Low molecular weight bicyclic thrombin inhibitors

Field of the Invention

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

Origin of the Invention

Unnecessary thrombus formation on vascular walls accelerates acute states of cardiovascular disease, which are the leading cause of death in economically advanced societies. Plasma proteins such as fibrinogen, proteases, and cellular receptors involved in hemostasis are important factors that play a role in acute and chronic coronary artery disease and cerebral artery disease; they are involved in the formation of thrombi or blood clots, which significantly reduce the normal flow and supply of blood. Vascular abnormalities resulting from primary pathological conditions such as hypertension, atherosclerotic plaque fractures, or ruptured endothelium activate biochemical cascades that serve as a response to the lesion and repair of the damaged site. In the coagulation cascade, thrombin is a key regulatory enzyme; it plays a pluralistic role both as a regulator of positive and negative feedback. However, in pathological states, the latter linkage is enhanced by the catalytic activation of cofactors required for thrombin generation as well as activation of factor XIII required for fibrin-binding and stabilization.

In addition to its direct effect on hemostasis, thrombin has a direct effect on various cell types that maintain and increase the pathogenesis of arterial thrombotic disease. This enzyme is the strongest activator of platelets, causing them to aggregate and release substances (eg ADP TXA₂ NE) that further promote the thrombotic cycle. Platelets within the fibrin network form the main framework of the white thrombus. Thrombin also has a direct effect on endothelial cells by causing the release of vasoconstrictor substances and translocation of adhesive molecules, which become binding sites for immune cells. In addition, this enzyme causes smooth muscle cell mitogenesis and fibroblast proliferation. From this assay, it becomes clear that inhibition of thrombin activity is an effective therapeutic approach to inhibiting thrombosis-related proliferation.

The major endogenous factor that neutralizes thrombin activity in mammals is antithrombin III (ATIII), a circulating plasma macroglobulin that has a low affinity for this enzyme. Heparin has clinical efficacy in venous thrombosis, enhancing ATIII / thrombin binding by catalytic effect. However, heparin also catalyzes the inhibition of other proteases of the coagulation cascade, and its efficacy in platelet-dependent thrombosis is greatly reduced or abolished due to its inaccessibility to the thrombus-bound enzyme. Undesirable effects such as thrombocytopenia, osteoporosis and triglyceridemia have been observed during long-term treatment with heparin.

Hirudin derived from glandular secretions of hirid medicinalis glands is one of the high molecular weight natural anticoagulant inhibitors of thrombin activity (Markvvardt F. Cardiovascular Drug Reviews, 10, 211, 1992). It is a pharmaceutical biopreparation with efficacy in experimental and clinical thrombosis. The major disadvantage of using hirudin as a therapeutic agent is its antigenicity and the lack of effective neutralization methods, particularly in terms of its highly expressed binding properties to thrombin. Its tremendous attraction to thrombin is unique and is attributed to its simultaneous interaction with the catalytic site as well as to the further anion binding of the enzyme 'anion.

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

Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for the thrombin catalytic site by inhibiting the proteolysis of this protein or other protein substrates such as thrombin receptor. The overall modeling strategy for enzyme-inhibiting compounds is based on the simulation of specificity inherent in the primary and secondary structure of the natural enzyme substrate. In this way, Blomback et al. were the first to develop a thrombin inhibitor which was modeled by a partial sequence of the fibrinogen A (LBi) α chain comprising its proteolytically accessible domain (Blomback, et al., J. Clin. Lab. Invest., 24, 59, 1969). This region of fibrinogen includes at least residues starting with phenylalanine:

Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-GlyGly-Gly-Val-Arg-Gly-Pro-Arg t Cleavable Link

Systemic amino acid substitution in this field has enabled optimization of the tripeptide inhibitor sequence exemplified by the (D) Phe-Pro-Arg peptide corresponding to interactions at the P3-P2-P1 local thrombin binding sites (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 Ine. Ann Arbor Ml, 1975, p.603).

Bajusz et al. also described congeners such as (D) PhePro-Arg- (CO) H (GYKI-14166) and (D) MePhe-Pro-Arg- (CO) H (GYKI-14766) (Peptides-Synthesis, Structure and Function; Proceedings of the Seventh American Peptide Symposium, Rich, DH & Gross, E. Eds., Pierce Chemical Company, 1981, p.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 considered to be highly involved in inhibitory activity due to its chemical reactivity with the thrombin catalytic Seri₉₅ with respect to the residue forming a semi-acetal intermediate.

Related work in the field of thrombin inhibitory activity has utilized a major recognition motif mediated by the (D) Phe-ProArg tripeptide by introducing various functional or reactive groups into a site corresponding to a commonly recognized cleavage linkage (i.e., P1-P1 ').

In U.S. Patent No. 4,318,904, Shaw describes chloromethyl ketones (PPACKs) which are reactive with Seri₉₅ and His₅7th with respect to. These two residues include part of the thrombin catalytic triad (Bode, W. et al., EMBO Journal, 8, 3467, 1989).

Other examples of thrombin inhibitors containing the (D) -Phe-Pro-Arg core moiety include inhibitors containing COOH-terminal borarginine variants such as boric acids or borates (Kettner, C. et al., J. Biol. Chem. , 268, 4734, 1993).

Still other related materials for this moiety are those containing phosphonate groups (Wang, CL. J., Tetrahedron Letters, 33, 7667, 1992) and α-ketoesters (lwanowicz, E. J. et al., Bioorganic and Medicinal Chemistry Letters, 12, 1607). , 1992).

Neises, B. et al. Described a trichloromethylketone-type thrombin inhibitor (MDL-73756) and Attenburger, J.M. et al. Described a related difluoraliclamide ketone (Tetrahedron Letters 32, 7255, 1991).

Maraganore et al (EP 0333356; WO 91/02750; US 5196404) describe several thrombin inhibitors containing the D-Phe-Pro-residue and suggest that this proper structure fits well into the "groove" adjacent to the thrombin active site. Various examples of such inhibitors include predominantly linear or cyclic peptides constructed on the basis of the D-Phe-Pro residue.

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

Still others have focused their attention on peptides, peptide derivatives, peptide alcohols, or cyclic peptides as antithrombotic agents (WO 93/22344, EP 0276014; EP 0341607; EP 0291982). Other authors have investigated amide sulfuric acid residues to obtain the same effect (US 4781866), while still others have studied para- or meta-substituted phenylalanine derivatives (WO 92/08709; WO 92/6549).

A number of Mitsubishi patents and patent applications describe clearly effective argininamide compounds for use as antithrombotic agents. The chemical structures described herein represent variations of the side groups of the argininamide compound (US 4173630; US 4097591; CA 1131621; US ​​4096255; US 4046876; US 4097472; CA 2114153).

Canadian Patent Applications 2076311 and 2055850 describe cyclic imine derivatives which exhibit inhibitory effects on cell aggregation.

Most of the examples quoted above eventually result in the maintenance of a linear acyclic tripeptide moiety containing an arginyl residue whose basic side chain is required for interaction with the carboxylate group at the bottom of the thrombin Pi specificity gap. Two contiguous hydrophobic groups are involved in the formation of an additional bond due to the beneficial interaction of Van der Walsh in the contiguous hydrophobic crack on the surface of the enzyme designated P3-P2.

It is an object of the present invention to provide thrombin inhibitors which have inhibitory activity against the target enzyme thrombin.

Another object of the present invention is to provide thrombin inhibitors which have inhibitory activity against the target enzyme thrombin and to provide them in a pharmacologically acceptable state.

Still another object of the present invention is to provide heterocyclic thrombin inhibitors and their formulations as anticoagulant and thrombin inhibiting agents.

Yet another object of the present invention is to provide heterocyclic thrombin inhibitors and their formulations for the treatment of various thrombotic disorders.

Yet another object of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The structure of the enzyme inhibitors of the present invention is represented by the general formula I.

Summary of the Invention

The present invention provides novel compounds having thrombin inhibitory activity of the general formula I:

Z

d)

<img file="LT4368B_D0001.tif" />

X wherein;

A is selected from (CH-Re) oi, S. SO, SO₂, O and NR₈, where R₈ is hydrogen, Ci.₆-alkyl, which may be optionally interrupted by 1 or 2 heteroatoms; C₆-i₆ -aryl, C₃.₇ -cycloalkyl or a heterocyclic ring or a hydrophobic group;

B is selected from S, SO₂, O, -N =, NH, -CH = and CR 6 R 2, wherein R 6 and R 6 are independently selected from hydrogen and Ci.₆-alkyl, provided that when A is S, SO, SO₂, O or NR₈, B is CR 6 R 7;

D is selected from (CH-R 8) 0-2 where R 8 is hydrogen, C 1-6 alkyl or C (O) R 1; and CH is double bonded to B when B is -N = or -CH =;

E is selected from CH₂ and CH substituted with -C (O) R 1, provided that only one of D and E has a -C (O) R 1 substituent;

X is selected from O, N-R 5 or CH-R₅;

Y is selected from O, S, SO, SO, NR₅ and CH-R₈, provided that when X is NR₅, then Y is CH-R₈ or O, and when X is O, then Y is CH-R₈;

Z is selected from O, S and H₂;

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

R₂ is selected from H and Ci.₆-alkyl which may be substituted such as C 6 -aryl, 6-membered heterocycle or C₃.7-cycloalic ring;

R₃ is selected from H, NR₆R7 and Ci.₆-alkyl; and

R₄ and R₅, independently of one another, are selected from H; NR₆R7; C₆-i6-aryl or C₃_7-cycloalkyl which may be substituted by Ci.₆-alkyl; Cne-alkyl which may be interrupted by one or more heteroatoms or a carbonyl group and may be substituted such as OH, SH, NR₆R7 or C₆-6-aryl, heterocycle or C₃.₇a -cycloalkyl group which may be substituted such as halogen, hydroxyl, C 1-6 alkyl; amino acid side chains; and hydrophobic groups.

As will become apparent from the following description, the molecules, compositions and methods of the present invention are useful as anticoagulants, or

Ί treatment and prevention of various diseases related to the undesirable effects of thrombin, as well as for diagnostic purposes.

Detailed Description of the Invention

The present invention relates to molecules that inhibit the enzyme thrombin. These molecules have the heterobicyclic residue shown in formula I:

<img file="LT4368B_D0002.tif" />

wherein X, Y, Z, A, B, D, E, and R 1 are R₄ are as described above.

The term "hydrophobic group (HG), as used herein, means any group that is not water related or displaces water. Hydrophobic groups include, but are not limited to, Ci.₂o-alkyl, C2-20 alkenyl (e.g. vinyl, allyl) or C₂-20-alkynyl (e.g., propargyl), which may be interrupted by a carbonyl group (e.g., an acyl group): C₆-16-aryl, C3-7-cycloalkyl, C6-20-aralkyl, C₆-20o-cycloalkyl, which may be substituted with C1-20 alkyl, the aliphatic portion of which may be interrupted by a carbonyl group (e.g., alkylation), and the ring portion may be substituted with, such as a C 1-10 alkyl, such as methyl , ethyl or t-butyl; or a side chain of a hydrophobic amino acid. Preferred hydrophobic groups are cyclohexyl, benzyl, benzoyl, phenylmethyl, phenethyl and para-t-butylphenylmethyl.

The term "arginyl residue" refers to an amino acid residue of an arginine, analogue or derivative thereof. For example, the analogue or derivative of the natural residue may have a longer or shorter methylene chain at the alpha carbon (i.e. ethylene or butylene chain): replacement of the guanidine group with a hydrogen bond donor or acceptor group (i.e., amino, amidino or methoxy); replacement of the methylene chain by a stationary group (ie aryl, cycloalkyl or heterocyclic ring); terminal carboxyl elimination (ie decarboxylation) or hydroxyl elimination (ie aldehyde group); or a combination of these changes.

The term "alkyl" means a straight or branched, saturated or unsaturated chain containing the total number of carbon atoms.

The term "aromatic or" aryl means an unsaturated carbocyclic ring (s) of 6 to 16 carbon atoms which may be substituted by one or two substituents such as OH, SH, amino (i.e., NReRz). halogen or C 1-4 alkyl. Aromatic rings include benzene, naphthalene, phenanthrene and anthracene. Preferred aromatic rings are benzene and naphthalene.

The term "cycloalkyl" means a saturated carbocyclic ring of 3 to 7 carbon atoms which may be substituted by one or two substituents such as OH, SH, amino (i.e. NReRz), halogen or Ci.₆-alkyl. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. A preferred cycloalkyl group is cyclohexyl.

The term "aralkyl" refers to a substituent containing an aryl moiety attached through an alkyl chain (e.g., benzyl, phenethyl) in which the total number of carbon atoms of the aryl moiety and the alkyl chain is as given. The aryl residue or chain may be substituted by one or two substituents such as OH, SH, amino (i.e. NReRz), halogen or₆-alkyl.

As used herein, the term "heteroatom" refers to oxygen, nitrogen, or sulfur (O, N, or S) as well as sulfoxyl or sulfonyl (SO or SO).₂) unless otherwise noted. It is understood that alkyl chains interrupted by one or more heteroatoms means that the carbon atom of the chain is replaced by heteroatoms having the appropriate valence. Preferably, the alkyl chains are interrupted by 0-4 heteroatoms and that two adjacent carbon atoms are not substituted.

The term "heterocycle" means a saturated or unsaturated mono- or polycyclic (i.e., bicyclic) ring containing one or more (i.e., 1-4) heteroatoms selected from N, O and S. It is understood that the heterocycle may be substituted with one or two such such as OH, SH, amino (i.e. NReRz), halogen, CF.₃, oxo or Cve-alkyl. Examples of suitable monocyclic heterocycles include, but are not limited to, pyridine, piperidine, pyrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran and thiophene. Examples of suitable bicyclic heterocycles include, but are not limited to, indole, quinoline, isoquinoline, purine and carbazole.

The term "hydrophobic amino acid" refers to an amino acid residue having an alkyl or aryl group attached to an a-carbon atom. Thus, glycine, which has no such group attached to the α-carbon atom, is not a hydrophobic amino acid. The alkyl or aryl group may be substituted, provided that the substituent or substituents do not diminish the overall hydrophobic character of the amino acid. Examples of hydrophobic amino acids include natural amino acids such as alanine, isoleucine, leucine, phenylalanine, and unnatural amino acids such as those described in The Peptides, Vol. 5,

1983, Academic Press, Chapter 6 by DC Roberts and F. Vellaccio. Suitable non-natural amino acids are cyclohexylalanine and 1 aminocyclohexanecarboxylic acid.

The term "amino acid side chain" refers to a substituent attached to a carbon at the α-position of an amino group. For example, the amino acid alanine side chain is a methyl group and the phenylalanine side chain is benzyl.

Preferably R₂ would be H or Ci.₆-alkyl. Even better when R₂ is H, methyl or ethyl, and preferably when R₂ is H.

Preferably R₃ would be H or Ci-₆-alkyl. Even better when R₃ is H, methyl or ethyl, and preferably when R₃ is H.

Preferably one of Rs₄ or R₅ would be a hydrophobic group such as a 5- or 6-membered saturated or unsaturated carbocycle which may be fused to another carbocyclic group and the second would be H, Ci.₆-alkyl which may be substituted, such as NR₆Rz or a carboxy group. The hydrophobic residue may be attached via a bridge such as Ci.₁₆-alkyl chain which may be interrupted by 1 or more (i.e. 1-4) heteroatoms, carbonyl or sulfonyl (SO₂) in groups. Preferably when one of Rs₄ and R₅ is phenyl, cyclohexyl, indole, thienyl, quinoline, tetrahydroisoquinoline, naphthyl or benzodioxolane attached via Cne-alkyl, which may be interrupted by a heteroatom or carbonyl, and the second is H, carboxymethyl or carboxyethyl.

Preferably, A is absent or CH2.

Preferably B is S or CH 2.

Preferably D is CH2.

Preferably E is CH substituted with -C (O) R_b wherein R 1 is as previously described.

Preferably X is CH-R₅ or N-R5.

Preferably Y is CH-Rs or S.

Preferably Z is O.

