766) (Peptides-Synthesis, Structure and Function: Proceedings of the Seventh American Peptide Symposium, Rich, DH & Gross, E. eds., Pierce Chemical Company, 1981, pp. 417). These tripeptidyl aldehydes are potent inhibitors of thrombin both in vitro and in vivo. For both GYKI-14166 and GYKI-14766, the aldehyde group is believed to contribute strongly to the inhibitory activity in terms of its chemical reactivity to the catalytic Ser thrombin residue by forming a hemiacetate intermediate.

Similar work in the field of thrombin inhibitory activities utilized the basic recognition binding motif provided by the tripeptide (D) Phe-Pro-Arg by incorporating various functional and reactive groups into the locus that corresponds to the putative cleavage (Pi-Pi).

In U.S. Pat. No. 4,318,904, Shaw discloses chloromethyl ketones (PPACKs) that are reactive against Ser and His. These two residues represent part of the thrombin catalytic triad (Bode, W et al., EMBO Joumal 8, 3467, 1989).

Other examples of thrombin inhibitors in which the underlying motif (D) of PhePro-Arg is found are those in which COOH-terminal boroarginine variants such as boric acid or borates are incorporated (Kettner, C. et al., J. Biol. Chem., 268, 4734 (1993).

Other related compounds of this motif are those containing phosphonates (Wang, CL.J., Tetrahedron Letters, 33, 7667, 1982) and α-keto esters (Iwanovicy, EJ et al., Bioorganic and Medicinal Chemistry Letters, 12, 1607). (1992).

-4 Neises, B. et al. have described a trichloromethyl ketone thrombin inhibitor (MDL-73756) and Attenburger, J.M. and company. have discovered a related difluoroalkyl amidoketone (Tetrahedron Letters, 32, 7255, 1991).

Maraganore et al. (European 0 333 356: WO 91/02750: USA 5 196 404) disclose a series of thrombin inhibitors that contain a D-Phe-Pro- residue and suggest that this proposed structure fits well in the furrow near the active site of thrombin. Variants of these inhibitors are essentially linear or cyclic peptides built on the remainder of D-Phe-Pro. Another series of patents and patent applications discloses attempts to develop effective thrombosis inhibitors using alpha-ketoamide and peptide aldehyde analogs. (EP 0333356: WO 93/15756: WO 93/22344: WO 94/08941: WO 94/17817).

Others have also focused their attention on peptides, peptide derivatives, peptide alcohols, or cyclic peptides as anti-thrombotic agents (WO93 / 22344, EP 0276014: EP 0341607: EP 0291982). Others have tried to use amidine sulfonic acid residues (US 4,781,866) for this purpose, while others have tried para- or meta-substituted phenylalanine derivatives (WO 92/08709: WO 92/6549).

A series of Mitsubishi patents and patent applications have apparently provided potent arginine amide compounds for use as antithrombotic agents. The chemical structures described in these documents represent side group variations on the arginine amide compound (US 4173 630: US 4 097 591: CA 1 131 621: US 4 096 255: US 4 046 876: US 4 097 472: CA 2 114 153 ).

Canadian patent applications 2,076,311 and 2,055,850 provide cyclic imino derivatives that have an inhibitory effect on cell aggregation.

Many of the examples above converge to maintain at least a linear acyclic tripeptide motif containing an arginyl unit, the side chain of which is required for interactions with a carboxyl group located based on the Pi specific thrombin graft. The two adjacent hydrophobic groups provide additional bonding through preferred Van der Waals interactions within the minor hydrophobic slit on the enzyme surface of the putative P-P site.

It is one object of the present invention to provide thrombin inhibitors that exhibit target enzyme inhibitory activity, thrombin.

It is another object of the present invention to provide thrombin inhibitors that exhibit inhibitory activity against the target thrombin enzyme and are provided in a pharmacologically acceptable form.

Yet another object of the present invention is to provide for the use of heterocyclic thrombin inhibitors, and their formulations as anticoagulants and thrombin inhibitors.

Yet another object of the present invention is to provide for the use of heterocyclic thrombin inhibitors and their formulations for therapeutic applications for various thrombotic diseases.

Another object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The enzyme inhibitors of the present invention are surrounded by the structure of Formula I.

SUMMARY OF THE INVENTION

The present invention provides four novel compounds that exhibit thrombin inhibitory activity as expressed in Formula I:

where:

A can be (CH-R 5) 0-1, S, SO, SO 2, O, and NR 5, wherein R 5 is hydrogen, C 1-6 alkyl optionally interrupted with 1 or 2 heteroatoms; A C 6-16 aryl, C 3-7 cycloalkyl or heterocyclic ring or a hydrophobic group;

B is selected from S, SO2, O, -N =, NH, -CH = and CR ₆ R ₇ wherein R $ and R ₇ may each independently H or Ci.g alkyl with the proviso that when A is S, SO, SO 2 , O or NR 5, then B is CR 1 R ₇

D can be (CH-R 9) 0-2 wherein R ₉ is hydrogen, C 1-6 alkyl or -C (O) R 1; and CH with a double bond to B when B is -N = or -CH =;

E may be CH 2 or CH substituted with -C (O) R 1, provided that only one D and E is substituted with -C (O) R 1;

X may be O, N-R 5, or CH-R 5;

-6Y may be O, S, SO, SO 2, N-R 5 or CH-R 8, with the proviso that when X is N-R 5, then Y is CH-R 8 or O, and when X is O then Y is CH -R ₈ ;

Z may be O, S and H2;

R 1 is a polar amino acid residue of arginyl or an analogue or derivative or derivative thereof optionally substituted with an amino acid, peptide or heterocycle;

R 2 can be H or C 1-6 alkyl optionally substituted with C 6 aryl, 6-membered heterocycle or C 3-7, cycloalkyl ring;

R 6 can be H, NR 6 R 7, or C 1-6 alkyl, and ^R 4 ^R 5 can be independently H; NR R ?; C 6-10 aryl or C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl;

C 1-6 alkyl optionally interrupted with one or more heteroatoms or carbonyl and optionally substituted with OH, SH, NR 5 R 7, or C 6-10 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted with halogen, hydroxyl, C 1-6 alkyl; an amino acid side chain; and a hydrophobic group.

As will be clear from the following findings, the molecules, compositions and methods of the invention are useful as anticoagulants, or in the treatment and prevention of various diseases attributed to thrombin as an adverse effect, as well as for diagnostic purposes.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to molecules that inhibit the enzyme thrombin. These molecules are characterized by heterocyclic residues as shown in Formula I:

wherein X, Y, Z, A, B, D, E, and R 1 to R 4 are as previously defined.

The term hydrophobic group (HG) as used hereinafter refers to any group that has no affinity for or displaces water. Hydrophobic groups include, but are not limited to, C 1-10 alkyl, C 2-20 alkenyl (e.g. vinyl, allyl), or C 2-20 alkynyl (e.g. propargyl C-C-CH-OH) optionally interrupted by a carbonyl group (e.g. acyl group); C 6-10 aryl, C 3-7 cycloalkyl, C 6-20 aralkyl, C 6-10 cycloalkyl substituted with C 1-20 alkyl, wherein the aliphatic moiety

-7 is optionally interrupted by a carbonyl group (e.g., forming an acyl group) and the ring moiety is optionally substituted with C 1-6 alkyl such as methyl, ethyl or tert-butyl; or a hydrophobic side amino acid chain. Preferred hydrophobic groups are cyclohexyl, benzyl, phenylmethyl, 2-phenylethyl, and para-tert-butylphenylmethyl.

The term "arginyl" represents an amino acid residue of arginine or an analogue, or a derivative thereof. For example, an analog or derivative of a natural residue may incorporate a longer or shorter alpha carbon chain (e.g., an ethylene or butylene chain); replacing the guanidine group with a hydrogen bond of a donor or acceptor group (e.g., amino, amidino or methoxy); replacing a methylene chain with a steric tension group (e.g., aryl, cycloalkyl, or heterocyclic ring); by eliminating the terminal carboxyl (e.g. decarboxy) or hydroxyl (e.g. aldehyde), or a combination thereof.

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

The term aromatic or aryl represents an unsaturated carbocyclic ring consisting of 6 to 16 carbon atoms, which are optionally mono- or di-substituted with OH, SH, amino (e.g. NR 6 R 7) halogen or C 1-6 alkyl. Aromatic rings include benzene, naphthalene, penantrene, and anthracene. Preferred aromatic rings are benzene and naphthalene.

The term cycloalkyl represents a saturated carbocyclic book consisting of 3 to 7 carbon atoms, which are optionally mono or di-substituted with OH, SH, amino (e.g. NR 6 R 6) halogen or C 1-6 alkyl. Cycloalkyl groups include cyclopropyl, -butyl, -pentyl, -hexyl and -heptyl. A preferred cycloalkyl group is cyclohexyl.

The term aralkyl represents a substituent containing an aryl moiety linked through an alkyl chain (e.g., benzyl, phenethyl) wherein the total number of carbon atoms of the aryl moieties and the alkyl chain is specified. The aryl or chain portion of the group is optionally mono- or di-substituted with OH, SH, an amine (e.g., NR 4 R 7), halogen or C 1-6 alkyl.

The term heteroatom, as used herein, represents oxygen, nitrogen or sulfur (O, N or S), sulfoxyl or sulfonyl (SO or SO2) unless otherwise indicated. It is understood that alkyl chains interrupted by one or more heteroatoms mean that the carbon atom of the chain is replaced by a heteroatom of the same valence. Preferably, alkyl

The -8 chain is interrupted with 0 to 4 heteroatoms and that two adjacent carbon atoms are not simultaneously replaced.

The term heterocyclic represents a saturated or unsaturated mono- or polycyclic (e.g. bicyclic) ring in which 1 or more (e.g. 1-4) heteroatoms, which may be N, O and S, are incorporated. It is understood that the heterocycle is optionally mono- or disubstituted with OH, SH, amino (e.g. (NR 1 R 3), halogen, CF 3, oxo or C 1-6 alkyl. Examples of suitable monocyclic heterocycles are pyridine, piperidine, pyrazine, imidazole, thiazole, oxazole, furan, pyran and Examples of suitable bicyclic heterocycles are indole, quinoline, isoquinoline, purine and carbazole.

The term hydrophobic amino acid represents an amino acid residue that carries an alkyl or aryl group attached to an α-carbon atom. Thus, a glycine that does not have such a group attached to the α-carbon atom is not a hydrophobic amino acid. The alkyl or aryl group may be substituted provided that the substituent or substituents do not decrease from the overall hydrophobic nature of the amino acid. Examples of hydrophobic natural amino acids are alanine, isoleucine, leucine, phenylalanine, and non-naturally occurring amino acids as described in The Peptides vol.5, 1983, Academic Press, Chapter 6, by D.C. Roberts and F. Vallaccio. Suitable non-naturally occurring amino acids are cyclohexylalanine and 1-aminocyclohexanecarboxylic acid.

By amino acid side chain is meant a substituent attached to the α-carbon relative to the amino group. For example, the amino acid side chain of alanine is a methyl group, while benzyl is the side chain of phenylalanine.

is preferably H or C 1-6 alkyl. More preferably, H is methyl or ethyl, and most preferably R2 is H.

Preferably R 1 is H or C 1-6 alkyl. More preferably, R 1 is H, methyl or ethyl, and most preferably R 3 is H.

It is preferred that one of R 4 or R 5 is a hydrophobic group, such as a saturated or unsaturated 5 or 6 carbon ring optionally linked to another carbon ring group, while the other is H, C 1-6 alkyl optionally substituted with NR 6 R 7 or carboxyl. The hydrophobic moiety may be attached via a linker such as a C 1-8 alkyl chain optionally interrupted with 1 or more (1-

4) heteroatoms, carbonyl or sulfonyl. More preferably, one of R 4 or R 5 is phenyl, cyclohexyl, indole, thienyl, quinoline, tetrahydroisoquinoline, naphthyl, or benzodioxalan bonded via C 1-10 alkyl optionally interrupted with a heteroatom or carbonyl, while the other is H, carboxymethyl or carboxyethyl.

It is preferred that A is absent or is CH2.

It is preferred that B is S or CH2.

It is preferred that D is CH2.

It is preferred that E is CH substituted with -C (O) R 1, wherein R 1 is as previously defined.

It is preferred that X is CH-R5 or N- R5.

It is preferred that Y is CH-R 8 or S.

It is preferred that Z is O.

In a more preferred embodiment, R1 is represented by one of formulas VIa to VId:

where :

R 11 is hydrogen or C 1-6 alkyl;

K is a bond or -NH-;

G is C 1-6 alkoxy; cyano; -NH 2; -CH 2 -NH 2; -C (NH) -NH ₂ ; -NH-C (NH) -NH ₂ ; -CH ₂ -NHC (NH) -NH ₂ ;

C 6 cycloalkyl or aryl substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH ₂ , -ΝΉC (NH) -NH 2 or -CH ₂ -NH-C (NH) -NH ₂ ; or a 5 or 6 membered, saturated or unsaturated heterocycle optionally substituted with cyano, -NH 2, -CH 2 -NH 2, -C (NH) -NH 2, -NH-C (NH) -NH _2, or -CH -NH-C (NH -NH ₂ ;

U is cyano, -NH ₂ , -C (NH) -NH _2, or -NH-C (NH) -NH ₂ .

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

x CH

OK

'CH

-10J is C 1-6 alkylene optionally substituted with OH, NH 2 and C 1-6 alkyl and optionally interrupted by a heteroatom, which may be O, S and N.

n is 0 or 1; and

T is H, OH, amino, a peptide chain, C 1-6 alkyl, C 1-6 alkoxy, C 6-20 aralkyl, or a heterocycle optionally substituted.

R 11 is preferably H or methyl, most preferably H.

K is preferably a bond.

G is preferably -NH-C (NH) -NH 2 attached through a methylene chain of 3-7 carbons, or phenyl substituted with -C (NH) -NH 2 attached through a methylene chain of 0 to 3 carbons. Most preferably G is -NH-C (NH) -NH 2 linked through a methylene chain of 3 atoms.

P is preferably -C (O) -.

J is preferably selected from: -CH 2 -S-CH 2 -CH 2; -CH2-O-CH2-CH2; -CH2-NH-CH2-CH2; and a bond when n is 0. Most preferably, when n is 0, and J is a bond,

In another embodiment of the invention, R may be one of the following ammonium acid derivatives prepared according to the procedure described in Bioorg. Med. Chem. 1995, 3: 1145

NH where n = 1-6, n 1 = 1-2, n 2 = 0-7 and T is as previously defined.

In a preferred embodiment of the invention, T is a peptide of 1 to 4 amino acids in length and A or B is preferably a fibrinogen chain or fragment thereof.

According to another embodiment of the invention, T is a heterocycle selected from the group consisting of:

wherein X ₅ , X ₁₀ and X 11 X 12 are independently selected from the group consisting of N or C-X 7, wherein X 7 is hydrogen, C 1-6 alkyl, or C 6-16 aryl;

X 6 and X 13 are each independently selected from the group consisting of C, O, N, S, N-X or CH-X 7;

R 'is hydrogen, C 1-10 alkyl optionally substituted with carboxyl, -C 0-16 alkyl-CO 2 - C 1-16 alkyl,

C6-20 aralkyl, C3-7 cycloalkyl, aryl or aromatic heterocycle.

- 15 Preferably T is selected from the group consisting of:

More preferably, T is selected from the group consisting of:

R

R '

Where R 'is as previously defined.

More preferably, T is selected from the group consisting of:

Where R 'is as previously defined.

Most preferably T is or

R '

wherein R 'is H or C 1-4 alkyl, such as methyl, ethyl, propyl or butyl, and most preferably R 1 is hydrogen. Another aspect of the invention is that T is 1,2 thiazole optionally substituted with R 'a, or is attached to J at the 2, 3, 4, or 5 position of the ring.

According to another embodiment of the invention, the compounds of the invention are represented by the formulas Π, ΠΙ, IV, and V, wherein X, Y, B, R to R 6 and R 8 are as previously defined.

A particularly preferred embodiment of the invention are compounds represented by formulas VII,

Wines, IX and X:

(IX) (IX)

where

B is O, S, -CH 2 -, or -NH-;

Y may be O, S, SO, SO 2, N-R 5, and CH-R 5;

-17R1 is an arginyl residue or analogue or derivative thereof optionally substituted with an amino acid, peptide or heterocycle;

R ₂ is H or C 1-6 alkyl;

R ₃ can be H, NR 6 R 7 and C 1-4 alkyl; and

R 1 and R 5 are independently selected from H; NR; C 3-7 aryl or C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; A C 1-6 alkyl optionally interrupted with one or more heteroatoms or a carbonyl group and optionally substituted with OH, SH, NR 6 R 7 or C 6-10 aryl, heterocycle or C 3-7 cycloalkyl optionally substituted with halogen, hydroxyl, C 1-6 alkyl; an amino acid chain; and a hydrophobic group,

R 5 is hydrogen, C 1-6 alkyl preferably interrupted with 1 or 2 heteroatoms, C 6-10 aryl, C 3-7 cycloalkyl or a heterocyclic ring and a hydrophobic group, and n is 1 or 2.

