Low molecular weight bicyclic thrombin inhibitors.

(57) Abstract

This invention relates to the discovery of heterocyclic competitive inhibitors of the enzyme thrombin having formula (I), their preparation, and pharnuiceutical compositions thereof. As well. this invention relates to the use of such compounds and compositions in vitro as anticoagulants and in vivo as agenis for the treatmeni and prophylaxis of thrombotíc disonders such as veneus thrombosis, pulmonary embolism and arterial thrombos »resulting in acute ischemic events such as myocardial infarction or cerebral infarction. Moreover, these compounds and compositions have thempeutic utility for the prevention and treatment of coagulopathies associated with coronary bypass operations as well as rcstcnotic events following transluminal angioplasty.

<img file="MX9704718A_D0001.tif" />

-1 BICYCLIC LOW MOLECULAR WEIGHT THROMBINE INHIBITORS

FIELD OF THE INVENTION

The present invention relates to compounds useful for the treatment of thrombotic disorders, and in particular to new heterocyclic inhibitors of thrombin enzyme.

BACKGROUND OF THE INVENTION

Thrombus formation does not order in the walls of the blood vessels precipitating states of acute cardiovascular disease that are the main cause of death in economically developed societies. Plasma proteins such as fibrinogen, proteases, and cellular receptors involved in hemostasis emerged as important factors that play a role in acute and chronic coronary heart disease, as well as diseases of the cerebral arteries, contributing to the formation of thrombi or blood clots. which effectively decrease normal blood flow and supply. Vascular aberrations arising from primary disease states such as hypertension, atherosclerotic plaque rupture, or stubborn endothelium activate biochemical cascades that function to respond to and repair the site of damage. Thrombin is the main regulatory enzyme in the cascade of. coagulation; It works as a positive and negative feedback regulator. However, under pathological conditions, the former is amplified through the catalytic activation of cofactors required for the generation of thrombin, as well as the activation of factor XIII

<td>necessary</td><td>for</td><td>the</td><td>interrelation</td><td>and stabilization of</td><td>the</td>

<td>fibrin.</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>also</td><td>of</td><td colspan="2">its direct effects on hemostasis,</td><td>the</td>

<td>thrombin</td><td>exercises</td><td colspan="2">direct effects</td><td>on various types</td><td>of</td>

cell that support and amplify the pathogenesis of arterial thrombus diseases. The enzyme is the strongest activator of platelets that cause and cause them to add and release substances (eg ADP TXA NE) that continue to propagate the thrombotic cycle. Platelets in a fibrin mesh comprise the main framework of a white thrombus. Thrombin also exerts direct effects on endothelial cells, causing the release of vasoconstrictor substances and the translocation of adhesion molecules that become sites for the binding of immune cells. Furthermore, the enzyme causes smooth muscle cell mitogenesis and fibroblast proliferation. From this analysis it is apparent that inhibiting thrombin 5 activity constitutes a viable therapeutic approach towards attenuation of proliterative events associated with thrombosis.

The main endogenous neutralizing factor for mammalian thrombin activity is antithrombin III (ATIII), a circulating plasma macroglobulin with little affinity for it. Heparin exerts clinical efficacy in venous thrombosis by increasing the ATIIl / thrombin binding through catalysis. However, heparin also catalyzes the inhibition of other proteases in the coagulation cascade and its efficacy in platelet-dependent thrombosis is mainly reduced due to the inaccessibility of the enzyme that binds to thrombi. Adverse side effects such as thrombocytopenia, osteoporosis, and triglyceridemia were observed after prolonged treatment with heparin.

Hirudin, derived from medicinal granular leech granular secretions, is one of the natural high molecular weight anticoagulant protein inhibitors of thrombin activity (Markwardt F.

Cardiovascular Drug Reviews, £ 1, 211, 1992). It is a biopharmaceutical whose efficacy was demonstrated in experimental and clinical thrombosis.

A potential disadvantage of the use of hirudin as a therapeutic agent is a possible antigenicity and a lack of an effective neutralization method, especially due to its extremely close binding characteristics with thrombin. The extremely high affinity for thrombin is unique and is attributed to the simultaneous interaction with the catalytic site, as well as the exosite binding to the above anions.

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

Thrombin activity can also be inhibited by low molecular weight compounds that compete with fibrinogen for the catalytic site of thrombin, thereby inhibiting proteolysis of the protein or other protein substrates such as the thrombin receptor. A common strategy for designing inhibitory compounds on top is to mimic the specificity inherent in the primary and secondary structures of the natural substrate above. Thus, Blomback et al., First designed a thrombin inhibitor modeled on the partial sequence of the A chain (LB1) a of fibrinogen that comprises its proteolytically susceptible region (Blomback, et al., J, Clin. Lab. Invest. , £ 2, 59, 1969). This fibrinogen region minimally includes residues that start with phenylalanine:

Ala-Asp-Ser-Gly-Glu-Gly ~ Asp-Phe-Leu-Ala-Glu-Gly -Gly-Gly-Val-Arg-Gly-Pro-Arg union detached you

Systematic amino acid substitution within this region resulted in optimization of the tripeptidyl inhibitory sequence exemplified by the (D) -Phe peptide

Pro-Arg that corresponds to interactions within local binding sites in thrombin (Bajusz S. et al. In Peptides: Chemistry Structure and Biology: Proceedings of the fourth American Peptide Symposium Walter R., Meienhofer J. Eds. Ann Arbor Science Publishers Inc., Ann Arbor MI, 1975, pp 603)

Bajusz et al. also reported related compounds such as (D) Phe-Pro-Arg- (CO) H (GYKI-14166) and (D) MePhe-Pro

Arg- (CO) H (GYKI-14766) (Peptides-Syñthesis, Structure and

Fuñetion:

Proceedings of the Seventh American Peptide

Symposium,

Rich, DH

Gross,

AND.

Pierce Chemical

Company, 1981, pp. 417).

These tripeptidil aldehydes are effective thrombin inhibitors both in vitro and in vivo. In the cases of

GYKI-14166 and GYKI-14766, it is presumed that the aldehyde group contributes strongly to the inhibitory in view of its chemical reactivity towards the catalytic Ser residue of thrombin, generating a hemiacetal intermediate.

Related research in the area of ​​thrombin inhibitory activity took advantage of the basic binding recognition motif engendered by the tripeptide (D) Phe-Pro-Arg, while incorporating various functional or reactive groups at the corresponding locus to the tear-off joint (ie Ρ_:-Ρ /).

In US Patent 4,318,904, Shaw reports chloromethyl ketones (PPACK) that are reactive towards Ser and His. These two residues comprise part of the catalytic triad of thrombin (Bode, W. et al., EMBO Journal 8, 3467, 1989).

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

Others with genes of this motif are those that have phosphonates (Wang, CL J., Tetrahedron Letters 33, 7667,1992) and aqueto esters (Iwanowicz, EJ et al., Bioorganic and Medicinal Chemistry Letters, 12, 1607, 1992 ).

Neises, B. et al. described a trichloromethyl ketone thrombin inhibitor (MDL-73756) and Attenburger, JM et al. revealed a related difluoroalkyl amide ketone (Tetrahedron Letters, 32, 7255, 1991).

Maraganore et al (European 0, 333, 356; WO 91/02750; US 5, 196, 404) reveal a series of thrombin inhibitors that incorporate the D-Phe-Pro- moiety and hypothesize that this preferred structure is consistent well within the sample adjacent to the active thrombin site. Variations of these inhibitors are essentially linear or cyclic peptides constructed in the D-Phe-Pro- half,

Other series of patents and patent applications described attempts to develop effective 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 other patents focused on peptides, peptide derivatives, peptide alcohols, or cyclic peptides as antithrombotic agents (WO 93/22344, EP 0276014; EP 0341607; EP 0291982). Others examined the amidine sulfonic acid halves to achieve this same purpose (US 4, 781, 866), while others examined phenylalanine derivatives derived from meta-substituted phenylalanine (WO 92/08709; WO 92/6549).

A series of Mitsubishi patents and patent applications revealed argininamide compounds apparently effective for use as antithrombotic agents. The chemical structures described in these documents represent variations of side groups of the argininamide compound (US 4,173,630; US 4,097,591; CA 1,131,621; US ​​4,096,255;

US 4,046,876; US 4,097,472; CA 2,114,153).

Cañada patent applications 2,076,311 and 2,055,850 reveal imino cyclic derivatives that have inhibitory effects on cell aggregation.

Many of the examples cited above are convergent in supporting at least one acyclic tripeptidyl linear motif consisting of an arginyl unit whose basic side chain is required for interaction with a carboxylate group located at the base of the P specificity overhang. thrombin. Two adjacent hydrophobic groups provide additional bonds through favorable Van der Waals interactions within an overhang

-1010 adjoining hydrophobic surface above designated as PP site.

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

A further object of the present invention is to provide thrombin inhibitors that exhibit inhibitory activity towards the target thrombin and that is delivered for a phacobiologically acceptable state.

Yet another objective of the present invention is to provide the use of heterocyclic thrombin inhibitors and formulas thereof as anticoagulant and thrombin inhibitory agents.

Yet another objective of the present invention is to provide the use of and formula for heterocyclic thrombin inhibitors for therapeutic treatments of various thrombotic diseases.

Still another objective of the present invention is a process for the synthesis of these low molecular weight thrombin inhibitors. The inhibitors above this

-1111 invention are encompassed by the structure of general formula 1.

SUMMARY OF THE INVENTION

The present invention provides novel compounds that have thrombin inhibitory activity as reflected in formula 1.

<img file="MX9704718A_D0002.tif" />

Where:

A. It is selected from (CH-R) S, SO, SO, O and NR where R_g is hydrogen C is alkyl optionally interrupted with 1 or 2 heteroatoms; C aryl, C cycloalkyl or heterocyclic ring or a hydrophobic group.

B. It is selected from S, SO, O, -N =, NH, -CH = and

CRR, where R₆ and R ₇, are independently selected from 1 hydrogen and are alkyl provided that A is S, SO, SO, O, or NR, and then B is CRR;

-1212

<td>D.</td><td>Is selected</td><td>of</td><td>(CH-R), where R</td><td>is</td>

<td>hydrogen, C</td><td>it is alkyl or -C (O) R; and</td><td>CH</td><td>with a double bond</td><td>with</td>

<td>B when B is</td><td>-N = or -CH =,</td><td></td><td></td><td></td>

<td>AND.</td><td>Is selected from</td><td>CH</td><td>and replaced with CH</td><td>with</td>

-C (O) R, provided that only one of D and Ξ is substituted with -C (O) R;

<td>X.</td><td>Is selected</td><td>of</td><td> 0,</td><td>NR, or</td><td>CH-R</td><td>ΐ</td><td></td>

<td>AND.</td><td>Is selected</td><td>of</td><td> 0,</td><td>s, SO,</td><td>SW,</td><td>NR and</td><td>CH-R,</td>

<td>on condition</td><td>that when X is NR,</td><td colspan="3">then Y</td><td>be</td><td>CH-R or</td><td>or, and</td>

<td>when X is</td><td>0 then let CH-R;</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Z.</td><td>Is selected</td><td>of</td><td> 0,</td><td>s and H;</td><td></td><td></td><td></td>

Rj_ is a residual arginyl moiety of a polar amino acid or analog or derivative thereof, optionally substituted with an amino acid, a peptide, or a heterocycle.

R₂ It is selected from H and C alkyl, optionally substituted with C aryl, a 6-membered heterocycle or a C-cycloalkyl ring;

R₃ It is selected from H, NR R and C alkyl; and

R₄ and ^r₅^are independently selected from H;

NR R; C aryl or C cycloalkyl optionally substituted with C

-1313 alkyl; C alkyl optionally interrupted by one or more heteroatoms or carbonyl groups and optionally substituted with OH, SH, NR R or a C aryl, heterocycle or Ccycloalkyl group optionally substituted with halogens, hydroxyl, 5 C alkyl, an amino acid side chain and a hydrophobic group.

As will be appreciated from the following disclosure, the molecules, compositions, and methods of the present invention are useful as anticoagulants, or in the treatment and prevention of various diseases attributed 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 thrombin enzyme. These molecules are characterized by a heterobicyclic moiety like the one illustrated in formula 1:

-1414

<img file="MX9704718A_D0003.tif" />

Where X, Y, Z, A, B, D, and R_r to R₄ they are like those previously defined.

The term hydrophobic group (HG) as used below, refers to any group that has no affinity for, or displaces water. Hydrophobic groups include, but are not limited to C alkyl, C alkenyl (eg, vinyl, allyl) or C alkynyl (eg, propargyl) occasionally interrupted by a carbonyl group, (eg, when forming an acyl group); C aryl, C cycloalkyl, C aralkyl, C cycloalkyl substituted with Calkyl, wherein the aliphatic portion is optionally interrupted by a carbonyl group (eg, forming an acyl group) and the ring portion is optionally substituted with C alkyl such such as methyl ethyl or t-butyl; or a hydrophobic amino acid side chain. The groups

-1515 hydrophobic include exyl, benzyl, benzoyl, phenyl methyl, phenethyl and para t-butyl-phenylmethyl.

The term "arginyl moiety" represents an arginine amino acid residue or an analogue or derivative thereof. For example, an analog or derivative of the natural residue may incorporate a longer or shorter methylene chain of the alpha carbon (eg, ethylene or butylene chains); substitution of the guanidino group with a hydrogen bond remained or accepted a group (eg amino, amidino or methoxy); replacing the methylene chain with a constrained group (an aryl, cycloalkyl, or heterocyclic ring); removal of the terminal carboxy (ie, decarboxy) or hydroxyl (eg, an aldheido); or a combination of these.

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

The term aromatic or aryl represents unsaturated carbocyclic rings of 6 to 16 carbon atoms that are optionally mono or di-substituted with OH, SH, amino

-1616 (eg NR R) halogen or C alkyl. The aromatic rings include benzene, naphthalene, phenanthrene, and anthracene. The preferred aromatic rings are benzene and naphthalene.

The term "cycloalkyl" represents a saturated carbocyclic ring of from 3 to 7 carbon atoms that are optionally mono or di-substituted with OH, SH, amino (eg NR R) halogen or C alkyl. Cycloalkyl groups include cyclopropyl, butyl, pentyl, hexyl, and heptyl. A preferred alkyl cycle group is the hexyl cycle.

The term aralkyl represents a substitute comprising an aryl moiety linked via an alkyl chain (eg benzyl, phenethyl) where the total sum of carbon atoms for the aryl moiety and the alkyl chain is as specified previously. The aryl or chain portion of the group is optionally mono or di-substituted with OH, SH, amino (eg NR R) halogen or C alkyl.

The term heteroatoms as used herein represents oxygen, nitrogen, or sulfide (O, No SO) as well as sulfoxyl or sulfonyl (SO or SO) unless otherwise indicated. It is understood that the alkyl chains

-1717 interrupted by one or more heteroatoms means that one carbon atom in the chain is replaced with a heteroatom with an appropriate valence. Preferably, an alkyl chain is interrupted by between 0 to 4 heteroatoms and that two adjacent carbon atoms are not simultaneously substituted.

The term heterocycles represents a saturated or unsaturated mono or polycyclic (eg bicyclic) ring incorporating one or more (eg 1-4) heteroatoms selected from N, O, and S. It is understood that a heterocycle is optionally mono or di- substituted with OH, SH, amino (for example NR R) halogen, CF, oxo or C alkyl. Examples of suitable monocyclic heterocyclics include, but are not limited to pyridine, piperidine, pyrazine, piperazine, pyrimidine, imidazole, thiazole, oxazole, furan, pyran, and thiophene. Examples of suitable bicyclic heterocyclics include, but are not limited to, quinoline, isoquinoline, purine, and carbazole.

The term hydrophobic amino acid represents an amino acid residue having an alkyl or aryl group

-1818 attached to the alpha-carbon atom. Therefore glycine, which does not have such a group attached to the alpha-carbon atom, is not a hydrophobic amino acid. The alkyl or aryl group may be substituted, provided that the substituent or substituents do not deviate from the general hydrophobic character of the amino acid. Examples of hydrophobic amino acids include natural amino acid residues such as alanine; isoleucine, - leucine; phenyl alanine; and non-naturally occurring amino acids, such as those described in The Peptides, vol 5, 1983, Academic Press, Chapter 6 by DC Roberts and F. Vellaccio. Suitable non-naturally occurring amino acids include cyclohexylalanine and 1-aminocyclohexane-carboxylic acid.

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

Preferably R₂ is H or C alkyl. Preferably R is H, methyl or ethyl and more preferably R is H.

-1919

Preferably R₃ is H or C alkyl. Preferably R is H methyl or ethyl and preferably R is H.