In a preferred embodiment of the present invention, R1 is represented by one of the formulas Vla-Vld:

<img file="LT4368B_D0003.tif" />

in which:

R 11 is hydrogen or C 1 -C 6 alkyl;

K is a bond or -NH-;

G is C 1-4 alkoxy; cyano, -NH₂; -CH₂NH₂; -C (NH) -NH₂; -NHC (NH) -NH₂; -CH₂-NH-C (NH) -NH₂, 'C₆-cycloalkyl or aryl substituted with substituents such as cyano, -NH₂; -CH₂NH₂; -C (NH) -NH₂; -NH-C (NH) -NH₂or -CH₂-NH-C (NH) -NH₂; or a 5- or 6-membered saturated or unsaturated heterocycle which may be substituted with cyano, NH₂; CH 2 NH 2: -C (NH) -NH₂; -NH-C (NH) -NH₂ or -CH₂-NH-C (NH) -NH₂;

U is a cyano group, -NH₂, -C (NH) -NH₂ or -NH-C (NH) -NH₂;

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

<img file="LT4368B_D0004.tif" />

J is Ci.₆-alkylene which may contain substituents such as OH, NH₂ and Ci.₆alkyl, and may be interrupted by a heteroatom selected from O, S and N;

n is 0 or 1; and

T is H, OH, amino, peptide chain, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C₆-₂o-aralkyl or heterocycle which may be substituted.

Preferably Rn is H or methyl, preferably H.

Preferably K is a connector.

Preferably G is -NH-C (NH) -NH₂, attached via a methylene chain of 3 to 7 carbon atoms, or phenyl having -C (NH) -NH₂ a substituent connected through a 0-3 carbon chain. Even more preferably, G is -NH-C (NH) -NH₂, connected through a chain of 3 carbon atoms.

Preferably P is -C (O) -.

Preferably J is selected from -CH₂-S-CH₂-CH₂-; -CH₂-OCH₂-CH₂-; -CH₂-NH-CH₂-CH₂-; and would be a bond when n is 0.

In certain embodiments of the present invention, Ri is selected from the following amino acid derivatives prepared according to Bioorg. Med. Chem., 1995, 3; Methods described in 1145:

<img file="LT4368B_D0005.tif" />

HN

<img file="LT4368B_D0006.tif" />

<img file="LT4368B_D0007.tif" />

<img file="LT4368B_D0008.tif" />

<img file="LT4368B_D0009.tif" />

wherein n = 1-6, n1 = 1-2, n2 = 0-7, and T is as previously described.

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

<img file="LT4368B_D0010.tif" />

in which

Xs, Χίο, Xn and Xi2 are independently selected from the group consisting of N or CX₇ , where X₇ is hydrogen, Ci.₄-alkyl or C₆-t6aryl;

Xe and Xi3 are independently selected from the group consisting of C, O, N, S, NX₇ or CH-X₇;

R 'is hydrogen; C 1 -C 6 alkyl which may be substituted, such as carboxyl; carboxyl; -C0-16 alkyl-CO₂-Ci-i₆-alkyl; C6-₂o-aralkyl C 3-7 cycloalkyl, aryl or aromatic heterocycle.

It is best to choose T from the group consisting of:

<img file="LT4368B_D0011.tif" />

<img file="LT4368B_D0012.tif" />

R '

<img file="LT4368B_D0013.tif" />

wherein R 'is as described above.

Even better, T is selected from the group consisting of:

<img file="LT4368B_D0014.tif" />

R '

<img file="LT4368B_D0015.tif" />

R '

<img file="LT4368B_D0016.tif" />

wherein R 'is as described above.

Even better, T is selected from the group consisting of;

<img file="LT4368B_D0017.tif" />

wherein R 'is as described above. Best of all, when T is:

R 'r

sarba

R 'wherein R' is H or C 1-4 alkyl such as methyl, ethyl, propyl or butyl, and preferably when R 'is hydrogen. In another embodiment, T is a 1,2-thiazole which may be substituted such as R 'and / or attached to J at the 2-, 3-, 4 or 5-position of the ring.

In certain embodiments, the compounds of the present invention are represented by Formulas II, III, IV and V wherein X, Y, B, Ri-R₄ and R₈ are as described previously.

<img file="LT4368B_D0018.tif" />

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

<img file="LT4368B_D0019.tif" />

<img file="LT4368B_D0020.tif" />

in which:

B is O, S, -CH₂- or -NH-;

Y is selected from O, S, SO, SO₂, NR₅ and CH-Re;

R 1 is an arginyl residue or an analogue or derivative thereof which may be substituted with an amino acid, a peptide or a heterocycle;

R₂ is H or Ci.₆-alkyl;

R₃ is selected from H, NR₆R₇ and C 1-6 alkyl; and

R₄ and R₅, independently of one another, are selected from H; NR6R7; C₆-i6-aryl or C3-₇-cycloalkyl which may be substituted, such as C 6 -alkyl; Cm₆alkyl which may be interrupted by one or more heteroatoms or a carbonyl group and may be substituted such as OH, SH, NR₆R₇ or Ce.i₆aryl, heterocycle or C₃.₇a -cycloalkyl group which may be substituted such as halogen, hydroxyl, Ci.₆-alkyl; amino acid side chains; and. · · 'hydrophobic groups;

Re is hydrogen, C1-6 alkyl, which may be interrupted by 1 or 2 heteroatoms; C₆-6-aryl, C3-₇-cycloalkyl or heterocyclic ring or hydrophobic group; and n is 1 or 2.

Suitable compounds of formula VII are:

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

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

0015 6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (propylcarbmethoxyquetoarginine)

0020 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide (benzyl ketoarginine)

0025 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (carbmethoxypropylcyclodithioketalarginine)

<img file="LT4368B_D0021.tif" />

<img file="LT4368B_D0022.tif" />

<img file="LT4368B_D0023.tif" />

HN '^^iKNH₂

<img file="LT4368B_D0024.tif" />

hn &lt;^ nh₂

O

<img file="LT4368B_D0025.tif" />

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide ((S) -Arg- (R) pipecolylic acid)

6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (carboxamidopropylcyclodithioketalarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide ((S) -Argnipecotamide)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide ((S) -Argisonone pecotamide)

6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (Carboxamidopentylcyclodithioketalarginine)

6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (carbmethoxypropylcyclodithioketalarginine)

<img file="LT4368B_D0026.tif" />

Ο^Η0 ^ ° οΕνηχΛΛ ^XNH

HN ^ NH, θ cf

ΓΛ

<img file="LT4368B_D0027.tif" />

NH,

<img file="LT4368B_D0028.tif" />

NH

A

HN

NH,

CUo

-NH

NH,

<img file="LT4368B_D0029.tif" />

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide (1-Carboxy-3-thiobutyl-ketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine 3R-Carboxamide (1-Carboxy-3-thiobutyl-ketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine 3R-carboxamide (1-Carboxy-2-methyl-3-thiobutyl-ketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine 3R-carboxamide ((3-Thiobutylsulfonic acid) ketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide (isoquinoline-methyl-ketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide (propylcarb

-siketoarginine)

<img file="LT4368B_D0030.tif" />

0090

0095

0100

0105

0110

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine3R-carboxamide ((propyl keto) ArgPhe-Arg-NH₂)

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

6S-Benzylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (Propyl Carb

-siketoarginine)

6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (abenzothiazole-ketoarginine)

6S-Cyclohexylpropylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine 3R-carboxamide (propylcarbmethoxy-ketoarginine) '6-benzyl-5-oxo-hexahydrothiazolo [3,2-a] pyridine-3-carboxylic acid [1-benzothiazole-2-carbonyl] 4-guanidinobutyl] amide

<img file="LT4368B_D0031.tif" />

0205 '

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

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

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

8a-Methyl-5-oxo-6-phenyl-1H-hexahydro-thiazolo [3,2a] pyridine-3-carboxylic acid [1-benzothiazole-2-carbonyl] -4-guanidinobutyl] -amide

8a-Methyl-5-oxo-6-phenethylhexahydro-thiazolo [3,2a] pyridine-3-carboxylic acid [1-benzothiazole-2-carbonyl] -4-guanidinobutyl] -amide

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

<img file="LT4368B_D0032.tif" />

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

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

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

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

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

H

<img file="LT4368B_D0033.tif" />

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

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

<img file="LT4368B_D0034.tif" />

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

6- (1,3-Dioxo-1,3-dihydro-isoindol-2-yl) -5-oxo-hexahydro-thiazolo [3,2a] pyridine-3-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

5-Oxo-6- (3-phenylpropionylamino) -hexahydro-thiazolo [3,2a] pyridine-3-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

<img file="LT4368B_D0035.tif" />

0315 5-Oxo-6- (3-phenylpropionylamino) -hexahydro-thiazolo [3,2a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

<img file="LT4368B_D0036.tif" />

More preferred compounds of formula VII are:

0085 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (propylcarbomethoxy-ketoarginine);

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

0095 6S-benzylhexahydro-5-oxo-5H-tazolo [3,2-a] pyridine-3R-carboxamide ((propanoic acid, ketoarginine);

0105 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (a-benzothiazole ketolargin);

0210 6-Benzyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide;

0220 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidino-butyl] -amide;

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

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

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

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

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

0315 5-Oxo-6- (3-phenyl-propionylamino) -hexahydro-thiazolo [3,2-a] pyridine-3-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide. Most preferred compounds of formula VII are;

0085 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (propylcarbomethoxy-ketoarginine); and

0105 6S-Cyclohexylmethylhexahydro-5-oxo-5H-thiazolo [3,2-a] pyridine-3R-carboxamide (α-benzothiazole ketoarginine).

Preferred compounds of formula VIII are 0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydro-pyrrolo [1,2-a] pyridine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidine-butyl] -amide.

<img file="LT4368B_D0037.tif" />

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

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

<img file="LT4368B_D0038.tif" />

NH HN ^ NH,

<img file="LT4368B_D0039.tif" />

H

<img file="LT4368B_D0040.tif" />

O

<img file="LT4368B_D0041.tif" />

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

4- Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (5-methylthiazole-2-carbonyl) butylamide

2- (3-Cyclohexylpropionyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidine-1- (2-thiazolecarbonyl) butylamide

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diaza-naphthalene-4-carboxylic acid- [4-guanidino-1- (thiazole-2-carbonyl) butylamide] 4-Oxo-2- (4-phenylbutyryl) octahydro-pyrrole [1,2-a] Pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] -amide

4-Oxo-2-phenylacetyl-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butylamide

<img file="LT4368B_D0042.tif" />

0375

0380

0385

0390

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

2- [2-Amino-3- (4-hydroxyphenyl) propionyl] -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butylamide]

2- [2-Amino-3- (4-fluorophenyl) propionyl] -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butylamide]

4-Oxo-2- (3-phenylpropyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-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

<img file="LT4368B_D0043.tif" />

<img file="LT4368B_D0044.tif" />

<img file="LT4368B_D0045.tif" />

<img file="LT4368B_D0046.tif" />

<img file="LT4368B_D0047.tif" />

0395

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

<img file="LT4368B_D0048.tif" />

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

<img file="LT4368B_D0049.tif" />

2- (3-1H-1H-imidazol-4-yl-propionyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid es [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

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

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

<img file="LT4368B_D0050.tif" />

<img file="LT4368B_D0051.tif" />

<img file="LT4368B_D0052.tif" />

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

2- (2-Hydroxy-3-phenylpropionyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

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

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

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

<img file="LT4368B_D0053.tif" />

NH

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

4- (6- [4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxo-hexahydro-pyrrolo [1,2-a] pyrazin-2-yl) -4-oxo-3- (2-propylpentanoylamino) ) -Butanecarboxylic acid methyl ester

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

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

<img file="LT4368B_D0054.tif" />

NH

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

<img file="LT4368B_D0055.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (5-ethylthiazole-2-carbonyl) -4-guanidinobutyl] -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidine-1- (5-methylthiazole-2-carbonyl) -butyl] -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidine-1- (4-methylthiazole-2-carbonyl) -butyl] -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (4-ethylthiazole-2-carbonyl) -4-guanidinobutyl] -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid (4-carbamimidoyl-phenylamide)

<img file="LT4368B_D0056.tif" />

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

4-Oxo-2- (3-phenylpropionyl) octahedro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (5-benzylthiazole-2-carbonyl) -4-guanidinobutyl] amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (4-carbamimidoylbenzyl) -2-oxo-2-thiazol-2-ylethyl] amide

<img file="LT4368B_D0057.tif" />

<img file="LT4368B_D0058.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (3-carbamimidoylbenzyl) -2-oxo-2-thiazol-2-ylethyl] amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (1-carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] amide

<img file="LT4368B_D0059.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (1-carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ethyl] amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (1-carbamimidoylpiperidin-2-ylmethyl) -2-oxo-2-thiazol-2-ethyl] amide [6- [4 -Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazin-3-yl] acetic acid [6- [4-Guanidino-1 - (Thiazole-2-carbonyl) -butylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazin-3-yl] -propane carboxylic acid [6- [1- (1-Carbamimidoylpiperidine-4) -ylmethyl) -2-oxo-2 • Thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] -pyrazin-3-yl] acetic acid

<img file="LT4368B_D0060.tif" />

<img file="LT4368B_D0061.tif" />

<img file="LT4368B_D0062.tif" />

<img file="LT4368B_D0063.tif" />

3- [6- [1- (1-Carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] pyrazine 3-yl] acetic acid [6- (1- (1-Carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] ] -pyrazin-3-yl] acetic acid [6- (3-Guanidinopropylcarbamoyl) -4-oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] -pyrazin-3-yl] acetic acid

3- [6- (3-Guanidinopropylcarbamoyl) -4-oxo-2- (3-phenylpropionyl) -octahydropyrrolo [1,2-a] -pyrazin-3-yl] propanoic acid

<img file="LT4368B_D0064.tif" />

<img file="LT4368B_D0065.tif" />

<img file="LT4368B_D0066.tif" />

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

<img file="LT4368B_D0067.tif" />

NH

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] methylamide [6- [1- (1-Carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] methylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] pyrazine - il] acetic acid

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ylethyl ] methylamide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid (3-guanidinopropyl) methylamide

<img file="LT4368B_D0068.tif" />

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

<img file="LT4368B_D0069.tif" />

o

0600 ,

0605

0610

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

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

2- (4-tert-Butylbenzoyl) -4-oxo-octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

<img file="LT4368B_D0070.tif" />

<img file="LT4368B_D0071.tif" />

0615

0620

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

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

<img file="LT4368B_D0072.tif" />

2- [2- (2-Methylbenzylidene) -but-3enoyl] -4-oxo-octahydro-pyrrolo [1,2-a] -pyrazirt-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2 -thiazol-2-ylethyl] amide

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

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

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid 4-carbamimidoyl-benzylamide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-imidazol-1-yl-1- (thiazol-2o)

<img file="LT4368B_D0073.tif" />

H

N

0650

0655

0670

0675

0680 carbonyl) butyl] amide 4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4- (2 amino-imidazol-1-yl) -1- (thiazole-2-carbonyl) butyl ] amide

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

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [3- (6-amino-6-chloro-pyridin-2-yl) -1 (thiazole-2-carbonyl) propyl] amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [3- (2aminopyridine-4-yl) -1- (thiazole-2-carbonyl) propyl] amide

<img file="LT4368B_D0074.tif" />

<img file="LT4368B_D0075.tif" />

<img file="LT4368B_D0076.tif" />

<img file="LT4368B_D0077.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [2- (2-aminopropyl-η-4-yl) -1- (thiazole-2-carbonyl) ethyl] amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [2- (6-amine-1-di-η-2-yl) -1 - (thiazole-2-carbonyl) ethyl] amide

2- [4-Oxo-2- (3-phenylpropionyl) octahydropyrrolo [1,2-a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) -1,2,3,4-tetrahydroisoquinoline- 6carboxamidine

<img file="LT4368B_D0078.tif" />

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

<img file="LT4368B_D0079.tif" />

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

<img file="LT4368B_D0080.tif" />

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

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

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

<img file="LT4368B_D0081.tif" />

<img file="LT4368B_D0082.tif" />

<img file="LT4368B_D0083.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid (2-oxo-1β-piperazin-4-yl-2-thiazol-2-yl-ethyl) -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid (2-oxo-1-piperidin-3-yl-2-thiazol-2-ethyl) amide

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

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (5-benzyl) -thiazole-2-carbonyl] -4-guanidinobutylamide.