Preferred compounds of formula VII are:

0005 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (propyl arginine)

0010 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (butyl arginine)

0015 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (propylcarbomethoxy arginine)

0020 6S-Cyclohexylmethyl-hexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (benzyl arginine)

- 180025 6S-Cyclohexylmethyl hexahydro-5-oxo-5H thiazolo [3,2-a] pyridine-3R-carboxamido (carbomethoxy propylcyclodithioketalarginine)

0030 6S-Cyclohexylmethyl hexaliydro-5-oxo-5H thiazolo [3,2-a] pyridine-3R-carboxamido ((S) -Arg- (R) Pipecolinic Acid

0035 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxyamido (carboxyamidopropyl cyclodithioketal arginine)

0040 6S-cyclohexylmethylhexahydro-

5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxyamido ((S) -Arg hexahydropyridine-

4-carboxylic acid

0045 6S-cyclohexylmethylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxyamido ((S) -Arg isohexahydropyridine-

4-carboxylic acid

0050 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (carboxyamidopentyl cyclodithioketal arginine)

I

H N

- 196S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (carbomethoxypropyl cyclodithioketal arginine)

S -cyclohexylmethylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (1-carboxy-3-thiobutyl ketoarginine)

S -cyclohexylmethylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (1-carboxy-3-thiobutyl ketoarginine)

6S-cyklohexylmetylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (1-carboxy-2-methyl-3-thiobutyl ketoarginine)

6S-cyklohexylmetylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido ((3-thiobutyl sulfonic acid) ketoarginine)

S -cyclohexylmethylhexahydro-

5-Oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (isoquinoline methyl ketoarginine)

with

-200085 6 S-cyclohexylmeths

5-oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (propylcarbomethoxy ketoarginine)

0090 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (propyl keto) ArgPhe-Arg-NH2)

0095 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido ((propanoic acid) ketoarginine)

0100 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (propyl carbomethoxy keto-arginine)

0105 6S-Cyclohexylpropylhexahydro 5-ΟΧΟ-5Η-thiazolo [3,2-a] pyridine-3R-carboxamido (abenzothiazolo ketoarginine) and

OHO 6S-cyclohexylpropylhexahydro 5-OXO-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (propylcarbomethoxy ketoarginine)

NH.

-21 0205 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0210 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0215 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0220 6-Benzyl-8α-methyl-5-oxohexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0225 8α-Methyl-5-oxo-6-phenylethyl-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

H

H

-22230 8α-Methyl-5-oxo-6-phenylethyl-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0240 8α-Methyl-5-oxo-6- (2-trifluoromethyl-quinolin-6-ylmethyl) -hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

0245 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0250 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0255 6-Benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4guanidino-1- (1-methyl-1H-imidazole-2-carbonyl) -butyl] -amide

H

-23 0260

0265

0275

0280

6-Benzyl-8α-methyl-5-oxohexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

5-Oxo-6- (3-cyclohexylpropyl) -hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

8α-Methyl-5-oxo-6- (3-phenyl-propyl-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

8α-Methyl-5-oxo-6- (3-phenyl-propyl-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

8-methyl-5-oxo-6- (2trifluórmetylchinolin-

6-Methyl-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0285

0295 6- (1,3-Dioxo-1,3-dihydroisoindol-2-yl) -5-oxohexahydro-thiazolo [3,2-a]

-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0305 5-Oxo-6- (3-phenylpropionyl amino) -hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

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

More preferred compounds of formula (VII) are:

6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2a] pyridine-3R-carboxamido (propylcarbomethoxyarginine);

6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2a] pyridine-3R-carboxamido (propyl keto) Arg-Phe-ArgNH ₂ );

6S-benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido ((propanoic acid) arginine);

6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2a] pyridine-3R-carboxamido (α-benzothiazolo-ketoarginine);

-250210 6-Benzyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide;

620-Benzyl-8α-methyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide;

0240 8α-Methyl-5-oxo-6- (2-trifluoromethyl-quinolin-6-ylmethyl) -hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1 (benzothiazole-2-carbonyl-4- guanidino-butyl] -amide;

0245 6-Benzyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide;

0260 6-Benzyl-8α-methyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-

2-carbonyl) -butyl] -amide;

0265 5-Oxo-6- (3-cyclohexyl-propyl) -hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-

2-carbonyl) -butyl] -amide;

0285 8α-Methyl-5-oxo-6- (2-trifluoromethyl-quinolin-6-ylmethyl) -hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid (4-guanidino-1- (thiazole-2-carbonyl) - butyl] -amide;

5-Oxo-6- (3-phenyl-propionyl amino) -hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidinol- (thiazole-2-carbonyl) -butyl] -amide;

The most preferred compounds of formula VII are:

6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2a] pyridine-3R-carboxamido (propylcarbomethoxy ketoarginine); and

6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2a] pyridine-3R-carboxamido (α-benzothiazoloketoarginine);

Preferred compounds of Formula VIH are:

0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo [1,2a] pyridine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -

4-guanidino-butyl] -amide

0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid f (benzothiazole-2-carbonyl) -4guanidino-butyl] -amide

0335 2-Benzoyl-4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanedino-1- (thiazole-2-carbonyl) butyl] -amide

0340 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-

2-carbonyl) -butyl] -amide

0345 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (5-methylthiazole-2-carbonyl) -butyl] -amide

about

2- (3-Cyclohexyl-propionyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (2-thiazolecarbonyl) -butyl] -amide

5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

4-Oxo-2- (4-phenyl-butyryl) octahydro-pyrrolofl, 2-

a] Pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

4-Oxo-2-phenylacetylooctahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

2- (2-Amino-3-phenylpropionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-

1- (Thiazole-2-carbonyl) -butyl] -amide

2- [2-Amino-3- (4-hydroxy);

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

NM

-282- [2-Amino-3- (4-fluorophenyl) -propionyl] -4-oxo-octahydro-pyrrolof 1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] - amide

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

2- (2-Amino-3- (1H-indol-3-yl) propionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl)] butyl] -amide

4-Oxo-2- (3-thiophen-3-yl-propionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

NH

NH

NH

4-Oxo-2- (3-thiophen-2-yl-propionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

-290410

2- (3-1H-Imidazol-4-ylpropionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

ABOUT

NM

0415 2- (2-Amino-3-thiophen-3-yl propionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

ABOUT

NM

0420 4-Oxo-2- (1,2,3,4-tetrahydroisoquinoline-3-carbonyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] ] amide

0425 2- (Hydroxy-phenyl-1-acetyl) -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0430 2- (2-Hydroxy-3-phenylpropionyl) -4-oxo-octahydropyrolof 1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

-300435 4-Oxo-2-phenoxyacetylooctahydro-pyrrolof 1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0440 4-Oxo-2- (3-phenoxy-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0445 4-Oxo-2- (2-phenylethanesulfonyl) -octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1 (thiazole-2-carbonyl) -butyl] amide

NH

° .i?

NH

0450 2- (Naphthalene-2-sulfonyl) -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0455 4- (6- [4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxohexahydro-pyrrolof1,2-a] pyrazin-2-yl) -4-oxo-3- (2-propylpentanoylamino) methyl butyrate

0460 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof1,2-a] pyrazine-6-carboxylic acid [4guanidino-1-butyl] -amide

NH

-31 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [3-guanidinopropyl] -amide

4- (6- [4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxohexahydro-pyrrolo [1,2-a] pyrazin-2-yl) -4-oxo-butyric acid

4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (5-ethyl-thiazole-2-carbonyl) 4-guanidino-butyl] -amide

4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (5-methyl-thiazole-2-carbonyl) -butyl] -amide

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

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

-320495 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (4-carbamimidoyl-phenyl) -amide

0500 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (4guanidino-1- (5-phenyl-thiazole-2-carbonyl) -butyl] -amide

0505 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (1 (5-benzyl-thiazole-2-carbonyl 4-guanidino-butyl) -amide

NH

0510 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [1 (4-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl-ethyl] - amide

0515 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl] -ethyl] amide

0520 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazole-2- ethyl] amide

-330525 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2 yl-

ethyl] -amide

0530 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidin-2-ylmethyl) -2-oxo-2-thiazole-2- ethyl] amide

0535 [6- [4-Guanidino-1- (thiazol-2-carbonyl) -butylcarbamoyl] -4-oxo2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazin-3-yl] -acetic acid

0540 3- [6- [4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxo-

2- (3-Phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-

3-yl] -propionic acid

0545 [6- [1- (1-Carbamimidoyl-piperin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) -octahydropyrrole [ 1,2-a] pyrazin-3-yl] acetic acid

-340550 3- [6- [1- (1-Carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrole 1,2 a] pyrazin

3-yl] -acetic acid

0555 [6- [1- (1-Carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] ] pyrazin

3-yl] -acetic acid

0560 [6- (3Guanidinopropylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1,2-a] pyrazin-3-yl] acetic acid

HTH

0565 3- [6- (3-Guanidinopropylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1,2-a] pyrazin-3-yl] propionic acid

0570 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -methyl-amide

-350575 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2 -yletylj-metybamid

0580 [6- [1- (1-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] -methylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydro- 1,2-a] pyrazine-

3-yl] -acetic acid

0585 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] -methyl amide

0590 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (3-guanidino-propyl) -methyl-amide

0595 2- (Naphthalene-2-carbonyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0600 2- (Naphthalene-1-carbonyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

-360605 2- (3-Naphthalen-1-ylpropionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0610 2- (4-tert-Butyl-benzoyl) -4-oxo-octahydro-pyrrolof 1,2a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0615 2- (Benzo [1,3] dioxole-5carbonyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0620 2- (3-Benzo [1,3] dioxol-5-yl propionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1 (thiazole-2-carbonyl) - butyl] -amide

0625 2- [2- (2-Methyl-benzylidene) -but-

3-enoyl] -4-oxo-octahydropyrrolo 1,2-a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] amide

0630 2- [2- (2-Methyl-benzylidene) -but-

3-enoyl] -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] amide

NK

ABOUT

NH.

-372- (2-Benzylidene-pent-3-enoyl) -oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (3-guanidinopropyl) -amide. . r. _:

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

6-Carboxylic acid 4-carbamimidoyl-benzyl amide

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

6-Carboxylic acid [4-imidazol-1-yl-1- (thiazole-2-carbonyl) butyl] -amide

) SNH

H. N

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

6-Carboxylic acid [4- (2-aminoimidazol-1-yl) -1- (thiazole-2-carbonyl) -butyl] -amide o

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

6-Carboxylic acid [3- (2-amino-6-methyl-pyrimidin-4-yl) -1 (thiazole-2-carbonyl) -propyl] -amide

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

ABOUT

me

-380675 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-

6-Carboxylic acid [3- (6-amino-pyridin-2-yl) -1- (thiazole-2-carbonyl) -propyl] -amide

0680 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-

6-Carboxylic acid [3- (2-amino-pyridin-4-yl) -1- (thiazole-2-carbonyl) -propyl] -amide

0685 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-

6-Carboxylic acid [2- (2-amino-pyridin-4-yl) -1- (thiazole-2-carbonyl) -ethyl] -amide

0690 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-

6-Carboxylic acid [2- (6-aminopyridin-2-yl) -1- (thiazole-2-carbonyl) -ethyl] -amide

0695 2- [4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) -1,2,3,4-tetrahydroisoquinoline- 6-carboxamidine

0700 2- [4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) -1,2,3,4-tetrahydroisoquinoline- 7-carboxamidine

-39N- [1- (4-Oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carbonyl] -5- (thiazole-2-carbonyl) -pyrrolidin-3-yl] guanidine

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

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

4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (4-amino-benzyl) -2-oxo-2-thiazol-2-ylethyl] - amide

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

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

ABOUT

-400735 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-ylethyl) -amide

0745 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (3-guanidino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] - amide

0750 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (4-guanidinocyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] - amide

0755 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (2-guanidinocyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] - amide

0760 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (5-benzyl-thiazole-2-carbonyl) -4guanidino-butyl] -amide

MN.

NM.

-41 0765 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidinol- (5-phenyl-thiazole-2-carbonyl) -butyl] -amide

0770 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidinol- (thiazole-2-carbonyl) -butyl] -amide

0775 5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0780 5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (4-carbamimidoyl-benzyl) -2oxo-2-thiazol-2-yl] ethyl] -amide

0785 5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2-oxo-2-thiazole-2- yl-ethyl] -amide

0790 5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-yl-ethyl)] - 2-oxo-2- thiazol-2-yl-ethyl] -amide

5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl-2-oxo-2-thiazol-2)] [4 (4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -5-oxo-ylethyl] -amide

7- (3-Phenyl-propions 1) octahydro-2-thia-4α, 7-thiazanaphthalen-6-yl] -acetic acid

5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-

2-carbonyl) butyl] -amide

3- [4- (4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -5-oxo-

7- (3-Phenyl-propenyl) octahydro-2-thia-4α, 7-thiazanaphthalen-6-yl] -acetic acid

5-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [3-guanidinopropyl] -amide

-Oxo-7- (3-phenyl-propionyl) octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl)] - 2-oxo-2-thiazole- 2-yl-ethyl] -amide

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

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

4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolof 1,2-a] pyrazine-6-carboxylic acid [4-guanidino-thiazol-2-yl-butyl] -amide

NM.

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

[6- [4-Methoxy-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxo-

2- (3-Phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-

3-yl] -acetic acid [2- [5-Methoxy-2 - ([4-oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carbonyl] -amino) -pentanoyl) -thiazole -5-yl] acetic acid

-440855 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-amino-1- (thiazole-2-carbonyl) -butyl] -amide

0860 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [5-amino-1- (thiazole-2-carbonyl) -pentyl] -amide

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

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

0875 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (1-methyl-1H-imidazole-2-carbonyl) -butyl] -amide

0880 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (1-thiazole-2-carbonyl) -butyl] amide

-45,085 8,8-Dimethyl-4-oxo- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1 (1-thiazole-2-carbonyl) -butyl] - amide

Preferred compounds of formula (IIH) are:

0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo [1,2a] pyridine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide

0330 3-Aminomethyl-4-oxo-2-phenylacetyl-octahydropyrrole 1,2-a] pyrazine-6-carboxylic acid [1 (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide

0515 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-yl- ethyl] -amide

0530 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (1-carbamimidoylpiperidin-2-ylmethyl) -2-oxo-2-thiazole- 2-yl-ethyl] -amide

0545 [6- [1- (1-Carbamimidoyl-piperin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrole [1,1, 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-pyrrole [ 1,2-a] pyrazin-3-yl] acetic acid

-460555 [6- (1- (1-Carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrole [1 2-a] pyrazin-3-yl] acetic acid

0560 (6- (3Guanidino-propylcarbamoyl) -4-oxo-2- (3-phenylpropionyl) -octahydro-pyrrolo [1,2-a] pyrazin-3-yl] -acetic acid

0565 3- [6- (3-Guanidinopropyl-carbamoyl) -4-oxo-2- (3-phenylpropionyl) -octahydro-pyrrolo [1,2-a] pyrazin-3-yl] propionic acid

0575 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazole- 2-yl-ethyl] -methyl-amide

0580 [6- (1- (1-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] -methylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrole [1,2-a] pyrazin-3-yl] acetic acid

0585 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (1- (1-carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazole- 2-yl-ethyl] -methyl-amide

0590 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1,2a) pyrazine-6-carboxylic acid (3-guanidinopropyl) methyl amide

0595 2- (Naphthalene-2-carbonyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

-470625 2- [2- (2-Methyl-benzylidene) -but-3-enoyl] -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidine-3) -ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] -amide

0630 2- [2- (2-Methyl-benzylidene) -but-3-enoyl] -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1 (1-carbamimidoyl-piperidine-4-) ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] amide

635 2- (2-Benzylidene-pent-3-enoyl) -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid (3-guanidinopropyl) -amide

0645 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-imidazol-1-yl-1- (thiazole-2-carbonyl) -butyl] - amide

0670 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3- (2-amino-6-chloropyrimidin-4-yl) -1- (thiazole) 2-carbonyl) -propyl] -amide

0675 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3- (6-aminopyridin-2-yl) -1- (thiazole-2-carbonyl) - propyl] -amide