Preferably one among R₄ or R₅ is. one hydrophobic group, such as a saturated or unsaturated carbocycle with from 5 to 6 members optionally attached to another carbocyclic group, while another is H, C alkyl optionally substituted by NR R or carboxy. The hydrophobic moiety may be attached via a spacer such as a C alkyl chain optionally interrupted with one or more (eg 1-4) heteroatoms, carbonyl, or sulfonyl (SO). Preferably one among R₄ and R₅ it is phenyl, cyclohexyl, nature, thienyl, quinoline, tetrahydroisoquinoline, naphthyl or benzodioxolane

<td>linked by a C-alkyl optionally interrupted with a</td>

<td>one carbonyl hetero atom while the other is H</td>

<td>carboxymethyl or carboxy ethyl.</td>

<td>Preferably, A is absent or is CH</td>

<td>Preferably B is S or CH</td>

<td>Preferably D is CH</td>

<td>Preferably E is CH substituted with -C (O) R</td>

where R is like what was defined above.

-2020

<td>Of</td><td>preference,</td><td>X</td><td>is</td><td>CH-R</td><td>or NR</td>

<td>Of</td><td>preference,</td><td>AND</td><td>is</td><td>CH-R</td><td>or S</td>

<td>Of</td><td>preference,</td><td>z</td><td>is</td><td> 0</td><td></td>

In a preferred embodiment, R_x It is represented by one of the Via a Vid formulas:

<img file="MX9704718A_D0004.tif" />

Where:

R_T1 it is hydrogen or C alkyl;

K is a bond or -NH-;

G is C alkoxy; cyano -NH, -CH -NH; -C (NH) -NH; -NHC (NH) -NH; -CH -NH-C (NH) -NH; a cycloalkyl or aryl substituted with cyano, -NH, -CH -NH, -C (NH) -NH, -NH-C (NH) -NH or -CH -NH-C (NH) -NH; or is a 5- or 6-membered heterocycle,

-2121 saturated or unsaturated, optionally substituted with cyano, NH, -CH -NH, -C (NH) -NH, -NH-C (NH) -NH or -CH -NH-C (NH) -NH;

U is cyano, -NH, -C (NH) -NH or -NH-C (NH) -NH;

P is a bond, -C (0) - or a bivalent group

Oh

<img file="MX9704718A_D0005.tif" />

J is C alkylene optionally substituted with OH,

NH and C alkyl and optionally interrupted by a hetero atom 10 selected from O, S and N;

n is o or 1; and

T is H, OH, amino, a chain of peptides, C alkyl, C alkoxy, C aralkyl or an optionally substituted heterocycle.

Preferably R_1X it is H or methyl, and more preferably H.

K is preferably a bond

Preferably G is -NH-C (NH) -NH linked by a 3-7 carbon methylene chain or phenyl substituted with

-2222

C (NH) -NH via a methylene chain of 0 to 3 carbons. Preferably G-NH-C (NH) -NH linked by a 3-atom methylene chain.

Preferably P is -C (O) Preferably J is selected from -CH -S-CH-CH-; -CH-O-CH-CH-; -CH-NH-CH-CH-; and a bond when n is 0. Preferably J is a bond as long as n is 0.

In particular embodiments of the present invention, R is selected from the following amino acid derivatives prepared in accordance with the procedures described in Bioorg. Med. Chem., 1995, 3: 1145.

-2323

<img file="MX9704718A_D0006.tif" />

<img file="MX9704718A_D0007.tif" />

<img file="MX9704718A_D0008.tif" />

<img file="MX9704718A_D0009.tif" />

<img file="MX9704718A_D0010.tif" />

<img file="MX9704718A_D0011.tif" />

<img file="MX9704718A_D0012.tif" />

<img file="MX9704718A_D0013.tif" />

<img file="MX9704718A_D0014.tif" />

-2424 χζ

<img file="MX9704718A_D0015.tif" />

<img file="MX9704718A_D0016.tif" />

<img file="MX9704718A_D0017.tif" />

-2525

<img file="MX9704718A_D0018.tif" />

-2626

<img file="MX9704718A_D0019.tif" />

-2727

<img file="MX9704718A_D0020.tif" />

OR

<img file="MX9704718A_D0021.tif" />

NH

OR

<img file="MX9704718A_D0022.tif" />

NH

Where n = l-6, nl = l-2, n2 = 0-7 and T is like what was previously defined.

In a preferred embodiment, T is a peptide of 1 to 4 amino acid residues in length, and preferably A or B chains or fragments of fibrinogens or derivatives thereof. In another preferred embodiment, T is a heterocycle selected from the group consisting of:

-2828

<img file="MX9704718A_D0023.tif" />

Where ^ 5 '* 10' Xll Y ^ 12 are each independently selected from the group consisting of N or CX where X₇ it is hydrogen, C alkyl, or C aryl;

X₆ and ^xi3 ^sorL each independently selected from the group consisting of C, O, N, S, NX or CH-X;

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

Preferably T is selected from the group consisting of:

-2929

<img file="MX9704718A_D0024.tif" />

<img file="MX9704718A_D0025.tif" />

<img file="MX9704718A_D0026.tif" />

<img file="MX9704718A_D0027.tif" />

<img file="MX9704718A_D0028.tif" />

where R 'is as defined above.

Preferably T is selected from the group consistency

-3030

<img file="MX9704718A_D0029.tif" />

<img file="MX9704718A_D0030.tif" />

<img file="MX9704718A_D0031.tif" />

Where R is like what was defined above

Preferably T is selected from the group consisting of

<img file="MX9704718A_D0032.tif" />

where R 'is as defined above

More preferably T is

R '

<img file="MX9704718A_D0033.tif" />

<img file="MX9704718A_D0034.tif" />

Where R is H or C_x_₄ alkyl such as methyl, ethyl, propyl or butyl, and preferably where R is hydrogen. In another embodiment, T is 1,2 thiazole optionally substituted with R and / or is attached to J at position 2, 3, 4 or 5 of the ring.

-3131

In particular modalities, the compounds of the invention of the formulas II

III, IV and V, where X, Y, B,

Rl to R₄ and R₅ they are like what was defined above.

In

<img file="MX9704718A_D0035.tif" />

<img file="MX9704718A_D0036.tif" />

<img file="MX9704718A_D0037.tif" />

In a particularly preferred embodiment, the compounds of the invention are represented by one of the

<img file="MX9704718A_D0038.tif" />

(VIII)

<img file="MX9704718A_D0039.tif" />

-3232

<img file="MX9704718A_D0040.tif" />

&lt;Χ)

<img file="MX9704718A_D0041.tif" />

Where

Β is 0, S, -CH-, or -NH-;

Y is selected from 0, S, SO, SO, NR₅ and CH-R_and it is an arginyl moiety or an analogue or derivative thereof, optionally substituted with an amino acid, a peptide or a heterocycle.

R₂ it is H or C alkyl;

R₃ is selected from H, NR R and C alkyl; and

R₄ Y are independently selected from H;

NR R; C aryl or C cycloalkyl optionally substituted with C alkyl; C alkyl optionally interrupted by one or more heteroatoms or carbonyl groups, and optionally substituted with OH, SH, NR R or a C aryl, heterocycle or C-cycloalkyl group optionally substituted with halogen,

-3333 hydroxyl, C alkyl; an amino acid side chain and a hydrophobic group.

R₈ it is hydrogen, C alkyl optionally interrupted with 1 or 2 heteroatoms; C aryl, C cycloalkyl or heterocyclic ring 5 or hydrophobic group, - and n is 1 or 2.

Preferred compounds according to formula VII include:

0005 6S benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3Rcarboxamide (propyl ketoarginine)

<img file="MX9704718A_D0042.tif" />

0010 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3Rcarboxamide (butyl ketoarginine)

-3434

<img file="MX9704718A_D0043.tif" />

0015 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine 3R5 carboxamide (propylcarbomethoxy ketoarginine)

<img file="MX9704718A_D0044.tif" />

0020 SS-cyclohexylmethyl hexahydro-5-oxo-5H-thiazolo (3,2-a) pyridine-3R-carboxamide (benzylketoarginine)

<img file="MX9704718A_D0045.tif" />

-3535

0025 6S-cyclohexyl methyl hexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine -3R-caboxamide (carbomethoxyl propyl cyclodithiochelarginine)

<img file="MX9704718A_D0046.tif" />

0030 6S-cyclohexylmethyl hexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3R-carboxamide ((S) -Arg- (R) -pipecolylic acid)

<img file="MX9704718A_D0047.tif" />

0035 6S-benzylhexahydro-5-oxo-5H-thiazolo (3₁ 2-a) pyridine-3Rcarboxamido (carboxamidopropyl cyclodithioketal arginine)

-3636

<img file="MX9704718A_D0048.tif" />

0040 6S-cyclohexylmethyl hexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3R-carboxamido ((s) -Arg nipecotamide)

<img file="MX9704718A_D0049.tif" />

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

<img file="MX9704718A_D0050.tif" />

-3737

0050 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3Rcarboxamido (carboxamidopentyl cyclodithioketal arginine)

<img file="MX9704718A_D0051.tif" />

0055 6S-benzylhexahydro-5-oxo-5H-thiazolo (3,2-a) pyridine-3Rcarboxamido (carbomethoxy propyl cyclodithioketal arginine)

<img file="MX9704718A_D0052.tif" />

0060 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3, 2a) pyridine-3R-carboxamido (l-carboxy-3-thiobutyl ketoarginine)

<img file="MX9704718A_D0053.tif" />

-3838

0065 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3, 2a) pyri dina-3 R-carboxami do (1-carboxy1-3 -1 i obut i1 ketoarginine)

<img file="MX9704718A_D0054.tif" />

0070 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,

2a) pyridine-3R-carboxamido ketoarginine) (l-carboxyl-2-methyl ~ 3-thiobutyl

<img file="MX9704718A_D0055.tif" />

-3939

0075 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,

2 acid ((3-thiobutyl

<img file="MX9704718A_D0056.tif" />

<img file="MX9704718A_D0057.tif" />

0080 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo

2a) pyridine-3R-carboxamido (iso-quinolinium methyl ketoarginine)

<img file="MX9704718A_D0058.tif" />

0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3, 2a) pyridine-3R-carboxamido (propylcarbomethoxy ketoarginine)

-4040

<img file="MX9704718A_D0059.tif" />

0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo ¢ 3, 2-a) pyridine-3R-carboxamido ((propylkete) Arg-Phe-Arg-NH)

<img file="MX9704718A_D0060.tif" />

0095 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3Rcarboxamido ((propanoic acid) ketoarginine)

<img file="MX9704718A_D0061.tif" />

-4141

0100 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2-a) pyridine-3Rcarboxamido (propyl carbomethoxy ketoarginine)

<img file="MX9704718A_D0062.tif" />

0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3, 2a) pyridine-3R-carboxamido (α-benzothiazolo keto arginine); and

<img file="MX9704718A_D0063.tif" />

0110 6S-cyclohexylpropylhexahydro-5-oxo-5H-thiazolo (3, 2

a) pyridine-3R-carboxamido (propylcarbomethoxy ketoarginine)

-4242

<img file="MX9704718A_D0064.tif" />

0205 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid (1- (benzothiazol-2-carbonyl) -4-guanidinobutyl) amide

<img file="MX9704718A_D0065.tif" />

0210 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3 carboxylic acid (1- (benzothiazol-2-carbonyl) -4-guanidino butyl) amide

-4343

<img file="MX9704718A_D0066.tif" />

0215 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid (1- (benzothiazol-2-carbonyl) -4-guanidino5 butyl) amide

<img file="MX9704718A_D0067.tif" />

0220 6-benzyl-8a-methyl-5-oxo-hexahydro-thiazolo (3, 2a) pyridine-3-carboxylic acid (1- (benzothiazol-2-carbonyl) -410 guani dino-buti1-ami da

<img file="MX9704718A_D0068.tif" />

-4444

0225 8a-Methyl-5-oxo-6-phenethyl-hexahydro-thiazolo (3, 2a) pyridine-3-carboxylic acid (l-benzothiazole-2-carbonyl) -4guanidino-butyl) amide

I

<img file="MX9704718A_D0069.tif" />

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

I

<img file="MX9704718A_D0070.tif" />

0240 8a-Methyl-5-oxo-6- (2-trifluoro methyl-quiniline-6-ylmethyl) 15 hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid ¢ 1 (benzothiazole-2-carbonyl) -4 -guanidino-butyl) -amide

-4545

<img file="MX9704718A_D0071.tif" />

0245 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2-carbonyl) butyl) -amide

<img file="MX9704718A_D0072.tif" />

0250 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2-carbonyl) butyl) -amide

H

<img file="MX9704718A_D0073.tif" />

0255 6-benzyl-5-oxo-hexahydro-thiazolo (3, 2-a) pyridine-3-carboxylic acid (4-guanidino-l- (l-methyl-lH-imidazole-2carbonyl) butyl) amide

-4646

Η

<img file="MX9704718A_D0074.tif" />

0260 6-benzyl-8a-methyl-5-oxo-hexahydro-thiazolo (3,

2a) pyridine-3-carboxylic acid (4-guanidino-1- (thiazole-25 carboni1) -buti1-amide

<img file="MX9704718A_D0075.tif" />

0265 5-ΟΧΟ-6- (3-cyclohexyl-propyl) -hexahydro-thiazolo (3, 210 a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) butyl) -amide

<img file="MX9704718A_D0076.tif" />

-4747

0275 8a-Methyl-5-οχο-6- (3-phenyl-propyl) -hexahydro-thiazolo (3,

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

<img file="MX9704718A_D0077.tif" />

0280 8a-Methyl-5-oxo-6- (3-phenyl-propyl) -hexahydro-thiazolo (3,

2-a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2 ~ carbonyl) -butyl) -amide

<img file="MX9704718A_D0078.tif" />

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

-4848

<img file="MX9704718A_D0079.tif" />

0295 6- (1,3-Dioxo-l, 3-dihydro-isoindole-2-yl) -5-oxo-hexahydro5 thiazolo (3, 2-a) pyridine-3-carboxylic acid (4-guanidino-l (thiazole- 2-carbonyl) -butyl) -amide

<img file="MX9704718A_D0080.tif" />

0305 S-oxo-6- (3-phenyl-propionyl amino) -hexahydro thiazolo (3,

2-a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0081.tif" />

-4949

0315 5-οχο-β- (3-phenyl-propionyl amino) -hexahydro thiazolo (3,

2-a) pyridine-3-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0082.tif" />

Most preferred compounds according to formula (VII) include

0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,2a] pyridine-3R-carboxamido (propylcarbomethoxyketoarginine);

0090 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,2a] pyridine-3R-carboxamido (propylkete) Arg-Phe-Arg-NH);

0095 6S-benzylhexahydro-5-oxo-5H-thiazolo (3, 2 ~ a) pyridine-3Rcarboxamido ((propanoic acid) ketoarginine)

0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,215 a] pyridine-3R-carboxamido (α-benzothiazolo keto.arginine);

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

-5050

0220 6-Benzyl-8a-methyl-5-oxo-hexahydro-thiazolo (3, 2a) pyridine-3-carboxylic acid (1- (benzothiazole-2-carbonyl) -4-guanidino-butyl) -amide; 0240 8a-Methyl-5 -oxo-6- (2trifluoromethyl-quinoline-6-ylmethyl) -hexahydro-thiazolo (3, 2a) pyridine-3-carboxylic acid (1- (benzothiazole-2-carbonyl) -4-guanidino-butyl) -amide;

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

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

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

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

0315 S-oxo-6- (3-phenyl-propionylamino) -hexahydro thiazolo (3, 2a) pyridine-3-carboxylic acid (4-guanidino-1- (thiazole-2carbonyl) -butyl) -amide.