<img file="LT4368B_D0084.tif" />

s

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [1- (5-benzyl) -thiazole-2-carbonyl] -4-guanidinobutyl] -amide

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

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrido [1,2-a] -pyrazine-6-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide

5-Oxo-2- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) butyl] amide

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (4-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-ylethyl] -amide

<img file="LT4368B_D0085.tif" />

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-ylethyl] -amide

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1 (1-carbamimidoyl-piperidin-3-yl-ethyl) -2-oxo-2-thiazol-2-ethyl]. amide

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1 (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ethyl] amide

<img file="LT4368B_D0086.tif" />

<img file="LT4368B_D0087.tif" />

<img file="LT4368B_D0088.tif" />

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

<img file="LT4368B_D0089.tif" />

o

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) butyl] amide

<img file="LT4368B_D0090.tif" />

o

3- [4- [4-Guanidino-1- (thiazole-2-carbonyl) butylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-6-ylpropionic acid.

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [3-guanidinopropyl] amide.

<img file="LT4368B_D0091.tif" />

5-Oxo-7- (3-phenylpropionyl) octahydro-2-thia-4a, 7-di-azanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ethyl]. amide

4-Oxo-7- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidine-1- (hydroxy-thiazol-2-ylmethyl) -butyl] -amide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2-ylmethyl) butyl] amide h₂ n ^ nh

<img file="LT4368B_D0092.tif" />

<img file="LT4368B_D0093.tif" />

nh₂

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2-yl] butylamide

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-methoxy (1-thiazole-2-carbonyl) butyl] -amide [6- [4-Methoxy-1 - (Thiol-2-carbonyl) -butylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazin-3-yl] acetic acid [2- [5-Methoxy-2 - ([4-oxo) -2- (3-phenylpropionyl] -octahydropyrrolo [1,2-a] -pyrazine-6-carbonyl] -amino) -pentanoyl) thiazol-5-yl] acetic acid

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

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

<img file="LT4368B_D0094.tif" />

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] -pyrazine-6-carboxylic acid [5-guanidine-1- (thiazole-2-carbonyl) pentyl] -amide

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

4-Oxo-2- (3-phenylpropionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidine-1- (1-methyl-1H-imidazole-2-carbonyl) -butyl] -amide 4-Oxo-2- ( 3-Phenylpropionyl) octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidine-1- (thiazole-2-carbonyl) -butyl] -amide.

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

<img file="LT4368B_D0095.tif" />

nh₂

<img file="LT4368B_D0096.tif" />

nh₂

<img file="LT4368B_D0097.tif" />

nh₂

<img file="LT4368B_D0098.tif" />

nh₂

Suitable compounds of formula VIII are:

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

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

4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2-oxo-2-thiazol-2-ylethylamide;

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

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

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

6- [1- (1-Carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2-thiazol-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) -octahydropyrrolo [1, 2a] pyrazin-3-yl) acetic acid;

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

3- [6- (3-guanidino-propylcarbamoyl) -4-oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1,2-a] pyrazin-3-yl) -acetic acid;

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

[6 - ([1-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] -methyl-carbamoyl) -4-oxo-2- (3-phenyl-propionyl) -octahydro- pyrrolo [1,2-a] pyrazin-3-ylacetic acid;

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

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

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

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

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

2- (2-Benzylidene-pent-3-enoyl] -4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid (3-guanidino-propyl) -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;

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;

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;

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;

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;

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

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

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

0705 N- [1- [4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carbonyl] -5- (thiazole-2-carbonyl) -pyrrolidin-3-yl ] guanidine;

0710 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (4-amino-cyclohexyl) -2-oxo-2-thiazol-2-yl] amide;

0730 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (3-aminomethyl-benzyl) -2-oxo-2-thiazol-2-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-ethylamide;

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

0795 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diaza-naphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-4-ylmethyl) -2-oxo- 2-Thiazol-2-yl-ethyl] -amide;

0800 [4- [4-Guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7-diaza-naphthalen-6-yl ] acetic acid;

0810 3- [4-guanidino-1- (thiazole-2-carbonyl) -butylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7-diaza-naphthalen-6-yl] propionic acid;

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

0820 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diaza-naphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidin-3-ylmethyl) -2-oxo-2- thiazol-2-yl-ethyl] -amide;

0830 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1-thiazol-2-ylmethyl-butyl] -amide;

0835 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 VIII are:

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

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

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

0715 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (4-amino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethyl] amide;

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

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

0735 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid (2-oxo-1-piperidin-4-ylmethyl) -2-thiazol-2-yl -ethyl) amide;

0740 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid (2-oxo-1-piperidin-3-yl-2-thiazol-2-yl-ethyl) amide;

0750 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (4-guanidino-cyclohexylmethyl) -2-oxo-2-thiazol-2-yl-ethylamide;

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

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

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

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

5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diaza-naphthalene-4-carboxylic acid [1- (4-carbamimidoyl-benzyl) -2-oxo-2-thiazole-2- ilethylamid;

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

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

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

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

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-phenylpropionyl) -octahydro-pyrrolo [1,2-a] pyrazin-3-yl] -acetate acid;

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

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

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

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

Preferred compounds of formula VIII are:

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

and

0340 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole); Preferred compounds of formula IX are:

0890 3-Amino-4-oxo-2-phenylhexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidinobutylamide.

0895 3-Amino-2-benzyl-4-oxohexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidinobutylamide

0900 3-Amino-2-cyclohexyl-4-oxo-hexahydro-pyrrolo [2,1-b] [1,3] thiazine-6-carboxylic acid [1 (benzothiazole-carbonyl) -4-guanidinobutyl] -amide: carbonyl) -butyl] -amide ;

<img file="LT4368B_D0099.tif" />

<img file="LT4368B_D0100.tif" />

<img file="LT4368B_D0101.tif" />

nh₂

Preferred compounds of formula X are:

7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [1- (benzothiazole-2-carbonyl) -4-guanidinobutyl] -amide

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

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

7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidino1 - (thiazole-2-carbonyl) -butyl] amide

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

<img file="LT4368B_D0102.tif" />

NHj

<img file="LT4368B_D0103.tif" />

NHj

<img file="LT4368B_D0104.tif" />

<img file="LT4368B_D0105.tif" />

O

<img file="LT4368B_D0106.tif" />

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

<img file="LT4368B_D0107.tif" />

o

LA, amid

0950 7-Benzyl-2,2,6-trioxyoctahydro-pyrido [2,1c] [1,4] thiazine-4-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) -butyl] H

<img file="LT4368B_D0108.tif" />

amide

Preferred compounds of formula X are:

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

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

Preferred compounds of formula III are:

0960 4-Oxo-1- (3-phenylpropionyl) - ° octahydropyrrolo [1,2] pyrimidine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butylamide

<img file="LT4368B_D0109.tif" />

0965 4-Oxo-1- (phenethylsulfonyl) octahydropyrrolo [1,2] pyrimidine-6-carboxylic acid [4-guanidino-1- (thiazole-2-carbonyl) butylamide

Various methods may be used to obtain compounds of formula VII, depending upon the starting materials and / or intermediates formed. One particular method is illustrated in the diagram below:

SCHEME

<img file="LT4368B_D0110.tif" />

stage stage:

Alkylation of compound a is carried out on the appropriate base according to Evans et al. (J. Am. Chem. Soc. 1981, 103, 2127; ibid, 1982, 104, 1737; Aldrichimica Act, 1982, 15, 23), and compound b is obtained.

stage:

Compound b, after hydroboration and oxidation under conditions reported in the literature (Synthesis, 1980,151), yields the aldehyde c.

stage:

The synthesis of adduct e from aldehyde c and compound d is carried out by mixing the reactants in aromatic solvents such as benzene or toluene in the presence of a catalytic amount of a suitable acid such as p-toluenesulphonic acid.

3 'stage and:

Conversion of aldehyde c to aldehyde g is readily accomplished by appropriate blocking-deprotection techniques available from T. Greene, Protective Groups in Organic Synthesis (John Wiley &amp; amp; Sons, 1981).

stage:

The cyclization of the adduct e to compound f can be readily accomplished using appropriate Lewis acids, such as trimethylaluminum, in suitable solvents, such as dichloromethane; the methodology can be found in T. Greene, supra.

Stage 4 ':

Alternatively, compound f may be obtained by treating aldehyde g with compound d in the presence of suitable aromatic solvents, such as benzene.

stage:

The ester group of the bicyclic intermediate of formula f (-C- (O) 0-R) is then hydrolyzed₂o) using a suitable agent such as HCl in a suitable solvent such as ethyl ether to give the free carboxylic acid. The resulting compound is then coupled with R) H using a peptide copolymer such as BOP in a suitable solvent such as DMF to give the bicyclic compound of Formula VII. Suitable peptide bonding conditions are well known in peptide chemistry. For example, see. Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, Nevv York, Tokyo 1984; and The Peptides, Analysis, Synthesis,

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

A variety of methods may be used to provide compounds of Formula VIII, depending upon the starting materials and / or intermediates formed. One particular method is illustrated in the diagram below:

<img file="LT4368B_D0111.tif" />

SCHEME

<img file="LT4368B_D0112.tif" />

<img file="LT4368B_D0113.tif" />

ozonolysis

<img file="LT4368B_D0114.tif" />

stage

<img file="LT4368B_D0115.tif" />

<img file="LT4368B_D0116.tif" />

stage

<img file="LT4368B_D0117.tif" />

where:

Pg is a nitrogen protecting group;

R20 and R21. independently of one another, is Cve-alkyl; and X, R 1, R 3, R 4 and R 8 are as previously described.

The method of obtaining depicted in the scheme is briefly described as follows:

stage:

The unsaturated amino and carboxy groups of the compound of formula a are protected with appropriate protecting groups. A wide variety of protecting groups suitable for protecting reactive functional groups, and suitable procedures for their blocking and deprotection, can be found in T. Greene, Protective Groups in Organic Synthesis (John Wiley &amp; Sons, 1981). The particular protecting group used in a particular synthesis scheme depends on many factors, including the presence of other reactive functional groups and the reaction conditions for its cleavage. The unsaturated compound of formula (a) can be readily obtained using methods and techniques known to one of ordinary skill in the art. The unsaturated blocked compound of formula a is cyclized under appropriate conditions using a suitable reagent such as mercury acetate in an inert solvent such as tetrahydrofuran (THF) to give the protected amino alcohol of formula b.

stage:

The blocked amino alcohol of formula b is oxidized using a suitable oxidizing agent such as sulfur trioxide-pyridine complex in a suitable solvent such as dichloromethane or dimethylformamide to give the blocked amino aldehyde of formula c. Alternatively, intermediate c may be prepared using the compound of formula a 'prepared according to Collado et al., J. Org. Chem., 1995, 60, 5011, ozonolysis,

The blocked aminoaldehyde of formula c is coupled with an alkyl ester of an amino acid of formula d, initially forming an imine which is then treated with an appropriate reagent such as sodium triacetoxyborohydride NaBH (OAc).₃, yields a cyclic intermediate of formula e.

stage:

The cyclic intermediate of formula e introduces an amino function and provides an amino-substituted cyclic intermediate of formula f.

The conditions suitable for such reactions are well known to those skilled in the art and depend on R₅ alternating in nature.

stage:

Under suitable conditions, the amino protecting group of the cyclic intermediate of formula I is removed and the resulting compound is then cyclized under appropriate conditions such as mild heating in an inert solvent to give the crude intermediate of formula g. This bicyclic intermediate of formula g may also be obtained by hydrolysis of the ester group of the cyclic intermediate of formula f (-C (O) 0 -R₂o) to the free carboxylic acid followed by standard peptide coupling in the presence of a suitable copolymer such as benzotriazol-1-yloxitrile (dimethylamino) phosphonium hexafluorophosphate (BOP) in an inert solvent such as dimethylformamide (DMF).

stage

The ester group (-C (O) OR 21) of the bicyclic intermediate intermediate is then hydrolyzed using an appropriate agent such as HCl in a suitable solvent such as ethyl ether to give the free carboxylic acid. The resulting compound is then coupled with R1H using a peptide copolymer such as BOP in a suitable solvent such as DMF to give the bicyclic compound of Formula VIII. Suitable peptide bonding conditions are well known to those skilled in the art of peptide chemistry. For example, see. Principles of Peptide Synthesis, Bodanszky M., SpringerVerlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol.1, edited by Gross E., and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

Various methods may be used to obtain compounds of formula IX, depending upon the starting materials and / or intermediates formed. One particular method is illustrated in the diagram below:

SCHEME

<img file="LT4368B_D0118.tif" />

where:

Pg is a sulfur or amino protecting group;

L is a leaving group;

R20 and R21 independently of one another are Ci.₆-alkyl; o R1, R₃, R₄ and Rs are as previously described.

The method of obtaining depicted in the scheme is briefly described as follows:

stage:

The carboxylic acid of formula a is coupled with a cyclic amino compound b using a peptide copolymer such as benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in a suitable solvent such as dimethylformamide (DMF) or dichloromethane (DCM) to give the amido compound of formula c. Suitable peptide bonding conditions are well known to those skilled in the art of peptide chemistry. For example, see. Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol.1, edited by Gross E., and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

stage:

The internal cyclization of compound c is carried out under suitable conditions to give the bicyclic intermediate of formula d. For example, acid-induced cyclization using p-toluene sulfuric acid or TFA in a suitable solvent such as dichloromethane can be performed.

stage:

Then, the ester group of the bicyclic intermediate of formula d (-C (O) 0 -R₂o) Hydrolyzing with an appropriate agent such as lithium hydroxide (UOH) in a suitable solvent such as tetrahydrofuran (THF) to give the free carboxylic acid. The resulting compound is then coupled with RiH using a peptide copulation agent such as BOP in a suitable solvent such as DMF to give compound e. Suitable peptide bonding conditions are well known to those skilled in the art of peptide chemistry. For example, see. Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol.1, edited by Gross E., and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

A variety of methods may be used to provide compounds of Formula X, depending upon the starting materials and / or intermediates formed. One particular method is illustrated in Scheme 4 below:

SCHEME

OO

<img file="LT4368B_D0119.tif" />

X which

R 20 and R 21 are independently C 1 -C 6 alkyl; o B, R_b R₃, R4 and R5_s are as described previously.

The method of obtaining depicted in the scheme is briefly described as follows:

stage:

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 about -25 to 0 ° C. The resulting mixture is treated under acidic conditions to give a halomethyl ketone of formula b.

stage:

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

stage:

The cyclic intermediate of formula d is treated under acidic conditions using an appropriate acid such as camphor sulfuric acid in a suitable solvent such as toluene to give the bicyclic intermediate of formula e.

stage:

Then the ester group of the bicyclic intermediate of formula e (-C (O) 0 -R₂(o) hydrolyze using a suitable agent such as lithium hydroxide (LiOH) to give the free carboxylic acid. The resulting compound is then coupled with RiH using a peptide copolymer such as BOP in a suitable solvent such as DMF to give a compound of formula X. Suitable peptide coupling conditions are well known to those skilled in the art of peptide chemistry. For example, see. Principles of Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Vol.1, edited by Gross E., and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

The compounds of the present invention are further characterized by their ability to inhibit thrombin catalytic activity as determined by the assay described below. For this test, solutions of the compounds of the present invention are prepared in buffer at concentrations ranging from 1 to 100 mM. To determine the inhibitory dissociation constant K of a particular compound, the thrombus chromogenic or fluorogenic substrate and thrombin must be added to the test compound solution and the resulting catalytic activity of the enzyme determined spectrophotometrically. This type of assay is well known to those skilled in the art.

The compounds of the present invention can be used as anticoagulants in the case of in vitro or ex vivo contact activation with foreign thrombogenic surfaces, such as those found in tubes used in extracorporeal shunts. The compounds of the present invention may also be used to coat the surfaces of such thrombogenic tubes. For this purpose, the compounds of the present invention are prepared as a lyophilized powder, redissolved in isotonic sodium chloride solution, and added in an amount sufficient to maintain the blood in an anticoagulant state.