0680 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3- (2-aminopyridin-4-yl) -1- (thiazole-2-carbonyl) - propyl] -amide

0685 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [2- (2-aminopyridin-4-yl) -1- (thiazole-2-carbonyl) - ethyl] -amide

0690 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [2- (6-aminopyridin-2-yl) -1- (thiazole-2-carbonyl) - ethyl] -amide

0695 2- [4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-] a] pyrazine-6-carbonyl] -3- (thiazole-2- carbonyl) -1,2,3,4-tetrahydro- isoquinolin-6-carboxamidine 0700 2- [4-oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [l, 2- a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) - rf 1,2,3,4-tetrahydro-isoquinoline-7-carboxamidine 0705 rf N- [1- (4-Oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1,2-a] pyrazine-6-carbonyl] -5- (thiazole-2-carbonyl) -pyrrolidin-3-yl] guanidine 0710 4-Oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [l, 2- a] pyrazine-6-carboxylic acid [1- (4-Aminocyclohexyl) - 2-oxo-2-thiazol-2-yl-ethyl] -amide 0730 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2- • a] Pyrazine-6-carboxylic acid [1- (3-aminomethylbenzyl) -2-oxo-2-thiazol-2-yl-ethyl] -amide 0745 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (3-guanidino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl] ethyl] -amide 0755 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (2-guanidinocyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl ] amide 0795 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylethyl)] - 2-oxo-2- thiazol-2-yl-ethyl] -amide

-490800 [4 (4-Guanidino-1- (thiazole-2-carbonyl) butylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazaphthalen-6-yl] -acetic acid

0810 3- [4- (4-Guanidino-1- (thiazole-2-carbonyl) butylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazaphthalen-6-yl acetic acid

0815 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α-diaza-naphthalene-4-carboxylic acid [3-guanidinopropyl] -amide

0820 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2- thiazol-2-yl-ethyl] -amide

0830 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-

a] Pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2-ylmethyl-butyl] -amide

083 5 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-

a] Pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2-yl-butyl] -amide

More preferred compounds of formula VRI are:

0335 2-Benzoyl-4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

0650 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4- (2-amino-imidazol-1-yl) -1- (thiazole-2- carbonyl) -butyl] -amide

0655 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3- (2-amino-6-methyl-pyrimidin-4-yl) -1- (thiazole) 2-carbonyl) -propyl] -amide

-500715 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-aminocyclohexylmethyl) -2-oxo-2-thiazol-2-yl] ethyl] -amide

0720 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-aminobenzyl) -2-oxo-2-thiazol-2-yl-ethyl] amide

0725 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-aminomethyl-benzyl) -2-oxo-2-thiazol-2-yl- ethyl] -amide

073 5 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid- (2-oxo-1-piperidin-4-ylmethyl-2-thiazol-2-yl-ethyl) -amide i

and

0740 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-

6-Carboxylic acid- (2-oxo-1-piperidin-3-yl-

2-thiazol-2-yl-ethyl) -amide

0750 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-guanidinocyclohexylmethyl) -2-oxo-2-thiazol-2-yl- ethyl] -amide

0760 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (5-benzylthiazole-2-carbonyl) -4-guanidino-butyl] -amide

0765 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidinothiazole-2-carbonyl] -butyl] -amide

0770 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

-51 0775 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] - amide

0780 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [1- (4-carbamimidoyl-benzyl) -2-oxo-2-thiazole-2- yl-ethyl] -amide

0785 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2-oxo-2-thiazole-2- yl-ethyl] -amide

0790 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylethyl)] - 2-oxo-2 -tiazol2-yl-ethyl] -amide

0805 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4α, 7-diaza-naphthalene-4-carboxylic acid [4-guanidino-1 (thiazole-2-carbonyl) -butyl] -amide

0825 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (hydroxytiazol-2-ylmethyl) -butyl] -amide

0840 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-methoxy-1- (thiazole-2-carbonyl) -butyl] -amide

0845 [6- [4-Methoxy-1- (thiazole-2-carbonyl) -butylcarbamoyl] -

4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazin-3-yl] -acetic acid

0850 [2- [5-Methoxy-2 - ([4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carbonyl] -amino) -pentanoyl) -thiazole- 5-yl] -acetic acid

0855 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-amino-1- (thiazole-2-carbonyl) -butyl] -amide

0860 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [5-amino-1- (thiazole-2-carbonyl) -pentyl] -amide

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

The most preferred compounds of formula VIII are:

0345 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (5-methylthiazole-2-carbonyl) -butyl] -amide and

0340 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

Preferred compounds of formula IX are:

0890 3-Amino-1-4-oxo-2-phenyl hexahydro-pyrrolo [2,1b [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4guanidinobutyl] amide

-530895 3-Amino-2-benzyl-4-oxohexahydro-pyrrolo [2,1-

b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4guanidinobutyl] amide

0900 3-Amino-2-cyclohexyl-4-oxohexahydro-pyrrolo [2,1-

b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] amide

Preferred compounds of formula X are:

0905 7-Benzyl-6-oxo-octahydropyrrolo [2,1-c] [1,4] thiazine-4-carboxylic acid [1 (benzothiazole-2-carbonyl) -4guanidino-butyl] -amide

0910 7 (4-tert-Butyl-benzyl) -Benzyl-6-ω-octahydro-pyrrolo [2,1-

c] [1,4] thiazine-4-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] amide

0915 6-Oxo-octahydro-pyrido [2,1-

c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

H

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-540925 7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidinol- (thiazole-2-carbonyl) butyl] amide

0935 7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidinol- (thiazole-2-carbonyl) butyl] amide

0940 6-Oxo-7-phenethyl-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

0950 7-Benzyl-2,2,6-trioxooctahydro-21,66-pyrido [2,1-cj [1,4] thiazine-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butyl] -amide

at

H

More preferred compounds of formula X are

0925 7-Benzyl-6-oxo-octahydro-pyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide and

0940 6-Oxo-7-phenethyl-octahydro-pyrido [2,1-c] [1,4] thiazine-4-carboxylic acid (4-guanidino-1- (thiazole-2-carbonyl) butyl) -amide

Preferred compounds of formula ΠΙ are:

-550960 4-Oxo- (3-phenyl-propionyl) octahydro pyrrolo [1,2] pyrimidine-

6-Carboxylic acid [4guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

0965 "4-Oxo-l- (fenetyIsulfonyl) octahydropyrrolo [l, 2] pyrimidine * 6-carboxylic acid [4-guanidino-1 - (thiazol-2 ^1-carbonyl) -butyl] -amide

Various procedures can be used to prepare the compounds of formula (VII) individually, depending on the starting material and / or intermediates formed. The following diagram is one of them.

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4 ..........

about

Step 1: Alkylation of α, is performed with a suitable base according to the procedure described by Evans et al. (J. Chem. Soc., 1981, 103, 2127 and 1982, 104, 1737; Aldrichimica Acta 1982, 15, 23) to obtain b.

Step 2: Compound b is subjected to hydroboration and oxidized according to conditions available in the literature (Synthesis, 1980, 151), resulting in an aldehyde.

Step 3: The formation of the product of addition e from aldehyde c with d is carried out by stirring the reactants in an aromatic solvent e.g. benzene or toluene, in the presence of a catalytic amount of a suitable acid, e.g. p-toluenesulfonic acid.

Step 3 ': Preventing the conversion of aldehyde c to aldehyde a is readily accomplished by protecting and removing protection according to the protocol found in T.Green: Protective Groups In Organic Synthesis. (John Wiley & Sons, 1981).

Step 4: Cyclization of the product of addition of e to f can be readily accomplished with a suitable Lewis acid e.g. trimethylalbumin in a suitable solvent e.g. dichloromethane, method found in T.Green, (see step 3 ') ·

Step 4 ': Alternatively, compound f can be formed by treatment of d with aldehyde and in the presence of a suitable aromatic solvent e.g. benzene.

Step 5: The ester function of the (-C (O) O-R) bicyclic intermediate of formula f is then subjected to hydrolysis, using a suitable reagent such as HCl, a suitable solvent such as ethyl ether to give the free carboxylic acid. The resulting compound is then coupled to R 1 H with the peptide using a coupling agent such as BOP in a suitable solvent such as DMF to form a bicyclic compound of formula (VHI). Conditions suitable for peptide bond formation are well known in peptide chemistry. E.g. see Principal of Peptide Synthesis, M. Bodanszky, Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academiec Press, New York, San Francisco, London, 1979.

Various procedures may be used to prepare the compounds of formula (VHI) depending on the starting material used and / or the intermediates formed. The following scheme is one possible preparation.

-58SCHÉMA2

Step 1 ozonolysis

g

VIII

-59where:

Pg is a nitrogen protecting group;

each R jq and R j j is independently C alkyl; and X, R 1, R R 1, and R 1 are as defined above.

In brief, the description of the process in Scheme 2 is as follows:

Step 1: The amino and carboxyl functional groups of the unsaturated compounds of formula (a) are protected with suitable protecting groups. A variety of protecting groups, for various reaction functional groups and suitable protection protocols and their removal are found in T.Green, Protective Groups In Organic Synthesis. (John Wiley & Sons, 1981). The use of suitable protecting groups in a given synthetic scheme will depend on many factors, including the presence of other reactants and the reaction conditions for their removal. The unsaturated compounds of the formula are readily obtainable by methods and protocols known in the chemical art. The protection of the unsaturated compounds of formula (a) is carried out under suitable conditions allowing cyclization using a suitable reagent such as mercuric acetate in an inert solvent such as tetrahydrofuran (THF) resulting in the protected aminoalcohol of formula (b).

Step 2: The protected aminoalcohol of formula (b) is oxidized using a suitable oxidizing agent such as pyridine-sulfur trioxide complex in a suitable solvent, such as dichloromethane or dimethylformamide, resulting in the protected aminoaldehyde of formula (c). Alternatively, intermediate (C) can be produced by ozonolysis of a compound of formula (a ') prepared according to Collado et al., J. Org. 1995, 60: 5011.

Step 3: The amino aldehyde of formula (c) is protected by coupling with an alkyl ester of the amino acid of formula (d), first forming the imine, then the imine obtained is added to a suitable reagent such as sodium triacetoxy borohydride NABH (OAc) to give the cyclic intermediate of formula )

Step 4: The cyclic intermediate of formula (e) is functionalized at the amino position, yielding an amino substituted cyclic intermediate of formula (f). Suitable conditions for such reactions are well known and will depend on the nature of the substituent R *.

Step 5: The amino protecting group of the cyclic intermediate of formula (f) is removed under suitable conditions, and the resulting compound is then subjected to internal ring closure under appropriate conditions, such as by heating at low temperature in an inert solvent or as a crude compound, resulting in a bicyclic intermediate of formula (g). The bicyclic intermediate of formula (g) may also be obtained by hydrolysis of the ester function (-QO 10 -R 7) of the cyclic ring.

An intermediate of formula (g) to release the carboxyl group, followed by coupling with the standard peptide using a suitable coupling agent such as benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (BOP), in an inert solvent such as dimethyl formamide ( DMF).

Step 6: The ester function of the (-CfO 3 O-R 6) bicyclic intermediate of formula (g) is then subjected to hydrolysis using a suitable reagent such as HCl in a suitable solvent such as ethyl ether resulting in the free carboxylic acid. The resulting compound is then coupled to R 1 H using a peptide coupling reagent such as BOP in a suitable solvent such as DMF resulting in a bicyclic compound of formula (VHI). Suitable conditions for peptide bond formation are well known in peptide chemistry. E.g. see Principal of Peptide Synthesis, M. Bodanszky, Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides Analvsis. Synthesis. Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academiec Press, New York, San Francisco, London, 1979.

Various methods may be used to prepare the compounds of formula (D <) depending on the starting material used and / or the intermediates formed. The following scheme is one possible preparation.

Scheme 3

b

STEP 1

---------- ►

MeO

Y = S-Pg NH-Pg

I

about

STEP 3

C

-61 where:

Pg is a sulfur or amine protecting group;

L is a leaving group, each R 20 ^and R 21 ^nezav ^ * ^ ^{p and} C alkyl; ^and R j> R y R j, and R 8 are as previously defined.

The procedure shown in Scheme 2 is briefly described as follows:

STEP 1:

The carboxylic compound (a) is coupled to the cyclic amine compound (b) by a peptide coupling reagent such as benzotriazol-1-yloxy-tris- (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in a suitable a solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to give the amide compound of formula (c). Suitable conditions for peptide bond formation are well known in peptide chemistry. E.g. see Principal of Peptide Synthesis, M. Bodanszkv, Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides Analysis, Synthesis, Biology, Vol. 1. edited by Gross E., and Meienhofer J., Academiec Press, New York, San Francisco, London, 1979.

STEP 2:

The compound of formula (c) is subjected to suitable conditions allowing internal cyclization resulting in a bicyclic intermediate of formula (d). E.g. acid mediated cyclization using p-toluenesulfonic acid, or TFA in a suitable solvent such as dichloroethane.

STEP 3:

The esterified functional group (-C (O) O-R) of the bicyclic intermediate of formula (d) is subjected to hydrolysis using a suitable reagent such as lithium hydroxide (LiOH) in a suitable solvent such as tetrahydrofuran (THF) resulting in the free carboxylic acid. The resulting compound is then coupled to P.H. using a peptide coupling agent such as BOP in a suitable solvent such as DMF to give compound (e). Suitable conditions for peptide bond formation are well known in peptide chemistry. E.g. see Principal of Peptide Synthesis, M. Bodanszkv, Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides Analysis, Synthesis. Biology. Vol. 1, edited by Gross E., and Meienhofer J., Academiec Press, New York, San Francisco, London, 1979.

Various procedures can be used to isolate the compounds of formula (IX) depending on the starting material and / or the intermediates formed.

-62 The following scheme 4 is one of the possible preparation methods.

SCHEME 4

STEP 1

STEP 4 ’

T

-63Kde:

Zaj ^much each independently alkyl; and B, R *, Rj, R *, and are defined as before. The procedure outlined in Scheme 4 is briefly described as follows:

KR0K1:

The halogenated compound of formula (a) is converted to a halomethyl ketone of formula (b) using a suitable reagent such as diazomethane in an inert solvent such as diethyl ether at a temperature of about -25 ° C to 0 ° C. The resulting mixture is then subjected to acidic conditions to form a halomethyl ketone of formula (b).

STEP 2:

The halomethyl ketone of formula (b) is coupled with an alkyl ester of the amino acid of formula (c) with a suitable base such as sodium cyanotetraborohydride in an organic solvent such as methanol (MeOH) to form the cyclic intermediate of formula (d).

STEP 3:

The cyclic intermediate of formula (d) is treated under acidic conditions using a suitable acid such as camphorsulfonic acid in a suitable solvent such as toluene to give the intermediate of formula (e)

STEP 4:

The ester function of the (-C (O) O-R 2q) bicyclic intermediate of formula (e) is then subjected to hydrolysis using a suitable reagent such as LiOH to give the free carboxylic acid. The resulting compound is then coupled to R 1g using a peptide coupling reagent such as BOP in a suitable solvent such as dimethylformamide resulting in a bicyclic coupled compound of formula (X). Suitable conditions for peptide bond formation are well known in peptide chemistry. E.g. see Principal of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides Analysis, Svnthesis, Biology, Vol.

1. edited by Gross E., and Meienhofer J., Academiec Press, New York, San Francisco, London, 1979.

The compounds of the present invention are further characterized by their ability to inhibit the catalytic activity of thrombin as explained in the following assay. The compounds of the present invention may be prepared for their determination by dissolving in a buffer with a final concentration of 1 to ΙΟΟμΜ. To determine the inhibitory dissociation constant of K-64 given compounds, a chromogenic or fluorogenic thrombin substrate is added to a solution containing the test compounds and thrombin; The resulting catalytic activity of the enzyme is determined spectrophotometrically. This type of assay is well known in the art for those skilled in the art.

The compounds of the present invention may be used as anticoagulants in vivo or ex vitro in the event of contact activation with a foreign thrombogenic surface that would be contained in a hose used for extracorporeal circulation. The compounds of the present invention may also be used as surface coatings for such thrombogenic hoses. Finally, the compounds of the present invention are obtained as a lyophilized powder, redissolved in isotonic saline and added in a sufficient amount to the blood maintained in the anticoagulant state.