-5151

The most preferable compounds according to the formula

VII include:

0085 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,2a) pyridine-3R-carboxamido (propylcarbon methoxy ketoarginine);

and

0105 6S-cyclohexylmethylhexahydro-5-oxo-5H-thiazolo (3,2a) pyridine-3R-carboxamido (α-benzothiazolo keto arginine)

Preferred compounds according to formula VIII include:

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

<img file="MX9704718A_D0083.tif" />

-5252

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

<img file="MX9704718A_D0084.tif" />

0335 2-Benzol-4 "OXO-octahydro-pyrrolo (1, 2-a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-2-carbonyl) -butyl) -amide

<img file="MX9704718A_D0085.tif" />

0340 4-OXO-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1, 2a) pyranine-6-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0086.tif" />

-5353

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

<img file="MX9704718A_D0087.tif" />

0350 2- (3-cyclohexyl-propionyl) -4-oxo-octahydro-pyrrolo (1, 210 a) pyrazine-6-carboxylic acid (4-guanidino-l- (2-thiazolecarbonyl) -butyl) -amide

<img file="MX9704718A_D0088.tif" />

-5454

0355 5-οχο-7 - (3-phenyl-propionyl) -octahydro-2-1ia-4a, 7-diazanaphthalene-4-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0089.tif" />

0365 4-OXO-2- (4-phenyl-butyryl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0090.tif" />

NH

0370 4-oxo-2-phenylacetyl-octahydro-pyrrolo (1, 2-a) pyrazine-6 ​​carboxylic acid (4-guanidino-l- (thiazole-2-carbonyl) -butyl) amide

-5555

<img file="MX9704718A_D0091.tif" />

0375 2- (2-Amino-3-phenyl-propionyl) -4-oxo-octahydro-pyrrolo (1,

2-a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-25 carbonyl) -butyl) -amide

<img file="MX9704718A_D0092.tif" />

0380 2- (2-Amino-3- (4-hydroxy ~ phenyl) -propionyl) -4-oxooctahydro-pyrrolo (1, 2-a) pyrazine-6-carboxylic acid (410 guanidino-1- (thiazole-2-carbonyl ) -butyl) -amide

<img file="MX9704718A_D0093.tif" />

NH

-5656

0385 2- (2-Amino-3- (4-fluoro-phenyl) -propionyl) -4-oxooctahydro-pyrrolo (1, 2-a) pyrazine-6-carboxylic acid (4guanidino-1- (thiazole-2-carbonyl) -butyl) -amide

<img file="MX9704718A_D0094.tif" />

0390 4-OXO-2- (3-phenyl-propyl) -octahydro-pyrrolo (1,

2a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0095.tif" />

NH

0395 2- (2-Amino-3- (lH-indole-3-yl) -propionyl) -4-oxo-octahydropyrrolo (1, 2-a) pyrazine-6-carboxylic acid (4-guanidino-l (thiazole-2 -carbonyl) -butyl) -amide

-5757

<img file="MX9704718A_D0096.tif" />

00 4-Oxo-2- (3-thiophene-3-yl-propionyl) -octahydro-pyrrolo (1,

2-a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-25 carbonyl) -butyl) -amide

<img file="MX9704718A_D0097.tif" />

0405 4-0xo-2- (3-thiophene-2-yl-propionyl) -octahydro-pyrrolo (1,

2-a) pyrazine-6-carboxylic acid (4-guanidino-l · - (tlazole-210 carbonyl) -butyl) -amide

<img file="MX9704718A_D0098.tif" />

-5858

0410 2- (3-1 H-Imidazol-4-yl-propionyl) -4-oxo-octahydropyrrolo (1, 2-a) pyrazine-6-carboxylic acid (4-guanidino-l (thiazole-2-carbonyl) -butyl )-amide

<img file="MX9704718A_D0099.tif" />

0415 2 (2-Amino-3-thiophene-3-yl-propionyl) -4-oxo-octahydropyrrolo (l, 2-a) pyrazine-6-carboxylic acid (4-guanidino-l (thiazole-2-carbonyl) -butyl )-amide

<img file="MX9704718A_D0100.tif" />

0420 4-oxo-2- (1,2,3,4-tetrahydro-isoquinoline-3-carbonyl) octahydro-pyrrolo (1,2-a) pyrazine-6-carboxylic acid (415 guanidino-1- (thiazole-2- carbonyl) -butyl) -amide

-5959

<img file="MX9704718A_D0101.tif" />

0425 2 - (Hydroxy-phenyl-acetyl) -4-oxo-octahydro-pyrrolo (1, 25 a) pyrazine-6-carboxylic acid (4-guanidino-1- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0102.tif" />

0430 2 - (2-Hydroxy-3-phenyl-propionyl) -4-oxo-octahydro10 pyrrolo (1,2-a) pyrazine-6-carboxylic acid (4-guanidino-1 (thiazole-2-carbonyl) -butyl) - amide

<img file="MX9704718A_D0103.tif" />

-6060

0435 4-oxo-2-phenoxyacetyl-octahydro-pyrrolo (1, 2-a) pyrazine-6 ​​carboxylic acid (4-guanidino-1- (thiazole-2-carbonyl) -butyl) amide

<img file="MX9704718A_D0104.tif" />

0440 4-0xo-2- (3-phenoxy-propionyl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-2carbonyl) -butyl) -amide

<img file="MX9704718A_D0105.tif" />

0445 4-Oxo-2- (2-phenyl-etasulfonyl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-215 carbonyl) -butyl) -amide

-6161

<img file="MX9704718A_D0106.tif" />

0450 2- (Naphthalene-2-sulfonyl) -4-oxo-octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-l- (thiazole-25 carbonyl) -butyl) -amide

<img file="MX9704718A_D0107.tif" />

0455 4- (6 - (4-Guanidino-1- (thiazole-2-carbonyl) butylcarbamoyl) -4-oxo-hexahydro-pyrrolo (1, 2-a) pyrazine-2-yl) 4-ΟΧΟ-3- (2 -propyl-pentanoylamino) -butyric acid methyl ester

-6262

<img file="MX9704718A_D0108.tif" />

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

<img file="MX9704718A_D0109.tif" />

0465 4-0xo-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1_t 2a) pyrazine-6-carboxylic acid (3-guanidino-propyl) - amide

<img file="MX9704718A_D0110.tif" />

-6363

0470 4- (6- (4-Guanidino-l- (thiazole-2-carbonyl) butylcarbamoyl) -4-oxo-hexahydro-pyrrolo (1, 2-a) pyrazine 2-yl) 4-oxo-butyric acid

<img file="MX9704718A_D0111.tif" />

0475 4-0xo-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (1- (5-ethyl-thiazole-2-carbonyl) 4-guanidino-butyl) -amide

<img file="MX9704718A_D0112.tif" />

0480 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-l- (5-methylthiazole-2-carbonyl) -butyl) -amide

-6464

<img file="MX9704718A_D0113.tif" />

0485 4-οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo (1, 2a) pyrazine-6-carboxylic acid (4-guanidino-1- (4-methyl5 thiazole-2-carbonyl) -butyl) -amide

<img file="MX9704718A_D0114.tif" />

0490 4-Oxo-2- (3-phenyl-propioiiil) -octahydro-pyrrolo [1, 210 a] pyrazine-6-carboxylic acid [l- (4-ethyl-thiazole-2-carbonyl) 4-guanidino-butyl] - amide

<img file="MX9704718A_D0115.tif" />

-6565

0495 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6-carboxylic acid (4-carbamimidoyl-phenyl ·) -amide

<img file="MX9704718A_D0116.tif" />

0500 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,

2a] pyrazine-6-carboxylic acid [4-guanidino-l- (5-phenylthiazole-2-carbonyl) - butyl] -amide

<img file="MX9704718A_D0117.tif" />

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

-6666

<img file="MX9704718A_D0118.tif" />

0510 4-Οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6-carboxylic acid [1- (4-carbamimidoyl-benzyl) -25 oxo-2-thiazol-2-yl- ethyl] -amide

<img file="MX9704718A_D0119.tif" />

0515 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,210 a] pyrazine-6-carboxylic acid [1- (3-carbamimidoyl-benzyl) -2oxo-2-thiazolo-2-yl-ethyl] -amide

-6767

<img file="MX9704718A_D0120.tif" />

0520 4-οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidine5 4-ylmethyl) -2-oxo-thiazolo-2- yl-ethyl] -amide

<img file="MX9704718A_D0121.tif" />

0525 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidine10 3-ylmethyl) -2-oxo-2-thiazolo- 2-yl-ethyl] -amide

-6868

<img file="MX9704718A_D0122.tif" />

0530 4-Oxo-2- ¢ 3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (1-carbamimidoyl-piperidine5 2-yl-ethyl] -amide

<img file="MX9704718A_D0123.tif" />

0535 [6- [4-Granadino-1- (thiazole-2-carbonyl) -butylcarbamoyl] 4-OXO-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2-a] pyrazine10 3-yl] - acetic acid

<img file="MX9704718A_D0124.tif" />

nh₂

-6969

0540 3- [6- [4-Guanidino-l- (thiazole-2-carbonyl) butylcarbamoyl] -4-OXO-2- (3-phenyl-propionyl) -octahydropyrrolo [l, 2-a] pyrazine-3-yl] -propionic acid

<img file="MX9704718A_D0125.tif" />

0545 [6- [1- (l-carbamimidoyl-piperidine-4-ylmethyl) -2-oxo-2-thiazolo-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1, 2-a] pyrazine-3-yl-acetic acid

<img file="MX9704718A_D0126.tif" />

0550 3- [6- [1- (l-carbamimidoyl-piperidine-4-ylmethyl) -2-oxo-2-thiazolo-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [ 1, 2-a] pyrazin-3-yl) -acetic acid

-7070

<img file="MX9704718A_D0127.tif" />

0555 [6- [1- (l-carbamimidoyl-piperidine-3-ylmethyl) -2-οχο-2-thiazolo-2-yl-ethylcarbamoyl] -4-oxo-2- (3-phenyl-propionyl) octahydro-pyrrolo [1, 2-a] pyrazin-3-yl) -acetic acid

<img file="MX9704718A_D0128.tif" />

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

-7171

<img file="MX9704718A_D0129.tif" />

0565 3- [6- (3-Guanidino-propylcarbamoyl) -4-oxo-2 - (- phenylpropionyl) -octahydro-pyrrolo [1,

2-a] pyrazin-3-yl) -propionic acid

<img file="MX9704718A_D0130.tif" />

0570 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6-carboxylic acid [4-guanidino-l- (thiazole-210 carbonyl) -butyl] -methyl-amide

-7272

<img file="MX9704718A_D0131.tif" />

0575 4-Οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1, 2a] pyrazine-6 ​​~ carboxylic acid [1- (1-carbamimidoyl-piperidine5 4-ylmethyl) -2-oxo-2-thiazolo- 2-yl-ethyl] -methyl-amide

<img file="MX9704718A_D0132.tif" />

0580 [6- ([l-Carbamimidoyl-piperidine-4-ylmethyl) -2-oxo-2-thiazolo-2-yl-ethyl] -methyl-carbamoyl) -4-oxo-2- (3-phenyl10 propionyl) -octahydro-pyrrolo [1,2-a] pyrazin-3-yl] -acetic acid

-7373

<img file="MX9704718A_D0133.tif" />

0585 4-Οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1_F 2a] pyrazine-6- carboxylic acid [1- (1-carbamimidoylpiperidine-3-ylmethyl) -2-oxo-2-thiazolo-2-yl-ethyl] -methyl-amide

<img file="MX9704718A_D0134.tif" />

0590 4-Oxo-2- (3 -phenyl-propionyl) -octahydro-pyrrolo [1,

2a] pyrazine-6-carboxylic acid (3-guanidino-propyl) -methyl10 amide

<img file="MX9704718A_D0135.tif" />

-7474

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

<img file="MX9704718A_D0136.tif" />

NH

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

<img file="MX9704718A_D0137.tif" />

0605 2- (3-Naphthalene-l-yl-propionyl) 4-oxo-octahydro-pyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-l- (thiazole-2carbonyl) -butyl] -amide

-7575

<img file="MX9704718A_D0138.tif" />

NHj

0610 2- (4-tert-butyl-benzoyl) -4-oxo-octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-1- (thiazole-25 carbonyl) -butyl] -amide

<img file="MX9704718A_D0139.tif" />

0615 2- (Benzo [1,3] dioxoles-5-carbonyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-110 (thiazole-2-carbonyl) -butyl] -amide

-7676

<img file="MX9704718A_D0140.tif" />

0620 2- (3-Benzo [1,3] dioxol-5-yl-propionyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [4-guanidino-l5 (thiazole-2-carbonyl ) -butyl] -amide

<img file="MX9704718A_D0141.tif" />

0625 2- [2- (2-Methyl-benzylidene) -but-3-enoyl] -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (ΙΙΟ carbamimidoyl-piperidine-3-ylmethyl ) -2-oxo-2-thiazol-2-yl-ethyl] amide

-7777

<img file="MX9704718A_D0142.tif" />

0630 2- [2- (2-Methyl-benzylidene) -but-3-enoyl] -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid [1- (15 carbamimidoyl-piperidine-4-ylmethyl) ) -2-oxo-2-thiazolo-2-ylethyl] -amide

<img file="MX9704718A_D0143.tif" />

0635 2- (2-Benzylidene-pent-3-enoyl) -4-oxo-octahydropyrrolo [1,2-a] pyrazine-6-carboxylic acid (3-guanidinopropyl) -amide

-7878

<img file="MX9704718A_D0144.tif" />

0640 4-0xo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-imidazol-l-yl-l- (thiazole-25 carbonyl) -butyl] -amide

<img file="MX9704718A_D0145.tif" />

0645 4-ΟΧΟ-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2 a] pyrazine-6-carboxylic acid [4} imidazol-l-yl-l- (thiazola-2carbonyl) -butyl] -amide .

<img file="MX9704718A_D0146.tif" />

-7979

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

<img file="MX9704718A_D0147.tif" />

0655 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3- (2-amino-6-methyl-pyrimidin4-yl) -1- (thiazole- 2-carbonyl) -propyl] -amide

<img file="MX9704718A_D0148.tif" />

0670 4-ΟΧΟ-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [3 - (2-amino-6-chloro-pyrimidin4-yl) -1- (thiazole- 2-carbonyl) -propyl] -amide

-8080

<img file="MX9704718A_D0149.tif" />

0675 4-Οχο-2- {3-phenyl-propionyl) -octahydro-pyrrolo [1-2a] pyrazine-6-carboxylic acid [3- (6-amino-pyridin-2-yl) -15 (thiazole-2-carboni1 ) -prop i1] -ami da

<img file="MX9704718A_D0150.tif" />

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

-8181

<img file="MX9704718A_D0151.tif" />

0685 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [2- (2-amino-pyridin-4-yl) -15 (thiazole-2-carbonyl ) -ethyl] -amide

<img file="MX9704718A_D0152.tif" />

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

-8282

<img file="MX9704718A_D0153.tif" />

0695 2- [4-Οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) -1,2,3,45 tetrahydro -isoquinoline-6-carboxamidine

<img file="MX9704718A_D0154.tif" />

0700 2- [4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,210 a] pyrazine-6-carbonyl] -3- (thiazole-2-carbonyl) -1,2,3,4tetrahydro-isoquinoline -7-carboxamidine

<img file="MX9704718A_D0155.tif" />

-8383

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

<img file="MX9704718A_D0156.tif" />

0710 4-0XO-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-amino-cyclohexyl) -2-oxo10 2-thiazolo-2-yl- ethyl] -amide

<img file="MX9704718A_D0157.tif" />

0725 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-amino-cyclohexylmethyl) -215 oxo-2-thiazolo-2-yl- ethyl] -amide

-8484

<img file="MX9704718A_D0158.tif" />

0720 4-oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-amino-benzyl) -2-oxo-25 thiazolo-2-yl- ethyl] -amide

<img file="MX9704718A_D0159.tif" />

0725 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,210 a] pyrazine-6-carboxylic acid [1- (4-aminomethyl-benzyl) -2-oxo2-thiazolo-2-yl-ethyl] -amide

-8585

<img file="MX9704718A_D0160.tif" />

0730 4-0χο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (3-aminomethyl-benzyl) -2-oxo5 ​​2-thiazolo-2-yl- ethyl] -amide

<img file="MX9704718A_D0161.tif" />

0735 4-0xo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid (2-oxo-l-piperidine-4-ylmethyl

2-thiazolo-2-yl-ethyl) -amide

<img file="MX9704718A_D0162.tif" />

-8686

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

<img file="MX9704718A_D0163.tif" />

0745 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (3-guanidino10 cyclohexylmethyl) -2-oxo-2-thiazolo-2-yl- ethyl] -amide

<img file="MX9704718A_D0164.tif" />

0750 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [1- (4-guanidino15 cyclohexylmethyl) -2-oxo-2-thiazolo-2-yl- ethyl] -amide

-8787

<img file="MX9704718A_D0165.tif" />

0755 4-Οχο-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,25 a] pyrazine-6-carboxylic acid [1- (2-guanidinocyclohexylmethyl) -2-oxo-2-thiazolo-2-yl- ethyl] -amide

<img file="MX9704718A_D0166.tif" />

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

<img file="MX9704718A_D0167.tif" />

NH

0765 4-Oxo-2- (3-phenyl-propionyl) -octahydro-pyrrolo [1,2a] pyrazine-6-carboxylic acid [4-guanidino-l- (5-phenyl5 thiazole-2-carbonyl) -butyl] -amide

<img file="MX9704718A_D0168.tif" />

NH

0770 4-0XO-2- (3-phenyl-propionyl) -octahydro-pyrido [1,2a] pyrazine-6-carboxylic acid [4-guanidino-l- (thiazole-210 carbonyl) -butyl] -amide

<img file="MX9704718A_D0169.tif" />

-8989

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

<img file="MX9704718A_D0170.tif" />

0780 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a,

7-diazanaphthalene-4-carboxylic acid [1- (4-carbamimidoylbenzyl) -2-oxo-2-thiazolo-2-yl-ethyl] -amide or

<img file="MX9704718A_D0171.tif" />

0785 5-0xo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7diazanaphthalene-4-carboxylic acid [1- (3-carbamimidoylbenzyl) -2-oxo-2-thiazolo-2-yl-ethyl ]-amide

-9090

<img file="MX9704718A_D0172.tif" />

0790 5-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl5 piperidine-3-ylmethyl) -2-oxo-2- thiazolo-2-yl-ethyl] -amide

<img file="MX9704718A_D0173.tif" />

nh₂

0795 B-Oxo-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (1-carbamimidoyl-piperidine10 4-ylmethyl) -2-oxo-2- thiazolo-2-yl-ethyl] -amide

<img file="MX9704718A_D0174.tif" />

Λ

HjN NH

-9191

0800 [4- [4-Guanadino-1- (thiazole-2-carbonyl) - butylcarbamoyl] 5-ΟΧΟ-7- (3-phenyl-propionyl) -octahydro-2-thia-4a, 7-diazanaftalen-6-yl] -acetic acid

<img file="MX9704718A_D0175.tif" />

0805 S-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7diazanaftalen-4-carboxylic acid- [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

<img file="MX9704718A_D0176.tif" />

0810 3- [4- [4-guanidin-l- (thiazol-2-carbonylbutylcarbamoyl] -5-oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a,

7-diazanaftalen-6-yl] -propionic acid.