The therapeutic agents of the present invention may be administered alone or in combination with pharmaceutically acceptable carriers. The proportion of each carrier is determined by the solubility and chemical nature of the compound, the route of administration, and standard pharmaceutical practice. For example, the compounds may be administered parenterally, i.e., intramuscularly, intravenously or subcutaneously. Compounds for parenteral administration should be prepared in the form of sterile solutions containing other dissolved substances, such as sufficient sodium chloride or glucose to make the solution isotonic. The compounds may be administered orally in the form of tablets, capsules or granules containing suitable excipients such as starch, lactose, white sugar and the like. These compounds may also be administered sublingually in the form of tablets or cachets, in which each active ingredient is mixed with sugar or cereal syrups, flavoring agents and dyes and then sufficiently dehydrated to compress the mixture into a solid form. The compounds may be administered orally and in the form of solutions which may contain coloring and / or flavoring agents.

Preferred dosages of therapeutic agents of the present invention will be determined by therapists.

Dosages may vary depending on the route of administration and the particular compound selected. In addition, the doses may be different for the patients being treated.

Generally, when the composition is administered orally, a greater amount of active agent is required to achieve the same effect than parenteral administration.

To further assist in understanding the invention, examples of such thrombin inhibiting compounds are provided below. It is understood that the following examples are not to be construed as limiting the invention; various variants of the present invention which are known or will be apparent to those skilled in the art are intended to be within the scope of the present invention as described herein. The preferred compounds of the present invention were synthesized using conventional preparative steps and isolation techniques known to those skilled in the art of organic and bio-organic synthesis, providing a novel unique combination of the overall synthesis of each compound. The preferred methods for synthesizing the intermediates involved in the synthesis and the anti-thrombotic compounds of the present invention are provided below.

EXAMPLES example

Boc och₃

I ch₃

1. Zn / Cu pair; ultrasound -&gt;

2. [(o-CH₃C₆H₄Py₂PdCl2

4-iodobenzonitrile

H ⁰ '&gt; i ch₃

A solution of tert-butyloxycarbonyl-iodalanine N, O-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) and dry N, N-dimethylacetamide (2.0 ml) was placed in a dry nitrogen purged round-bottom flask containing zinc-copper alloy (0.90 g). The mixture is sonicated, purging with nitrogen until no starting material remains (TLC). Bis (tri-o-tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) was added followed by 4-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added and the mixture was filtered into a separatory funnel. After washing successively with aqueous HCl (50 mL; 0.1 N), distilled H₂O (3 x 50 mL) after drying over Na₂SO₄after filtration and concentration under reduced pressure, the crude product is obtained. Express chromatography on silica gel (light petroleum ether / ethyl acetate gradient) affords the purified product.

Boc

<img file="LT4368B_D0120.tif" />

2. [(o-CH₃C₆H₄Py₂PdCl₂

I CH.

3-iodine nzonitrile

H

O

1-Zn / Cu pair; ultrasound

<img file="LT4368B_D0121.tif" />

OCH₃

A solution of tert-butyloxycarbonyl-iodalanine-NO-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) and dry N, N-dimethylacetamide (2). (0 ml) was poured into a dry nitrogen purged round bottom flask containing zinc-copper alloy (0.90 g). The mixture is sonicated, purging with nitrogen until no starting material remains (TLC). Bis (tri-o-tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) was added followed by 3-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added, and the mixture was filtered into a separatory funnel. After washing successively with aqueous HCl (50 mL; 0.1 N), distilled H₂O (3 x 50 mL) after drying over Na₂SO₄after filtration and concentration under reduced pressure, the crude product is obtained. Purification by silica gel chromatography (gradient of light petroleum ether and ethyl acetate) affords the purified product.

Boc o

IEA

OCH

<img file="LT4368B_D0122.tif" />

A solution of tert-butyloxycarbonyl-iodalanine-N, O-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) and dry N, N-dimethylacetamide (2.0 ml) was placed in a dry nitrogen purged round-bottom flask containing zinc-copper alloy (0.90 g). The mixture is sonicated, purging with nitrogen until no starting material remains (TLC). Bis (tri-o-tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added, and the mixture was filtered into a separatory funnel. After washing successively with aqueous HCl (50 mL; 0.1 N), distilled H₂O (3 x 50 mL) after drying over Na₂Filtration and concentration under reduced pressure of SO4 yields a crude product. Purification by silica gel chromatography (gradient of light petroleum ether and ethyl acetate) affords the purified product.

<img file="LT4368B_D0123.tif" />

To a solution of tert-butyloxycarbonyl-para-cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxylamine hydrochloride (0.416 g, 6.0 mmol) and diisopropylethylamine (1.02 g). ml, 6.0 mmol). The mixture is refluxed and then cooled. The precipitate is filtered off, washed with cold ethanol, diisopropyl ether and the solution is dried over MgSO4.₄, is concentrated under reduced pressure and used in the downstream stage. The semi-solid is suspended in a mixture of acetic acid (20 mL) and dry ethanol (40 mL) under heating. Pd / C catalyst (0.30 g, 10% Pd) is then added and hydrogen is bubbled through the mixture while heating. The hydrogenation is continued until the starting material is no longer detectable by TLC. The catalyst is filtered off, the solution is concentrated under reduced pressure (50 mL), HCl (50 mL, 1N) is added and the mixture is again concentrated to 50 mL. This solution is cooled overnight to give the desired compound.

<img file="LT4368B_D0124.tif" />

c H, To a solution of tert-butyloxycarbonyl-methacyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxylamine hydrochloride (0.416 g, 6.0 mmol) and diisopropylethylamine ( 1.02 mL, 6.0 mmol). The mixture is refluxed and then cooled. The precipitate is filtered off, washed with cold ethanol, diisopropyl ether and the solution is dried over MgSO4.₄, is concentrated under reduced pressure and used in the downstream stage. The semi-solid is suspended in a mixture of acetic acid (20 mL) and dry ethanol (40 mL) under heating. Pd / C catalyst (0.30 g, 10% Pd) was then added and hydrogen was bubbled through the mixture with heating. The hydrogenation is continued until the starting material is no longer detectable by TLC. The catalyst is filtered off, the solution is concentrated under reduced pressure (50 mL), HCl (50 mL, 1N) is added and the mixture is again concentrated to 50 mL. This solution is cooled overnight to give the desired compound.

Boc

<img file="LT4368B_D0125.tif" />

/ OCH₃

1. NH₂OH, DIEA / EtOI ^

2. H2, Pd / C / EtOH; HOAc

<img file="LT4368B_D0126.tif" />

To a solution of tert-butyloxycarbonyl-ortho-cyanophenylalanine-NO-dimethylamide (1.33 g, 4.0 mmol) in dry ethanol (20 mL) was added hydroxylamine hydrochloride (0.416 g, 6.0 mmol) and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture is refluxed and then cooled. The precipitate is filtered off, washed with cold ethanol, diisopropyl ether and the solution is dried over MgSO4.₄, is concentrated under reduced pressure and used in the downstream stage. The semi-solid is suspended in a mixture of acetic acid (20 mL) and dry ethanol (40 mL) under heating. Pd / C catalyst (0.30 g, 10% Pd) was then added and hydrogen was bubbled through the mixture with heating. The hydrogenation is continued until the starting material is no longer detectable by TLC. The catalyst is filtered off, the solution is concentrated under reduced pressure (50 mL), HCl (50 mL, 1N) is added and the mixture is again concentrated to 50 mL. This solution is cooled overnight to give the desired compound.

<img file="LT4368B_D0127.tif" />

A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added dropwise to n-BuLi (1.6 m / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution stirring. A solution of tert-butyloxycarbonyl-para-amidinophenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and evaporation under reduced pressure.

<img file="LT4368B_D0128.tif" />

A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added dropwise to n-BuLi (1.6 m / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution stirring. A solution of tert-butyloxycarbonyl-methamidophenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and evaporation under reduced pressure.

<img file="LT4368B_D0129.tif" />

A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added dropwise to n-BuLi (1.6 m / hexane, 8.9 mL, 13.9 mmol) at -78 ° C, and the solution stirring. A solution of tert-butyloxycarbonyl-ortho-amidinophenylalanine-N, O-dimethylamide (1.15 g, 3.3 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and evaporation under reduced pressure.

NC

Boc

<img file="LT4368B_D0130.tif" />

H

<img file="LT4368B_D0131.tif" />

/ OCH₃ p

ch₃

Tert-Butyloxycarbonyl-para-cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL) and porous Renee nickel (100 mg) was added. Shake the solution with H₂ (pressure 40 psi) at room temperature. The solution is filtered through zeolite and concentrated under reduced pressure to give a clear residue. This residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL) and concentrated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure.

<img file="LT4368B_D0132.tif" />

Tert-Butyloxycarbonyl-methacyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL) and porous Renee nickel (100 mg) was added. Shake the solution with H₂ (pressure 40 psi) at room temperature. The solution is filtered through zeolite and concentrated under reduced pressure to give a clear residue. This residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL) and concentrated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure.

<img file="LT4368B_D0133.tif" />

Tert-Butyloxycarbonyl-ortho-cyanophenylalanine-N, O-dimethylamide (1.33 g, 4.0 mmol) was dissolved in ethanol saturated with ammonia (30 mL) and porous Renee nickel (100 mg) was added. Shake the solution with H₂ (pressure 40 psi) at room temperature. The solution is filtered through zeolite and concentrated under reduced pressure to give a clear residue. This residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL) and concentrated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure.

<img file="LT4368B_D0134.tif" />

NZ

Tert-butyloxycarbonyl-para-aminomethylphenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled and N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl were added.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the pure compound.

<img file="LT4368B_D0135.tif" />

<img file="LT4368B_D0136.tif" />

ch₃

<img file="LT4368B_D0137.tif" />

oh₃

Tert-butyloxycarbonyl-meta-aminomethylphenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution is cooled and N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl are added.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the pure compound.

<img file="LT4368B_D0138.tif" />

ZN

Tert-Butyloxycarbonyl-ortho-aminomethylphenylalanine-N, O-dimethylamide (1.00 g, 3.1 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution is cooled and N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl are added.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. After chromatography on silica gel

<img file="LT4368B_D0139.tif" />

A solution of NZ thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) was added dropwise at -78 ° C to n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol), and the solution is stirred. A solution of the blocked amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and concentration under reduced pressure.

<img file="LT4368B_D0140.tif" />

<img file="LT4368B_D0141.tif" />

. A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol), and stirring. A solution of the blocked amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and concentration under reduced pressure.

<img file="LT4368B_D0142.tif" />

A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol), and stirring. A solution of the blocked amino acid (1.36 g, 3.3 mmol) in THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and concentration under reduced pressure.

OCH,

Boc ^

N i

CH / OCH₃

1. Ζη / Cu pair, ultrasound

2. [(o-CH₃C_eH₄)₃P]₂PdCI₂2-amino-5-bromopyridine

Boc ^

<img file="LT4368B_D0143.tif" />

I

CH,

A solution of tert-butyloxycarbonyl-iodalanine-NO-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) and dry N, N-dimethylacetamide (2). (0 ml) was poured into a dry nitrogen purged round bottom flask containing zinc-copper alloy (0.90 g). The mixture is sonicated, purging with nitrogen until no starting material remains (TLC). Bis (tri-o-tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) was added followed by 2-amino-5-bromopyridine (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added and the mixture was filtered into a separatory funnel. After washing successively with aqueous HCl (50 mL; 0.1 N), distilled H₂O (3 x 50 mL) after drying over Na₂SO₄after filtration and concentration under reduced pressure, the crude product is obtained. Purification by silica gel chromatography (gradient of light petroleum ether and ethyl acetate) affords the purified product.

<img file="LT4368B_D0144.tif" />

<img file="LT4368B_D0145.tif" />

A solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol), and the solution stirring. A solution of the amino acid N, O-dimethylamide (1.07 g, 3.3 mmol) in anhydrous THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4._4l filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

H

Boc-f '

N l

CH, .OCH,

1. Zn / Cu pair, ultrasound

2. [(o-CH₃C₆H₄)₃P]₂PdCr₂2-cyano-5-bromopyridine

NC

O

<img file="LT4368B_D0146.tif" />

A solution of tert-butyloxycarbonyl-iodalanine-NO-dimethylamide (2.68 g, 7.5 mmol) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 mL) and dry Ν, Ν-dimethylacetamide (2 (0 ml) was placed in a dry nitrogen purged round bottom flask containing zinc-copper alloy (0.90 g). The mixture is sonicated under nitrogen until no starting material is present (by TLC), bis (tri-o-tolylphosphine) palladium dichloride (0.35 g, 0.40 mmol) is added followed by 2-cyano-5-bromopyridine (1.72 g, 7.5 mmol). The resulting mixture was stirred under nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added, and the mixture was filtered into a separatory funnel. After washing successively with aqueous HCl (50 mL; 0.1 N), distilled H₂O (3 x 50 mL) after drying over Na₂SO₄after filtration and concentration under reduced pressure, the crude product is obtained. Purification by silica gel chromatography (gradient of light petroleum ether and ethyl acetate) affords the purified product.

<img file="LT4368B_D0147.tif" />

To a solution of tert-butyloxycarbonyl- (4-cyano) 3-pyridylalanine-N, O-dimethylamide (1.34 g, 4.0 mmol) in dry ethanol (20 mL) was added N, hydroxylamine hydrochloride (0.416 g, 6.0 mmol) ) and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture is refluxed and then cooled. The precipitate is filtered off, washed with cold ethanol, diisopropyl ether and the solution is dried over MgSO4.₄, is concentrated under reduced pressure and the resulting material is used in the next step. The semi-solid is suspended in a mixture of acetic acid (20 mL) and dry ethanol (40 mL) under heating. Pd / C catalyst (0.30 g, 10% Pd) is then added and hydrogen is bubbled through the mixture while heating. The hydrogenation is continued until the starting material is no longer detectable by TLC. The catalyst is filtered off, the solution is concentrated under reduced pressure (50 mL), HCl (50 mL, 1N) is added and the mixture is again concentrated to 50 mL. This solution is cooled overnight to give the desired compound.

<img file="LT4368B_D0148.tif" />

n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) was added dropwise to a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL) at -78 ° C, and the solution stirring. A solution of the amino acid N, O-dimethylamide (1.16 g, 3.3 mmol) in anhydrous THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and concentrated under reduced pressure.

<img file="LT4368B_D0149.tif" />

Tert-Butyloxycarbonyl-3- (4-pyridyl) alanine-N, O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL) and PtO was added.₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. Filtration of the solution through zeolite and concentration under reduced pressure afforded tert-butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide. The residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), concentrated sodium chloride solution (2 x 50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure to give the desired product.

o ^ / OCHa -i _{H? |} ptoyĄcOH I

<img file="LT4368B_D0150.tif" />

oh₃

BoA

<img file="LT4368B_D0151.tif" />

Tert-Butyloxycarbonyl-3- (3-pyridyl) alanine-N, O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL) and PtO was added.₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. Filtration of the solution through zeolite and concentration under reduced pressure afforded tert-butyloxycarbonyl-3- (3-piperidyl) alanine-N, O-dimethylamide. The residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), concentrated sodium chloride solution (2 x 50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure to give the desired product.

<img file="LT4368B_D0152.tif" />

Tert-Butyloxycarbonyl-3- (2-pyridyl) alanine-N, O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL) and PtO was added.₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. Filtration of the solution through zeolite and concentration under reduced pressure afforded tert-butyloxycarbonyl-3- (2-piperidyl) alanine-N, O-dimethylamide. The residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), concentrated sodium chloride solution (2 x 50 mL), dried over MgSO 4₄, filtered and concentrated under reduced pressure to give the desired product.