The therapeutic agents of the present invention may be administered alone or with a suitable pharmaceutically acceptable carrier. The proportion of each carrier is determined by the solubility and chemical nature of the compound by which it is administered and standard pharmaceutical practice. E.g. the compounds may be parenterally injected, and it may be intramuscular, intravenous or subcutaneous. For parenteral administration, the compounds may be used in the form of sterile solutions containing other solutions, e.g. appropriate saline or glucose to make the solution isotonic. The compounds may be administered orally in the form of tablets, capsules, or granules containing a suitable vehicle such as starch, lactose, white sugar and the like. The compounds may also be administered sublingually in the form of tablets, pills or lozenges in which each active ingredient is mixed with sugar or rye syrups, flavoring agents and coloring agents, and then a sufficiently dehydrated mixture suitable for compression into solid form is made. The compounds may be administered orally in the form of a solution, which may contain coloring and / or flavoring agents.

Physicians will determine the dose of the active substance that is most appropriate. Dosages may vary depending on the route of administration and the particular compound selected. In addition, the dosage may vary according to the conditions of the particular patient.

If the composition is administered orally, more active substance is required to have the same effect as when administered parenterally.

To further explain the present invention, it follows to provide an unlimited number of examples of such thrombin inhibiting compounds. The following examples, of course,

They will not limit the scope of the present invention, variations currently known or later developed will be considered to be within the scope of the present invention described previously, given the skill of the art. Preferred compounds of the invention are synthesized using conventional preparative steps and procedures known to those skilled in the art of organic and bioorganic synthesis, while providing novel and unique combinations for the overall synthesis of each compound. Preferred synthetic routes for intermediates include, in addition, the resulting antithrombotic compounds of the present invention in the syntheses.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.65 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in anhydrous benzene (30 mL), and anhydrous N, The dimethylacetamide (2.0 mL) was placed on the bottom of a round, nitrogen-purged flask filled with a zinc / copper pair (0.9 g). The resulting mixture is sonicated under nitrogen until no starting material remains! (as determined by thin-layer chromatography). Palladium (II) bis- (tri-o-tolylphosphine) (0.35 g, 0.40 mmol) was then added followed by 4-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen with heating followed by cooling, and then ethyl acetate (100 mL) was added and the mixture was filtered through a separatory funnel. It is further washed with HCl (50 mL, 0.1 N), distilled water (3 x 50 mL, and dried over Na 2 SO 4, filtered, concentrated under reduced pressure to give a crude product.) Flash chromatography over silica gel (petroether-ethyl acetate gradient) ) provides the purified compound.

-66Tert solution. butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.65 g, 7.5 mmol) (J.Org.Chem. 1992, 57, 3397-3404) in anhydrous benzene (30 mL), and anhydrous N, N -dimethylacetamide (2.0 mL) was placed on the bottom of a round, nitrogen purged flask filled with zinc / copper (0.9 g). The resulting mixture was sonicated under nitrogen until no starting material remained (detected by thin layer chromatography). . Palladium (II) bis (tri-o-tolylphosphine) chloride (0.35 g, 0.40 mmol) was then added followed by 3-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen with heating followed by cooling, and then ethyl acetate (100 mL) was added and the mixture was filtered through a separatory funnel. It was further washed with HCl (50 mL, 0.1 N), distilled water (3 x 50 mL), and dried over IS 2 SO 4, filtered, concentrated under reduced pressure to give a crude product. Flash chromatography over silica gel (petroleum ether-ethyl acetate) gradient) provides the purified compound.

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.65 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in anhydrous benzene (30 mL), and anhydrous N, N-dimethylacetamide (2.0 mL) was placed on the bottom of a round, nitrogen-purged flask filled with a zinc / copper pair (0.9 g). The resulting mixture was sonicated under nitrogen until no starting material remained (detected by thin-layer chromatography). layer chromatography). Then, bis- (tri-o-tolylphosphine) palladium (II) chloride (0.35 g, 0.40 mmol) is added, followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen with heating followed by cooling, and then ethyl acetate (100 mL) was added and the mixture was filtered through a separatory funnel. Next, the mixture was washed with HCl (50 mL, 0.1 N), distilled water (3 x 50 mL, and dried over 1 SO 4, filtered, concentrated under reduced pressure to give a crude product. Flash chromatography over silica gel (petroether) (ethyl acetate gradient) provides the purified compound.

-67K To a solution of tert-butyloxycarbonyl-β 7 R -7-cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) in anhydrous ethanol (20 mL) was added hydroxylamine 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, diisopropyl ether, dried with MgSO 4, concentrated under reduced pressure and used directly for the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL) and anhydrous ethanol (40 mL) with heating. . Subsequently, Pd / C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture while heating. Hydrogenation was continued until no starting material was detectable by thin layer chromatography. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL) and HCl (50 mL) was added, and the mixture was again concentrated to 50 mL. The solution was frozen overnight to give the title compound.

To a solution of tert-butyloxycarbonyl-m -cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) in anhydrous ethanol (20 mL) was added hydroxylamine 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, diisopropyl ether, dried with MgSO 4, concentrated under reduced pressure and used directly for the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL) and anhydrous ethanol (40 mL) with heating. Subsequently, Pd / C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture while heating. Hydrogenation was continued until no starting material was detectable by thin layer chromatography. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL) and HCl (50 mL) was added, and the mixture was again concentrated to 50 mL. The solution was frozen overnight to give the title compound.

-68K. A solution of tert-butyloxycarbonyl-η 7 -cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) in anhydrous ethanol (20 mL) was added hydroxylamine 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, diisopropyl ether, dried with MgSO 4, concentrated under reduced pressure and used directly for the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL) and anhydrous ethanol (40 mL) with heating. Subsequently, Pd / C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture while heating. Hydrogenation was continued until no starting material was detectable by thin layer chromatography. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL) and HCl (50 mL) was added, and the mixture was again concentrated to 50 mL. The solution was frozen overnight to give the title compound.

To a solution of the thiazole (1.28 g, 15.0 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. Then tert-butylcarbonyl-N- (7H-amidino-phenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The mixture was quenched with a saturated aqueous ammonium chloride solution, diluted with ethyl acetate (150 mL), and the organic layer was washed with a saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

A -69K solution of thiazole (1.28 g, 15.0 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise to n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. Then tert-butylcarbonyl- / -eta-amidino-phenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The mixture was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

To a solution of the thiazole (1.28 g, 15.0 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. Then tert-butylcarbonyl-orro-amidino-phenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The mixture was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

- 70Terc. Butyloxycarbonyl-para-cyano-phenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and porous Raney nickel (100 mg) was added. The solution was stirred on a shaker under room temperature (40 psi). The solution was filtered through celite and concentrated under reduced pressure to give 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 4, filtered, and concentrated under reduced pressure.

T. butyloxycarbonyl-meta-cyano-phenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and porous Raney nickel (100 mg) was added. The solution was stirred on a shaker under room temperature (40 psi). The solution was filtered through celite and concentrated under reduced pressure to give 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 4, filtered, and concentrated under reduced pressure.

T. butyloxycarbonyl-ortho-cyano-phenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL), and porous Raney nickel (100 mg) was added. The solution was stirred on a shaker under H ₂ at room temperature (40 psi). The solution was filtered through celite and concentrated under reduced pressure to give 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 4, filtered, and concentrated under reduced pressure.

tert-Butyloxycarbonyl-N-O-aminomethyl-phenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in anhydrous tetrahydrophirane (10 mL) with stirring under nitrogen. The solution was cooled. N, N'-Bis- (benzyloxycarbonyl) -S-methyl-isothiourea (1.14 g, 3.2 mmol) and HgCl 2 (0.95 g, 3.5 mmol) were added. The solution was concentrated under reduced pressure, the 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) gave the pure compound.

tert-Biityloxycarbonyl-ethanaminomethyl-phenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) with stirring under nitrogen. The solution was cooled. N, N'-Bis- (benzyloxycarbonyl) -S-methyl-isothiourea (1.14 g, 3.2 mmol) and HgCl 2 (0.95 g, 3.5 mmol) were added. The solution was concentrated under reduced pressure, the obtained 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) gave the pure compound.

OCH

HgCK / THF

-72 tert-Butyloxycarbonyl-η 7o-aminomethyl-phenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) with stirring under nitrogen. The solution was cooled. N, N'-Bis- (benzyloxycarbonyl) -S-methyl-isothiourea (1.14 g, 3.2 mmol) and HgCl 2 (0.95 g, 3.5 mmol) were added. The solution was concentrated under reduced pressure, the obtained 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) gave the pure compound.

To a solution of thiazole (1.28 g, 15 mmol) in anhydrous tetrahydrofuran (30 mL) was added n BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was mixed. The protected amino acid (1.36 g, 3.3 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

To a solution of the thiazole (1.28 g, 15 mmol) in anhydrous tetrahydrofuran (30 mL) was added n BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. The protected amino acid (1.36 g, 3.3 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting

The mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

1. Lithium thiazole / THF

To a solution of the thiazole (1.28 g, 15 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. The protected amino acid (1.36 g, 3.3 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO ₄ , filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

OCH

I

CH,

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.65 g, 7.5 mmol) (J.Org.Chem. 1992, 57, 3397-3404) in anhydrous benzene (30 mL), and anhydrous N, N dimethylacetamide (2.0 mL) was placed on the bottom of a round, nitrogen purged flask filled with zinc / copper pair (0.9 g). The resulting mixture was sonicated under nitrogen until no starting material remained (detected by thin-layer analysis). chromatography). Then, bis- (tri-o-tolylphosphine) palladium (II) chloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture is stirred under a nitrogen atmosphere with heating followed by cooling, and then ethyl acetate (100 mL) is added and the mixture is filtered through a separatory funnel. It is further washed

HCl (50 mL, 0.1N), distilled water (3 x 50 mL, and dried over Na ₂ SO ₄ , filtered, concentrated under reduced pressure to give a crude product. Flash chromatography over silica gel (petroleum ether - ethyl acetate gradient) ) provides the purified compound.

To a solution of thiazole (1.28 g, 15 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. Amino acid Ν, Ο-dimethylamide (1.36 g, 3.3 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

1. Zn / Cu pair; ultrasound

2. IO CH., C ₆ H _{4) 2} 3PliPdCl

2-cyano-S-bromopyridine

ABOUT

A solution of tert-butyloxycarbonyl-iodo-alanine-N, O-dimethylamide (2.65 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in anhydrous benzene (30 mL), and anhydrous N, N-dimethylacetamide (2.0 mL) was placed on the bottom of a round, nitrogen-purged flask filled with a zinc / copper pair (0.9 g). The resulting mixture is sonicated under nitrogen until no starting material remains (as determined by thin layer chromatography). Palladium (II) bis (tri-o-tolylphosphine) chloride (0.35 g, 0.40 mmol) is then added, followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture is stirred under a nitrogen atmosphere with heating followed by cooling, and then ethyl acetate (100 mL) is added and the mixture is filtered through a separatory funnel. It is further washed with HCl (50 mL, 0.1 N), distilled water (3 x 50 mL, and dried over Na ₂ SO 4, filtered, concentrated under reduced pressure to give a crude product. Flash chromatography over silica gel (petroleum ether - ethyl acetate gradient) provides the purified compound.

To a solution of tert-butyloxycarbonyl- (4-cyano) -3-pyridylalanine-N, O-dimethylamide (1.34 g, 4.0 mmol) in anhydrous ethanol was added Ν, Ο-hydroxylamine 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 and washed with cold ethanol, diisopropyl ether, dried with MgSO 4, concentrated under reduced pressure, and used directly for the next step. The semi-solid was suspended in a mixture of acetic acid (20 mL) and anhydrous ethanol (40 mL) with heating. . Subsequently, Pd / C catalyst (0.30 g, 10% Pd) was added and hydrogen was bubbled through the mixture while heating. Hydrogenation was continued until no starting material was detectable by thin layer chromatography. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 mL) and HCl (50 mL) was added, and the mixture was again concentrated to 50 mL. The solution was frozen overnight to give the title compound.

To a solution of the thiazole (1.28 g, 15 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution was stirred. Amino acid Ν, Ο-dimethylamide (1.16 g, 3.3 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

l.Hj.ľtOj / AcOH

tert-Butyloxycarbonyl-3- (4-pyridyl) alanine-N, O-diethylamide (4.5 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under H _? to his rt gas saturation. The solution was filtered through celite, and concentrated under reduced pressure to give 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 over MgSO 4, filtered and concentrated under reduced pressure to give the title compound.

1. H _; . PtO./AcOH

tert-Butyloxycarbonyl-3- (3-pyridyl) alanine-N, O-dimethylamide (4.5 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under H 2 until saturated with gas. The solution was filtered through celite, and concentrated under reduced pressure to give tert-butyloxycarbonyl-3- (3-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 MgSO 4. filtered and concentrated under reduced pressure to give the title compound.

1. H _; . PtO./AcOH

-

Tert-Butyloxycarbonyl-3- (2-pyridyl) alanine-N, O-dimethylamide (4.5 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under to gas saturation. The solution was filtered through celite, and concentrated under reduced pressure to give 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 over MgSO 4, filtered and concentrated under reduced pressure to give the title compound.

tert-Butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), with stirring under nitrogen. The solution was cooled. N, N'-bis- (benzyloxycarbonyl) -S-methylthiourea (1.14 g, 3.2 mmol), and HgCl 2 (0.95 g, 3.5 mmol) were added. The solution was concentrated under reduced pressure, the residue obtained 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) gave the title compound.

tert-Butyloxycarbonyl-3- (3-piperidyl) amino-N, O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), with stirring under nitrogen. The solution was cooled. N, N'-bis- (benzyloxycarbonyl) -S-methylthiourea (1.14 g, 3.2 mmol), and HgCl 2 (0.95 g, 3.5 mmol) were added. The solution was concentrated under reduced pressure, the obtained 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) gave the title compound.

ΝΖ

HgCl / THF

tert-Butyloxycarbonyl-3- (2-piperidyl) alanine-N, O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), with stirring under nitrogen. The solution was cooled. N, N'-bis- (benzyloxycarbonyl) -S-methylthiourea (1.14 g, 3.2 mmol), and HgCl _? (0.95 g, 3.5 mmol). The solution was concentrated under reduced pressure, the residue obtained 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) gave the title compound.

To a solution of the thiazole in anhydrous tetrahydrofuran (1.23 g, 14.4 mmol) was added dropwise n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) at -78 ° C, and the solution was stirred . The guanidized 4-piperidylalanine derivative (2.0 g, 3.2 mmol) in anhydrous tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure.

1. Lithium thiazole / 1H1

A 79K solution of thiazole in anhydrous tetrahydrofuran (1.23 g, 14.4 mmol) was added dropwise to n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) at -78 ° C, and the solution was mixed. The guanidized 3-piperidyl-alanine derivative (2.0 g, 3.2 mmol) in anhydrous tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure.

about

1. Lithium thiazole / THF

To a solution of the thiazole in anhydrous tetrahydrofuran (1.23 g, 14.4 mmol) was added dropwise n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) at -78 ° C, and the solution was stirred . The guanidized 2-piperidylalanine derivative (2.0 g, 3.2 mmol) in anhydrous tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure.

tert-Butyloxycarbonyl- N -ra-nitro-phenylalanine-N, O-dimethylarnide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under to gas saturation. The solution was filtered through celite, and concentrated under reduced pressure, taken up in H 2 O (150 mL) and lyophilized. to give tert-butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide. The semi-solid was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH

-80 (3 x 50 mL), brine (3 x 50 mL), dried over MgSO _4, filtered and concentrated under reduced pressure to give the title compound.

/ CH

N I OCH,

1 _H. 1ΊΟ _: /Α<.ΌΗ

/ CH, N

I

OCH, tert-Butyloxycarbonyl-neta-nitro-phenylalanine-N, O-dirylamine (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under to gas saturation. The solution was filtered through celite, and concentrated under reduced pressure, transferred with ΡΕ, Ο (150 mL) and lyophilized. to give tert-butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide. The semi-solid was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (3 x 50 mL), brine (3 x 50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure to give the title compound.

1. H ,. PtOi / AcOH

---------------------->.

tert-Butyloxycarbonyl-η 7o-nitro-phenylalanine-N, O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was shaken under EL until saturated with gas. The solution was filtered through celite, and concentrated under reduced pressure, transferred to ΕΕ, Ο (150 mL) and lyophilized. to give tert-butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide. The semi-solid was dissolved in ethyl acetate (250 mL), washed with 1 N NaOH (3 x 50 mL), brine (3 x 50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure to give the title compound.

1. tert-Butyloxycarbonyl-3- (N, N- (4-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in a saturated aqueous solution of sodium bicarbonate and tetrahydrofuran (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 sodium bicarbonate was added. THF was removed under reduced pressure and the resulting aqueous phase was poured into ethyl acetate (250 mL), and thoroughly stirred. The aqueous phase was removed and the resulting solution was washed with saturated aqueous sodium bicarbonate solution (2 x 50 mL), 4 N aqueous sodium hydrogen sulfate (2 x 50 mL). The solution was dried with MgSO 4, filtered and concentrated under reduced pressure. The semi-solid was chromatographed over silica gel (ethyl acetate / hexane).