-9292

<img file="MX9704718A_D0177.tif" />

0815 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a_/ 75 diazanaphthalen-4-carboxylic acid [3-guanidinpropyl] -amide.

<img file="MX9704718A_D0178.tif" />

08020 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 710 diazanaftalen-4-carboxylic acid [1- (l-carbamimidolpiperidin-

3-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

<img file="MX9704718A_D0179.tif" />

-9393

0825 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid [4-guanidin-l- (hydroxythiazol-2ylmethyl) -butyl] -amide.

<img file="MX9704718A_D0180.tif" />

NHj

0830 4-0xo-2- (3-phenylpropionyl) -octahydropyrrol [1,2

a] pyrazine-6-carboxylic acid (4-guanidin-l-thiazol-2-ylmethyl) butyl] -amide.

<img file="MX9704718A_D0181.tif" />

0835 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid (4-guanidin-l-thiazol-2-ylbutyl) butyl] -amide.

-9494

<img file="MX9704718A_D0182.tif" />

0840 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid (4-methoxy-l- (thiazol-2-carbonyl) 5 butyl] -amide.

<img file="MX9704718A_D0183.tif" />

0845 [6- [4-Methoxy-l- (thiazol-2-carbonyl) -butylcarbamoyl] 10 4-OXO-2- (3-phenylpropionyl) -octahydropyrrol [1,2-a] pyrazin-3-ylacetic acetic acid.

<img file="MX9704718A_D0184.tif" />

-9595

0850 [2- (5-Methoxy-2- (| 4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazine-6-carbonyl] -amino) -pentanoi1) thiazol-5-yl] - acetic acid.

<img file="MX9704718A_D0185.tif" />

0855 4-Oxo-2- (3-phenylpropionyl) -octahydropyrolol [1,2a] pyrazine-6-carboxylic acid [4-amino-1- (thiazol-2-carbonyl) butyl] -amide.

<img file="MX9704718A_D0186.tif" />

0860 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid [5-amino-l- (thiazol-2-carbonyl) pentyl] -amide.

-9696

<img file="MX9704718A_D0187.tif" />

0865 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,25 a] pyrazine-6-carboxylic acid [5-guanidin-l- (thiazol-2carbonyl) -pentyl] -amide.

<img file="MX9704718A_D0188.tif" />

0870 2- (3-Naphthalen-2-yl-propionyl) -4-oxo-octahydropyrrol [1,2-a] pyrazine-6-carboxylic acid [4-guanidin-l (thiazol-2-carbonyl) -butyl] -amide .

-9797

<img file="MX9704718A_D0189.tif" />

0875 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-5-carboxylic acid [4-guanidin-l- (methyl-lH5 imidazol-2-carbonyl) -butyl] -amide.

<img file="MX9704718A_D0190.tif" />

NH,

0880 4-0XO-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid [4-guanidin-1- (thiazol-210 carbonyl) -butyl] -amide.

<img file="MX9704718A_D0191.tif" />

-9898

0885 8,8-Dimethyl-4-oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2-a] pyrazine-6-carboxylic acid [4-guanidin-1 (thiazol-2-carbonyl) -butyl] -amide .

<img file="MX9704718A_D0192.tif" />

Preferred compounds according to Formula (VIII) include:

0325 3-Aminomethyl-2-benzoyl-4-oxo-octahydropyrrol [1,2a] pyridine-6-carboxylic acid [1- (benzothiazol-2-carbonyl) guanidinbutyl] -amide.

0330 3-Aminomethyl-4-oxo-phenylacetyl-octahydropyrrol [1,215 a] pyridine-6-carboxylic acid [1- (benzothiazol-2-carbonyl) guanidinbutyl] -amide.

-9999

0515 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid [1- (3-carbamimidoylbenzyl) -2oxo-2-thiazol-2-ylethyl] -amide.

0530 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazine-6-carboxylic acid [1- (l-carbamimidoylpiperidin-2ylmethyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

0545 [6- [1- (l-Carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylthylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin-3- il) -acetic acid.

0550 3- | 6 - [1- (l-Carbamimidoylpiperidin-4-ylmethyl) -2-oxo2-thiazol-2-ylthylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin- 3-yl) -acetic acid.

0555 [6- [1- (l-Carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ylethylcarbamoyl] -4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin-3- il) -acetic acid.

-100100

0560 [6- (3-Guanidinpropylcarbamoyl) -4-oxo-2- (3phenylpropionyl) -octahydropyrrol [1,2-a] pyrazin-3-yl) -acetic acid.

0565 3- | 6- (3-Guanidinpropylcarbamoyl) -4-oxo-2- (3phenylpropionyl) -octahydropyrrol [1,2-a] pyrazin-3-yl) -propionic acid.

0575 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid | 1- (l-carbamimidoyl-piperidin-4ylmethyl) -2-oxo-2-thiazol-2-ylmethyl] - methylamide.

0580 [6 - ([l-Carbamimidoyl-piperidin-4-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] -methylcarbamoyl) -4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin -6-acetic acid.

0585 4-0ΧΟ-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (1-carbamimidoyl-piperidin-3ylmethyl) -2-oxo-2-thiazol-2-ylmethyl] - methylamide.

-101101

0590 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid (3-guanidinpropyl) -methylamide.

0595 2- (Naphthalen-2-carbonyl) -4-oxo-octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

0625 2- [2- (2-Methylbenzylidene) -but-3-enoyl] -4-oxooctahydropyrrol [1,2-a] pyrazin-6-carboxylic acid [1- (1carbamimidoyl-piperidin-3-ylmethyl) -2-oxo -thiazol-2-ylethyl] amide.

0630 2- [2- (2-Methylbenzylidene) -but-3-enoyl] -4-oxooctahydropyrrol [1,2-a] pyrazin-6-carboxylic acid [1- (1carbamimidoyl-piperidin-4-ylmethyl) -2-oxo -thiazol-2-ylethyl] amide.

0635 2- (2-Benzyliden-pent-3-enoyl) -4-oxo-octahydropyrrol [1,2-a] pyrazin-6-carboxylic acid (guanidinpropyl) amide.

-102102

0625 2- [2- (2-Methylbenzylidene) -but-3-eneyl] -4-oxooctahydropyrrol [1,2-a] pyrazin-6-carboxylic acid [1- (1carbamimidoyl-piperidin-3-ylmethyl) -2-oxo -thiazol-2-ylethyl] 5 amide.

0630 2- [2- (2-Methylbenzylidene) -but-3-enoyl] -4-oxooctahydropyrrol [1,2-a] pyrazin-6-carboxylic acid [1- (1carbamimidoyl-piperidin-4-ylmethyl) -2-oxo -thiazol-2-ylethyl] 10 amide.

0635 2- (2-Benzylidene-pent-3-enoyl) -4-oxo-octahydropyrrole [1,2-a] pyrazin-6-carboxylic acid (guanidinpropyl) amide.

0645 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-imidazol-l-yl-l- (thiazol-2carbonyl) -butyl] -amide.

-103103

0670 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [3- (2-amino-6-chloro-pyrimidin-4yl) -1- (thiazol-2-carbonyl) -propyl] -amide.

0675 4-0XO-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [3- (6-aminopyridin-2-yl) -1 (thiazol-2-carbonyl) -propyl] -amide .

0680 4-0xo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [3- (2-aminopyridin-4-yl) -1 (thiazol-2-carbonyl) -propyl] -amide .

0685 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [2- (2-aminopyridin-4-yl) -1 (thiazol-2-carbonyl) -ethyl] -amide .

0690 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [2- (6-aminopyridin-2-yl) -1 (thiazol-2-carbonyl) -ethyl] -amide .

-104104

0695 2- [4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carbonyl] -3- (thiazol-2-carbonyl) -1,2,3,4tetrahydro-isoquinolin-6 - carboxamidine.

0700 2- [4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carbonyl] -3- (tlazol-2-carbonyl) -1,2,3,4t et rahidro-i soqui no1i n-7 -carboxami dina.

0705 N- [1- [4-Oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin-6-carbonyl] -5- (thiazol-2carbonyl) -pyrrolidin-3-yl] -guanidine.

0710 4-0ΧΟ-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (4-aminocyclohexyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

0730 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (3-aminomethylbenzyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

-105105

0745 4-0ΧΟ-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (3-guanidincyclohexylmethyl) 2-oxo-2-thiazol-2-ylethyl] -amide.

0755 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (2-guanidincyclohexylmethyl) 2-oxo-2-thiazol-2-ylethyl] -amide.

0795 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 710 diazanaphthalen-4-carboxylic acid [1- (1-carbamimidoylpiperidin-4-ylmethyl) -2-oxo-2-thiazol-2- iletil] -amide.

0800 [4- [4-Guanidin-l- (thiazol-2-carbonyl) butylcarbamoyl) -5-oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 15 7-diazanaftalen-6-yl] - acetic acid.

0810 3- [4- [4-Guanidin-1- (thiazol-2-carbonyl) -butylcarbamoyl) -5-ΟΧΟ-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7diazanaftalen-6-yl] - acetic acid.

-106106

0815 5-Οχο-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7diazanaphthalen-4-carboxylic acid [3-guanidinpropyl] -amide.

0820 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 75 diazanaftalen-4-carboxylic acid [1- (1-carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2- iletil] -amide.

0830 4-Oxo-7- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [(4-guanidin-l-thiazol-210 ylmethylbutyl) -amide.

0835 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l-thiazol-2ylmethylbutyl] -amide.

More preferably the compounds according to the

Formula VIII include:

-107107

0335 2-Benzoyl-4-οχο-2-octahydropyrrol [1,2-a] pyrazin-6 carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) butyl] amide.

0650 4-0xo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4- (2-aminoimidazol-l-yl) -l (thiazol-2-carbonyl) -butyl] amide.

0655 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [3- (2-amino-6-methylpyrimidin-4yl) -1- (thiazol-2-carbonyl) -propyl ]amide.

0715 4-0xo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (4-amino-cyclohexylmethyl) -2oxo-2-thiazol-2-ylethyl] -amide.

0720 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (4-aminobenzyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

-108108

0725 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (4-aminomethylbenzyl) -2-oxo-2-thiazol-2-ylethyl] -amide.

0735 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid (2-oxo-l-piperidin-4-ylmethyl-2-thiazol-2-ylethyl] -amide.

0740 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid (2-oxo-l-piperidin-3-yl-2-thiazol-2-ylethyl] -amide.

0750 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,215 a] pyrazin-6-carboxylic acid [1- (4-guanidincyclohexylmethyl) 2-oxo-2-thiazol-2-ylethyl] -amide.

0760 4-0XO-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [1- (5-benzyl-thiazol-2-carbonyl) 20 4-guanidinbutyl] -amide.

-109109

0765 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l- (5-phenylthiazol-2carbonyl) -butyl] -amide.

0770 4-0XO-2- (3-phenylpropionyl) -octahydropyrido [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

0775 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

0780 S-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (4-carbamimidoylbenzyl) 2-oxo-2-thiazol-2-ethyl] -amide.

0785 S-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7diazanaphthalene-4-carboxylic acid [1- (3-carbamimidoylbenzyl) 2-oxo-2-thiazol-2-ylethyl] -amide.

-110110

0790 S-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7-diazanaphthalene-4-carboxylic acid [1- (1carbamimidoylpiperidin-3-ylmethyl) -2-oxo-2-thiazol-2-ylethyl] 5 amide.

0805 5-Oxo-7- (3-phenylpropionyl) -octahydro-2-thia-4a, 7diazanaphthalene-4-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) butyl] -amide.

0825 4-0ΧΟ-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l- (hydroxythiazol-2ylmethyl) -butyl] -amide.

0840 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-methoxy-l- (thiazol-2-carbonyl) butyl] -amide.

-111111

0845 [6- [4-Methoxy-l- (thiazol-2-carbonyl) -butylcarbamoyl] -

4-oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2-a] pyrazin-3yl] -acetic acid.

0850 [2- [5-Methoxy-2 - ([4-oxo-2- (3-phenylpropionyl) octahydropyrrol [1,2-a] pyrazin-6-carbonyl] -amino) -pentanoyl) thiazol-5-yl] - acetic acid.

0855 4-Oxo-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-amino-l- (thiazol-2-carbonyl) butyl] -amide.

0860 4-Oxo-2 - (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [5-amino-l- (thiazol-2-carbonyl) pentyl] -amide.

0865 4-Oxo-2 - (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [5-guanidin-l- (thiazol-2 carbonyl) -pentyl] -amide.

-112112

The most preferred compounds according to

Formula VIII include:

0345 4-0XO-2- (3-phenylpropionyl) -octahydropyrrol [1,2a] pyrazin-6-carboxylic acid [4-guanidin-l- (5-methylthiazol-25 carbonyl) -butyl] -amide; and

0340 4-oxo-2- (3-phenylpropionyl) -octahydropyrrole [1,2-

a] pyrazin-6-carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) -butyl] -amide.

Compounds according to Formula IX include:

0890 3-Amino-4-oxo-2-phenylhexahydropyrrol [2,1-

b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazol-2- * carbonyl) -4-guanidinbutyl] -amide.

<img file="MX9704718A_D0193.tif" />

-113113

0895 3-Amino-2-benzyl-4-oxo-hexahydropyrrol [2,1b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazol-2carbonyl) -4-guanidinbutyl] -amide.

<img file="MX9704718A_D0194.tif" />

0900 3-Amino-2-cyclohexyl-4-oxo-hexahydropyrrol [2,1b] [1,3] thiazine-6-carboxylic acid [1- (benzothiazol-2carbonyl) -4-guanidinbutyl] -amide.

<img file="MX9704718A_D0195.tif" />

Preferred compounds in accordance with

Formula X include:

-114114

0905 7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4 carboxylic acid [1- (benzothiazol-2-carbonyl) -4guanidinbutyl] -amide.

<img file="MX9704718A_D0196.tif" />

<img file="MX9704718A_D0197.tif" />

0910 7- (Terbutylbenzyl) -6-oxo-octahydropyrido [2, Ιο] [1,4] thiazine-4-carboxylic acid [1- (benzothiazol-210 carbonyl) -4-guanidinbutyl] -amide.

<img file="MX9704718A_D0198.tif" />

<img file="MX9704718A_D0199.tif" />

0915 6-Oxo-octahydropyrido [2,1-c] [1,4] thiazine-4-carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) -butyl] -amide.

-115115

<img file="MX9704718A_D0200.tif" />

0925 7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4 carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) -butyl] amide.

<img file="MX9704718A_D0201.tif" />

0935

7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4 carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) -butyl] amide.

<img file="MX9704718A_D0202.tif" />

-116116

0940 6-Oxo-7-phenethyl-octahydropyrido [2,1-c] [1,4] thiazine4-carboxylic acid [4-guanidin-1- (thiazol-2-carbonyl) -butyl] amide.

<img file="MX9704718A_D0203.tif" />

0950 7-Benzyl-2,2,6-trioxo-octahydro-21&gt; 6_pirido [2,1-

c] [1,4] thiazine-4-carboxylic acid [4-guanidin-1- (thiazol-2-carbonyl) -butyl] -amide.