<img file="LT4368B_D0153.tif" />

Tert-Butyloxycarbonyl-3- (4-piperidyl) alanine-N, O-dimethylamide (1.00 g,

3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled and added N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

<img file="LT4368B_D0154.tif" />

<img file="LT4368B_D0155.tif" />

Tert-Utiloxycarbonyl-3- (3-piperidyl) alanine-N, O-dimethylamide (1.00 g,

3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled and N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl were added.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

<img file="LT4368B_D0156.tif" />

<img file="LT4368B_D0157.tif" />

Tert-Butyloxycarbonyl-3- (2-piperidyl) alanine-N, O-dimethylamide (1.00 g,

3.2 mmol) was dissolved in dry THF (10 mL) under nitrogen with stirring. The solution was cooled and N, N'-bis (benzyloxycarbonyl) -methylisothiourea (1.14 g, 3.2 mmol) and HgCl were added.₂ (0.95 g, 3.5 mmol). The solution is concentrated under reduced pressure, the remaining residue is suspended in ethyl acetate (200 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

<img file="LT4368B_D0158.tif" />

NZ

To a solution of thiazole (1.23 g, 14.4 mmol) in anhydrous THF at -78 ° C was added dropwise n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) and the solution was stirred. A solution of guanidylated 4-piperidylalanine derivative (2.00 g, 3.2 mmol) in anhydrous THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure.

<img file="LT4368B_D0159.tif" />

A solution of thiazole (1.23 g, 14.4 mmol) in anhydrous THF at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) and the solution was stirred. A solution of guanidylated 3-piperidylalanine derivative (2.00 g, 3.2 mmol) in anhydrous THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4._4l filtered and concentrated under reduced pressure.

H i?

I och₃

K ^ nhz ZN

Li-thiazole / THR

<img file="LT4368B_D0160.tif" />

Boc

<img file="LT4368B_D0161.tif" />

A solution of thiazole (1.23 g, 14.4 mmol) in anhydrous THF at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) and the solution was stirred. A solution of guanidylated 2-piperidylalanine derivative (2.00 g, 3.2 mmol) in anhydrous THF (15 mL) was then added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure.

<img file="LT4368B_D0162.tif" />

Tert-Butyloxycarbonyl-para-nitrophenylalanine-N, O-dimethylamide (13.88 g,

39.3 mmol) was dissolved in acetic acid (100 mL) and PtO was added₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. The solution is filtered through zeolite, concentrated under reduced pressure, and the resulting material is dissolved in H₂O (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL) and concentrated sodium chloride solution (3 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure to give the title compound.

<img file="LT4368B_D0163.tif" />

<img file="LT4368B_D0164.tif" />

Tert-Butyloxycarbonyl-meta-nitrophenylalanine-N, O-dimethylamide- (13.88 g,

39.3 mmol) was dissolved in acetic acid (100 mL) and PtO was added₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. The solution is filtered through zeolite, concentrated under reduced pressure, and the resulting material is dissolved in H₂O (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL) and concentrated sodium chloride solution (3 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure to give the title compound.

<img file="LT4368B_D0165.tif" />

Tert-Butyloxycarbonyl-ortho-nitrophenylalanine-N, O-dimethylamide (13.88 g,

39.3 mmol) was dissolved in acetic acid (100 mL) and PtO was added₂ (100 mg). Shake the solution with H₂ in the atmosphere until gas absorption stops. The solution is filtered through zeolite, concentrated under reduced pressure, and the resulting material is dissolved in H₂O (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL) and concentrated sodium chloride solution (3 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure to give the title compound.

<img file="LT4368B_D0166.tif" />

1. Tert-Butyloxycarbonyl-3- (cis / trans -4-aminocyclohexyl) alanine N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium carbonate and THF with stirring [60 mL, (1: 1)]. . After cooling the solution, a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) is added dropwise. Excess solid sodium carbonate was added, THF was evaporated under reduced pressure, the remaining aqueous phase was poured into ethyl acetate (250 mL) and mixed well. The aqueous phase is separated and the remaining solution is washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4N aqueous sodium bisulfate (2 x 50 mL) and saturated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4_4lfiltered and concentrated under reduced pressure. The semi-solid is chromatographed on silica gel (ethyl acetate / hexane).

2. | A solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) and the solution was stirred. A solution of the above blocked amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is chromatographed on silica gel (ethyl acetate / hexane) and the solution is evaporated under reduced pressure.

<img file="LT4368B_D0167.tif" />

<img file="LT4368B_D0168.tif" />

1. Tert-Butyloxycarbonyl-3- (cis / trans-3-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium carbonate and THF with stirring [60 mL, (1: 1)]. . After cooling, a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium carbonate was added, THF was evaporated under reduced pressure, the remaining aqueous phase was poured into ethyl acetate (250 mL) and mixed well. The aqueous phase is separated and the remaining solution is washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4N aqueous sodium bisulfate (2 x 50 mL) and saturated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure. The semi-solid is chromatographed over silica gel (ethyl acetate / hexane).

2. A solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) and the solution was stirred. A solution of the above blocked amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is chromatographed on silica gel (ethyl acetate / hexane) and the solution is evaporated under reduced pressure.

<img file="LT4368B_D0169.tif" />

OCH,

1. Z-Cl, NaHCO₃/ THF: HjQ ^

2. L i -thiazole / THF

<img file="LT4368B_D0170.tif" />

1. Tert-Butyloxycarbonyl-3- (cis / trans-2-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium carbonate and THF with stirring [60 mL, (1: 1)]. . After cooling, a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Excess solid sodium carbonate was added, THF was evaporated under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL) and mixed well. The aqueous phase is separated and the remaining solution is washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4N aqueous sodium bisulfate (2 x 50 mL) and saturated sodium chloride solution (2 x 50 mL). The solution was dried over MgSO4₄, filtered and concentrated under reduced pressure. The semi-solid is chromatographed on silica gel (ethyl acetate / hexane).

2. A solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF at -73 ° C was added dropwise to n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) and the solution was stirred. A solution of the above blocked amino acid amide (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is chromatographed on silica gel (ethyl acetate / hexane) and the solution is evaporated under reduced pressure.

<img file="LT4368B_D0171.tif" />

<img file="LT4368B_D0172.tif" />

1. Tert-Butyloxycarbonyl-3- (cis / trans-4-aminocyclohexyl) alanine-N, Odimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring. The solution was cooled to 0 ° C and N, N'bis (benzyloxycarbonyl) -S-methylisothiourea (2.18 g, 6.1 mmol) and HgCl were added.₂(1.81 g, 6.7 mmol). The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

2. n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) was added dropwise to a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF at -78 ° C and the solution was stirred. A solution of the above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel. (ethyl acetate / hexane) and the solution was concentrated under reduced pressure.

<img file="LT4368B_D0173.tif" />

Ch₃N ³

I och₃

NZ

X

1. ch₃s nhz

HgClj / THF 2. Li-thiazole / THF

<img file="LT4368B_D0174.tif" />

ZHN

1. Tert-Butyloxycarbonyl-3- (cis / trans-3-aminocyclohexyl) alanine-N, O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring. The solution was cooled to 0 ° C and N, N'bis (benzyloxycarbonyl) -S-methylisothiourea (2.18 g, 6.1 mmol) and HgCl were added.₂(1.81 g, 6.7 mmol). The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

2. A solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF at -78 ° C was added dropwise to n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) and the solution was stirred. A solution of the above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and the solution is concentrated under reduced pressure.

<img file="LT4368B_D0175.tif" />

1. Tert-butyloxycarbonyl-3- (cis / trans-2-aminocyclohexyl) alanine-N, O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring. The solution was cooled to 0 ° C and N, N'bis (benzyloxycarbonyl) -S-methylisothiourea (2.18 g, 6.1 mmol) and HgCl were added.₂(1.81 g, 6.7 mmol). The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL) and filtered through zeolite. The filtrate is concentrated under reduced pressure. Express chromatography on silica gel (hexane / ethyl acetate gradient) gives the title compound.

2. n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) was added dropwise to a solution of thiazole (2.32 g, 27.3 mmol) in anhydrous THF at -78 ° C and the solution was stirred. A solution of the above guanidylated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction is quenched with a saturated aqueous solution of ammonium chloride. 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), saturated sodium chloride solution (50 mL), dried over MgSO4.₄, filtered and concentrated under reduced pressure. The crude material is purified by chromatography on silica gel (ethyl acetate / hexane) and the solution is concentrated under reduced pressure.

<img file="LT4368B_D0176.tif" />

example

Synthesis of intermediates

<img file="LT4368B_D0177.tif" />

(4S, 5R) -3- (1-Oxo-3-phenylpropyl) -4- (phenyl) -5- (methyl) -2-oxazolidone (2). into a solution of 10.0 g (1 eq, 56.4 mmol) of (4S, 5R) -4-phenyl-5-methyl-2-oxazolidone (1) in 250 mL of dry THF stirred at -78 ° C under argon, n-butyl lithium (1.6M in hexane, 1.1 eq., 38.8 mL) was added dropwise. After stirring for 30 min, 8.4 ml (1.0 eq, 56.4 mmol) of hydrocinnamoyl chloride are added dropwise over 10 minutes. The resulting mixture was warmed to 0 ° C, stirred for an additional hour and quenched with a saturated ammonium chloride solution. The solvent was evaporated in vacuo and the resulting white solid was dissolved in ethyl acetate and ddH.₂O. The aqueous phase is separated off and extracted with two further portions of ethyl acetate. The extracts are combined, washed with a saturated sodium chloride solution, dried over sodium sulfate and evaporated in vacuo to give a white solid as a white solid (2). Yield 91%.

Lyd. temp .: 95-96.6; [α] D = -35.8 (c = 1, CH₂CI₂); [a]_D = -26.6 (c = 1.018 CH₂CI₂);

¹1 H-NMR (CDCl 3) d 0.89 (d, 3H, CH₃, J = 6.6 Hz), 3.00-3.05 (m, 2H), 3.26-3.34 (m, 2H) 4.73-4.78 (m, 1H), 5.64 (d, 1H, J = 7.4 Hz), 7.22-7.46 (m,

10H).

<img file="LT4368B_D0178.tif" />

<img file="LT4368B_D0179.tif" />

A solution of 5.0 g (1.00 eq., 16.2 mmol) of (2) in 100 mL of dry THF is cooled to -78 ° C. Enolization is carried out with 17.8 ml (1.1 eq, 17.8 mmol) of lithium bistrimethylsilylamide administered by syringe. The solution is stirred for 30 minutes, after which 4.45 ml (3.0 eq., 48.5 mmol) of oleic iodide are added and the mixture is warmed to -15 ° C. After 1 hour, quench the reaction with saturated ammonium chloride solution and extract with ethyl acetate (3x). The organic phase is washed with sodium metabisulphite, dried over sodium sulphate and evaporated in vacuo to give a colorless oil. Purify by silica gel flash chromatography using a step gradient (15: 1, 12: 1, 10: 1) to give a colorless oil (95%).

[a]_D = 47.5 (c = 3.12, CH₂CI₂);

¹1 H-NMR (CDCl 3) δ 0.82 (d, 3H, CH 8, J = 6.6 Hz), 2.31-2.40 (m, 1H, RCH = CHCH₂), 2.49-2.57 (m, 1H, RCH = CHCH)₂), 2.84-3.00 (m, 2H, Ph-CHa), 4.32-4.37 (m, 1H, CH- (N) CO), 4.53-4.58 (m, 1H , CH 3 -CH-), 5.03-5.13 (m, 2H,

ΑΒΧ, CH = CH₂), 5.21 (d, 1H, Ph-CH, J = 7.1 Hz), 5.81-5.89 (m, 1H, CH = CH)₂), 7.20-7.42 (m, 10H, ArH);

¹³C-NMR (CDCl 3) δ 14.4, 36.2, 38.2, 43.9, 54.7, 78.7, 117.1, 125.4, 126.3, 128.2, 128.5 , 129.0, 133.1, 134.8, 138.9, 152.4, 174.9.

<img file="LT4368B_D0180.tif" />

A sample of the allyl compound (3) (4.75 g, 13.6 mmol) in THF (100 mL) was treated with 13.6 mL (1.0 eq., 13.6 mmol) of 1.0M borohydrofuran complex, 0 ° With stirring at C for 2 hours. Evaporate the solvent and inject chloroform (100 ml) using a syringe. The oxidation of the organo-boron compound was accomplished by adding 4.7 g (2.0 eq, 27.8 mmol) of 3-chloroperoxybenzoic acid at 0 ° C, warming to room temperature and stirring for another hour. The organic phase is washed with 5% Na₂CO₃, ddH₂Oh and dried over sodium sulfate. Due to the volatility of the alcohol, rapid column chromatography is carried out to remove additional polar and non-polar substances arising from 3-chloroperoxybenzoic acid. The alcohol (4) is obtained in a yield of 65%.

[a]_D = 39.3 (c = 1.038, CH₂CI₂);

¹1 H-NMR (CDCl 3) δ 1.07 (d, 3H, .delta., J = 6.5 Hz), 1.81-1.93 (m, 3H, CH₂-CH-H), 2.10-2.19 (m, 1H, CH)₂-CH-H), 3.10-3.17 (m, 2H, Ph-CH5), 3.87-3.90 (m, 2H, CH5-OH), 4.43-4.49 (m, 1H, CH-CO), 4.70-4.75 (m, 1H,

CHα-CH), 5.36 (d, 1H, Ph-CLC ± J = 7.1 Hz), 7.41-7.63 (m, 10H, ArH).

<img file="LT4368B_D0181.tif" />

O

O

O

<img file="LT4368B_D0182.tif" />

876 mg (1.5 eq., 4.1 mmol) of pyridine chlorochromate and 1.0 g of 4 angstroms were added; the mixture changes color from bright orange to j black. The reaction is monitored by TLC and after 30 min, if the starting material is still present, molecular mesh is added again. The solution is filtered through zeolite and the solvent is evaporated. The residue is dissolved in ethyl acetate and washed with a saturated sodium chloride solution. If the organic phase still has an orange color, it is filtered again through a layer of zeolite. Quantitatively yielded aldehyde (5) which is a clear colorless oil.

¹1 H-NMR (CDCl 3) δ 0.84 (d, 3H, CH 2 -CH, J = 6.6 Hz), 1.87-1.94 (m, 1H, CH₂-CH (H) -CHO), 2.04-2.13 (m, 1H, CH)₂-CH (H) -CHO), 2.45-2.50 (m, 2H, PhCH₂), 2.79-2.85 (dd, 1H, CH2-CHO, J = 13.3 &amp; amp; J = 6.6), 2.92-2.99 (dd, 1H, CH2-CHO, J = 13 , 2 &amp; amp; J = 8.8), 4.19-4.22 (m, 1H, CH-CO), 4.46-4.51 (m, 1H, CH₃-CH), 5.13-5.25 (m, 1H, Ph-CH), 7.20-7.39 (m, 10H, ArH), 9.69 (s, 1H, CHO);

¹³C-NMR (CDCl 3) δ 14.2, 23.8, 39.0, 41.2, 43.8, 54.9, 78.6, 125.3, 126.4, 128.2, 128.4 , 128.5, 128.9, 132.8, 138.4, 152.4, 174.9, 201.1.

<img file="LT4368B_D0183.tif" />

<img file="LT4368B_D0184.tif" />

The aldehyde (5) (2.6 g, 7.10 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 mmol) of L- of cysteine ​​ethyl ester and 4 angstroms molecular networks. The reaction mixture was stirred overnight at room temperature and then the solvent was evaporated in vacuo. The residue is dissolved in chloroform, washed with saturated sodium chloride solution, double-distilled water and dried over sodium sulfate. Evaporation of the solvent in vacuo gives a sticky solid (6).

<img file="LT4368B_D0185.tif" />

<img file="LT4368B_D0186.tif" />

In 2.0 M triethylaluminium hexane (2.4 mL, 4.8 mmol, 3 equiv.), Slowly add the starting material (6) (800 mg, 1.61 mmol) in anhydrous dichloromethane under argon atmosphere using a furnace. After stirring overnight, HPLC indicates that the reaction is complete. The mixture was partitioned between excess methanol and then filtered through a short silica gel column (washed with 10% excess methanol in ethyl acetate). Evaporation gives 784 mg of crude material which is purified by silica gel chromatography using 2: 1 hexane: EtOAc to give 258 mg (0.81 mmol, 50% yield) of pure compound (7) - 6S-benzylhexahydro-5-oxo -5H-thiazolo [3,2-a] pyridine-3R-ethyl ester which is a yellowish solid.