2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous tetrahydrofuran was added dropwise n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) at -78 ° C, and the solution was stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

1. Z-Cl. NuHCO / THF: H _: O -------------: ------------>

- Lithium thiazole / THF

1. tert-Butyloxycarbonyl-3-Cc (trans-5- (3-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in a saturated aqueous solution of sodium bicarbonate and tetrahydrofuran (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 sodium bicarbonate was added. THF was removed under reduced pressure and the resulting aqueous phase was poured into ethyl acetate (250 mL), and thoroughly stirred. The aqueous phase was removed and the resulting solution was washed with saturated aqueous sodium bicarbonate solution (2 x 50 mL), 4 N aqueous sodium hydrogen sulfate (2 x 50 mL). The solution was dried with MgSO 4, filtered and concentrated under reduced pressure. The semi-solid was chromatographed over silica gel (ethyl acetate / hexane).

2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous tetrahydrofuran was added dropwise n BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) at -78 ° C, and the solution was mixed. The above protected amino acid amide (1.41 g, 3.0 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

1. Z-C1. NuHCO / THF: H. O.

2. Lithium thiazole / THF

1. tert-Butyloxycarbonyl-3- (η 5 -N-75-2-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in a saturated aqueous solution of sodium bicarbonate and tetrahydrofuran ( 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 sodium bicarbonate was added. THF was removed under reduced pressure and the resulting aqueous phase was poured into ethyl acetate (250 mL), and thoroughly stirred. The aqueous phase was removed and the resulting solution was washed with saturated aqueous sodium bicarbonate solution (2 x 50 mL), 4 N aqueous sodium hydrogen sulfate (2 x 50 mL). The solution was dried with MgSO 4, filtered and concentrated under reduced pressure. The semi-solid was chromatographed over silica gel (ethyl acetate / hexane).

2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous tetrahydrofuran was added dropwise n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) at -78 ° C, and the solution was stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in tetrahydrofuran (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

1. terc. Butyloxycarbonyl-3-trans-4-aminocyclohexyl) alanine-N, O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in anhydrous tetrahydrofuran (THF) (20 mL) under nitrogen, with stirring. The solution was cooled to 0 ° C, and N, N'-bis- (benzyloxycarbonyl) -S-methyl-isomeric (2.18 g, 6.1 mmol) was added. The solution was concentrated under reduced pressure, and the obtained 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) gave the purified product

2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous tetrahydrofuran was added dropwise n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) at -78 ° C, and the solution was stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with a saturated aqueous ammonium chloride solution (2 x 50 mL),

NZ brine (50 mL), dried over MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

och ₃

HgCl / THF

2. Lithium thiazole / THF

1. tert-Butyloxycarbonyl-3- (η 5 / trara-3-aminocyclohexyl) alanine-N, O-dimerylamide (2.0 g, 6.1 mmol) was dissolved in anhydrous tetrahydrofuran (THF) (20 mL) under nitrogen, with stirring. The solution was cooled to 0 ° C, and N, Ni-bis- (benzyloxycarbonyl) -S-methyl-isourea product (2.18 g, 6.1 mmol) was added. The solution was concentrated under reduced pressure, and the obtained 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) yielded purified

2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous tetrahydrofuran was added dropwise n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) at -78 ° C, and the solution was stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

HgCl / THF

2. Lithium thiazole / THF

1. tert-Butyloxycarbonyl-3-7 (5-tert-2-aminocyclohexyl) alanine-N, O-dimethylamide (2.0 g,

6.1 mmol) was dissolved in anhydrous tetrahydrofuran (THF) (20 mL) under nitrogen, with stirring. The solution was cooled to 0 ° C, and N, N 1 -bis- (benzyloxycarbonyl) -S-methylisourea (2.18 g, 6.1 mmol) was added. The solution was concentrated under reduced pressure, and the obtained 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) gave the purified product

2. To a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous tetrahydrofuran was added dropwise n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) at -78 ° C. , and the solution was stirred. The above guanidylated amino acid (3.88 g, 6.1 mmol) in tetrahydrofuran (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonium chloride solution. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonium chloride solution (2 x 50 mL), brine (50 mL), dried with MgSO 4, filtered, and concentrated under reduced pressure. The crude material was purified on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

Example 2

Synthesis of intermediates.

THF / -7g9c (4S, 5R) -3- (1-oxo-3-phenylpropyl) -4- (phenyl) -5- (methyl) -2-oxazolidone (2).

To a solution of (4S, 5R) -4-phenyl-5-methyl-2-oxazolidone (10.0 g, 1.0 eq., 56.4 mmol) in 250 mL of anhydrous THF, stirred at -78 ° C. n-BuLi (1.6 M in hexane, 1.1 eq., 38.8 mL) was added dropwise under argon. After 30 min. with stirring, hydrocinamoyl chloride was added dropwise at 10 minute intervals. 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 was dissolved in ethyl acetate and in dd? O. The aqueous phase was removed with two additional additions of ethyl acetate. The extracts were combined, washed with saturated sodium chloride, dried over sodium sulfate, and the solvent was removed in vacuo to give a white crystalline solid (2) (91% yield).

bt 95-96.5 0C; [a] _D = -35.8 (c = _{CH 2} C 1); [α] _β = -26.6 (c = ₂ C 1 018 _CH?);

1 HNMR (CDCl ₃ ) δ 0.89 (d, 3H, CH ₃ J = 6.6 Hz), 3.00-3.05 (m, 2H), 3.26-3.34

(m, 2H), 4.73-4.78 (m, 1 H), 5.64 (d, 1H, J = 7.4 Hz), 7.22-7.46 (m, 10H) what / A T A 1. LHMDS - J Me 1 - ‘‘ Ί Ph Third THF / 7S L ⁱⁿ 7 X Me 2 3

A solution of 5.0 g (1 eq., 16.2 mmol) of (2) in 100 mL of anhydrous THF was cooled to 78 ° C. Enolization was achieved with lithium bis-trimethylamide, which was added dropwise with stirring. The solution was stirred for 30 min. before 4.45 mL (3 eq., 48.5 mmol) of allyl iodide was added, and the reaction was warmed to -15 ° C. After 1 hour the reaction was quenched with saturated ammonium chloride and extracted (3x) with ethyl acetate. The organic phase was washed with sodium metabisulfite, dried over sodium sulfate, and the solvent was removed in vacuo to give a colorless oil. Purification was achieved by flash chromatography on

-86 silica gel using a stepwise gradient (15: 1, 12: 1, 10: 1) to give (3), a colorless oil (95%).

[α] _D = 47.5 (c = 3.12 CH ₂ Cl ₂ );

1 HNMR (CDCl ₃ ) δ 0.82 (d, 3H, CH ₃ J = 6.6 Hz), 2.31-2.40 (m, 1H

RCH = CHCH ₂ ), 2.49-2.57 (m, 1H RCH = CHCH ₂ ), 2.84-3.00 (m, 2H, Ph-CH ₂ ) 4.32-4.37 (m, 1H CH (N) CO), 4.53 to 4.56 (m, 1HCH ₃ -CH), 5.03-5.13 (m, ΤΗ, ΑΒΧ,

CH = CH ₂ ), 5.21 (d, 1H, Ph-CH, J = 7.1 Hz), 5.81-5.89 (m, 1H, CH = CH ₂ ), 7.20- 7.42 (m, 10H, ArH);

13 C (CDC1 ₃₎ δ 14.4, 36.2, 38.2, 43.9, 54.7, 78.4,

133.1, 134.8, 138.9, 152.4, 174.9.

117.1, 125.4, 126.3, 128.2, 129.0,

Allyl compound (3), (4.75g, 13.6 mmol) in THF (100 mL), was treated with 13.6 mL (1 eq. 13.6 mmol) of 0.1 M tetrahydrofuran borane complex at 0 ° C and stirred for 2h. The solvent was evaporated and chloroform (100 mL) was added with stirring. Oxidation of the organoborane was accomplished by the addition of 4.7 g (2 eq., 27.2 mmol) of 3-chloroperoxybenzoic acid at 0 ° C, warming to ambient temperature and stirring for an additional hour. The organic phase was washed with 5% Na ₂ CO ₃ , ddH ₂ O and dried over sodium sulfate. Due to the great instability of the alcohol, a column was removed quickly to remove the non-polar material that was derived from 3-chloroperoxybenzoic acid. The alcohol (4) was obtained in 65% yield.

[α] _D 39.3 (c = 1.936 CH ₂ Cl ₂ );

1 NMR (CDC1 ₃₎ δ l, 07 (d, 3H, CH ₃ CH J = 6.5 Hz), 1.81-1.92 (m, 3 H CH ₂ -CHH), 2.10-2.19 (m, 1HR, _ΟΗ? CH-H), 3J0-3,17 (m, 2H, Ph-CH ₂₎ 3.87 to 3.90 (m, 2H

CH ₂ OH), 4.43-4.49 (m, 1H CH-CO), 4.70-4.75 (m, fH, CH ₃ -CH), 5.36 (d, 1H, Ph) -CH, J = 7.1 Hz), 7.41-7.63 (m, 10H, ArH);

To a solution of alcohol (4) (1.0 g, 2.7 mmol) dissolved in dichloromethane (27 mL) was added 876 mg (1.5 eq, 4.1 mmol) pyridine chloromethane and 1.0 g 4. angstrom molecular sieve and the mixture changed from orange to black. The reaction was monitored by thin layer chromatography and after 30 min, when there was no more starting material, an additional amount of molecular sieve was 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 solution. If the orange color persisted in the organic phase, further filtration through celite was performed. The aldehyde was obtained in quantitative yield as a transparent colorless oil (5).

1 H NMR (CDCl 3 δ 0.84 (d, 3H, CH 3 CH- J = 6.6 Hz), 1.87-1.94 (m, 1H CH ₂ CH (H) -CHO), 2.04-2, 12 (m, 1H CH ₂ -CH (H 2 -CHO), 2.45-2.50 (m, 2H, Ph-CH 3), 2.79-2.85 (dd, 1HCH ₂ -CHO J = 13, 3 and J = 6.6), 2.92-2.99 (dd, 1HCH ₂ -CHO J = 13.2 and J = 8.8), 4.19-4.22 (m, 1H CH-CO ), 4.46 to 4.51 (m, 1 H, CH ₃ -CH-), 5.13 to 5.25 (m., 1 H, Ph), 7.20-7.39 (m, 10 H, ArH), 9.69 (s, 1H, CHO);

13 C (CDC1 ₃₎ δ 14,2,23,8, 39.0, 41.2, 43.8, 54.9, 78.6, 125.2, 126.4, 128.2, 128.4, 128.5.

:

Aldehyde (5) (2.6 g, 7.0 mmol) was dissolved in benzene (70 mL) and a catalytic amount of p-toluenesulfonic acid was added, followed by 1.58 g (1.2 eq, 8.52). mmole) of L-cysteine ethyl ester and a 4 A molecular sieve. The reaction was allowed to stir overnight at room temperature, followed by removal of the solvent in vacuo. The residue was dissolved in chloroform, washed with saturated sodium chloride solution, ddH ₂ O, and dried over sodium sulfate. The solvent was removed in vacuo to give a gummy solid (6).

2.0 M trimethylaluminium in hexane (2.4 mL, 4.8 mm -1, 3 eq) was added slowly to the starting material (6) (800 mg, 1.61 mmol) with stirring in dichloromethane under argon, when using a dryer. After stirring overnight, HPLC showed that the reaction was complete.

The mixture was quenched with excess methanol, then filtered on a short silica gel column (washed with excess 10% methanol in ethyl acetate). Evaporation gave 784 mg of crude material, which was purified using a silica gel column eluted with 2: 1 hexane: ethyl acetate to afford 258 mg (0.81 mmol, 50% yield) of pure (7), 6S-benzylhexahydro-5-oxo. 5H-thiazolo [3,2-a] pyridine-3R-ethyl ester as a yellow-white solid.

1 HNMR (CDCl ₃ ) δ 1.28-1.31 (M, 3H), 1.72-1.81 (m, 3H), 2.10-2.13 (m, 1H), 2.66 (dd) 1 H, J = 11.5 and 6.0 Hz), 3.29-3.34 (m, 2H), 4.19-4.29 (M, 2H), 4.88 (dd, 1H, J = 9.0 and 5.0 Hz), 5.22 (dd, 1H J = 8.0 and 6 Hz), 7.18-7.23 (M, 3H), 7.28-7.31 (m, 2H).

8

LiOH .40 (48 mg, 1.12 mmol) in 10 mL water was added to the starting material (7) (240 mg, 0.76 mmol) dissolved in 10 mL dioxane. After 1 hour, thin layer chromatography in hexane: ethyl acetate 1: 1 showed no starting material. The reaction mixture was quenched with 10% citric acid, then extracted twice with dichloromethane. By drying and evaporation, the combined organic layers gave 354 mg of crude product. This was dissolved in dichloromethane and then precipitated with excess hexane. The product was filtered to give 200 mg (0.68 mmol, 90% yield) of an off-white solid (8), also known as 6S-benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxylic acid acid.

-89H NMR (CD3OD) d 1.71-1.82 (m, 3H), 2.12-2.17 (m, 1H), 2.67 (dd, 1H, J = 14 and 11 Hz), 2 77-2.81 (m, 1H), 3.30-3.40 (m, 3H), 4.81 (dd, 1H J = 8.5 and 4.9 Hz), 5.16 (t, 1H (J = 7.5 Hz), 7.18-7.31 (m, 5H).

OH (COCI) ₂

---------- ►

C

Oxalyl chloride (9) (25 g, propionic acid (20 mL, 0.14 mol)) was stirred overnight The resulting mixture was distilled in 84% yield of a colorless liquid (10), cyclohexane propionic acid chloride.

ABOUT

0.197 mol) was cooled to 0 ° C and cyclohexane was added

1. n-BuLi (THF) -78 ° C

HN O

CH ₃ Ph

Second

CH ₃ Ph

Cl

Auxiliary chiral (11) (13.6 g 76.7 mmol, 1 eq) was dissolved in THF and cooled to -78 ° C. Then n-BuLi (52.8 mL, 84.4 mmol, 1.2 eq.) Was added and left for 30 min (dark orange solution). Acid chloride (10) (13.4 g, 76.6 mmol, 1 eq) was then added and allowed to stir overnight. It was further worked up by rapid cooling with ethyl acetate saturated with ammonium chloride. The extracts were washed with water and brine, dried over sodium sulfate and concentrated. A solid product (hexane: ethyl acetate 6: 1) eluted the pure product. This gave a white solid (12) which was recrystallized from ether and hexane to give the title compound in 78% yield.

[α] _D = -20.1 (c = 1, EtOH), MP / BP, mp = 90.5-91.5 ° C

1 HNMR (CDCl ₃ ) δ 0.86-1.10 (M, 5H), 1.18-1.30 (m 4H), 1.54-1.75 (m, 7H), 2,862.97 (m, 2H), 4.70-4.76 (m, 1H), 5.65 (d, 1H, J = 7.2 Hz), 7.28-7.42 (m, 5H).

The starting material (12) (9.13 g, 29 mmol, 1 eq.) Was dissolved in THF and cooled to 78 ° C, then LiHMDS (31.9 mL, 31.9 mmol) was added dropwise over 40 min. 1.1 eq). Then 30 min later was slowly over 10 min. allyl bromide (7.5 mL, 86.9 mmol, 3 eq.) was added. The mixture was allowed to warm overnight. The work up was quenched with saturated ammonium chloride, extracted with ethyl acetate, washed with 10% sodium thiosulfate, and decolourised with charcoal, dried over sodium sulfate and concentrated in vacuo. The product was obtained as a yellow oil (13) in 96% yield.

[α] _D = +9.5 (c = 1, EtOH),

1 HNMR (CDCl ₃ ) δ 0.92-1.10 (M, 5H), 1.10-1-35 (m, 5H), 1.63-1.75 (m, 6H), 2.272.42 (m 2H, 4.01-4.14 (m, 1 H), 4.76-4.85 (m, 1 H), 5.00-5.07 (m, 2 H, 5.65 (d, 1 H, J = 7 Hz, 5.64-5.88 (m, 1H, 7.27-7.46 (M, 5H)).