<img file="MX9704718A_D0204.tif" />

Among the most preferred compounds according to the Formula

X include:

-117117

925 7-Benzyl-6-oxo-octahydropyrido [2,1-c] [1,4] thiazine-4 carboxylic acid [4-guanidin-l- (thiazol-2-carbonyl) -butyl] amide; and 940 6-Oxo-7-phenethyl-octahydropyrido [2.15 c] [1,4] thiazine-4-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

Preferred compounds according to the Formula

III include:

0960 4-Oxo-l- (3-phenylpropionyl) -octahydropyrrol [1,2] pyrimidin-6-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

<img file="MX9704718A_D0205.tif" />

-118118

0965 4-Oxo-l- (3-phenethylsulfonyl) -octahydropyrrol [1,2] pyrimidin-6-carboxylic acid [4-guanidin-l- (thiazol-2carbonyl) -butyl] -amide.

<img file="MX9704718A_D0206.tif" />

J = NH

M₃N

For the preparation of the compounds of Formula (VII) various methods can be used, depending on the particular starting materials and / or intermediates that are used. The following scheme is a particular method of:

-119119

SC HEME 1

<img file="MX9704718A_D0207.tif" />

STEP-1

-------------&gt;

<img file="MX9704718A_D0208.tif" />

S1EP-2

<img file="MX9704718A_D0209.tif" />

<img file="MX9704718A_D0210.tif" />

<img file="MX9704718A_D0211.tif" />

-120120

Step 1:

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

Step 2 j

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

Step 3:

Formation of the adduct £ of the aldehyde £ with d is carried out by stirring the reagent in aromatic solvents, for example benzene or toluene, in the presence of a suitable catalytic amount of acid, for example ptoluenesulfonic acid.

Step 3':

-121121

The interconversion of the aldehyde £ to the aldehyde g is easily carried out by means of the appropriate protection and deprotection protocols that appear in T. Greene, Protective Groups in Organic Synthesis (John Wiley &amp; Sons, 5 1981).

Step 4:

Cyclization of the £ to £ adduct can be easily accomplished by appropriate Lewis acids, eg, trimethylaluminum in suitable solvents, eg, dichloromethane, where the methodology appears in T. Greene, supra.

Step 4¹:

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

Step 5:

The ester function (-C (O) OR) of the bicyclic intermediate of formula £ is then hydrolyzed using an appropriate agent such as HC1 in an appropriate solvent, such as ethyl ether to produce the free carboxylic acid. The resulting compound is then coupled

-122122 with RiH with a peptide coupling agent, such as BOP in an appropriate solvent such as DMF to produce a coupled bicyclic compound of Formula (VIII). The conditions suitable for the formation of peptide bonds are well known 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, Biologv, Volume 1, edited by Gross E. and Meienhofer J., Academic Press, New York, San 10 Franc is co, London s, 1979.

Various methods may be used for the preparation of the compounds of Formula (VIII), depending on the particular starting and / or intermediate materials involved. The following scheme is a particular method of preparation.

-123123

SCHEME 2

<img file="MX9704718A_D0212.tif" />

<img file="MX9704718A_D0213.tif" />

S1EP 1

Oh

<img file="MX9704718A_D0214.tif" />

O ba

<img file="MX9704718A_D0215.tif" />

-124124 where:

pg is a nitrogen protecting group;

^R2nd and ^R2i ^are independently a C alkyl; and x, R_lf R₃, R₄ and R₅ they are as defined above.

The process in Scheme 2 is briefly described as follows:

STEP 1

The amino and carboxylic functions of the unsaturated compound in formula (a) are protected with appropriate protecting groups. A variety of protection groups known as reactive functional groups and suitable protection and deprotection protocols can be found in T. Greene, Protective Groups in Organic Synthesis. (John Wiley &amp; Sons, 1981). The appropriate protecting group that can be used in a particular synthesis scheme will depend on many factors, including the presence of other reactive functional groups and the desired reaction conditions for removal. The unsaturated compound of the formula is easily obtained by and protocols known to the chemist

-125125 knowledgeable in the art. The protected unsaturated compound of formula (a) is subjected to the appropriate conditions to allow cyclization using an appropriate reagent such as mercury acetate in an inert solvent, such as tetrahydrofuran (THF) to produce a protected amino alcohol of formula ( b).

STEP 2.

The protected amino alcohol of formula (b) is oxidized using an appropriate oxidizing trioxide agent, such as the sulfur complex in a suitable solvent, such as dichloromethane, dimethylformamide to produce an aminoaldehyde of formula (c). Alternatively, the intermediate (C) may of a compound of formula (a ') prepared according to Collado et a,

J.

Org. Chem., 1995, 60: 5011.

STEP 1 with a forming

The protected aminoaldehyde of formula (c) amino acid alkyl ester of formula (d) the imine followed by contacting is coupled, first the imine obtained with an appropriate reagent, such as borohydride of

-126126 sodium triacetoxy NaBH (OAc) to produce a cyclic intermediate of formula (e).

STEP 4

The cyclic intermediate of formula (e) is functionalized in the amino position to produce the amino substituted cyclic intermediate of formula (f). The appropriate conditions for these reactions are well known in the art, and will depend on the nature of the R substitute.₅.

STEP 5

The amino protecting group of the cyclic intermediate of formula (f) is removed under appropriate conditions, and the resulting compound is then subjected to an appropriate condition for internal ring closure such as low temperature in an inert solvent, or as a primary compound to produce a bicyclic intermediate of formula (g). The bicyclic intermediate of formula (g) can also be obtained by hydrolyzing the ester function (-C (O) OR₂₀) of the cyclic intermediate of formula (g) with the free carboxylic acid, followed by a conventional peptide coupling using a coupling reagent such as benzotriazol-l-yloxy-tris- (dimethylamino) phosphonium hexafluoro

-127127 phosphate (BOP) in an inert solvent such as dimethylformamide (DMF).

STEP 6.

The ester function (~ C (O) O ~ R₂₁) of the cyclic intermediate of formula (g) is then hydrolyzed using an appropriate agent, such as HC1, in an appropriate solvent such as ethyl ether to produce the free carboxylic acid. The resulting compound is then coupled to R₂H with a peptide coupling agent, such as BOP, in an appropriate solvent, such as DMF, to produce a coupled bicyclic compound of Formula (VIII). Suitable conditions for the formation of peptide bonds are well known 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, Analvsis, Synthesis. Biology, Volume 1, edited by Gross E. and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

For the preparation of the compounds of Formula (IX) various methods can be used, depending on the

-128128 initial and / or particular intermediate materials that are involved.

-129129

<img file="MX9704718A_D0216.tif" />

SCHEME 3

SCHEME 3

<img file="MX9704718A_D0217.tif" />

<img file="MX9704718A_D0218.tif" />

b

<img file="MX9704718A_D0219.tif" />

Y = S-Pg

NH-Pg

<img file="MX9704718A_D0220.tif" />

and

<img file="MX9704718A_D0221.tif" />

<img file="MX9704718A_D0222.tif" />

d where:

Pg is a sulfur or amino protecting group;

L is an outgoing group;

-130130

R₂₀ AND ^R2i ^are independently a C alkyl; and R_x yr_3/ R₄ and ^Rs are as defined above.

The process shown in Scheme 3 is briefly described as follows:

STEP 1

A carboxylic acid compound (a) is coupled to the cyclic amino compound with a peptide coupling agent such as benzotriazol-l-yloxy-tris- (dimethylamino) phosphonium hexafluoro-phosphate (BOP reagent) in the presence of a base such as n-methylmorpholine in an appropriate solvent such as dimethylfor-mamida (DMF) or dichloromethane (DCM) to produce an amido compound of formula (c). Suitable conditions for the formation of peptide bonds are known in the art of peptide chemistry. For example see, Principies of Peptide Synthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis .__ Biology, Volume 1, edited by Gross E. and

-131131

Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

STEP 2

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

STEP 3

The ester function (-C (O) -R) of the bicyclic intermediate of formula (d) is subjected to hydrolysis using an appropriate agent, such as lithium hydroxide (LiOH) in an appropriate solvent, such as tetrahydrofuran (THF) to produce carboxylic acid. The resulting compound is then coupled to R_XH with a peptide coupling agent, such as BOP in an appropriate solvent such as DMF, to produce compound (e). Suitable conditions for the formation of peptide bonds are known in the art of peptide chemistry. For example, see Principles of

-132132

Peptide Synthesis, Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, Biology, Volume 1, edited by Gross E. and Meienhofer J., Academic Press, New York, San Francisco, 5 London, 1979.

Various methods may be used for the preparation of the compounds of Formula (X), depending on the particular starting and / or intermediate materials involved in the preparation. Scheme 4 below is a particular method of preparation.

-133133

SCHEME 4

<img file="MX9704718A_D0223.tif" />

-134134 where:

Rjg and R₂₁ they are independently a C alkyl; and B, R_x, R₃, R₄ and R_s they are as previously defined.

The process shown in Scheme 4 is briefly described as follows:

STEP 1

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

STEP 2

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

-135135

PASQ-1

The cyclic intermediate of formula (d) is treated under acidic conditions using an appropriate acid, such as camphor sulfonic acid in an appropriate solvent such as toluene to produce a bicyclic intermediate of formula (e).

PASPA

The ester function (-C (O) OR₂₀) of the bicyclic intermediate of formula (e) is hydrolyzed using an appropriate reagent such as LiOH to produce the free carboxylic acid. The resulting compound is then coupled with R₁H with a peptide coupling agent such as BOP in an appropriate solvent such as dimethylformamide to produce a coupled bicyclic compound of formula (X). Suitable conditions for the formation of peptide bonds are known in the art of peptide chemistry. For example, see Principles of Peptide Svnthesis. Bodanszky M., Springer-Verlag, Berlin, Heidelberg, New York, Tokyo 1984; and The Peptides, Analysis, Synthesis, __ BíolQgy, Volume 1, edited by Gross E. and Meienhofer J., Academic Press, New York, San Francisco, London, 1979.

-136136

The compounds of the present invention are further characterized by their ability to inhibit the catalytic activity of thrombin, which is demonstrated in the following assay. The compounds of the present invention can be prepared for testing by dissolving in a buffer whose concentrations vary from 1 to 100 µΜ. In an assay to determine the constant inhibitory dissociation, K, for a given compound, a fluorogenic or chromogenic thrombin substrate was added to a solution containing a test compound and thrombin; the resulting catalytic activity of the enzyme was determined spectrophotometrically. This type of test is familiar to those skilled in the art.

The compounds of the present invention can be used as anticoagulants in vitro or ex vivo, as in the case of activation by contact with foreign thrombogenic surfaces, such as those that appear in tubes used in extracorporeal probes. The compounds of the present invention can also be used to coat the surface of these thrombogenic ducts. For this end,

-137137 The compounds of the invention are obtained as lyophilized powders, redissolved in isotonic saline and added in sufficient quantity to keep the blood in an anticoagulated state.

The pharmaceutically acceptable therapeutic agents of the present invention. The ratio of each vehicle is determined by the chemical nature of the compound, the route of administration, and conventional pharmaceutical practice.

The compounds can be injected parentherically;

This can be intramuscular, intravenous or subcutaneous. For parenteral administration, the compound in the form of a sterile solution containing other solutes, eg, enough saline or glucose to make it isotonic. The compounds can be administered orally in the form of tablets, capsules, or granules containing suitable excipients, such as starch, lactose, white sugar, and the like. The compounds can also be administered sublingually in tablet form, where each active ingredient is mixed with sugar syrup or

-138138 corn, flavoring agents and dyes, and then dehydrated enough to make the mixture suitable for compressing into solid form. The compounds can be administered orally in the form of solutions containing coloring and / or flavoring agents.

Physicians will determine the dosage of the present therapeutic agents that are most suitable. Doses may vary with the mode of administration and the particular compound chosen. Furthermore, the dose may vary with the particular patient under treatment.

When the composition is administered orally, typically a larger amount of active agent will be required to produce the same effect caused by a smaller amount administered parenterally.

To further aid in understanding the present invention, the following non-limiting examples of thrombin inhibitory compounds are provided. Of course, the following examples are not to be construed as specifically limiting the present invention, and variations which are known or developed herein.

-139139 thereafter, which would be within the scope of the person skilled in the art, and are regarded as being within the present invention as described herein. The preferred compound of the present invention is synthesized using conventional preparation steps and recovery methods familiar to those known in the art of organic and bioorganic synthesis, while providing a new and one-of-a-kind combination for general synthesis. of each compound. The preferred synthetic pathways for the intermediates involved in the synthesis, as well as the resulting antithrombotic compounds of the present invention are set out below.

EXAMPLE 1

<img file="MX9704718A_D0224.tif" />

<img file="MX9704718A_D0225.tif" />

-140140

A solution of tert-butoxycarbonyl-iodo-alanineN, O-dimethylamide (2.68 grams, 7.5 millimoles) (J. Org. Chem.

1992, 57, 3397-3404) in dry benzene (30 milliliters), and dry N, N ~ dimethylacetamide (2.0 milliliters) were added to a nitrogen purged dry round flask charged with zinc-copper coupling (0.90 grams). The resulting mixture was sonicated under nitrogen until no starting material remained (judged by TLC). Bis (trio-tolylphosphine) palladium dichloride (0.35 grams, 0.40 millimoles) was added followed by 4-iodobenzonitrile (1.72 grams, 7.5 millimoles). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 milliliters) was added, and the mixture was filtered through a separating funnel. A subsequent wash with aqueous HC1 (50 milliliters; 0.1N), H₂Or distilled (3 x 50 milliliters), dried in Na₂S0₄Filtration and concentration under reduced pressure produced the primary product. Gel chromatography

-141141 Silica (light petroleum-ethyl acetate gradient) allowed the purified compound.

<img file="MX9704718A_D0226.tif" />

<img file="MX9704718A_D0227.tif" />

A solution of tert-butoxycarbonyl-iodo-alanineN, O-dimethylamide (2.68 grams, 7.5 millimoles) (J. Org. Chem. 1992, 57, 3397-3404) in dry benzene (30 milliliters), and N, Ndimethylacetamide dry (2.0 milliliters ) were added to a nitrogen purged dry round flask charged with zinc-copper coupling (0.90 grams). The resulting mixture was sonicated under nitrogen until no starting material remained (judged by TLC). Bis (trio-tolylphosphine) palladium dichloride (0.35 grams, 0.40 millimoles) was added followed by 3-iodobenzonitrile (1.72 grams, 7.5 millimoles). Mix

The resulting -142142 was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 milliliters) was added, and the mixture was filtered through a separating funnel. A subsequent wash with aqueous HC1 (50 5 milliliters; 0.IN), H₂Or distilled (3 x 50 milliliters), dried in Na₂SW₄Filtration and concentration under reduced pressure produced the primary product. Chromatography with silica gel (light petroleum-ethyl acetate gradient) allowed the purified compound.

<img file="MX9704718A_D0228.tif" />

<img file="MX9704718A_D0229.tif" />

.och₃

I ch₃

A solution of tert-butoxycarbonyl-iodo-alanineN, O-dimethylamide (2.68 grams, 7.5 millimoles) (J. Org. Chem.

1992, 57, 3397-3404) in dry benzene (30 milliliters), and dry N, Ndimethylacetamide (2.0 milliliters) were added to a

-143143 nitrogen purged dry round flask, charged with zinc-copper coupling (0.90 grams). The resulting mixture was sonicated under nitrogen until no starting material remained (judged by TLC). Bis (tri5 o-tolylphosphine) palladium dichloride (0.35 grams, 0.40 millimoles) was added followed by 2-iodobenzonitrile (1.72 grams, 7.5 millimoles). The resulting mixture was stirred under a nitrogen atmosphere with heating, allowed to cool, ethyl acetate (100 milliliters) was added, and the mixture was filtered through a separating funnel. A subsequent wash with aqueous HC1 (50 milliliters; 0.IN), H₂0 distilled (3 x 50 milliliters), dried in Na₂SW₄Filtration and concentration under reduced pressure produced the primary product. Chromatography with silica gel (light petroleum-ethyl acetate gradient) 15 allowed the purified compound.

-144144

<img file="MX9704718A_D0230.tif" />

<img file="MX9704718A_D0231.tif" />

^ och₃ hl

I CH₃

To a solution of tert-butoxycarbonyl-para5-cyanophenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimoles) in dry ethanol (20 milliliters) was added hydrochloride and hydroxylamine (0.416 grams, 6.0 millimoles), and diisopropylethylamine (1.02 milliliters, 6.0 millimoles) . The mixture was refluxed and then cooled. The precipitate was filtered off, washed with cold ethanol, diisopropylether, dried with MgSO₄, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 milliliters) and dry ethanol (40 milliliters) with heating. Subsequently, Pd / C catalyst (0.30 grams, 10% Pd) was added and hydrogen bubbles were passed through the mixture with

-145145 heating. Hydrogenation was continued until starting material could not be detected, according to TLC. The catalyst was removed by filtration, the solution was concentrated, under reduced pressure (50 milliliters), HC1 (50 5 ml, 1N) was added, and the mixture was concentrated again to 50 milliliters. The solution was cooled overnight, yielding the title compound.