¹1 H-NMR (CDCl 3) δ 1.28-1.31 (m, 3H), 1.72-1.81 (m, 3H,), 2.10-2.13 (m,

1H), 2.66 (dd, 1H, J = 11.5 and 6.0 Hz), 3.29-3.34 (m, 2H), 4.19-4.29 (m, 2H),

4.88 (dd, 1H, J = 9.0 and 5.0 Hz), 5.22 (dd, 1H, J = 8.0 and 6.0 Hz), 7.18-7.23 (m,

3H), 7.28-7.31 (m, 2H).

<img file="LT4368B_D0187.tif" />

<img file="LT4368B_D0188.tif" />

to the starting material (7) (240 mg, 0.76 mmol) dissolved in 10 mL of dioxane was added LiOH.H₂O (48 mg, 1.12 mmol) in 10 mL of water. After 1 or. TLC (1: 1 Hexane: EtAc) does not show the starting material. The reaction was quenched with 10% citric acid, then extracted twice with dichloromethane. Drying and evaporation of the combined organic layers gives 354 mg of crude product; This product is dissolved in dichloromethane and precipitated by the addition of excess hexane. The product is filtered off to give 200 mg (0.68 mmol, 90% yield) of an off-white material (8) which is 6-benzylhexahydro-5-oxo-5H-thiazole [3,2-a] pyridine-3R-carboxylic acid.

¹1 H-NMR (CD₃OD) δ 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).

<img file="LT4368B_D0189.tif" />

<img file="LT4368B_D0190.tif" />

no

The oxalyl chloride (9) (25 g, 0.197 mol) was cooled to 0 ° C and cyclohexanepropionic acid (20 mL, 0.14 mol) was added. Stir overnight. The resulting mixture is distilled to give a colorless liquid (10) cyclohexylpropionic acid chloride (84% yield).

O

X

HN O

<img file="LT4368B_D0191.tif" />

Ph h₃c

The chiral auxiliary (11) (13.6 g, 76.7 mmol, 1 equiv) was dissolved in dry THF and cooled to -78 ° C. N-BuLi (52.8 mL, 84.4 mmol, 1.2 eq.) Was then added and left for 30 min. (dark orange solution). Acid chloride (10) (13.4 g, 76.6 mmol, 1 equiv.) Was then added and stirred overnight. Treatment with saturated NH₄CI, extracting with ethyl acetate, washing the extracts with water and brine, drying over sodium sulfate, and concentrating. The product was purified using a fast flowing, dry packed column (6: 1 hexane: ethyl acetate). A white solid (12) is obtained which is recrystallized from ether and hexane. The desired compound is obtained in 78% yield.

Lyd. 90.5-91.5 ° C; [a]_D = -20.1 (c = 1, EtOH);

¹1 H-NMR (CDCl 3)₃δ 0.86-1.10 (m, 5H), 1.18-1.30 (m, 4H,), 1.54-1.75 (m, 7H), 2.86-2.97 (m, 2H), 4.70-4.76 (m, 1H), 5.65 (d, 1H, J = 7.2Hz), 7.28-7.42 (m, 5H).

O

<img file="LT4368B_D0192.tif" />

Me

<img file="LT4368B_D0193.tif" />

The starting material (12) (9.13 g, 29 mmol, 1 equiv) was dissolved in dry THF and cooled to -78 ° C, followed by 40 min. LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise. After 30 min, allyl bromide (7.5 mL, 86.9 mmol, 3 equiv) was added slowly over 10 min. The mixture is left to warm overnight. Work-up includes quenching with saturated ammonium chloride solution, extraction with ethyl acetate, washing with 10% sodium thiosulfate, decolorization with activated carbon, drying over sodium sulfate and evaporation in vacuo. Yield: 96% (yellow oil).

[a]_D = +9.5 (c = 1, EtOH);

¹1 H-NMR (CDCl 3) δ 0.92-1.10 (m, 5H), 1.10-1.39 (m, 5H), 1.63-1.75 (m, 6H), 2.27 -2.42 (m, 2H), 4.01-4.14 (m, 1H), 4.76-4.85 (m, 1H), 5.00-5.07 (m, 2H), δ , 65 (d, 1H, J = 7Hz), 5.64-5.88 (m, 1H), 7.27-7.46 (m, 5H).

<img file="LT4368B_D0194.tif" />

<img file="LT4368B_D0195.tif" />

o

<img file="LT4368B_D0196.tif" />

o

Ph

2-Methyl-2-butene is added dropwise to the boron-dimethylsulfide complex at -12 ° C. The reaction mixture was maintained at this temperature for 15 minutes, then warmed to 0 ° C and stirred for 2 hours. Dissamylborane is then added to the solution of the starting material (13) in THF using a double-ended needle at 0 ° C. The mixture is stirred for 2 hours, after which the solvents are evaporated and the residue is dissolved in dichloromethane. Carefully place the residue in a suspension of pyridine chlorochromate in dichloromethane in a refluxed flask. When the initial exothermic reaction subsides, the mixture is refluxed at 50 ° C for 1 hour. The dark brown liquid was dissolved in ethyl acetate and filtered through Florisil. The black PCC residue was extracted with ethyl acetate and filtered again through the same Florisil layer. Evaporation of the filtrates gives a yellow sticky product (14) with a yield of 78%.

[a]_D = -17.8 (O = 1.245, EtOH);

¹1 H-NMR (CDCl 3)₃? d 0.89-1.18 (m, 5H), 1.20-1.47 (m, 8H), 1.60-1.74 (m,

6H), 1.83-2.00 (m, 1H), 2.48-2.53 (m, 2H), 3.90-4.10 (m, 1H), 4.12-4.16 (m,

1H), 4.76-4.80 (m, 1H), 5.67 (d, 1H, J = 7Hz), 7.27-7.46 (m, 5H), 9.77 (s, 1H ).

<img file="LT4368B_D0197.tif" />

<img file="LT4368B_D0198.tif" />

The resulting aldehyde (14) (7.7 g of crude, 20.8 mmol, 1 equiv) is dissolved in 75 mL of toluene. | to this solution was added a catalytic amount of p-toluenesulfonic acid (50 mg), 10 g of 4 angstroms molecular grids and L-cysteine ​​ethyl ester (3.78 g, 20.8 mmol, 1 eq). The mixture was stirred overnight at room temperature, filtered and concentrated. The residue was chromatographed on silica gel (6: 1 hexane: ethyl acetate) to give 6.36 g of product (15) in 61% yield.

[a]₀ = -48.3 (c = 1.095, EtOH);

¹1 H-NMR (CDCl 3) δ 0.84-0.98 (m, 4H), 1.11-1.38 (m, 7H), 1.50-1.90 (m, 10H), 2.80- 2.99 (m, 1H), 3.24-3.34 (m, 1H), 3.77-4.29 (m, 4H), 4.46-4.81 (m,

7.27-7.46 (m, 5H).

2H), 5.66 (d, 1H, J = 7Hz)

<img file="LT4368B_D0199.tif" />

Me, Al / DCM ---&gt;

0 ° C RT

<img file="LT4368B_D0200.tif" />

The starting material (15) (1.97, 3.9 mmol, 1 equiv.) Is dissolved in 20 mL of dry dichloromethane and cooled to 0 ° C. Trimethylaluminium (5.9 mL, 11.8 mmol, 3 equiv.) Was added dropwise and the mixture was stirred overnight. Upon completion of the reaction (by HPLC), methanol was added until a solid yellow mass was formed. Dissolve the solid in dichloromethane, stir for 15-30 minutes and filter. After concentration in vacuo, the residue is chromatographed on a fast-passing column (6: 1 hexane: ethyl acetate) to remove the excipients and as many polar degradation products as possible, yielding a yellow oil (16) with a yield of 50%.

¹1 H-NMR (CDCl 3)₃? d 0.83-0.98 (m, 2H), 1.09-1.38 (m, 10H), 1.57-2.00 (m, 11H), 1.83-2.00 ( m, 1H), 2.12-2.18 (m, 1H), 2.49-2.54 (m, 1H), 3.10 (dd, 1H, J = 11 and 6 Hz), 3, 27 (dd, 1H, J = 11.5 and 8.0 Hz), 4.11-4.25 (m, 2H), 4.88 (dd, 1H, J = 11.0 and 5.0 Hz) , 5.14 (dd, 1H, J = 10 and 6 Hz).

<img file="LT4368B_D0201.tif" />

<img file="LT4368B_D0202.tif" />

The starting material (16) (0.95 g, 2.9 mmol, 1 equiv) is dissolved in 10 mL of dioxane. The solution was cooled to 10 ° C and LiOH.H was added₂O (0.123 g, 2.9 mmol, 1 equiv) dissolved in 10 mL water. The cooling bath is removed and the mixture is stirred at room temperature for 1 hour. TLC indicates complete reaction; the solvent is then evaporated off under vacuum. The remaining aqueous layer was washed with ether (2x), acidified with 10% citric acid and extracted with dichloromethane (3x). The combined extracts were dried over sodium sulfate and evaporated to give a white solid which was recrystallized from ether. Concentrate the filtrate and purify by silica gel column chromatography (2: 1 hexane; ethyl acetate) to give the product (17).

Lyd. mp: 198.2-199 ° C;

¹H-NMR (DMSC-d₆δ d 0.78-0.93 (m, 2H), 1.11-1.27 (m, 5H), 1.34-1.36 (m, 1H), 1.51-1.56 (m) , 1H), 1.60-1.75 (m, 1H), 1.82-1.87 (m, 1H), 2.15-2.18 (m, 1H), 2.37-2.41 (m, 1H), 3.03 (dd, 1H, J = 11.5 and 5.5 Hz), 3.35-3.38 (m, 2H), 4.83 (dd, 1H, J = 9 and 4 Hz), 4.95 (dd, 1H, J = 8 and 5.5 Hz).

100

<img file="LT4368B_D0203.tif" />

Cbz Cbz Cbz

19 20

BOC-DiCbz-Arg (18) (7.6 g, 14.0 mmol) was dissolved in anhydrous THF (40 mL) and cooled to 0 ° C. Triethylamine (2.2 mL) is added and 14.5 mmol of a 1M solution of isopropyl chloroformate in toluene is added via syringe. The reaction mixture was stirred at 0 ° C for 30 minutes and then filtered rapidly. A white solid is separated off and the filtrate is bubbled through freshly prepared diazomethane until the solution turns yellow. The reaction mixture is left overnight under well-ventilated draft, which facilitates removal of excess diazomethane. Dry ether is added to precipitate the diazoketone. The product was filtered off and dried in vacuo to give a slightly yellowish, lumpy solid (4.6 g, 58%). Diazoketone (19) (1 g, 1.77 mmol) was dissolved in THF (20 mL) and to this solution was added 1M HCl in ether (20 mL) at 0 ° C. The reaction mixture was stirred overnight at room temperature: a white precipitate formed during this time. More precipitate is obtained by the addition of ether. Filtration and drying of the solid afforded the product (20) (1.02 g, 100%).

¹1 H-NMR (DMSO-d 6) δ 1.65-1.77 (m, 3H), 2.06-2.50 (m, 1H), 3.86-3.90 (m, 2H), 4, 29 (m, 1H), 4.76 (d, 1H, J = 18Hz), 4.95 (d, 1H, J = 18Hz), 7.35 (s, 2H), 7.36 (s, 2H), 7.35-7.41 (m, 10H), 8.71 (m.s, 13H), 10.1 (m.s, 2H).

¹³C-NMR (DMSO-d 6) δ 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3, 128.6, 128.7, 128.8, 128 , 9, 135.2, 135.9, 153.4, 157.4, 198.9.

101

O

<img file="LT4368B_D0204.tif" />

To a simulator (17) (0.422 g, 1.42 mmol) in THF (50 mL) at 0 ° C in the presence of N-methylmorpholine (0.19 mL) was added slowly 1M isopropyl chloroformate in toluene (1.71 mL). The reaction mixture was stirred at 0 ° C for 30 min then treated with small portions of aminochloromethyl ketone (20). After the addition of the aminochloromethyl ketone, the reaction mixture was stirred for an additional 15 minutes, after which N-methylmorpholine (0.19 ml) was added. After stirring at room temperature for 3 hours, the mixture is extracted with ethyl acetate, followed by washing with saturated sodium chloride solution and 10% aqueous citric acid. Evaporation of the organic solvent gave a white foam (21) (1.03 g, 96%) which was used without further purification.

¹1 H-NMR (CDCl 3) δ 0.07-0.97 (m, 1H), 1.15-1.41 (m, 7H), 1.62-1.91 (m, 10H), 2.10-. 2.16 (m, 1H), 2.43-2.48 (m, 1H), 2.74-2.80 (m, 1H), 3.01-3.07 (m, 1H), 3, 87-3.94 (m, 1H), 4.11-4.19 (m, 2H), 4.60-4.66 (m, 1H), 4.74-4.86 (m, 2H), 5.09-5.24 (m, 4H), 7.30-7.39 (m, 10H), 7.95 (d, 1H, J = 8Hz), 9.4 (ss, 1H) , 9.56 (pl.s, 1H).

BOC

<img file="LT4368B_D0205.tif" />

example

<img file="LT4368B_D0206.tif" />

H

N \ / \ / \ /

NH / J

HN

NH, (Nt-BOC-N-tosyl) butyrylketoarginine (240 mg, 0.515 mmol) was deprotected using 30% TFA in dichloromethane. The deprotected arginine derivative is coupled to simulator (8) (100 mg, 0.343 mmol) in DMF

102 in basic conditions {Et₃N, pH 8-9) using BOP reagent (228 mg,

0.52 mmol) as a dehydrating agent. The reaction usually occurs within 2-4 hours. Extraction with ethyl acetate, washing with saturated sodium chloride solution and 10% aqueous citric acid gives a crude product. This product was purified by column chromatography to give 180 mg (76%) of pure product. This product is then exposed to HF to remove the tosyl group. Purification of the isolated deprotected product by HPLC gives BCH-2737.

<img file="LT4368B_D0207.tif" />

HN = &lt;

, Example of NCbz

<img file="LT4368B_D0208.tif" />

HN = &lt;

u

NH,

Chloromethyl ketone (21) (0.188 g, 0.245 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. Extraction of the reaction mixture with ethyl acetate followed by washing with saturated sodium chloride solution and 10% aqueous citric acid and evaporation of the organic solvent gave a crude product which was purified by column chromatography to give a foamy solid (0.125 g, 62%).

This blocked precursor (0.125 g, 0.154 mmol) was dissolved in DCM (5 mL) and cooled to -78 ° C. Slowly add 1M BBr₃ (1.54 mL, 1.54 mmol) in DCM. The reaction mixture was stirred at room temperature for 5 hours, then cooled again to -78 ° C and treated with anhydrous methanol (2 mL). The reaction mixture was warmed to room temperature and stirred for a further 2 hours. The solvents were evaporated under reduced pressure and the residue partitioned between ether and water. The aqueous layer is collected, lyophilized to give the final product (23) as a powder, purified by HPLC and lyophilized.

103

The products of the above reaction may be isolated in free form or in the form of salts. In addition, the products may be prepared in the form of pharmaceutically acceptable acid addition salts with one of the free acid bases. Similarly, the product may be obtained in the form of its pharmaceutically acceptable salts by treating one of the free carboxylic acids with a base. Also, the free amide is regenerated by treating the salts with base or acid.

example

A general method for the synthesis of compounds of formula II or III is:

<img file="LT4368B_D0209.tif" />

CBzNH

NH₂CBz nh₂

<img file="LT4368B_D0210.tif" />

<img file="LT4368B_D0211.tif" />

example

104 synthesis

<img file="LT4368B_D0212.tif" />

Step 2: Synthesis of 2-Benzyloxycarbonylamino-4-hydroxybutanecarboxylic acid tert-butyl ester

NH 2 (Cl k in salmon hexyl) NH · HOOC (D o

οΆι

NMM.THF

CO₂t-Bu 2) NaBH₄, MeOH

NHZ

CO₂tBu (2) into a solution of blocked aspartic acid (1) (Bachem, 2.50 g, 4.95 mmol) in 50 mL dry tetrahydrofuran (THF) at -10 ° C, N₂ N-methylmorpholine (109 mL, 0.2 equiv.) and isopropyl chloroformate (1.0 M / toluene: 384 mL, 1.1 equiv.) were added under atmosphere. The solution was stirred at -10 ° C for 60 minutes. In another flask, NaBH is suspended₄ (375 mg, 2 equiv.) In dry THF / MeOH 5: 1 (50 mL) at -78 ° C₂ in the atmosphere. This suspension was stirred at -78 ° C for 30 minutes. Then to NaBH₄• The suspension is added dropwise via a tube to the mixed anhydride solution and the final solution is stirred at -78 ° C. 3 hours. Acetic acid (2.8 mL, 10 equiv.) Was then added and the solution was warmed to room temperature (30 min). The solvents were evaporated, the residue was dissolved in EtOAc and washed with saturated aqueous NaHCO3₃ (2x) and saturated sodium chloride solution. The organic layer was dried over MgSO4₄After filtration and evaporation of the solvent, 1.53 g (4.95 mmol, 100%) of alcohol (2) is obtained which is a clear oil.