To the borane dimethylsulfide complex was added 2-methyl-2-butene at -12 ° C. The reaction was maintained at this temperature for 15 min. and then warmed to 0 ° C, and then allowed to stir for 2 hours. The disilamyl borane was then added to the mixture of the starting material (13) in THF using a double sided needle at 0 ° C. The mixture was allowed to stir for 2 hours, after which the solvent was removed and the residue was dissolved in dichloromethane. This was carefully added to a suspension of pyridinium chlorochromate in dichloromethane which was in a reflux flask. After the initial exothermic reaction decreased, the mixture was refluxed for 1 hour. at 50 ° C. The dark brown liquid was dissolved in ethyl acetate and filtered through Florisil. The black PCC residue was extracted with ethyl acetate and filtered through the same florisil. Concentration of the filtrate gave a 78% yield of a yellow gum (14).

[α] _D = -17.8 (¢ = 1.245, EtOH)

-91 1 HNMR (CDCl ₃ ) δ 0.89-1.18 (m, 5H), 1.20-1.47 (m, 8H), 1.60-1.74 (m, 6H) m, 1H) 2.48-2.53 (m, 2H), 3.90-4.10 (m, 1H, 4.12-4.16 (m, 1H), 4.76-4.80 (m, 1H) m, 1H, 5.67 (d, 1H, J = 7 Hz), 7.27-7.46 (m, 5H), 9.77 (s, 1H).

CO ₂ Et

The obtained aldehyde (14) (7.7 g crude, 20.8 mmol, 1 eq) was dissolved in 75 mL toluene. To the solution was added a catalytic amount of p-toluenesulfonic acid (50 mg), 10 g of 4 A molecular sieve and L-cysteine ethyl ester (3.87 g, 20.8 mmol, 1 eq.) The mixture was allowed to stir overnight, filtered and concentrated. The residue was purified by silica gel chromatography (hexane: ethyl acetate 6: 1) to give 6.36 g of the product (15) in 61% yield.

[α] _D = -48.3 (c = 1.095, EtOH)

1 HNMR (CDCl ₃ ) δ 0.84-0.98 (m, 4H), 1.11-1.38 (m, 7H), 1.50-1.90 (m, 1OH), 2.802.99 (m 1H, 3.24-3.34 (m, 1H), 3.77-4.29 (m, 4H), 4.46-4.81 (m, 2H), 5.66 (d, 1h) J = 7 Hz), 7.27-7.46 (m, 5H).

ABOUT

i.

WITH

NH

CH

ph

Me ₃ Al / DCM

P *

2 ° C RT

CO ₂ Et

The starting material (15) (1.97 g, 3.9 mmol, 1 eq) was dissolved in 20 mL dry dichloromethane and cooled to 0 ° C. Trimethylaluminum (5.9 mL, 11.8 mmol, 3 eq) was added dropwise, and the mixture was allowed to stir overnight. After complete reaction, proved by

-92HPLC, methanol was added until a yellow solid formed. Dichloromethane was added to dissolve and the whole was stirred for 15-30 min. and then filtered. The residue after concentration in vacuo was passed through a flash column (hexane: ethyl acetate 6: 1) to remove by-products and as many polar decomposition products as possible in 50% yield of a yellow oil (16).

1 HNMR (CDCl ₃ ) δ 0.83-0.98 (m, 2H), 1.09-1.38 (m, 1OH), 1.57-2.00 (m, 11H), 2.122.18 (m (IH), 2.49-2.54 (m, IH), 3.10 (dd, IH,> 11 and 6 Hz), 3.27 (dd, IH,> 11.5 and 8.0 Hz) 4.11-4.25 (m, 2H), 4.88 (dd, 1H, > 11.0 and 5.0 Hz), 5.14 (dd, 1H > 10 and 6 Hz).

The starting material (16) (0.95 g, 2.9 mmol, 1 eq) was dissolved in 10 mL dioxane. The solution was cooled to 10 ° C and LiOH was added. F 2 O (0.123 g, 2.9 mmol, 1 eq) dissolved in 10 mL water. The bath was removed and the mixture was stirred at room temperature for 1 hour. Thin layer chromatography showed the reaction was complete and the solvent was evaporated under vacuum. The resulting aqueous layer was washed (2X) with ether, acidified with 10% citric acid, and extracted (3X) with dichloromethane. The combined extracts were dried over sodium sulfate and concentrated to give a white solid, which was recrystallized from ether. Concentration of the filtrate and purification by silica gel column chromatography (hexane: ethyl acetate 2: 1) gave more product (17) with m.p. 198.2-199 ° C.

1 HNMR (DMSO-d 6) d 0.78-0.93 (m, 2H), 1.11-1.27 (m, 5H), 1.34-1.36 (M, 1H), 1.511.56 ( m, 1H), 1.60-1.75 (m, 1H), 1.82-1.87 (m, 1H), 2.15-2.18 (m, 1H), 2.37-2, 41 (m, 1H), 3.03 (dd, IH,> 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.83 (dd, IH,> 9 and 4 Hz) .

4.95 (dd, 1H,> 8 and 5.5 Hz).

C3Z /

N

HN N i-.

which

HN NH

-93BOC-DiCBz Arg (18) (17.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0 ° C. Then triethylamine (2.2 ml) was added followed by 14.5 mmol of IM toluene solution of isopropyl chloroformate via syringe. The reaction was stirred at 0 ° C for 30 min and then quickly filtered. The white solid was removed. Freshly prepared diazomethane was bubbled through the filtrate until a yellow color appeared. The reaction mixture was allowed to stand overnight in a well ventilated evaporator enclosure, which was taken up by excess diazomethane. Dry ether was added to precipitate the diazoketone. The product was filtered and dried under vacuum to give a pale yellow fluffy solid (4.6 g, 58%).

Diazoketone (19) (1g, 1.77 mmol) was dissolved in THF (20 mL) and this solution was added at 0 ° C, 1M HCl in ether (20 mL). The reaction was allowed to stir overnight at room temperature, and during this time a white precipitate formed. Precipitation was achieved by addition of ether. Filtration and drying of the product gave 20 (1.02 g, 100%).

1 HNMR (DMSO-d 6) δ 1.65-1.77 (m, 3H), 2706-2.50 (m, 1H), 3.86-3.90 (m, 2H),

4.76 (d, 1H, J = 18.7Hz), 4.95 (d, 1H, J = 18Hz), 7.35 (s, 2H), 7.35-7.41 (m, 10H) 8.71 (br s, 3H), 10.1 (br s, 2H).

C NMR (DMSO-d 6) δ 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3, 128.6, 126.7, 128.8, 128, 9, 135.2, 135.9, 153.4, 157.4, 198.9.

Q

HN NC5Z

N

NH

1.42 mmol) in THF (50 mL) at 0 ° C, and in the presence of N-methyl

WITH?

To compound (17) (0.422 morpholine) was slowly added 1M toluene solution of isopropyl chloroformate (1.71 mL) The reaction was stirred with stirring at 0 ° C, 30 min, and then treated in small portions with aminochloromethylketone (20). The reaction was allowed to stir at room temperature for 3 hours, then extracted with ethyl acetate, followed by washing with brine and 10% citric acid. gave a white foam (21) (1.03 g, 96%) which was used without further purification.

1 HNMR (CDCl ₃ ) δ 0.07-0.97 (m, 1H), 1.15-1.41 (m, 7H), 1.62-1.91 (m, 1OH), 2,102.16 (m 1 H, 2.43-2.48 (m, 1 H), 2.74-2.80 (m, 1 H), 3.01-3.07 (m, 1 H), 3.87-3.94 (m,

-941Η), 4.11-4.19 (m, 2H), 4.60-4.66 (m, 1H), 4.74-4.86 (m, 2H), 5.09-5.24 (m, 4H)

47.30-7.39 (m, 1H), 7.95 (d, 1H, J = 8 Hz), 9.4 (br s, 1H), 9.56 (br s, 1H).

EXAMPLE 3

From (N-t-BOC-N-tosyl) butyryl ketoarginine (240 mg, 0.525 mmol), the protecting group was removed using 30% trifluoroacetic acid in dichloromethane. After deprotection, the arginine derivative, coupled with the smimetic (8) (100 mg, 0.343 mmol) in DMF under alkaline conditions (EtN, pH = 8-9), was used BOP (228 mg, 0.52 mmol) as dehydrating agent. . The reaction was complete in 3-4 hours. Extraction with ethyl acetate was followed by successive washing with brine and 10% aqueous citric acid solution to give the crude product. The crude product was purified by column chromatography to give 181 mg (76%) of pure product. HF was used to remove the tosyl group from this product. Purification of the isolated, deprotected product by HPLC gave BCH-2737.

EXAMPLE 4

1. HS-CH2-CO2H

NHCO 3 (NMM / THF / RT ncs:

2. BBR 3 / DCM

ABOUT

Chloromethylketone (21) (0.188 g, 0.254 mmol) was dissolved in THF (10 mL), treated with NMM (0.036 mL) followed by mercaptoacetic acid (0.02 mL, 0.299 mmol). The reaction mixture was stirred at room temperature overnight. After reaction of the reaction mixture with ethyl acetate, followed by successive washing with brine and 10% citric acid, and evaporation of the organic

A -95 solvent gave the crude product which was purified by column chromatography to give a foamy solid (0.125 g, 62%).

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

The products of the reactions described above can be obtained in free form or in the form of their salts. In addition, the products obtained in a pharmaceutically acceptable acid may be added by the addition of salts reacting one of the free bases with an acid. Similarly, the product can be obtained as a pharmaceutically acceptable salt by reacting one of the free carboxylic acids with a base. Similarly, by treating the salt with a base or acid, the free amide is regenerated.

EXAMPLE 5

General method for the synthesis of compounds of formula Π or ΙΠ.

ch ₃ cn

NaCN8H ₃

RT 16 hi

KHMDS / THF

Tosyl Ng

as in Example 3

-97Example 6

Synthesis of compound:

(16>

STEP 1:

Synthesis of tert-butyl 2-benzyloxycarbonylamino-4-hydroxybutanoate

1) \ A) —o Cl

N MM. THF

2) NaBH ₄ . MeOH

NHZ (Cv-hexvi). NH.HOOC _XzZ Jx _co

HO

NHZ

CO.t-Bu

To a solution of protected group aspartic acid (1) (Bachem) (2.5 g, 4.95 mmol) in 50 mL of anhydrous tetrahydrofuran (THF), at -100C, under nitrogen, was added N-methylmorpholine (109 μΐ, 0, 2 equiv.) And isopropyl chloroformate (1.0M / toluene: 384 μΐ, 1.1 equiv.) The solution was stirred at -10 ° C for 60 min. In another flask was suspended NaBH 4 (375 mg, 2 equiv.) In Anhydrous mixture (THF / MeOH (50ml), at -78 ° C, under nitrogen. This suspension was stirred at -78 ° C, 30 min. The mixed anhydride solution was then added dropwise to NaBH 4 via cannula, and the resulting solution was After stirring at -78 ° C for 3 h, acetic acid (2.8 mL, 10 eq.) was added and the solution warmed to room temperature (30 min), then the solvents were evaporated and the residue was taken up in ethyl acetate and washed with sat. NaHCO 3 solution (2x) and brine The organic layer was dried over MgSO 4, the solid was filtered and the solvent evaporated to give 1.53 g (4.95 mmol, 100%) a1 methoxybenzyl alcohol (2), as a clear oil. 1 H NMR (CDC1 ₃ 400 MHz): delta 7.40-7.31 (m, 5H, Ar-H), 5.63 (d, 1 H, J = 7.3, NH), 5 13 (AB system, 2H, J = 12.2, C-Ph), 4.43 (m, 1H, H-2), 3.69 (m, 2H, H-4), 2.17 (m 1H, H-3), 1.63 (m, 1H, H-3), 1.48 (s, 9H, t-Bu).

STEP 2

2-Benzyloxycarbonylamino-4-iodobutanoic acid tert-butyl ester

NHZ Ph, P. I NHZ HO ^ (2) Imidazole. -IO ° C CHjCN / EUO I '^ ^{S * X X} _CO; t-Bu (2)

A -98K solution of alcohol (2) (1.53 g, 4.95 mmol) in CH 2 CN / Et ₂ O (50 mL) at -10 ° C, under nitrogen, was subsequently added imidazole (607 mg, 1 mL). , 8 eq) and phenol phosphate (2.21 g, 1.7 eq). Then iodine (2.14g, 17 eq) was added in small portions over 15 min. Upon complete addition, a white precipitate formed and the solution was brown. This was stirred for 45 min. at - 10 ° C. Then it was poured into ethanol, and the organic phase was washed with saturated sodium sulfite solution, saturated copper sulfate solution, water and dried over magnesium sulfate. The solids were filtered and the solvent was evaporated to give a yellow oil which was purified by flash chromatography (silica gel, 5 to 20% ethyl acetate / hexane). Iodide (3) was obtained in 83% yield (1.71 g) as a clear oil

1 H NMR (CDCl _3, 400 MHz): δ 7.41-7.31 (m, 5H, ArH), 5.35 (bd, 1H, J = 7.3, NH), 5.13 (s, 2H, CH) ₂ Ph), 4.30 (m, 1H, H-2), 3.22-3.12 (m, 2H, H-4), 2.42 (m, 1H, H-3), 2.20 (m, 1H, H-3), 1.48 (s, 9H, t-Bu).

STEP 3

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

NHZ MgBr. Cul NHZ I ^^^^ CO.t-Bu THF. 78 C (3) (4)

To a suspension of CuJ (2.27 g, 5 eq) in anhydrous THF (20 mL) at -78 ° C, under nitrogen, was slowly added 1.0 M magnesium vinyl bromide in THF (23.4 mL, 9.8 eq). .). The solution was then warmed to 10 ° C (mixed) and cooled back to -78 ° C. A solution of iodide (3) (1.0 g, 2.39 mmol) in anhydrous THF (3.5 mL) was then slowly added to the cupric solution. The reaction mixture was allowed to stir at -78 ° C, 2.5 h. To the mixture was added saturated ammonium chloride solution (50 mL), and the mixture was brought back to room temperature with vigorous stirring. It was then poured into ethanol and stirred for 5 min. The dark suspension was filtered through a separatory funnel and the phases were separated. The aqueous phase was extracted twice with ethanol, and the combined organic extracts were dried over magnesium sulfate. The solid was filtered off, the solvent was evaporated and the crude oil was purified by flash chromatography (silica gel, 5% ethyl acetate / hexane) to give 0.51 g (67%) of pure alkene (4).

1 H NMR (CDCl _3, 400 MHz): δ 7.37-7.31 (m, 5H, ArH), 5.80 (m, 1H H-5), 5.33 (d, 1H,

J = 7.8, NH), 5.12 (s, 2H, _CH2 Ph), 5.05 (d, 1 H, J = 17.2, H-6), 5.01 (d, 1 H, J = 10.4, H-6),

-994.30 (q, 1H, J = 17.2, H-2), 2.16-2.08 (m, 2H, H-4), 1.92 (m, 1H, H-3), 1.74 (m, 1 H,

H-3), 1.48 (s, 9H, t-Bu).

STEP 4

Synthesis of tert-butyl ester-benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinecarboxylic acid.

(4)

To a solution of alkene (4) (50 mg, 0.157 mmol) in anhydrous THF (3.1 mL), at room temperature, under nitrogen, was added mercuric acetate (75 mg, 1.5 eq). The solution was stirred at room temperature for 18 hours, then cooled to 0 ° C. Saturated NaHCO 3 solution (2 mL) was then added, and the mixture was stirred at 0 ° C, 30 min. KBr (0.11 g, 6 eq.) Was then added and the mixture was stirred at room temperature for 2 h. It was then poured into O0 / 10 ^0 and the phases were separated. The aqueous phase was extracted with Et 2 O (2x) and the combined organic phases were dried over MgSO 4. The solid phase was filtered off and the solvents were evaporated. Oxygen (C 1) was bubbled through a suspension of NaBH 4 (3.3 mg, 0.55 eq) in dry DMF (0.4 mL) for 1 hour when added dropwise (syringe, 3ml / hour) and a solution of organo-mercuric bromide in DMF (1.1 mL) in the presence of oxygen. Bubbling continued for 1 hour. and Et 2 O (5 mL) was added. The gray suspension was filtered through celite and the filtrate was evaporated. The residue was chromatographed (silica gel, ethyl acetate: hexane, 4: 6) to give pyrrolidinol (5) (30 mg, 57%) as a clear oil.

1 HNMR (CDCl ₃ 400 MHz): δ 7.37-7.28 (m, 5H, ArH), 5.22-5.09 (m, 2H, C 1 H 3 O), 4.30 (dd, 1H, J = 1.4, 8.3, 2-H), 4.24 (m, 1H, H-5), 3.70-3.57 (m, 3H, CH ₂ OH), 2.25 (m 1H, 2.13 (m, 1H), 1.92 (m, 1H), 1.34 (s, 9H t-Bu).