<img file="MX9704718A_D0232.tif" />

To a solution of tert-butoxycarbonyl-meta-cyanophenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimoles) in dry ethanol (20 milliliters) was added hydroxylamine hydrochloride (0.416 grams, 6.0 millimoles), and diisopropylethylamine (1.02 milliliters, 6.0 millimoles) ). The

-146146 mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO₄, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 milliliters) and dry ethanol (40 milliliters) with heating. Subsequently, Pd / C catalyst (0.30 grams, 10% Pd) was added and hydrogen bubbles were passed through the mixture with heating. Hydrogenation was continued until starting material could not be detected, according to TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 milliliters), HC1 (50 milliliters, 1N) was added, and the mixture was concentrated again to 50 milliliters. The solution was cooled overnight, yielding the title compound.

-147147

<img file="MX9704718A_D0233.tif" />

To a solution of tert-butoxycarbonyl-ortho-cyano5 phenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimoles) in dry ethanol (20 milliliters) was added hydroxylamine hydrochloride (0.416 grams, 6.0 millimoles), and diisopropylethylamine (1.02 milliliters, 6.0 millimoles). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO₄, concentrated under reduced pressure, and used directly in the next step. The semi-solid was suspended in a mixture of acetic acid (20 milliliters) and dry ethanol (40 milliliters) with heating. Subsequently, 15 Pd / C catalyst (0.30 grams, 10% Pd) was added and bubbles of

-148148 hydrogen through the mixture with heating. Hydrogenation was continued until starting material could not be detected, according to TLC. The catalyst was removed by filtration, the solution was concentrated under reduced pressure (50 5 milliliters), HC1 (50 milliliters, 1N) was added, and the mixture was again concentrated to 50 milliliters. The solution was cooled overnight, yielding the title compound.

<img file="MX9704718A_D0234.tif" />

<img file="MX9704718A_D0235.tif" />

To a solution of thiazole (1.28 grams, 15.0 millimoles) in anhydrous THF (3.0 milliliters), n-BuLi (1.6 M / hexane, 8.9 milliliters, 13.9 millimoles) was added dropwise at -78 ° C, and the solution was stirred . Then tert-butoxycarbonyl-para-amidino-phenylalanine-N, 0-dimethylamide (1.15 grams, 3.3 mmol) in THF (15 milliliters) was added dropwise, and the resulting mixture was stirred. The reaction was quenched with chloride

-14914 9 saturated aqueous ammonia. The mixture was diluted with ethyl acetate (150 milliliters), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 milliliters), brine (50 milliliters), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0236.tif" />

millimoles) in anhydrous THF (30 milliliters) n-BuLi (1.6 M / hexane, 8.9 milliliters, 13.9 millimoles) was added dropwise at -78 ° C, and the solution was stirred. Then tert-butoxycarbonyl-meta-amidino-phenylalanine-N, O-dimethylamide (1.15 grams, 3.3 mmol) in THF (15 milliliters) was added dropwise, and the mixture was stirred

-150150 resulting mixture. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 milliliters), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 milliliters), brine (50 5 milliliters)_t dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0237.tif" />

<img file="MX9704718A_D0238.tif" />

To a solution of tlazole (1.28 grams, 15.0 millimoles) in anhydrous THF (30 milliliters), n-BuLi (1.6 M / hexane, 8.9 milliliters, 13.9 millimoles) was added dropwise at -78 ° C, and the solution was stirred. Then tert-butoxy-151151 carbonyl-ortho-amidino-phenylalanine-N, O-dimethylamide (1.15 grams, 3.3 mmol) in THF (15 milliliters) was added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with 5-ethyl acetate (150 milliliters), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 milliliters), brine (50 milliliters), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0239.tif" />

Terbutyloxycarbonyl-para-cyanophenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimoles) was dissolved in ethanol saturated with ammonia, and Raney Ni sponge (100 milligrams) was added. The solution was stirred under H₂ to

-152 152 ambient temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to produce a clear residue. The residue was dissolved in ethyl acetate (250 milliliters), and washed with 1N NaOH (2 x 50 5 milliliters) and brine (2 x 50 milliliters). The solution was dried with MgS0₄, filtered and concentrated under reduced pressure.

<img file="MX9704718A_D0240.tif" />

Terbutyloxycarbonyl-meta-cyano 10 phenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimols) was dissolved in ethanol saturated with ammonia, and Raney Ni sponge (10 0 milligrams) was added. The solution was stirred under H₂ at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to produce

-153153 a transparent residue. The residue was dissolved in ethyl acetate (250 milliliters), and washed with 1N NaOH (2 x 50 milliliters) and brine (2 x 50 milliliters). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure.

<img file="MX9704718A_D0241.tif" />

<img file="MX9704718A_D0242.tif" />

Terbutyloxycarbonyl-ortho-cyanophenylalanine-N, O-dimethylamide (1.33 grams, 4.0 millimoles) was dissolved in ethanol saturated with ammonia, and Raney Ni 10 sponge (100 milligrams) was added. The solution was stirred under H₂ at room temperature (40 psi). The solution was filtered through celite, and concentrated under reduced pressure to produce a clear residue. The residue was dissolved in ethyl acetate (250 milliliters), and washed with 1N NaOH (2 x 50 15 milliliters) and brine (2 x 50 milliliters). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure.

-154154

<img file="MX9704718A_D0243.tif" />

<img file="MX9704718A_D0244.tif" />

T erbut i1oxy c arboni1-para-aminome t i1phenylalanine-N, O-dimethylamide (1.00 grams, 3.1 millimoles) was dissolved in

Dry THF (10 milliliters) under nitrogen with stirring. The solution was cooled, N, N'-bis- (benzyloxycarbonyl) -S-methylisothiourea (1.14 grams, 3.2 millimoles), and HgCl (0.95 grams,

3.5 millimoles). The solution was concentrated under reduced pressure, where the resulting residue was suspended in ethyl acetate (200 milliliters), and filtered through celite. Filtrate 10 was concentrated under reduced pressure. Evaporation chromatography on silica gel (hexane / ethyl acetate gradient) produced the purified compound.

-155155

<img file="MX9704718A_D0245.tif" />

Terbutyloxycarbonyl-meta-aminomethylphenylalanine-N, O-dimethylamide (1.00 grams, 3.1 millimoles) was dissolved in

Dry THF (10 milliliters) under nitrogen with stirring. The solution was cooled, N, N'-bis- (benzyloxycarbonyl) -S-methylisothiourea (1.14 grams, 3.2 millimoles), and HgCl (0.95 grams,

3.5 millimoles). The solution was concentrated under reduced pressure, where the resulting residue was suspended in ethyl acetate (200 milliliters), and filtered through celite. Filtrate 10 was concentrated under reduced pressure. Evaporation chromatography on silica gel (hexane / ethyl acetate gradient) produced the purified compound.

-156156

<img file="MX9704718A_D0246.tif" />

<img file="MX9704718A_D0247.tif" />

HgClj / THF

<img file="MX9704718A_D0248.tif" />

<img file="MX9704718A_D0249.tif" />

Terbutyloxycarbonyl-ortho-aminomethylphenylalanine-N, O-dimethylamide (1.00 grams, 3.1 millimoles) was dissolved in dry THF (10 milliliters) under nitrogen with stirring. The solution was cooled, N, N'-bis- (benzyloxycarbonyl) -S-methylisothiourea (1.14 grams, 3.2 millimoles), and HgCl (0.95 grams,

3.5 millimoles). The solution was concentrated under reduced pressure, where the resulting residue was suspended in ethyl acetate (200 milliliters), and filtered through celite. Filtrate 10 was concentrated under reduced pressure. Evaporation chromatography on silica gel (hexane / ethyl acetate gradient) produced the purified compound.

-157157

<img file="MX9704718A_D0250.tif" />

To a solution of thiazole ¢ 1.28 grams, 15.0 millimoles) in anhydrous THF (30 milliliters), n-BuLi (1.6 M / hexane, 8.9 milliliters, 13.9 millimoles) was added dropwise at -78 ° C, and the solution was stirred. The protected amino acid (1.36 grams,

3.3 millimoles) in THF (15 milliliters) was added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 milliliters), and the organic layer 10 was washed with saturated aqueous ammonia chloride (2 x 50 milliliters), brine (50 milliliters), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

-158158

<img file="MX9704718A_D0251.tif" />

<img file="MX9704718A_D0252.tif" />

To a solution of thiazole (1.28 grams, 15.0 millimoles) in anhydrous THF (30 milliliters), n-BuLi (1.6 M / hexane, 8.9 milliliters, 13.9 millimoles) was added dropwise at -78 ° C, and the solution was stirred. The protected amino acid (1.36 grams,

3.3 millimoles) in THF (15 milliliters) was added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 milliliters), and the organic layer 10 was washed with saturated aqueous ammonia chloride (2 x 50 milliliters), brine (50 milliliters), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material is

-159159 purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0253.tif" />

one. Lithium thiazole / THF

--------►

<img file="MX9704718A_D0254.tif" />

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL), n-Buli (1.6 M / hexane, 8.9 mL, 13.9 mmol) was added dropwise at -78 ° C, and the solution was successful. The protected amino acid (1.36 g, -3.3 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 mL), brine (50 mL), and dried with magnesium sulfate, painted and it was concentrated under reduced pressure. The primary material was purified on silica gel (15-ethyl acetate / hexane), and concentrated under reduced pressure.

-160160

<img file="MX9704718A_D0255.tif" />

one. Zn / Cu couple; ultrasound

2. | o-CH₃C₆H₄)₃P]₂PdCI₂

2-amine-5-bromopyridinc

H₂I

OR

<img file="MX9704718A_D0256.tif" />

A solution of tert-butoxycarbonyl-iodo-alanineN, O-dimethylamine (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) were added to a dry, nitrogen purged round flask charged with a zinc-copper coupler (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (according to TLC). Bis (tri-o-toliphosphine) palladium dichloride (0.35 g,

0.40 mmol) followed by 2-iodobenzonitrile (1.72 g, 7.5 mmol).

The resulting mixture was stirred under a heated nitrogen atmosphere, allowed to cool, ethyl acetate (100 mL) was added, and the mixture was filtered through a separating funnel. Sequential washing with aqueous HC1 (50 mL; 0.IN), 15 H₂Or distilled (3 x 50 mL), it was carried out with Na₂SW₄, filtration and concentration under reduced pressure produced the product

-161161 primary. Evaporation chromatography on silica gel (light petroleum / ethyl acetate gradient) yielded the purified compound.

<img file="MX9704718A_D0257.tif" />

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

-162162

OR

<img file="MX9704718A_D0258.tif" />

\ ch₃

<img file="MX9704718A_D0259.tif" />

A solution of treto-butyloxycarbonyl-iodo-alanineN, 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) were added to a dry, nitrogen purged round flask loaded with a zinc-copper coupler (0.90 g). The resulting mixture was sonicated under nitrogen until no starting material remained (according to TLC). Bis (tri-o-toliphosphine) palladium dichloride (0.35g, 0.40mmol) was added followed by 2-iodobenzonitrile (1.72g, 7.5mmol). The resulting mixture was stirred under nitrogen atmosphere with heating, filtered distilled (3 x 50 the mixture in a separating funnel.

with aqueous HC1 mL), dried in

The (50 mL; 0. IN), h₂or

Na₂SW₄, filtration concentration under reduced pressure produced the product

-163163 primary. Evaporative chromatography on silica gel (light petroleum / ethyl acetate gradient) produced the purified compound.

<img file="MX9704718A_D0260.tif" />

^ och₃

N

I ch₃

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, O-hydroxylamine hydrochloride (0.416 g, 6.0 mmol) , and diisopropylethylamine (1.02 mL, 6.0 mmol). The mixture was refluxed and then cooled. The precipitate was filtered, washed with cold ethanol, diisopropylether, dried with MgSO, concentrated under reduced pressure and the next step was used directly. The semi-solid was suspended in a mixture of acetic acid (20 mL) and dry ethanol (40 mL) with heating. Subsequently,

-164164 added Pd / C catalyst (0.30 g, 10% Pd), and bubbled hydrogen was passed through the mixture with heating. Hydrogenation was continued until no starting material could be detected with TLC. The catalyst was removed by filtration, and the solution was concentrated under reduced pressure (50 mL), HC1 (50 mL, 1N), and the mixture was again concentrated to 50 mL. The solution was cooled overnight to produce the title compound.

<img file="MX9704718A_D0261.tif" />

To a solution of thiazole (1.28 g, 15.0 mmol) in anhydrous THF (30 mL), n-BuLi (1.6 M / hexane, 8.9 mL, 13.9 mmol) was added dropwise at -78 ° C, and the solution was stirred. The amino acid N, 0-diethylamide (1.16 g, 3.3 mmol) in anhydrous THF (15 mL) was then added dropwise, and the resulting mixture was stirred. The reaction was stopped with ammonia chloride

-165165 aqueous and saturated. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 mL), brine (50 mL), dried with

MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on a silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0262.tif" />

one. H₂, PtO₂/ AcOH

<img file="MX9704718A_D0263.tif" />

<img file="MX9704718A_D0264.tif" />

<img file="MX9704718A_D0265.tif" />

Tert-butyloxycarbonyl-3- (4pyridyl) alanine-N, O-dimethylamide (4.50 g, 14.4 mmol) was dissolved in 10 acetic acid (100 mL) _t and PtO (100 mg) was added. The solution was filtered under H₂ until gas absorption ceased. The solution was filtered through celite, and concentrated under reduced pressure producing tert-butyloxycarbonyl-3- (4piperidyl) alanine-N, O-dimethylamide. The 15 residues were dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgSO₄, leaked and

-166166 was concentrated under reduced pressure to produce the title compound.

<img file="MX9704718A_D0266.tif" />

Terto-butyloxycarbonyl-3- (35 pyridyl) alanine-N, O-dimethylamide (4.50 g, 14.4 tnmol) was dissolved in acetic acid (100 mL) _t and PtO (100 mg) was added. The solution was stirred under H₂ until gas absorption ceased. The solution was filtered through celite, and concentrated under reduced pressure producing tert-butyloxycarbonyl-3 - (3 10 piperidyl) alanine-N, O-dimethylamide. The residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure to produce the title compound.

-167167

<img file="MX9704718A_D0267.tif" />

one. H₂, PtO₂/ AcOH

<img file="MX9704718A_D0268.tif" />

<img file="MX9704718A_D0269.tif" />

Tert-Butyloxycarbonyl-3- (3pyridyl) alanine-N, 0-dimethylamide (4.50 g, 14.4 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was stirred under H₂ until gas absorption ceased. The solution was filtered through celite, and concentrated under reduced pressure, yielding tert-butyloxycarbonyl-3 - (3 piperidyl) alanine-N, O-dimethylamide. The residue was dissolved in ethyl acetate (250 mL), washed with 1N NaOH (2 x 50 mL), brine (2 x 50 mL), dried with MgS0₄, filtered and concentrated under reduced pressure to produce the title compound.

-168168

<img file="MX9704718A_D0270.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 or with stirring. The solution was cooled, and the remaining isothiourea filtered solution was added (1.14 g, 3.2 concentrated

N, N'-bis- (benzyloxycarbonyl) -S-methylmmol), and HgCl under pressure was suspended in acetate through reduced pressure. La (0.95 g, 3.5 mmol). The reduced one. The ethyl residue (200 mL), and celite. The filtrate was concentrated under chromatography by evaporation on silica gel (hexane / ethyl acetate gradient) produced the title compound.

-169169

<img file="MX9704718A_D0271.tif" />

<img file="MX9704718A_D0272.tif" />

HgCl₂/ THF

<img file="MX9704718A_D0273.tif" />

<img file="MX9704718A_D0274.tif" />

Tert-butyloxycarbonyl-3- (3 piperidyl) alanine-N, O-dimethylamide (1.00 g, 3.2 mmol) was dissolved in cooled THF, remaining solution was added filtered to concentrated mmol), and HgCl under pressure was suspended in acetate of through reduced pressure. La (0.95 g, 3.5 mmol). The reduction, the re duo ethyl (200 mL), and celite. The filtrate was concentrated under chromatography by evaporation on silica gel (hexane / ethyl acetate gradient) produced the title compound.