¹1 H-NMR (CDCl 3, 400 MHz): δ 7.40-7.31 (m, 5H, ArH), 5.63 (d, 1H, J = 7.3, NH), 5.13 (AB system, 2H, J = 12.2, CH₂Ph), 4.43 (m, 1H, H-2), 3.69 (m, 2H, H-4), 2.17 (m, 1H, H-3), 1.63 (m, 1H, H-3), 1.48 (s, 9H, t-Bu).

105 Step 2: Synthesis of 2-Benzyloxycarbonylamino-4-iodobutanecarboxylic acid tert-butyl ester

HO

NHZ

Ph P, I 'CO₂t-Bu (2) CH₃CN / Et₂O

NHZ

CO₂t-Bu (3)

Of a solution of alcohol (2) (1.53 g, 4.95 mmol) in CH₃CN / Et₂O in a 1: 1 mixture (50 mL) at -10 ° C N₂ imidazole (607 mg, 1.8 equiv.) was added to the atmosphere followed by Ph₃P (2.21 g, 1.7 eq). Iodine (2.14 g, 1.7 eq.) Was then added in small portions over 15 minutes. The addition of iodine gives a white precipitate and the solution turns brown. It was stirred at -10 ° C for 45 min. The solution is then poured into Et₂Oh, the organic phase is washed with saturated aqueous Na₂SO₃, saturated aqueous CuSO₄, H₂Oh and dried over MgSO4₄. The solids were filtered off and the solvent evaporated to give a yellow oil which was purified by flash chromatography (silica gel, 5% to 20% EtOAc / hexane). Iodide (3) is obtained as a clear oil (1.71 g) in 83% yield.

¹1 H-NMR (CDCl 3, 400 MHz): δ 7.41-7.31 (m, 5H, ArH), 5.35 (ppm, 1H, J = 7.3, NH), 5.13 (s , 2H, CH₂Ph), 4.30 (m, 1H, H-2), 3.22-3.12 (m, 2H, H-4), 2.42 (m, 1H, H-3), 2.20 ( m, 1H, H-3), 1.48 (s, 9H, t-Bu).

Step 2: Synthesis of 2-Benzyloxycarbonylamino-4-hexanoic acid tert-butyl ester

<img file="LT4368B_D0213.tif" />

MgSO 4, Cul THF, -78 ° C

NHZ

<img file="LT4368B_D0214.tif" />

(4)

CO₂t-Bu j Cul (2.27 g, 5 equiv.) in dry THF (20 mL) at -78 ° C.₂ a 1.0 M solution of vinylmagnesium bromide in THF (23.4 mL, 9.8 eq) was added slowly to the atmosphere. The solution is then warmed to -10 ° C for 30 minutes (turns black), then cooled again to -78 ° C. To this hump solution was slowly added a solution of iodide (3) (1.00 g, 2.39 mmol) in dry THF (3.5 mL).

106

The reaction mixture was stirred at -78 ° C for 2.5 hours. Saturated aqueous NH is added₄CI (50 mL) was allowed to warm to room temperature with vigorous stirring. The mixture is then poured into Et₂Oh and stir for 5 minutes. Filter the dark suspension through a sintered glass funnel and separate the phases. The aqueous phase is extracted with Et₂O (2x), and the combined organic extracts were dried over MgSO 4₄. The solids are filtered off, the solvents are evaporated and the crude oil is purified by flash chromatography (silica gel, 5% AcOEt / hexane) to give 0.51 g (67%) of pure alkene (4).

¹1 H-NMR (CDCl 3, 400 MHz): δ 7.37-7.31 (m, 5H, ArH), 5.80 (m, 1H, H5), 5.33 (d, 1H, J = 7.8). , NH), 5.12 (s, 2H, J = 12.2, CH)₂Ph), 5.05 (d, 1H, J = 17.2, H-6), 5.01 (d, 1H, J = 10.4, H-6). 4.30 (kv, 1H, J = 7.4, H-2), 2.162.08 (m, 2H, H-4), 1.92 (m, 1H, H-3), 1.74 (m , 1H, H-3), 1.48 (s, 9H, t-Bu).

Step 1: Synthesis of 1-Benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinecarboxylic acid tert-butyl ester

HO

NHZ

<img file="LT4368B_D0215.tif" />

1) Hg (O Ac) 2, THF

2) NaHCO3.KBr_&gt;

3) NaBH4, O2

<img file="LT4368B_D0216.tif" />

Of a solution of alkene (4) (50 mg, 0.157 mmol) in dry THF (3.1 mL) at room temperature under N₂ mercury acetate (75 mg, 1.5 eq.) was added in the atmosphere. The solution is stirred at room temperature for 18 hours, then cooled to 0 ° C. Saturated aqueous NaHCO is then added₃ (2 mL) and the mixture was stirred at 0 ° C for 30 min. KBr (0.11 g, 6 equiv.) Was added and the mixture was stirred at room temperature for 2 hours. It is then poured into H₂O / No₂Oh, and the phases are separated. The aqueous phase is extracted with Et₂O (2x), and the combined organic extracts were dried over MgSO 4₄. The solids are filtered off and the solvents are evaporated to NaBH₄ (3.3 mg, 0.55 equiv.) In dry DMF (0.4 mL) was bubbled with oxygen (O) for 1 h.₂) and a solution of mercuric organobromide in DMF (3.1 ml) is added dropwise (syringe pump, 3 ml / h),₂. Oxygen was allowed to flow for another 1 h and Et was added₂O (5 mL). The gray suspension is filtered through zeolite, and

107 the filtrate was evaporated. The residue was chromatographed (silica gel, 6: 4 hexane / EtOAc) to give pyrrolidinol (5) (30 mg, 57%) as a clear oil.

¹1 H-NMR (CDCl 3)₃, 400 MHz): d 7.37-7.28 (m, 5H, ArH), 5.22-5.09 (m, 2H, CH₂Ph), 4.30 (dd, 1H, J = 1.4 and 8.3, H-2), 4.24 (m, 1H, H-5), 3.70-3.57 (m, 3H, CH₂-OH), 2.25 (m, 1H), 2.13 (m, 1H), 1.92 (m, 1H), 1.70 (m, 1H), 1.34 (s, 9H, t- Bu).

Step 1: Synthesis of 1-Benzyloxycarbonyl-5-carboxy-2-pyrrolidinecarboxylic acid tert-butyl ester

<img file="LT4368B_D0217.tif" />

of alcohol (5) (50 ng, 0.149 mmol) and Et₃N (62 mL, 3 equiv.) In dry CH₂CI₂ (0.8 mL) N₂ of the SCb-pyridine complex (71 mg, 3 equiv.) in dry DMSO was slowly added to the atmosphere at 0 ° C. The solution was stirred at 0 ° C for 30 min and 10% citric acid (2 mL) was added. 1M NaOH is adjusted to pH = 4 and the aqueous phase is extracted with Et₂O (3x). The combined organic extracts were dried over MgSO4₄. The solids are filtered off and the solvents are evaporated to give a crude oil which is purified by flash chromatography (silica gel, 7: 3 hexane / EtOAc). The pure aldehyde (6) was obtained as an oil (45 mg, 90%).

¹1 H-NMR (CDCl 3, 400 MHz): δ 9.68 + 9.56 (ds, 1H, CHO), 7.36-7.29 (m, 5H, ArH), 5.23-5.11 (m , 2H, CH₂Ph), 4.57-4.39 (m, 2H, H-2, H-5), 2.30-1.97 (m, 4H, H-3, H-4), 1.47 + 1 , 36 (2s, 9H, t-Bu).

stage:

108

<img file="LT4368B_D0218.tif" />

<img file="LT4368B_D0219.tif" />

Pyrrolidine-aldehyde (6) is coupled with a protected diaminoaminopropanoic acid (7), primarily to form imine (8) (MgSO₄, CH₂CI₂). the solid (8) is isolated by filtration with MgSO₄ and evaporation of the solvent. The crude imine is then exposed to NaBH (OAc)₃ and acetic acid (AcOH) in THF for 15 hours, and extraction and treatment of the extract gives the amine (8).

<img file="LT4368B_D0220.tif" />

OBut

<img file="LT4368B_D0221.tif" />

OBut

The protecting group of the amine (8) CB7 (7) is removed by hydrogenation using 10% palladium on activated carbon in methanol (MeOH) as the catalyst. The catalyst is filtered off and the MeOH evaporated to give the crude diamine (9) which can be used without further purification.

<img file="LT4368B_D0222.tif" />

OBut

<img file="LT4368B_D0223.tif" />

OBut

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

109 stage:

<img file="LT4368B_D0224.tif" />

The secondary amino group of the bicyclic lactam (10) is blocked by converting it into an amide using benzoyl chloride in pyridine. Removal of the pyridine and extraction procedures gives the bicyclic lactamide (11).

stage:

<img file="LT4368B_D0225.tif" />

The BOC and t-butyl ester protecting groups of the bicyclic lactamide (11) are removed under acidic conditions (HCl in ethyl ether (Et₂O)). The solution precipitates the amine salt (12), which is collected by filtration.

stage:

<img file="LT4368B_D0226.tif" />

c₆h₅ch₂ococi K₂CO₃, CH₃'CN

<img file="LT4368B_D0227.tif" />

The primary amino group of compound (12) is blocked by the CBZ group in the presence of benzyl chloroformate in acetonitrile (CH₃CN) with K₂CO₃, as a base. The extraction procedures give a fully blocked carboxylic acid (13) which can be used in Step 12 without further purification.

110

<img file="LT4368B_D0228.tif" />

<img file="LT4368B_D0229.tif" />

HN ^ NH2

The carboxylic acid (13) is coupled with benzothiazole ketoarginine (14) in DMF using BOP as a copolymer in the presence of diisopropylethylamine (EtNiPr₂ ). Extraction with ethyl acetate (EtOAc) gives a white solid (15) which is purified by chromatography.

stage:

<img file="LT4368B_D0230.tif" />

Both CBZ (Z) protecting groups of compound (15) are removed by catalytic hydrogenation with Pd / C 10% as catalyst. Filtration of the catalyst and evaporation of the solvent afforded the aminoguanidine (16).

Example Synthesis of compound (10)

111

<img file="LT4368B_D0231.tif" />

<img file="LT4368B_D0232.tif" />

<img file="LT4368B_D0233.tif" />

At room temperature, the carboxylic acid (2) (1.7 g, 4.9 mmol, 1.0 equiv), 4-hydroxyproline (3) (5.39 mmol, 1.1 equiv) and BOP reagent (2.17 g, 4.9 mmol, 1.0 eq.) in anhydrous THF (10 mL) was added 4-methylmorpholine (NMM). The reaction mixture was stirred at room temperature overnight / quenched with saturated sodium chloride solution (50 mL) and ethyl acetate (100 mL). The organic layer was washed with aqueous citric acid solution (10%, 2 x 50 mL), sodium bicarbonate (10%, 2 x 50 mL) and saturated sodium chloride solution (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered and the solvent evaporated. The residue was purified by flash chromatography (5: 4: 1, ethyl acetate-hexane-methanol) to give 1.1 g of pure product (4) in 48% yield.

stage:

112

<img file="LT4368B_D0234.tif" />

(4) (5)

To a solution of the 4-hydroxyproline derivative (4) (115 mg, 240 mm, 1 eq.) In dichloromethane (anhydrous, 10 mL) at 0 ° C was added triethylamine (72 mg, 720 mmol, 3.0 eq.) And methanesulfonyl chloride (28). mg, 240 mmol, 1.0 eq.) and the reaction mixture was stirred at room temperature. The reaction was then quenched with aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was washed with 10% citric acid and saturated sodium chloride solution, dried, filtered and evaporated to dryness to afford the compound (5).

stage:

<img file="LT4368B_D0235.tif" />

<img file="LT4368B_D0236.tif" />

Enamine (5) (1.0 eq) was treated with mercury acetate (1.1 eq) in THF. The solvent is evaporated to dryness and the residue is dissolved in methanol. The resulting organic mercury compound is reductively cleaved with sodium borohydride (1.3 eq.). The resulting crude lactam thioether is purified by silica gel chromatography to give compound (6).

stage:

113

<img file="LT4368B_D0237.tif" />

<img file="LT4368B_D0238.tif" />

\

To a solution of lactam thioether (6) (1.0 eq.) In dry dichloromethane at 0 ° C was added N-chlorosuccinimide (1.0 eq.). When no starting material is seen in the reaction mixture, the solid is filtered off and the solvent is evaporated to dryness. The crude material (7) is used in step 5 without further purification.

<img file="LT4368B_D0239.tif" />

O \

stage·.

<img file="LT4368B_D0240.tif" />

O \

To a solution of alpha-chlorothioether (7) (1.0 equiv.) In THF (anhydrous) is added a solution of phenyl cuprate (1.0 equiv.) (Prepared according to the literature) at low temperature. When the reaction mixture no longer shows the starting chlorothioether, saturated sodium chloride solution and ethyl acetate are added. The organic layer is dried, filtered and evaporated to dryness to give the desired product (8). '

<img file="LT4368B_D0241.tif" />

stage:

<img file="LT4368B_D0242.tif" />

114

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

stage:

The crude carboxylic acid (9) is coupled to the benzothiazole ketoarginine in DMF using BOP as a copolymer in the presence of diisopropylethylamine. Extraction with EtOAc yields a solid which is purified by chromatography on silica gel to give the protected amide. The CBZ protecting group is removed by BBr3 in dichloromethane at room temperature, and finally a bicyclic benothiazole ketoarginine inhibitor is obtained (10).

The following compounds are prepared in the same manner, except that the corresponding substituents are used in the reactions to produce the final products.

compound

115

<img file="LT4368B_D0243.tif" />

compound

<img file="LT4368B_D0244.tif" />

example stage:

OO

I.CH.₂= N = N, THF / Ether, -15 ° C &lt;-j

2.HC1 / Ether j

O ^ OCHj O ^ -OCHj

Commercially available glutaric acid monomethyl ester chloride (1) (20 mL, 0.144 mmol) was dissolved in 40 mL dry tetrahydrofuran (THF) and cooled to -15 ° C. To this solution is added a freshly prepared solution of diazomethane in 300 ml of ether through a tube at -15 ° C. The mixture was allowed to warm to room temperature overnight. Remove excess diazomethane from the flask with a stream of argon. To complete the reaction, 75 mL of 1N HCl in ether was added at 0 ° C and the mixture was allowed to warm to room temperature over 5 hours. The solvent volume is reduced and then washed

116

2x5% NaHCO₃, dried Na₂CO₃ and evaporation to give crude methyl methyl ketone (20.46 g, 79%) which was used in the next step without further purification.

¹1 H-NMR (CDCl 3)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).

<img file="LT4368B_D0245.tif" />

stage:

<img file="LT4368B_D0246.tif" />

The crude chloromethyl ketone (2) (10.04 g, 56.15 mmol) was dissolved in 300 mL of dry MeOH. Sodium acetate (2 eq, 9.21 g, 112.3 mmol) was added followed by L-cysteine ​​ethyl ester hydrochloride (1.3 eq, 13.55 g, 72.98 mmol) and sodium cyanoborohydride 1, 4 eq., 4.9 g, 78.59 mmol). The heterogeneous mixture was stirred at room temperature for 2.5 hours. Then 200 mL of methanol (MeOH) is added to dissolve all solids and the pH is adjusted to 2 with 1N HCl. The mixture is then basified with saturated NaHCO₃ to pH = 8. Evaporate methanol and wash the remaining aqueous solution with ethyl acetate and dichloromethane. The solvents are mixed, dried over Na₂SO₄ and evaporated. The crude residue is purified by silica gel chromatography using an ethyl acetate / hexane gradient of 3: 7, 5: 5, 6: 4, 7: 3. The cyclic compound (3) is obtained.