STEP 5

Synthesis of 1-benzyloxycarbonyl-5-carboxy-2-pyrrolidinecarboxylic acid tert-butyl ester.

SO, Pyr. Et, N

DMSO / CHjClj. 0 ° C

Ot-Bu

(6)

- a 10 ° C solution of alcohol (5) (50 mg, 0.149 mmol) and Et 2 N (62 μΐ, 3 eq.) in dry CH 2 Cl 2 (0.8 mL) was slowly added under nitrogen at 0 ° C, a solution of SOypyridine ( 71 mg, 3 eq) in dry DMSO. The solution was stirred at 0 ° C for 30 min, and then 10% aq. citric acid. The pH was adjusted to 4 with 1M NaOH, and the aqueous phase was extracted with Et ₂ O (3x). The combined organic extracts were dried over MgSO 4. The solid phase was filtered off and the solution evaporated to give a crude oil which was purified by flash chromatography (silica gel, ethyl acetate: hexane, 3: 7). Pure aldehyde (6) was obtained as a clear oil (45 mg, 90%).

1 H NMR (CDCl _3, 400 MHz): δ 9.63-9.56 (ds, 2H, CHO), 7.36-7.29 (m, 5H, ArH), 5.23-

5.11 (m, 2H, CH ₂ Ph), 4-57-4,19 (m, 2H, H-2, H-5), 2.30 to 1.97 (m, 4 H, H-3, H-4), 1.47 (2s, 9H t-Bu).

STEP 6 (7)

MgSO ₄ , CH ₂ c: 2

2) Na 3 H (OAC) _;

AcOH, TH?

(about)

The pyrrolidine-aldehyde (6) was coupled to the protected diamino-propanoic acid (7) by first forming the imine (8) (MgSO 4 CH ₂ Cl ₂ ). Isolation of the imine (8) was accomplished by filtration of MgSO 4 and evaporation of the solution. The crude imine treated with NaBH (OAc) and acetic acid in THF, 15 h, after extraction to give the amine (8).

STEP 7

U * ·>

Hi!

U -M- / IH (?

The CB7 amine protecting group (8) was removed by hydrogenation with palladium on 10% activated carbon in methanol used as catalyst. The catalyst was filtered off and the methanol was evaporated, yielding the crude diamine (9) which can be used without further purification.

- 101 STEP 8

OBUCA

Heat

--------->

The cyclization was carried out by heating the crude oil (9), from step 7, slightly above the boiling point of methanol. The bicyclic lactam (10) was purified by flash chromatography.

STEP 9

8CC. (10;

G H

8CC (12)

The bicyclic lactam secondary amine (10) was protected as an amide using chloride in pyridine. Evaporation of pyridine and treatment with extraction afforded the bycyclic lactam-amide (11).

STEP 10

9OC (i: · < ^{1 - 2!}

The protecting groups of the bicyclic lactam amide (11), BOC and tert-butyl ester, were removed under acidic conditions (HCl in ethyl ether (Et 2 O). The amine salt (12) precipitated from the solution by filtration.

STEP 11

(12)

The primary amine compound (12) was protected with a CBZ group formed by the reaction of benzyl chloroformate in acetonitrile with K 2 CO 2 as the base. The extraction work-up gave the fully protected carboxylic acid (13), which can be used for step 12 without further purification.

STEP 12

The carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF, using the BOP coupling agent in the presence of diisopropylethyleneamine (EtNiPr). Elution with ethyl acetate (EtOAc) gave compound 15 as a solid which was chromatographically purified.

STEP 13

Catalytic hydrogenation was used to remove the two CBZ (Z) protecting groups from compound (15) using 10% Pd / C as a catalyst. The catalyst was filtered off and the solvent removed by evaporation to give amino-guanidine (16).

Example 7

Compound synthesis (10)

STEP 1

(2)

4-Methylmorpholine (NMM) was added to a solution of the carboxylic acid (2) (1.7g, 4.9 mmol, 1.0 eq), 4-hydroxyproline (3) (5.39 mmol, 1.1 eq), and BOP reagents (2.17g, 4.9mmol, 1.0eq) in anhydrous DMF (10ml) at room temperature. The reaction mixture was stirred at room temperature overnight, cooled with brine (50 mL) and ethyl acetate (100 mL). The organic layers were washed with aqueous citric acid (10%, 2 x 50 mL), sodium bicarbonate (10%, 2 x 50 mL), and brine (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered and the solvent was evaporated. The crude residue was purified by flash chromatography (5: 4: 1, ethyl acetate-hexane-methanol). 1.1 g of pure product were obtained in 48% yield.

1) triaylamine

2) MsO

CPC

- 104 To a solution of the 4-hydroxyproline derivative (4) (115 mg, 240 pmol, 1.0 eq.) In dichloromethane (10 mL, anhydrous) at 0 ° C was added triethylamine (72 mg, 720 pmol, 3 eq.) and methanesulfonyl chloride (28 mg, 240 pmol, 1 eq) and the reaction mixture was stirred at room temperature. The mixture was then cooled with aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was washed with 10% citric acid and brine, dried, filtered and the solvent was evaporated to give compound (5).

STEP 3

(l) Hg (CAc) ₂

CF3CCCH / CPC

2) Nc ₃ H ₄

The enamine (5) (1 eq) was treated with mercuric acetate (1.1 eq) in THF. The solvent was evaporated by drying and the residue was dissolved in methanol. The resulting organo-mercuric compound was reductively cleaved with sodium borohydride (1.3 eq). The resulting crude lactam thioether was purified by flash chromatography on silica gel to afford compound (6).

STEP 4

To a solution of thioether lactam (6) (1.0 eq) in dry dichloromethane was added N-chlorosuccinimide (1.0 eq) at 0 ° C. The reaction mixture was warmed to room temperature. if

The starting material is no longer detected in the reaction, so the solid is filtered and the solvent is evaporated by drying. The crude product (7) was used without further purification for step 5.

STEP 5

(7)

PhjClli

THF

(8)

To a solution of alpha-chlorothioether (7) (1.0 eq) in THF (anhydrous) was added a solution of phenyl copper (1.0 eq) (prepared according to literature) at low temperature. When the reaction mixture does not show the starting chlorothioether, ethyl acetate and brine are added. The organic layer is dried, filtered and evaporated by drying to give the desired product (8).

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

STEP 7

ABOUT)

(10)

The untreated carboxylic acid (9) is coupled with benzylthiazole ketoarginine in DMF using BOP as coupling agent in the presence of diisopropylethylamine. Extraction with ethyl acetate gave a solid which was purified on silica to give the protected amide. The CBZ protecting group is removed with BBr in dichloromethane at room temperature to finally give the bicyclic benzothiazole ketoarginine inhibitors (10).

In addition, the following compounds were prepared by making appropriate product substitutions to obtain the final compounds.

Compound # 11

(11) Lnh 'H N

Compound # 12

(12)

-107 -

Example 8

The commercially acceptable monoethyl ester of glutaric acid chloride (1) (20 mL, 0.144 mol) was dissolved in 40 mL of dry tetrahydrofuran (THF) and cooled to -15 ° C. An excess of freshly prepared diazomethane in 300 mL of ether was cannulated at -15 ° C into the solution. The mixture was allowed to warm to room temperature overnight. Excess diazomethane was evacuated from the vessel with a stream of argon. To quench the reaction, add 75 mL of 1 N HCl in ether at 0 ° C and allow the mixture to warm to room temperature for 5 hours. The solvent volume was reduced and then washed with 2 x 5% NaHCO 3. dried over Na ₂ CO 3 and evaporated to give the crude chloromethyl ketone (20.46 g, 79%), which was used in the next step without further purification.

1 HNMR (CDCl ₃ , 400 MHz, d 1.16-1.2 (t, 1H), 1.83-1.9 (m, 2H), 2.27-2.35 (m, 2H), 2 6-2.64 (t, 1H); 3.6 (s, 3H); 4.04 (s, 2H).

STEP 2

¹ HC.H-Cs-CtT / CH.COCNa / NcCNBH.

/ --------------- ------------ 3, _the MeCH.RT ^ _OCH3

Untreated chloromethyl ketone (2) (10.04 g, 56.15 mmol) was dissolved in 300 mL dry methanol. Sodium acetate (2 eq., 9.21 g, 112.3 mmol) was added, followed by L-cysteine ethyl ester hydrochloride (1.3 eq, 13.55 g, 72.98 mmol) and sodium cyanoborohydride (1.4 eq.). eq., 4.9 g, 78.59 mmol). The heterogeneous mixture was stirred at room temperature for 2.5 hours. Then 200 mL of methanol was added to dissolve all solids and the pH was maintained with 1N HCl. The mixture was then basified with saturated sodium bicarbonate until pH 8. Metano, evaporated

-108 and the remaining aqueous solution was washed with ethyl acetate and dichloromethane. The solutions were combined, dried over sodium sulfate and evaporated. The crude residue was purified by flash column chromatography on silica gel using ethyl acetate / hexane gradient eluents in the following ratios: (3: 7, 5: 5, 7: 3) to give cyclic compound (3).

1 H NMR (CDCl ₃ , 400 MHz) of compound (3) d 1.21-1.27 (t, 3H), J = 7.06 Hz), 1.41-1.48 (m, 2H), 1 65-1.73 (m, 2H), 2.28-2.39 (m, 4H), 2.57-2.63 (t, 1H, J = 10.9), 2.72-2, 76 (dd, 1H, J = 10.7 Hz), 2.8-2.86 (m, 1H), 3.6-3.64 (d, 4H, J = 2.55 Hz), 3.63 (s, 3H), 4.13-4.2 (m, 2H) ^{, 3} C NMR (CDCl ₃ , 400 MHz), 13,078,19,888, 28,326, 31,133, 32,741, 35,277, 50,462, 56,394, 59,149, 60,188, 69.713, 170.182.172.52.

STEP 4

O.CHCH

Toluene / H ⁺

RefliK

Cyclic compound (3) (913 mg, 3.32 mmol) was dissolved in 50 mL dry toluene. (1S) - (+) - 10-Camphorsulfonic acid (92 mg, 0.39 mmol) was added and the mixture was refluxed for 4 days. When all of the starting material (TLC) was consumed, the mixture was treated by evaporation of the solvent, the residue was dissolved in ethyl acetate and washed with 2 x 5% sodium bicarbonate. The ethyl acetate layer was dried over sodium sulfate and evaporated. The crude residue was purified by flash column chromatography on silica gel using 60% ethyl acetate / 40% hexane followed by 70% ethyl acetate / 30% hexane to give 62.5% bicyclic compound (4).

1 H NMR (CDCl 3, 400MHz) Compound (4) d 1.27-1.31 (t, 3H, J = 7Hz), 1.5-1.6 (m, 1H), 1.72-1.87 (m, 2H), 2.02-2.1 (m, 1H), 2.33-2.46 (m, 2H), 2.52-2.59 (m, 2H), 2.83-2 .88 (dd, 1H, J = 14.4 Hz), 3.14-3.18 (d, 1H), 3.78-3.85 (m, 1H), 4.2-4.27 (Q) (2H, J = 3.9 Hz), 5.9-5.92 (t, 1H, J = 3.4 Hz).

-109 STEP 5

ΗίΟΗ.Η ₂ Ο.ΤΗΡ / Η ₂ Ο

0 ° C + G.T

5

The bicyclic compound (4) (3.66 mg, 1.5 mmol) was dissolved in 25 mL THF and 5 mL water. Lithium hydroxide monohydrate (1.1 eq., 7.05 mg, 1.68 mmol) in 2.3 mL of water at 0 ° C was added and the mixture was allowed to stir at this temperature for 1 hour and at room temperature for 3 hours. The THF was then evaporated and the remaining aqueous mixture was acidified by addition of citric acid to pH 2. Extraction with 2 x CH 2 Cl 2 and 2 x ethyl acetate, drying the combined organic layer with sodium sulfate and evaporation gave a crude residue which was purified by flash column chromatography on silica gel using 70% ethyl acetate / 30% hexane followed by 4.7% HOAC / ethyl acetate to give pure acid (5) in 5% yield, 16% starting material recovered.

1 HNMR (MeOD, 400 MHz) of compound (5) d 1.57-1.69 (m, 1H), 1.70-1.80 (m, 1H), 1.81-1.89 (m, 1H) 2.05-2.12 (m, 1H), 2.35-2.5 (m, 2H), 2.51-2.66 (m, 2H), 2.86-2.91 (dd) 1H, J = 13.8 (4 Hz), 3.12-3.17 (d, 1H), 3.3-3.32 (m, 1H), 3.78-3.84 (m, 1H) , 5.76-5.78 (t, 1H, J = 3.53 Hz).

¹³ C NMR (MeOD, 400 MHz) d 17.052, 27.7, 28.928, 31.382, 32.096, 51.016, 55.138, 170.088, 171.24.

STEP 6

ή

To a solution of lithium bis (trimethylsilyl) amide (5 mL of 1 M THF solution, 5 mmol) in THF (10 mL was added at -78 ° C a solution of carboxylic acid (5) (500 mg, 2.32 mmol).

The 110 solution was stirred at -78 ° C for 1 hour. Benzyl bromide (0.26 mL, 2.22 mmol) was then added and the mixture was left at room temperature and stirred for 15 hours. The mixture was then poured into 10% HCl (50 mL) and extracted with dichloromethane (4 x 60 mL). The combined organic phase was dried with magnesium sulfate and the solvent was evaporated to give the crude alkylated amide (6).

The crude alkylated amide (6) was coupled with benzylthiazole ketoarginine in DKF using BOP as coupling agent in the presence of diisopropylethylamine. Extraction with ethyl acetate gave a solid which was purified on silica gel to afford the protected amide. The CBZ protecting group was removed with BBr in dichloromethane at room temperature to finally obtain the bicyclic benzthiol keto-arginine inhibitors.

The following compounds were prepared in the same way, with suitable substitutions being made to obtain the final compounds.

Compound # 8

^X NH 4 NH HN

Example 9

Determination of K value of heterocyclic compounds

The affinity of thrombin inhibitors was measured according to the methods described in (DiMaio et al. J.Bio.

Chem., 1990, 265: 21698). The inhibition of amidolytic activity of human thrombin was measured

-111 fluorometrically using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50 mM TrisHCl buffer (pH 7.52 at 37 ° C) containing 0.1 M NaCl and 0.1% polyethylene glycol 8000 at room temperature (Szewczuk et al., Biochemistry, 1992 31: 9132).

Substrate hydrolysis by thrombin was monitored on a Varian-Cary 2000 spectrometer with fluorescence mode (λεΧ = 383 nm, λem = 455 nm) or Hitachi F2000 ^Ti ^ fluorescence spectrometer (λ = 383 nm, λ = 455 nm), and the fluorescence intensity was calibrated using AMC. The reaction became steady state within 3 minutes after mixing of thrombin with the substrate and inhibitor. The steady state speed was then measured for a few minutes. The compounds of the invention were also preincubated with thrombin for 20 minutes at room temperature before adding the substrate. Steady state was reached within 3 minutes and measured for several minutes. Kinetic data (steady state velocity at different substrate concentrations and inhibitors) of competing inhibitors were analyzed using the methods described by Segel (1975). The nonlinear PNLIN regression program in the IMSL library (IMSL, 1987), the LMDER in the MINPACK library (More et al., 1980) or Microsoft ™ Excell ™ were used to calculate kinetic parameters (K, V and K).

dTT test

The fibrin clotting assay was performed in 50 mM Tris HCl buffer (pH 7.52 at 37 ° C) containing 0.1 M NaCl and 0.1% polyethylene glycol 8000 with 9.0 x 10-10 M (0.1 NIH units / ml) and 0.03% (w / v) final concentrations of human thrombin and cattle fibrinogen, respectively as described elsewhere (Szewczuk et al., supra). Clotting time was plotted against inhibitor concentration and IC 50, calculated as inhibitor concentration required for two clotting times relative to control. The results are summarized in Tables 1 and 2 below.

Fibrin clotting test

The fibrin clotting assay was performed essentially as described by Krtenansky et al., FEBS, 1987, 211: 10. Dilute inhibitor solution was prepared in 50 mM Tris HCl buffer (pH 7.8 at 23 ° C) containing 0.1 M NaCl and 0.1% (w / v) polyethylene glycol 8000. Human plasma (60 μΐ, collected in sodium citrate) , blood / anticoagulant 9: 1) was added to a microtitre vessel (Falcon plate) containing 100 μΐ of various inhibitor dilutions. The solution was stirred after adding 50 μΐ of human thrombin (1 nM, final horse) and stirred for 15 seconds. The turbidity of clotting was immediately monitored by a microplate reader (Dynateck MR 5000) at 405

-112 nm and recorded every 3 minutes. Maximum turbidity in the absence of inhibitors was achieved within 60 minutes. IC values were calculated at 30 minutes as the concentration of inhibitor that gave half the optical density of the control.