-170170

<img file="MX9704718A_D0275.tif" />

<img file="MX9704718A_D0276.tif" />

HgCl₂/ THF

<img file="MX9704718A_D0277.tif" />

<img file="MX9704718A_D0278.tif" />

Tert-butyloxycarbonyl-3- (2piperidyl) 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, added

N, N'-bis- (benzyloxycarbonyl) -S-methyl solution was concentrated, remaining mmol), and HgCl under pressure was suspended in acetate (0.95 g, 3.5 rnmol). The reduced, ethyl residue (200 mL), and was filtered through reduced pressure. Silica gel vaporization chromatography (hexane / ethyl acetate gradient) produced the title compound.

-171171

<img file="MX9704718A_D0279.tif" />

<img file="MX9704718A_D0280.tif" />

To a solution of thiazole in anhydrous THF (1.23 g,

14.4 mmol) n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) was added dropwise at -78 ° C, and the solution was stirred. The guanidylated derivative of 4-piperidylalanine (2.00 g, 3.2 mmol) in anhydrous THF (15 mL), was then added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous 10 ammonia chloride (2 x 50 mL), brine (50 mL), dried with MgSO₄, filtered, and concentrated under reduced pressure. The primary material was purified on a silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

-172172

<img file="MX9704718A_D0281.tif" />

<img file="MX9704718A_D0282.tif" />

To a solution of thiazole in anhydrous THF (1.23 g,

14.4 mmol) n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) was added dropwise at -78 ° C, and the solution was stirred. The mixture was stirred at

-78 ° C for 1 hour. The guanidylated 3-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x mL), brine (50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure.

-173173

<img file="MX9704718A_D0283.tif" />

one. Lithium thiazole / THF

-

<img file="MX9704718A_D0284.tif" />

To a solution of thiazole in anhydrous THF (1.23 g,

14.4 mmol) n-BuLi (1.6 M / hexane, 8.4 mL, 13.4 mmol) was added dropwise at -78 ° C, and the solution was stirred. The mixture was stirred at

-78 ° C for 1 hour. The guanidylated 2-piperidylalanine derivative (2.00 g, 3.2 mmol) in THF (15 mL) was then added dropwise, and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x mL), brine (50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure.

-174174

<img file="MX9704718A_D0285.tif" />

<img file="MX9704718A_D0286.tif" />

Tert-butyloxycarbonyl-para-nitrophenylalanine-N, O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was stirred under H₂ until gas absorption ceased. The solution was filtered through celite, concentrated under reduced pressure, added to H₂O (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL), and brine (3 x 50 mL). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure, yielding the title compound.

-175175

<img file="MX9704718A_D0287.tif" />

^ ch₃

N

I och₃

one. H₂, PtO₂/ AcOH

-

<img file="MX9704718A_D0288.tif" />

Tert-butyloxycarbonyl-meta-nitrophenylalanine-N, O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was stirred under H₂ until gas absorption ceased. The solution was filtered through celite, concentrated under reduced pressure, added to H₂0 (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL), and brine (3 x 50 mL). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure, yielding the title compound.

-176176

<img file="MX9704718A_D0289.tif" />

one. H₂, PtORAcOH

-

<img file="MX9704718A_D0290.tif" />

Tert-butyloxycarbonyl-ortho-nitrophenylalanine-N, O-dimethylamide (13.88 g, 39.3 mmol) was dissolved in acetic acid (100 mL), and PtO (100 mg) was added. The solution was stirred under H₂ until gas absorption ceased. The solution was filtered through celite, concentrated under reduced pressure, added to H₂O (150 mL) and lyophilized. The semi-solid was dissolved in ethyl acetate (350 mL), washed with 1N NaOH (3 x 50 mL), and brine (3 x 50 mL). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure, yielding the title compound.

-177ΙΊΊ

<img file="MX9704718A_D0291.tif" />

<img file="MX9704718A_D0292.tif" />

dissolved tert-butyloxycarbonyl-3 (cis / trans-4-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) in saturated aqueous sodium bicarbonate, and THF 5 [60 mL, (1: 1)] with agitated. The solution was cooled and a solution of benzyl chloroformate (10 mL) in THF was added dropwise.

Surplus solid sodium bicarbonate was added, the

THF under reduced pressure and the remaining aqueous phase was poured into ethyl acetate (250 mL) and carefully mixed. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4N aqueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgS0₄, filtered and concentrated under reduced pressure. The semisolid was chromatographed on silica gel (ethyl acetate / hexane).

-178178

<td></td><td> 2.</td><td>To a solution</td><td>thiazole (1.16</td><td>g.</td><td colspan="2"> 13.7</td>

<td>mmol)</td><td>in THF it</td><td>added n-BuLi (1.</td><td>6 M / hexane, 8.0</td><td>mL,</td><td> 12</td><td> .8</td>

<td>mmol)</td><td>in drops to</td><td>"78 ° C, and stirred</td><td>the solution. The</td><td colspan="2">amide</td><td>of</td>

Protected amino acid above (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate {150 mL), and the organic layer was washed with aqueous ammonia chloride (2 x 10 50 mL), brine (50 mL), dried with MgS0₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0293.tif" />

/ CH₃r eight₃

one. Z-CI, NaHCO₃/ THF: H₂OR

-----------------»

2. Lithium thiazole / THF

<img file="MX9704718A_D0294.tif" />

-179179

one. Tert-butyloxycarbonyl-3 (cis / trans-3-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) was dissolved in saturated aqueous sodium bicarbonate, and THF [60 mL, (1: 1)] with agitated. The solution was cooled and a solution of benzyl chloroformate (0.43 mL, 3.0 mmol) in THF (10 mL) was added dropwise. Surplus solid sodium bicarbonate was added, THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL) and carefully mixed. The aqueous phase was discarded and the remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4N aqueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure. The semisolid was chromatographed on silica gel (ethyl acetate / hexane).

<td> 15</td><td> 2.</td><td>To a thiazole solution (1.16</td><td> 9, 13</td><td> . 7</td>

<td>mmol)</td><td>in THF it</td><td>added n-BuLi (1.6 M / hexane, 8.0</td><td>mL, 12</td><td> .8</td>

<td>mmol)</td><td>in drops to</td><td>-78 ° C, and the solution was stirred. The</td><td>amide</td><td>of</td>

Protected amino acid above (1.41 g, 3.0 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with aqueous ammonia chloride and

-180 180 saturated. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 mL), brine (50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material 5 was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

<img file="MX9704718A_D0295.tif" />

one. Z-Cl, NaHCCh / THF: H₂OR

2. Lithium thiazole / THF

<img file="MX9704718A_D0296.tif" />

dissolved tert-butyloxycarbonyl-3 (cys / trans-2-aminocyclohexyl) alanine-N, O-dimethylamide (1.00 g, 3.0 mmol) in saturated aqueous sodium bicarbonate, and THF [60 mL, (1: 1)] with agitated. The solution was cooled and a solution of benzyl chloroformate (10 mL) in THF (0.43 mL, 3.0 mmol) was added dropwise. Surplus solid sodium bicarbonate was added, THF was removed under reduced pressure, and the remaining aqueous phase was poured into ethyl acetate (250 mL) and carefully mixed. The aqueous phase was discarded and the

-181181 Remaining solution was washed with saturated aqueous sodium bicarbonate (2 x 50 mL), 4 Ñ aqueous sodium bisulfate (2 x 50 mL), and brine (2 x 50 mL). The solution was dried with MgSO₄, filtered and concentrated under reduced pressure. The semisolid was chromatographed on silica gel (ethyl acetate / hexane).

2. To a solution of thiazole (1.16 g, 13.7 mmol) in anhydrous THF, n-BuLi (1.6 M / hexane, 8.0 mL, 12.8 mmol) was added dropwise at -78 ° C, and the solution was stirred. The above protected amino acid amide (1.41 g, 3.0 mmol) in THF (15-10 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with aqueous ammonia chloride and

<td>saturated</td><td>(2 x</td><td colspan="2">50 mL), brine</td><td>(50 mL),</td><td colspan="3">dried with MgSO₄, I know</td>

<td>15 leaked and</td><td>I know</td><td>concentrated</td><td>low</td><td>Pressure</td><td>reduced.</td><td>The</td><td>material</td>

<td>primary</td><td>I know</td><td>purified</td><td>on</td><td>gel</td><td>silica</td><td colspan="2">(acetate</td>

ethyl / hexane), and concentrated under reduced pressure.

-182182

<img file="MX9704718A_D0297.tif" />

<img file="MX9704718A_D0298.tif" />

one. Tert-butyloxycarbonyl-3 (cis / trans-4-aminocyclohexy1) 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 Ν, Ν'-bis (benzyloxycarbonyl) -S-methyl-isothiurea (2.18. G, 6.1 mmol), and HgCl (1.18 g, 6.7 mmol) were added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Silica gel vaporization chromatography (hexane / ethyl acetate gradient) produced the purified product.

2. To a solution of thiazole (2.32 g, 23.7 mmol) in anhydrous THF, n-BuLi (1.6 M / hexane, 15.9 mL, 15 25.4 mmol) was added dropwise at -78 ° C, and the solution was stirred. I know

-183183 added the above guanilinated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) dropwise and the resulting mixture was stirred.

The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 mL), brine (50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane)_F and concentrated under reduced pressure.

<img file="MX9704718A_D0299.tif" />

<img file="MX9704718A_D0300.tif" />

HgCh / THF

2. Lithium thiazole / THF

<img file="MX9704718A_D0301.tif" />

one. Tert-butyloxycarbonyl-3 (cus / trans-3-aminocyclohexyl) alanine-N, O-dimethylamide (2.0 g, 6.1 mmol) was dissolved in dry THF (20 mL) under nitrogen with stirring.

-184184

The solution was cooled to 0 ° C, and Ν, Ν'-bis (benzyloxycarbonyl) -S-methyl-isothiurea (2.18, g, 6.1 mmol) were added, and

HgCl (1.18 g, 6.7, mmol). The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Silica gel vaporization chromatography (hexane / ethyl acetate gradient) produced the purified product.

2. To a solution of thiazole (2.32 g, 23.7 mmol) in anhydrous THF was added n-BuLi (1.6 M / hexane, 15.9 mL,

25.4 mmol) dropwise at -78 ° C, and the solution was stirred. The above guanilinated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred.

The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with saturated aqueous ammonia chloride (2 x 50 mL), brine (50 mL), dried with MgSO₄, filtered and concentrated under reduced pressure. The primary material was purified on silica gel (ethyl acetate / hexane), and concentrated under reduced pressure.

-185185

<img file="MX9704718A_D0302.tif" />

<img file="MX9704718A_D0303.tif" />

one. 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-methyl-isothiurea (2.18. G, 6.1 mmol), and HgCl (1.18 g, 6.7 mmol) were added. The solution was concentrated under reduced pressure, the remaining residue was suspended in ethyl acetate (300 mL), and filtered through celite. The filtrate was concentrated under reduced pressure. Silica gel vaporization chromatography (hexane / ethyl acetate gradient) produced the purified product.

-186186

2. To a solution of thiazole (2.32 g, 23.7 mmol) in anhydrous THF, n-BuLi (1.6 M / hexane, 15.9 mL, 25.4 mmol) was added dropwise at -78 ° C, and the solution was stirred. The above guanilinated amino acid (3.88 g, 6.1 mmol) in THF (15 mL) was added dropwise and the resulting mixture was stirred. The reaction was quenched with saturated aqueous ammonia chloride. The mixture was diluted with ethyl acetate (150 mL), and the organic layer was washed with aqueous ammonia chloride and

<td></td><td>saturated</td><td>(2 x</td><td colspan="2">50 mL), brine</td><td>(50 mL),</td><td>dried up</td><td>with</td><td>MgSO₄, I know</td>

<td> 10</td><td>leaked and</td><td>I know</td><td>concentrated</td><td>low</td><td>Pressure</td><td>reduced.</td><td>The</td><td>material</td>

<td></td><td>primary</td><td>I know</td><td>purified</td><td>on</td><td>gel</td><td>silica</td><td colspan="2">(acetate</td>

ethyl / hexane), and concentrated under reduced pressure.

Example 2

Synthesis of Intermediate

-187187

<img file="MX9704718A_D0304.tif" />

one. n-BuLi

--------------► ²-Cr ^^cl

THF / -7§C

<img file="MX9704718A_D0305.tif" />

(4S, 5R) -3- (l-oxo-3-phenylpropyl) -4- (phenyl) -5- (methyl) -2oxazolidone (2). A solution of 10.0 g (1.0 equiv., 56.4 mmol) of 4S, 5R) -4-phenyl-5-methyl-2-oxazolidone (1) in 205 mL of dry THF, stirred at -78 ° C under argon, was Drop treated with n-butyl lithium (1.6M in hexane, 1.1. eq., 38.8 mL). After stirring for 30 minutes, 8.4 mL (1.0 equiv., 56.4 mmol) of hydrocinamoyl chloride was added dropwise over a period of 10 minutes. The resulting mixture was heated to 0 ° C, stirred for an additional hour, and quenched with saturated ammonia chloride. The solvent was removed in vacuo and the resulting white solid was dissolved in ethyl acetate and ddH₂O. The aqueous phase was extracted with two additional portions of ethyl acetate. The extracts were combined, washed with saturated sodium chloride, dried over

-188188

<td>sulfate</td><td>sodium and the</td><td>solvent</td><td>in</td><td>empty for</td>

<td>produce a</td><td>white crystalline solid</td><td>(2 in</td><td>a</td><td>performance</td>

<td>9%).</td><td></td><td></td><td></td><td></td>

mp 95-96.5;

(c = 1,018 CH₂C1₂) .

¹HNMR (CDC1₃) δ 0.89 (d, 3H, CH ^

3.00-3.05 (m, 2H), 3.26-3.34

4.73-4.78

1H), 5.64, d, 1H, J-7.4 Hz), 7.22-7.46 (m, 10H).

<img file="MX9704718A_D0306.tif" />

A 5.0 g solution (1.00 equiv.

16.2 mmol) of (2) in 100 mL of dry THF was cooled to -78 ° C. Enolization was achieved with 17.8 mL (1.1 equiv., 17.8 mmol) of lithium bis15 trimethylsilylamide which was added dropwise with a syringe. The solution was stirred for 30 minutes, the solution was stirred 30 minutes before 4.45 mL (3.0

-189189 equiv., 48.5 mmol) of allyl iodide and the reaction was heated to -15 ° C. After 1 hour, the reaction was quenched with saturated ammonia chloride and extracted (3X) with ethyl acetate. The organic phase was washed with sodium metadisulfite, dried over sodium sulfate and the solvent was removed in vacuo to obtain a light colored oil. Purification was achieved by evaporation chromatography on silica gel using a step gradient (15: 1, 12: 1, 10: 1) to produce (3), a colorless oil (95%).

[to]_D 47.5 (c = 3.12, CH₂C1₂) .

^XHNMR (CDC1₃) δ 0.82 (d, 3H, CH ^ J = 6.6 Hz), 2.31-2.40 (m, IH, RCH = CHCH₂), 2.49-2.57 (m, IH, RCH = CHCH₂), 15 2.84-3.00 (m, 2H, Ph-CH₂), 4.32-4.37 (m, IH, CH- (N) CO), 4.53-

4.58 (m, IH, CH₃-CH-), 5.03-5.13 (m, 2H, ABX, CH = CH₂), 5.21 (d, IH, Ph-CH, J = 7.1 Hz), 5.81-5.89 (m, IH, CH = CH₂), 7.20-7.42 (m, 10H, ArH);

-190190

13_c (CDC1₃) δ 14.4, 36.2, 38.2, 43.9, 54.7,

78.4, 117.1, 125.4, 126.3, 128.2, 128.5, 129.0, 133.1, 134.8,

138.9, 152.4, 174.9.

<img file="MX9704718A_D0307.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 equiv., 13.6 mmol) of a 1.0 M solution of borane10 tetrahydrofuran complex at 0 ° C and stirred for 2 hours. Solvent was evaporated and chloroform (100 mL) was added with a syringe.

Organoborane oxidation was achieved. by adding

4.7 g (2.0 equiv., 27.2 mmol) 3-chloroperoxybenzoic acid a

0 ° C, warming to room temperature and stirring for an additional hour. The organic phase was washed with 5% Na₂CO₃, ddH₂Or, and dried over sodium sulfate. Due to the instability of the alcohol, a fast column was performed to extract the

-191191 extreme polar and non-polar material that originated from 3-chloroperoxybenzoic acid. Alcohol (4) was obtained in a yield of 65%.

[&lt;x]_D 39.3 (c = 1.38, CH₂C1₂) .