(3) of the compound ¹1 H-NMR (CDCl 3)₃, 400 MHz): 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 Hz), 2.72-2.76 (dd, 1H, J = 10 , 7Hz), 2.8-2.86 (m, 1H), 3.6-3.64 (d, 4H, J = 2.55Hz), 3.63 (s, 3H), 4.13 -4.2 (m, 2H).

¹³C-NMR (CDCl_3i 400 MHz): d 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.

117

<img file="LT4368B_D0247.tif" />

<img file="LT4368B_D0248.tif" />

The cyclic compound (3) (913 mg, 3.32 mmol) was dissolved in 50 mL of dry toluene. (1S) - (+) - 10-Camphor sulfuric acid (92 mg, 0.39 mmol) was added and the mixture was refluxed for 4 days. When all starting material is consumed (by TLC), the mixture is treated by evaporation of the solvent, dissolving the residue in ethyl acetate and washing with 2x5% NaHCO₃. The ethyl acetate layer was dried over Na₂SO₄ and is suppressed. The crude residue was purified by silica gel chromatography using 60% EtOAc / 40% hexane followed by 70% EtOAc / 30% hexane. 62.5% of the bicyclic compound (4) is obtained.

(4) of the compound ¹1 H-NMR (CDCl 3)₃, 400 MHz): 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), 2,522.59 (m, 2H), 2.83-2.88 (dd, 1H, J = 14 and 4 Hz), 3.14-3.18 (d, 1H), 3.78-3.85 (m, 1H), 4.2-4.27 (kv, 2H, J = 3.9 Hz) , 5.92 (t, 1H, J = 3.4 Hz).

stage:

<img file="LT4368B_D0249.tif" />

<img file="LT4368B_D0250.tif" />

The bicyclic compound (4) (366 mg, 1.5 mmol) was dissolved in 25 mL of THF and 5 mL of H₂O. Add a solution of lithium hydroxide monohydrate (1.1 eq, 7.05 g, 1.68 mmol) in 2.3 mL of H₂O at 0 ° C, and the mixture was stirred at 0 ° C for 1 hour and at room temperature for 3 hours. Then

118 evaporated with THF and the remaining aqueous mixture acidified to pH = 2 with citric acid. After extraction of the aqueous mixture with 2x CH₂CI₂ and 2x EtOAc after drying the combined organic layers over Na₂SO₄ and evaporation yields a residue which is purified by silica gel column chromatography using 70% EtOAc / 30% hexane followed by 4.7% HOAc / ethyl acetate. Pure acid (5) is obtained with a yield of 54%. 16% of the starting material is also isolated (4).

(5) of the compound ¹1 H-NMR (MeOD, 400 MHz): d 1.57-1.69 (m, 1H), 1.70-1.80 (m, 1H), 1.81-1.89 (m, 1H), 2.02-2.1 (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 and 4 Hz), 3.12-3.17 (d, 1H),

3.3-3.32 (m, 1H), 3.78-3.84 (m, 1H), 5.76-5.78 (t, 1H, J = 3.53 Hz).

¹³C-NMR (MeOD, 400 MHz): d 17,052, 27.07, 28,928, 31,382, 32,096, 51,016, 55,138, 170,088, 171.24.

<img file="LT4368B_D0251.tif" />

<img file="LT4368B_D0252.tif" />

A solution of carboxylic acid (5) (500 mg, 2.32 mmol) was added to a solution of lithium bis (trimethylsilyl) amide (5 mL of 1M solution in THF, 5 mmol) at -78 ° C and the resulting solution was stirred for 1 hour. at this temperature. Benzyl bromide (0.26 mL, 2.22 mmol) was then added and the mixture was allowed to reach room temperature and stirred for 15 h. The mixture was poured into 10% HCl (50 mL) and extracted with dichloromethane (4x60 mL). The combined organic phases were dried over MgSO4₄and evaporation of the solvent afforded the crude alkylated amide (6).

stage:

119

<img file="LT4368B_D0253.tif" />

<img file="LT4368B_D0254.tif" />

2) BBr₃ Z CH₂CI₂

<img file="LT4368B_D0255.tif" />

The crude alkylated amide (6) is coupled with benzothiazoleto ketoarginine in DMF using BOP as a copolymer in the presence of diisopropylethylamine. Extraction with EtOAc yields a solid which is purified by chromatography on silica gel to give the protected amide. The CBZ protecting group is removed in BBr₃ dichloromethane at room temperature, and finally a bicyclic benothiazoleto ketoarginine inhibitor is obtained (7).

The following compound is prepared in the same manner except that the corresponding compounds are substituted in the reactions to give the final product.

<img file="LT4368B_D0256.tif" />

example

Determination of the Heterocyclic Compound K The inhibitory pull on thrombin was measured according to DiMaio et al., J. Biol.

Chem., 1990, 265: 21698. Inhibition of human thrombin amidolytic activity was measured fluorimetrically,

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

Thrombin-catalyzed hydrolysis of the substrate was controlled with a Varian-Cary 2000 ™ spectrophotometer in its fluorescence variant (l_ex = 383 nm, km = 455 nm) or with a Hitachi F2000 ™ fluorescence spectrophotometer (l_ex= 383 nm, km = 455 nm), and fluorescence intensity was calibrated using AMC. After mixing thrombin and substrate with inhibitor, the reaction reaches steady state within 3 minutes. The steady state speed is then measured for several minutes. The compounds of the present invention were also incubated before the thrombin assay for 20 minutes at room temperature before adding substrate. Steady state is reached within 3 minutes and measured in minutes. The kinetic data (steady state velocity at different concentrations of substrate and inhibitors) of competitive inhibition were analyzed using the methods described by Segei (1975). The kinetic parameters (Km, V_max and K) a nonlinear regression program RNLIN from the IMSL library (IMSL, 1987), LMDER from the MINPACK library (More et al, 1980) or Microsoft ™ Excell ™ was used for estimation.

dTT test

The fibrin coagulation assay was performed in 50 mM Tris-HCl buffer (pH 7.52, 37 ° C) containing 0.1 M NaCl and 0.1% polyethylene glycol 8000 at 9.0 x 10 ', respectively.¹° M (0.1 NIH units / ml) and 0.03% (w / v) for final concentrations of human thrombin and bovine fibrinogen as described by Szevvczuk et al., Supra. Plot a plot of clotting time versus inhibitor concentration and determine IC₅o, as the concentration of inhibitor required to double the clotting time compared to the control. The results are given in Tables 1 and 2.

Fibrin clot test

121

The fibrin clot assay was performed essentially as described by Krtenansky et al., FEBS, 1987, 211: 10. A series of inhibitor dilutions were made in 50 mM Tris-HCl buffer (pH 7.8, 23 ° C) containing 0.1 M NaCl and 0.1% (w / v) polyethylene glycol 8000 microtiter wells (microtiter plate, Falcon). containing 100 ml of various dilution inhibitor solutions, human plasma (60 ml, collected in 3.8% sodium citrate, blood / anticoagulant ratio 9: 1) was added. The solution was mixed and then added with 50 ml human thrombin (final concentration 1 nM) and stirred again for 15 seconds. The clot turbidity was immediately determined using a plate maker (Dynateck MR 5000) at 405 nm and measured every 3 minutes. Maximum turbidity in the absence of inhibitor was achieved after 60 min. The ICs were deducted₅o values ​​at 30 min which indicate the concentration of inhibitor which gives half the value of the optical density of the control sample.

Platelet aggregation and secretion

Rat blood was collected by ACD (6/1 v / v) cardiac puncture. Washed platelet suspensions were prepared according to Ardlie et al. (Br. J. Haematol. 1970, 19: 7 and Proc. Soc. Exp. Biol. Med. 1971, 136: 1021). The final suspension medium was modified Tyrode solution (NaCl 138 mM, KCl 2.9 mM, HEPES 20 mM, NaH₂PO₄ 0.42 mM, NaHCO₃ 12 mM, CaCl₂ 1 mM, MgCl2₂ 2 mM, 0.1% glucose, 0.35% albumin, 1 ml / ml apyrase; pH 7.4). Platelet counts were adjusted to 5,000,000 / ml.

Platelets were labeled in the first washing solution to measure the release rate of dense granules. ¹⁴Of C-serotonin (5-HT) (1 mCi / 10 mL of washing liquid) and was assayed ¹⁴C-serotonin release according to Holmsen et al. (Enzymology, 1989, 169: 206). Inipramine (final concentration 5mM) was added to provide release of released serotonin.

Platelet aggregation was recorded at 37 ° C on an aggregate meter (BioData PAP-4) at 1100 rpm. mixing speed as measured by light

122 bandwidth changes. Aggregation percentages determined after 3 min. after the addition of a stimulating agent (human thrombin, final concentration 0.1 lU / ml). Inhibitors were incubated for 1 min before addition of the stimulating agent. At 37 ° C. IC50 values ​​represent the concentration required to inhibit platelet aggregation or secretion at 50% of control.

Model of arterial thrombosis

FeCIg-induced carotid artery injury model

FeCI₃ rat carotid artery injury was induced by Kurtz, KD, Main, RW, Sandusky, GE, Thromosis Research, 60, 269280 (1990) and Schumacher, WA, et al., J. Pharmacology and Experimental Therapeutics, 267, 1237-1242 ( 1993) described the methodology.

Male Sprague-Dawley rats (375-410 g) were anesthetized with urethane (1500 mg / kg ip). The animals were placed on a heated (37 ° C) tray. The carotid artery is opened through the middle neck. The blood vessel is separated from the carotid artery by a careful incision with a blunt needle. Using tweezers, the artery was raised to allow enough space for two small pieces of polyethylene tube (PE-205) underneath. A temperature gauge is inserted between one of the pieces and the artery (Physitemp MT23 / 3). The damage is caused by the use of Watman's No. 1 filter paper pre-soaked in 35% FeCl₃ local application of a small disc (3 mm diameter) on the carotid artery above the temperature gauge. The cut area is covered with aluminum foil to make FeCI₃ protected from light-induced degradation. Vascular temperature was monitored 60 minutes after FeCI₃ applications as an indicator of blood circulation. Changes in blood vessel temperature are recorded with a thermister (Cole-Palmer Model 0853341).

Time between FeCI₃ applications and the time when the blood vessel temperature drops sharply (&gt; 2.4 ° C) are accepted as the time of the blood vessel occlusion. Inhibitory compounds were administered as an IV dose followed by an IV infusion (mg / kg / min via the femoral vein). Determining

123 the dose of inhibitor required to double the occlusion time compared to control animals in which the lesion was induced in the absence of inhibitor.

table

<td rowspan="2">The compound</td><td rowspan="2">K, μΜ³</td><td colspan="2">Antiplatelet activity, μΜ</td><td rowspan="2">dTT IC50, μΜ</td><td rowspan="2">Plasma Fibrin Coagulation Test IC₅o, μΜ</td>

<td>Aggregation *</td><td>5-HT secretion</td>

<td> 0005</td><td> 4</td><td>not set</td><td>not set</td><td> 47</td><td> &gt;450</td>

<td> 0010</td><td> 4,6</td><td> 21</td><td> 19</td><td> 89,5</td><td> &gt;450</td>

<td> 0015</td><td> 16</td><td> &gt;100</td><td> &gt;100</td><td> 162</td><td> &gt;450</td>

<td> 0020</td><td> 2,2</td><td> 18</td><td> 14,2</td><td> 22</td><td> &gt;450</td>

<td> 0025</td><td> 53</td><td> &gt;100</td><td> &gt;100</td><td> &gt;625</td><td> &gt;450</td>

<td> 0030</td><td> 8,6</td><td> &gt;100</td><td> &gt;100</td><td> 67</td><td> 320</td>

<td> 0035</td><td> 34</td><td> &gt;100</td><td> &gt;100</td><td> 319</td><td> &gt;450</td>

<td> 0040</td><td> 19</td><td> &gt;100</td><td> &gt;100</td><td> 207,5</td><td> &gt;450</td>

<td> 0045</td><td> 74</td><td>not set</td><td>not set</td><td> 415</td><td> &gt;450</td>

<td> 0050</td><td> 62</td><td>not set</td><td>not set</td><td></td><td> &gt;450</td>

<td> 0065</td><td> 32,7</td><td> 47,5</td><td> 52</td><td> 42</td><td> 200</td>

<td> 0070</td><td> 4,4</td><td> 22</td><td> 2,1</td><td> 25</td><td> 78</td>

<td> 0080</td><td> 0,048</td><td> 0,4</td><td> 0,38</td><td> 0,375</td><td>not set</td>

<td> 0090</td><td> 0,031</td><td>not set</td><td>not set</td><td> 0,33</td><td>not set</td>

<td> 0095</td><td> 26</td><td>not set</td><td>not set</td><td></td><td></td>

<td> 0100</td><td> 19</td><td>not set</td><td>not set</td><td> 165</td><td>not set</td>

<td>* mouse mice</td><td colspan="5">autothrombocyte suspension</td>

^a Inhibitory dissociation constant for human cc-thrombin.

table

<td>The compound</td><td>K, nM</td><td>dTT, nM</td><td>Jvedimo way, ivb + inf</td><td>MOT, min ± hr fall</td>

<td> 0220 0225</td><td> 18 550</td><td></td><td> 0,75-50</td><td> 23±7</td>

<td> 0245</td><td> 235 5</td><td></td><td> 0,5-30</td><td> 27±3</td>

<td> 0250</td><td> 8 40.</td><td> 350</td><td> 0,75-50 0,25-20</td><td> 22,6±2,6 23±8</td>

<td>0295a</td><td> 1500</td><td></td><td> 0,75-50 0,75-50</td><td> 22±3 20±1</td>

<td>0295b 0240</td><td> 18</td><td> 5000 520</td><td> 0,75-50 0,75-50</td><td> 19±2,7 17±2,6</td>

<td> 0210</td><td> 8</td><td></td><td> 0,75-50</td><td> 20,13±3,4</td>

<td> 0255 0260</td><td> 500 16</td><td></td><td> 0,75-50</td><td> 14,78±0,2</td>

<td>0305a</td><td> 220</td><td></td><td></td><td></td>

124

<td>0305b 0265a 0265b 0285 0315a + b 0315b 0335</td><td> 12000 4 18 10 150 45 10 25</td><td> 138</td><td> 0,75-50 0,75-50 0,75-50 0,75-50 0,75-50</td><td> 21,5±10 14,83±2,3 11,33±1,34 30,33±8,4 45,8±14,2</td>

<td></td><td></td><td></td><td> 0,5-30</td><td> 41,5±7,27</td>

<td></td><td></td><td></td><td> 0,25-20</td><td> 27,5±11,3</td>

<td> 0340</td><td> 0,6</td><td></td><td> 0,25-20</td><td> 36±9,6</td>

<td></td><td></td><td></td><td> 0,75-50</td><td> 42,25±11,9</td>

<td> 0345</td><td> 2</td><td></td><td> 0,75-50</td><td> 50±5,86</td>

<td> 0915</td><td> 1600</td><td></td><td> 0,75-50</td><td> 15±1,3</td>

<td> 0935</td><td> 120</td><td></td><td></td><td></td>

<td>0925a + b</td><td> 10</td><td></td><td> 0,75-50</td><td> 19,6±0,2</td>

<td>0925b</td><td> 30</td><td></td><td></td><td></td>

<td>0925a</td><td> 7</td><td></td><td> 0,75-50</td><td> 20,3±3,5</td>

<td>0940a</td><td> 16</td><td></td><td> 0,75-50</td><td> 15,2±0,82</td>

<td>0940b</td><td> 160</td><td></td><td></td><td></td>

<td>0950a</td><td> 150</td><td></td><td></td><td></td>

<td>0950b</td><td> 1000</td><td></td><td></td><td></td>

<td colspan="2">a = RP of early leaching</td><td colspan="3">HPLC single isomer</td>

b = individual isomer of subsequent HPLC HPLC a + b = mixture

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