Platelet aggregation and secretion

Rat blood was collected by ACD (6/1 v / v) cardiac puncture. The washed platelet suspension was prepared according to the procedure described by Ardlie et al. (Br. J. Haematol. 1970, 19: 7 and Proc. Soc. Exp. Biol. Med., 1971, 136: 1021). The final suspending medium was modified with Tyrode solution ((NaCl 138 mM, KCl 2.9 mM, HEPES 20 mM, Na2HPO4 0.42 mM, NaHCCh 12 mM, CaCh, MgCl 2.0.1% glucose, 0.35% albumin, apyrase 1 μΙ / ml pH 7.4) The resulting platelet count was adjusted to 5,000,000 / μ1.

To allow measurement of the extent of release of the dense round bodies, platelets were labeled when first washed with a solution of ¹⁴ C-serotonin (5-HT) (1 pCi / 10 ml wash liquid) and the released ¹⁴ C-serotonin was determined according to Holmsen et al. (Enzymology, 1989, 169: 206). Inipramine (5 μΜ final horse) was added to detect the reabsorption of serotonin released.

Platelet aggregates were recorded at 37 ° C in an aggregometer (BioData PAP-4) at a stirring speed of 1,100 rpm by measuring changes in light transmittance. Aggregation percentages were determined 3 minutes after addition of stimulator (human thrombin 0.1 IU / ml final horse.) Inhibitors were preincubated for 1 minute at 37 ° C before addition of stimulator. IC values represent the concentration required to suppress platelets or secretions relative to 50% of the control samples.

Model of arterial thrombosis

FeCE Model of induced carotid arterial injury

FeCE-induced carotid artery (artery) damage in rats was induced according to the procedure described by Kurz, K.D., Main, R.W., Sandusky, G.E., Trombosis Research 60, 269-280, 1990 and Schumacher W.A. and others J. Pharmacology and Experimental Therapeutics 267. 12371242, 1993. Male, Sprague-Dawley rats (375-410 g) were anesthetized with urethane (1500 mg / kg ip). The animals were placed on a plate heated to 37 ° C. The carotid artery was exposed

-113 effect across the midline of the cervical incision. Careful blunt dissection was used to isolate the vessel from the carotid vagina. Using the tweezers, the artery was lifted, and two small pieces of polyethylene tubes (PE-205) were inserted under it to effect removal. A temperature probe (Physitemp MT23 / 3) was placed between one piece of tube and the artery. The disease was induced by topical application to the carotid artery of the above small disk temperature probe (3 mm dia). Whatman č. 1 wherein the filter paper was first immersed in a 36% FeCl3 solution. The incision region was covered with aluminum foil to protect FeCh from light degradation. The vessel temperature was monitored for 60 minutes after FeCl2 administration as an indication of blood flow. Changes in vessel temperature were recorded on a thermistor (Cole-Palmer Model 08533-41).

The time between application of FeCl 3 and the time at which the vessel temperature drops sharply (> 2.4 ° C) was recorded as the time to vessel occlusion. The inhibitor compounds were administered iv as a bolus (pill) (mg / kg) followed immediately by iv infusion (pg / kg / min via the thigh vein). It was determined that the dose of inhibitor required twice the time for occlusion compared to a control animal in which injury was induced in the absence of inhibitor.

114 Table 1

Antithrombocyte activity 'μΜ

CMPD Ki μΜ clustering 5-HT secretion dnic _with μΜ Plasma fibrin clotting test ic, μΜ 0005 4 ND ND 47 > 450 0010 4.6 21 19 89.5 > 450 0015 Ίό > 100 > 100 162 > 450 0020 2.2 18 14.2 22 > 450 0025 53 > 100 > 100 > 625 > 450 0030 8.6 > 100 > 100 67 320 0035 34 > 100 > 100 319 > 450 0040 19 > 100 > 100 207.5 > 450 0045 74 ND ND 415 > 450 0050 62 ND ND > 450 0065 32.7 47.5 52 42 200 0070 4.4 22 2.1 25 78 0080 0048 0.4 00:38 0.375 ND 0090 0.031 ND ND 00:33 ND 0095 26 ND ND 0100 19 ND ND 165 ND

´ Suspension of washed platelets from rats Inhibitory dissociation constant of human α-thrombin

-115Table 2

compound ki (NM) dTT (NM) Path ivb + inf M. O. (Min) +/- sem 0220 18 0225 550 235 • 0.75-50 23 +/- 7 0245 5 8 0.5-30 0.75-50 27 +/- 3 22.6 +/- 2.6 0250 40 350 0.25-20 0.75-50 23 +/- 8 22 +/- 3 0295 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 0.75-50 20:13 +/- 3.4 0255 500 0260 16 0.75-50 14.83 +/- 0.2 0305a 220 0305b 12000 0265 4 0.75-50 21.5 +/- 10 0265b 18 0.75-50 14.83 +/- 2.3 0285 10 150 0.75-50 11:33 +/- 1.34 0315-b 45 0315b 10 0.75-50 30.33 +/- 8.4 0335 25 138 0.75-50 0.5-30 0.25-20 45.8 +/- 14.2 41.5 +/- 27.7 27.5 +/- 11.3 0340 0.6 0.25-20 0.75-50 36 +/- 9.6 42.25 +/- 11.9 0345 2 0.75-50 50 +/- 5.86 0915 1600 0.75-50 15 +/- 1.3 0935 120

-116Table 2 (continued)

compound ki (NM) dTT (NM) Path ivb + inf M. O. (Min) +/- sem 0925-b 10 0.75-50 19.6 +/- 0.2 0925b 30 0925 7 0.75-50 20.3 +/- 3.5 0940 16 0.75-50 15.2 +/- 0.82 0940b 160 0950 150 0950b 1000

a = early elution of single isomer on RP HPLC b = later elution of single isomer on RP HPLC a + b - mixture

7 /

- 117PATENT CLAIMS

Claims (20)

1. Compound of formula (I): Where A can be (CH-R 5) 0-1, S, SO, SO 2, O, and NR 5, wherein R 5 is hydrogen, C 1-6 alkyl optionally interrupted with 1 or 2 heteroatoms; A C 6-16 aryl, C 3-7 cycloalkyl or heterocyclic ring or a hydrophobic group; B may be S, SO 2, O, -N =, NH, -CH = and CR 6 R 7 wherein R 6 and R 7 may independently be hydrogen or C 1-6 alkyl with the proviso when A is S, SO, SO 2, O or NR 5 , then B is CR $ R7 D can be (CH-R 9) 0-2 wherein R 9 is hydrogen, C 1-6 alkyl or -C (O) R 1; and CH with a double bond to B when B is -N = or -CH =; E may be CH 2 or CH substituted with -C (O) R 1, provided that only one D and E is substituted with -C (O) R 1; X may be O, N-R 5, or CH-R 5; Y may be O, S, SO, SO 2, N-R 5 or CH-R 5, with the proviso that when X is N-R 5, then Y is CH-R 5 or O, and when X is O then Y is CH-R 5 ; Z may be O, S and H2; R 1 is a polar amino acid residue of arginyl or an analogue or derivative or derivative thereof optionally substituted with an amino acid, peptide or heterocycle; R ₂ may be H or C 1-6 alkyl optionally substituted with C 6 aryl, 6-membered heterocycle or C 3-7 cycloalkyl ring; R ₃ can be H, NR 6 R 7, or C 1-6 alkyl and R ₄ and R _g may be each independently H; NROR? ; C 6-14 aryl or C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; C 1-6 alkyl optionally interrupted with one or more heteroatoms or a carbonyl group and optionally substituted with OH, SH, NR 3 R 7, or C 6-10 aryl, heterocycle or A-118 C 3-7 cycloalkyl group optionally substituted with halogen, hydroxyl, C 1-6 alkyl; an amino acid side chain; and a hydrophobic group. Compound according to claim 1, characterized in that Vla to VId: R j is represented by one of the formulas R 11 is hydrogen or C 1-6 alkyl; K is a bond or -NH-; G is C 1-4 alkoxy; cyano; NH ?; CH? NH ?; -C (NH) -NH ₂ ; -NH-C (NH) -NH ₂ ; -CH ₂ -NHC (NH) -NH 2; C 6 cycloalkyl or aryl substituted with cyano, -NH ₂ , -CH ₂ -NH ₂ , -C (NH) -NH 2, -NH-C (NH) -NH ₂ or -CH ₂ -NH-C (NH) - NH ?; or a 5 or 6 membered, saturated or unsaturated heterocycle optionally substituted with cyano, -NH 2, -CH 2, or CH 2; -NH 2, -C (NH) -NH 2, -NHC (NH) -NH 2? or -CH -NH-C (NH) -NH 2; U is cyano, -NH ₂ ,. C (NH) NH? or -NH-C (NH) -NH 2. P is a bond, -C (O) - or a divalent group: OH J is C 1-6 alkylene optionally substituted with OH, NH? C 1-6 alkyl and optionally interrupted by a heteroatom, which may be O, S and N. n is 0 or 1; and T is H, OH, amino, a peptide chain, C 1-6 alkyl, C 6 alkoxy, C 6-10 aralkyl, or a heterocycle optionally substituted. . A compound according to claim 2, wherein T is a heterocycle selected from the group consisting of: -119kde X 5, X 10 and X 11 X 12 are independently selected from the group consisting of N or C-X 7, wherein X 7 is hydrogen, C 1-6 alkyl, or C 6-10 aryl; X ₆ and X 13 are each independently selected from the group consisting of C, O, N, S, NX or CH-X 7; R 1 is hydrogen, C 1-16 alkyl optionally substituted with carboxyl, -C 0-10 alkyl-C 1 -C 6 alkyl, C 6-20 aralkyl, C 3-7 cycloalkyl, aryl or an aromatic heterocycle. 3-F WITH· - 120where R is hydrogen, Ci_i ₆ alkyl optionally substituted by carboxyl, C ₀ .i6 alkyl-CO ₂ - ₆ m alkyl, Cg-20 aralkyl, C _{3. 7 is} cycloalkyl, aryl or aromatic heterocycle. 4 and R ₅ is a hydrophobic group selected from C 1-20 alkyl, C 2-20 alkenyl or C 2-20 alkynyl optionally interrupted by a carbonyl group; ₆ .i C ₆ aryl, C _{3. 7} cycloalkyl, C ₆ -2o aralkyl, C _{6. 20} cycloalkyl substituted with C 1-20 alkyl, wherein the aliphatic moiety is optionally interrupted by a carbonyl group and the ring moiety is optionally substituted with C 1-6 alkyl; and a hydrophobic amino acid side chain. A compound according to claim 3, wherein T is selected from the group consisting of: where R 'is hydrogen, C 1-6 alkyl optionally substituted with carboxyl, -C 0-16 alkyl-CC 2 - C 6 alkyl, C 6-20 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle. 5-Oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamido (abenzothiazolo ketoarginine) - 124- 5-oxo-5H-thiazolo [3,2-a] pyridine-3-R-carboxyamido (propylcarbomethoxy ketoarginine) 0105 6S-Cyclohexylpropylhexahydro A compound according to claim 4, wherein T is selected from the group consisting of: R ' 6. A compound according to claim 1, wherein one R &apos; A compound according to claim 6, wherein R ₃ is hydrogen. A compound according to claim 1 wherein Z is O. A compound according to claim 1 wherein R ₃ is hydrogen A compound of formula VH wherein R is a portion of arginyl or an analog or derivative or derivative thereof optionally substituted with an amino acid, peptide or heterocycle; R 2 can be H or C 1-6 alkyl; R ₃ can be H, NR 8 R ₇ , or alkyl and ^R 4 and ^R ₈ can independently be H; NR R ?; C ₆ aryl, or C -i6 _{third 7} cycloalkyl optionally substituted with alkyl; C 1-6 alkyl optionally interrupted with one or more heteroatoms or a carbonyl group and optionally substituted with OH, SH, NR 4 R 6, or C 6-16 aryl, heterocycle or C ₃ . _A cycloalkyl group optionally substituted with halogen, hydroxyl, C 1-4 alkyl; an amino acid side chain; and a hydrophobic group. - 121 - A compound according to claim 10, wherein R 1 is one of formulas VIa to VId: wherein: R 11 is hydrogen or C 1-6 alkyl K is a bond or -NH-; G is C 1-6 alkoxy; cyano; -NH ₂ ; CH? -NH ₂ ; -C (NH) -NH ₂ ; -NH-C (NH) -NH ₂ ; -CH ₂ -NHC (NH) -NH ₂ ; C 6 cycloalkyl or aryl substituted with cyano, -NH ₂ , -CH ₂ -NH ₂ . -C (NH) NH ₂ , -NH-C (NH) -NH _2, or -CH ₂ -NH-C (NH) -NH ₂ ; or a 5 or 6 membered, saturated or unsaturated heterocycle optionally substituted with cyano, -NH ₂ , -CH ₂ -NH ₂ , -C (NH) -NH 2, -NH-C (NH) -NH _2, or -CH -NH -C (NH) -NH 2; U is cyano, -NH ₂ , -C (NH) -NH _2, or -NH-C (NH) -NH 2, P is a bond, -C (O) - or a divalent group: OH J is alkylene optionally substituted with OH, NH? and C 1-6 alkyl and optionally interrupted by a heteroatom, which may be O, S and N. n is 0 or 1; and T is H, OH, amino, a peptide chain, C 1-6 alkyl, C 6 alkoxy, C 6-10 aralkyl, or a heterocycle optionally substituted. The compound of claim 11, wherein T is a heterocycle selected from the group consisting of: I - 122where X _a, X _n Χίο and X 12 are independently selected from the group consisting of N or C _7, wherein X ₇ is hydrogen, alkyl, or C _{6. I6} aryl; X ₆ and X 11 are each independently selected from the group consisting of C, O, N, S, N- X ₇ or CH-X ₇ ; R is hydrogen, C _W6 alkyl optionally substituted by carboxyl, -C s alkyl-CO ₂ - C. _C1-6 alkyl, C1-10 aralkyl, C3. _{7 is} cycloalkyl, aryl or aromatic heterocycle. A compound according to claim 12, wherein T is a heterocycle selected from the group consisting of: and R 'is hydrogen, C 1-8 alkyl optionally substituted with carboxyl, -C 0-6 alkyl-CO ₂ -C 1-6 alkyl, C 6-20 aralkyl, C 3-7 cycloalkyl, aryl or aromatic heterocycle. A compound according to claim 13, wherein T is selected from: WITH - 123 and R is H, m ₆ alkyl optionally substituted by carboxyl, C ₀ .i6 alkyl-CO ₂ - a C ₁₆ alkyl, CFI-20 aralkyl, C _{3. 7 is} cycloalkyl, aryl or aromatic heterocycle. A compound according to claim 10, wherein R ₂ and R ₃ are H. A compound according to claim 10, wherein R 4 is C 1-10 alkyl optionally interrupted with a heteroatom or carbonyl, and optionally substituted with a C 1-10 aromatic hydrocarbon, C ₃ . _{7 is a} cycloalkyl or heterocyclic ring wherein the ring is optionally substituted with CF ₃ or oxo. 17. The compound according to claim 10, characterized in that R _p is hydrogen. A compound according to claim 12, characterized in that R ₃ is hydrogen; R 4 is Calkyl optionally interrupted with a heteroatom or a carbonyl, and optionally substituted with C ₆ _i ₆ aromatic hydrocarbon, C _{3. A} cycloalkyl or heterocyclic ring wherein the ring is optionally substituted with CF ₃ or oxo; and R ₅ is hydrogen. A compound according to claim 12, characterized in that it is selected from: 0085 6S-cyclohexylmethylhexahydro- A compound according to claim 1, of formula VIII wherein R 1 is an arginyl residue or analog or derivative thereof optionally substituted with an amino acid, peptide or heterocycle; R ₂ is H or alkyl; R a can be H, NR 1 R ₇ and C 1-6 alkyl; and R 4 and R 5 are independently selected from H; NR6R ₇ ; C 6-6 aryl or C 3-7 cycloalkyl optionally substituted with C 1-4 alkyl; C 1-6 alkyl optionally interrupted with one or more heteroatoms or carbonyl and optionally substituted with OH, SH, NR 6 R ₇ or C ₆ . ₁₆ aryl, heterocycle or C _{3. 7} cycloalkyl optionally substituted with halogen, hydroxyl, C 1-6 alkyl; an amino acid side chain; and a hydrophobic group,