¹HNMR (CDClj) δ 1.07 (d, 3H, CH3-CH, J = 6.5 Hz), 1.81-1.93 (m,

3H, CH₂-CH-H), 2.10-2.19 (m, 1H, CH₂-CH-H), 3.10-3.17 (m, 2H,

Ph-CH₂), 3,873.90 (m, 2H, CH₂OH), 4.43-4.49 (m, 1H, CH-CO),

4.70-4.75 (m, 1H, CH₃-CH), 5.36 (d, 1H, Ph-CH, J-7.1 Hz),

7.41-7.63 (m, 10H, ArH);

<img file="MX9704718A_D0308.tif" />

<td>TO</td><td>a solution</td><td>of</td><td colspan="2">alcohol</td><td>(4) (1.0 g,</td><td> 2.7</td><td>mmol),</td>

<td>dissolved in</td><td>dichloromethane</td><td> (27</td><td>mL),</td><td>I know</td><td>they introduced</td><td> 876</td><td>mg (1.5</td>

<td>equiv._t 4.1</td><td colspan="3">mmo 1.) de c 1 oroc rornato</td><td>of</td><td>pyridinium and</td><td> 1.0</td><td>g of 4</td>

-192192 4 angstrom molecular meshes, and the mixture changed from bright orange to black.

Reaction was monitored

<td></td><td>by TLC</td><td>and</td><td>then</td><td>of</td><td colspan="3">30 minutes, if material remained</td>

<td></td><td>initial,</td><td>I know</td><td colspan="2">added</td><td>molecular filters</td><td>traditional.</td><td>The</td>

<td> 5</td><td>solution</td><td>I know</td><td>filter</td><td>to</td><td>through celite</td><td>and it evaporated</td><td>the</td>

<td></td><td>solvent.</td><td>The</td><td>residue</td><td>I know</td><td>dissolved in acetate</td><td>ethyl and it</td><td>Washed</td>

with saturated sodium chloride. If the orange color persisted in the organic phase, additional filtrations were carried out through celite mesh. Aldehyde was obtained in a quantitative yield as a clear, colorless oil (5).

¹HNMR (CDC1₃) δ 0.84 (d, 3H, CHA₃-CH, J = 6.6Hz),

1.87-1.94 (m,

1H, CH₂-CH (H) -CHO), 2.04-2.13 (m, 1H, CH₂-CH (H) -CHO), 2.4515 2.50 (m, 2H, Ph-CH₂), 2.79-2.85 (dd, 1H, CH₂-CHO, J = 13.3 &amp;

J = 6.6), 2.92-2.99 (dd, 1H, CH₂-CHO, J = 13.2 &amp; J-8.8), 4.19-4.22 (m, 1H, CH-CO), 4.46-4.51 (m_F 1H, CH₃-CH), 5.13-5.25 (m, 1H,

Ph-CH), 7.20-7.39 (m, 10H, Arfi), 9.69 (s, 1H, CHO);

-193193

13_c (CDC1₃) 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="MX9704718A_D0309.tif" />

<img file="MX9704718A_D0310.tif" />

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 equiv., 8.52 mmol) of L- ethyl ester cysteine ​​and 4A molecular filters. The reaction was allowed to stir overnight at room temperature followed by removal of solvent in vacuo. The residue was dissolved in chloroform, washed with saturated sodium chloride, ddH₂Or, and dried over sodium sulfate. Solvent was removed in vacuo to obtain a rubber textured solid (6).

-194194

<img file="MX9704718A_D0311.tif" />

CO₂Et

2. OM of trimethyl aluminum in hexane (2.4 mL, 4.8 mmol, 3 equivalents) was slowly added to the starting material (6) (800 mg, 1.61 mmol) by stirring in anhydrous dichloromethane under argon, using oven drying equipment. After stirring overnight, HPLC indicated that the reaction was complete.

The mixture was quenched with surplus methanol, then filtered over a short column of silica gel (washing with a surplus of 10% methanol in ethyl acetate. Evaporation produced 784 mg of primary material which was purified using 2-.1 hexane. : EtOAc on a gel column

-195195 silica producing 258 mg (0.81 mmol, which is a 50% yield) of pure compound (7), an ester of 6Sbenzylhexahydro-5-oxo-5H-thiazole [3,2-a] pyridine-3R- Ethyl as a yellowish white solid.

<td colspan="2">^lH NMR (CDC13) d 1.28-1</td><td>.31 (τη, 3H), 1.</td><td> .72-1.81</td><td>(m, 3H), 2</td><td> .10-</td>

<td> 2.13</td><td>(m, 1H), 2.66 (dd.</td><td>1H, J = 11.5 and</td><td>6.0 Hz),</td><td> 3.29-3.34</td><td>(m,</td>

<td>2H),</td><td>4.19-4.29 (m, 2H),</td><td>4.88 (dd, 1H,</td><td>J = 9.0 and</td><td>5.0 Hz),</td><td> 5.22</td>

<td>(dd,</td><td>1H, J = 8.0 and 6.0 H</td><td>z), 7.18-7.23</td><td>(m, 3H),</td><td> 7.28-7.31</td><td>(m,</td>

<td>2H).</td><td></td><td></td><td></td><td></td><td></td>

<img file="MX9704718A_D0312.tif" />

<img file="MX9704718A_D0313.tif" />

LÍOH.H2O (48 mg, 1.12 mmol) in 10 mL of water was added to the starting material (7) (240 mg, 0.76 mmol), dissolved in mL of dioxane. After 1 hour, TLC at 1: 1 Hexane: EtOAc showed no starting material. The reaction was quenched with 10% citric acid, and extracted twice with

-196196 dichloromethane. Drying and evaporation of the combined organic layers yielded 354 mg of the primary product. This was redissolved in dichloromethane, and precipitated by adding excess hexane. The product was filtered to yield 5 200 mg (0.68 mmol, 90% yield) of an off-white solid, (8), also known as 6S-benzylhexahydro-5oxo-5H-thiazolo [3, 2-a] pyridine- 3R-carboxylic acid.

<td></td><td colspan="2">¹H NMR (CD3OD) d 1.71-1.82</td><td>(m,</td><td>3H), 2.12-2.17 (m, 1H), 2.67</td>

<td> 10</td><td>(dd,</td><td>1H, J = 14 and 11 Hz), 2.</td><td> 77-2</td><td>.81 (m, 1H9, 3.30-3.40 (, 3H),</td>

<td></td><td> 4.81</td><td>(dd, 1H, J = 8.5 and 4.9</td><td>Hz),</td><td>5.16 (t, 1H, J = 7.5 Hz), 7.18-</td>

<td></td><td> 7.31</td><td>(m, 5H).</td><td></td><td></td>

<img file="MX9704718A_D0314.tif" />

<img file="MX9704718A_D0315.tif" />

Oxalyl chloride (9) (25 g, 0.197 mol) was cooled to

0 ° C and propionic acid cyclohexane (20 ml, 0.14 mol) was added. This was left stirring overnight. Mix

The resulting -197197 was distilled to yield an 84% yield of colorless liquid (10) cyclohexyl propionic acid chloride.

<img file="MX9704718A_D0316.tif" />

The chiral auxiliary (11) (13.6 g, 76.7 mmol, 1 eq) was dissolved in dry THF and cooled to -78 ° C. Then nBuLi (52.8 mL, 84.4 mmol, 1.2 equiv.) Was added and left for 30 minutes (dark orange solution). The acidic chlorudo (10) (13.4 g, 76.6 mmol 1 eq) was then added and allowed to stir overnight. Maneuvers were performed interrupting it with NH₄C1 saturated by extracting it with ethyl acetate, washing the extracts with water and brine, drying it over sodium sulfate and with concentration. A fast dry load column (6: 1 ethyl acetate / hexane) was passed to purify the product. This produced a white solid (12)

-198198 which was recrystallized from ether and hexane to produce the title compound in 78% yield.

[a] D = -20.1 (c = 1, EtOH); MP / BP mp = 90.5-91.5oC

1H NMR (CDC13) d 0.86-1.10 (m, 5H), 1.18-1.30 (m, 4H), 1.541.75 (m 7H), 2.86-2.97 (m, 2H0, 4.70-4.76 (m, 1H), 5.65 (d,

1H, J = 7.2 Hz), 7.28-7.42 (m, 5H).

<img file="MX9704718A_D0317.tif" />

<img file="MX9704718A_D0318.tif" />

The starting material (12) (9.13 g, 29 mmol, leq) was dissolved in dry THF and cooled to -78 ° C, after which LiHMDS (31.9 mL, 31.9 mmol, 1.1 eq) was added dropwise over 40 minutes . Then, 30 minutes later, alili bromide (7.5 mL, 86.9 mmol, 3 eq) was added slowly, and over 10 minutes.

The mixture was allowed to warm overnight. Maneuvers included stopping the reaction with saturated ammonium chloride, extracting it with ethyl acetate, washing it with

-199199 10% sodium thiosulfate, discolor with charcoal, dry over sodium sulfate and concentrate in vacuo. The product was obtained as a yellow oil (13) in 96% yield.

[a] D = + 9.5 (c = 1.0, EtOH)

<td></td><td>H NMR (CDC13)</td><td>d 0.92-1.10</td><td>(τη, 5H9, 1.10-1.39</td><td>(m,</td><td>5H), 1.63-</td>

<td></td><td>1.75 (m, 6H),</td><td> 2.27-2.42 (,</td><td>2H), 4.01-4.14 (m,</td><td>1 HOUR)</td><td> , 4.76-4.85</td>

<td> 10</td><td>(m, 1H), 5.00-</td><td>• 5.07 (m, 2HO,</td><td>5.65 (d, 1H, J = 7</td><td>Hz)</td><td> , 5.64-5.88</td>

(m, 1H0, 7.27-7.46 (m, 5H).

<img file="MX9704718A_D0319.tif" />

2-Methyl-2-butene was added dropwise to borane dimethylsulfide complex at -12 ° C. The reaction was held at this temperature for 15 minutes and then heated to 0 ° C,

-200200 after which it was stirred for 2 hours. Then disiamyl borane was added to a mixture of the starting material (13) in THF using a double-ended needle at 0 ° C. The mixture was then stirred for 2 hours, after which solvents were removed and the residue was dissolved in dichloromethane. A suspension of pyridinium chlorochromate in dichloromethane contained in a flask equipped with a reflux condenser was carefully added. After the initial exothermic reaction was reduced, the mixture was refluxed at 50 ° C for 1 hour. The dark brown liquid was dissolved in ethyl acetate and filtered through Florisil. The black PCC residue was extracted with ethyl acetate and was also filtered through the same Florisil pad. The concentration of the filtrates resulted in a yield of 15 78% of a yellowish product gum (14).

[a] D = -17.8 (c = l.245, EtOH) ^XH NMR (CDC13) d 0.89-1.18 (m, 5H), 1.20-1.47 (m, 8H), 1.6020 1.74 (m, 6H), 1.83-2.00 (m, 1H), 2.48-2.53 (m, 2H), 3.90-4.10

-201201 (m, 1HO, 4.12-4.16 (m, 1H)

4.76-4.80 (m, 1HO, 5.67 (d, 1H, J =

7Hz), 7.27-7.46 (m, 5H), 9.77 (s, 1H).

<img file="MX9704718A_D0320.tif" />

Toluene / p-TSA / mol. if you see

HS NH,

CO₂Et

<img file="MX9704718A_D0321.tif" />

The resulting aldehyde (14) (7.7 g crude, 20.8 mmol equiv.) Was dissolved in 75 mL of toluene. To the solution was added a catalytic amount of p-toluenesulfonic acid (50 mg), 10 g of 4A molecular filter, and L10 cysteine ​​ethyl ester (3.87 g, 20.8 mmol, 1 equiv.). The mixture was stirred overnight, filtered, and concentrated. The residue was then purified with silica gel chromatography (6: 1 hexane: ethyl acetate) and 6.36 g of the product (15) was produced in a yield of 61%.

-202202 [a] D = -48.3 (c = 1.095, EtOH) * Η NMR (CDC13) d 0.84-0.98 (τη, 4Η), 1.11-1.38 (m, 7Η), 1.50-

1.90 (m,

4.46-4.81 (τη,

4H),

5H).

(τη, (τη,

2H), 5.66 (d

7Ηζ), 7.27-7.46

<img file="MX9704718A_D0322.tif" />

<img file="MX9704718A_D0323.tif" />

<img file="MX9704718A_D0324.tif" />

The starting material (15) (1.97 g, 3.9 mmol, 1 equiv.) Was dissolved in 20 mL of dry dichloromethane and cooled to 0 ° C. Trimethyl aluminum (5.9 mL, 11.8 mmol, 3 equiv.) Was added dropwise, and the mixture was allowed to stir overnight.

I

After obtaining the complete reaction according to HPLC checks, methanol was added until a yellow solid mass formed, dichloromethane was added to dissolve the solid and

-203203 The entire mixture was stirred for 15-30 minutes and then filtered. The residue after concentration in vacuo was passed through a fast column (6: 1 hexane: hexyl acetate), to extract the auxillary and as many polar decomposition products as possible, producing a 50% yield of a yellow oil ( 16).

^lH NMR (CDC13) d 0.83-0.98 (m, 2H), 1.09-1.38 (m, 10H), 1,572.00 (m, 11H), 2.12-2.18 (m, IH), 2.49-2.54 (m, IH) , 3.10 (dd, IH, J = 11 and 6 Hz), 3.27 (dd, IH, J = 11.5 and 8.0 Hz), 4.114.25 (m, 2H), 4.88 (dd, IH, J = 11.0 and 5.0 Hz ), 5.14 (dd, IH, J = 10 and 6 Hz).

The starting material (16) (0.95 g, 2.9 mmol, 1 equiv.) Was dissolved in 10 mL of dioxane. The solution was cooled to 10 ° C, and LiOH HO (0.123 g, 2.9 mmol, 1 eq.) Was added to it.

-204204 dissolved in 10 mL of water. The bath was removed and the mixture was stirred at room temperature for 1 hour, TLC showed a complete reaction and the solvent was evaporated in vacuo. 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 concentrated to produce a white solid that was recrystallized from ether. Concentration of the filtrate and purification by silica gel column chromatography (2: 1 hexane: ethyl acetate) resulted in more product (17) with a mp of 198.2-199 C.

<td colspan="2">^XH NMR (DMSO-d6) d 0.78-0.93</td><td>(m,</td><td colspan="2">2H), 1.11-1.27</td><td colspan="2">(m, 5H), 1.34-</td>

<td>1.36 (m, 1H), 1.51</td><td>-1.56 (m,</td><td>1 HOUR) ,</td><td> 1.60-1.75</td><td>(m,</td><td>1H), 1.82-</td><td> 1.87</td>

<td>(m, 1H), 2.15-2.18</td><td>(m, 1H),</td><td> 2.37</td><td>-2.41 (m,</td><td>1 HOUR) ,</td><td>3.03 (dd,</td><td>1 HOUR,</td>

<td>J = 11.5 and 5.5 Hz),</td><td> 3.35-3.38</td><td>(m,</td><td>2H), 4.83</td><td>(dd₁</td><td>_r 1H, J = 9</td><td>and 4</td>

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

-205205

<img file="MX9704718A_D0325.tif" />

Cbz

BOC-DiCbz Arg (18) (7.6 g, 14.0 mmol) was dissolved in

THF anhydrous (40 mL) and cooled to 0 ° C. Triethylamine (2.2 mL) was added followed by 14.5 mmol of a 1M toluene solution of isopropyl chloroformate via syringe. The reaction was stirred at 0 ° C for 30 minutes, then rapidly cooled. The white solid was discarded. Freshly prepared diazomethane bubbles were passed into the filtrate until the color of the solution turned yellow. The mixture was allowed to stand overnight in a well ventilated fireplace, and discharge of excess diasomethane was facilitated.

Dry ether was added to precipitate the diazoquetone. The product was dried and filtered under vacuum to produce a yellowish solid (4.6g, 58%).

-206206

Diazoquetone (19) (1 g, 1.77 mmol) was dissolved in THF (20 mL) and 1M HC1 in ether (20 mL) was added to this solution at 0 ° C. The reaction was stirred at room temperature overnight, after which a white precipitate formed. Yet another precipitation was achieved by adding ether. Filtration and drying of solid produced the product (20) (1.02 g, 100%).

<td></td><td>^XH. NMR (DMSO-d6) d 1.65-1.77 (m, 3H),</td><td>2.06-2.50 (m,</td><td>1 HOUR) ,</td>

<td> 10</td><td>3.86-3.90 (m, 2H), 4.29 (τη, 1H), 4.76 (d,</td><td>1H, J = 18Hz),</td><td> 4.95</td>

<td></td><td>(d, 1H, J = 18 Hz), 7.35 (s, 2H), 7.36 (s.</td><td>2H), 7.35-7.41</td><td>(m,</td>

<td></td><td>10H), 8.71 (br s, 3H), 10.1 (br s, 2H).</td><td></td><td></td>

C NMR (DMSO-d6) d 23.7, 26.4, 47.2, 47.9, 56.2, 68.0, 69.3,

128.6, 128.7, 128.8, 128.9, 135.2, 135.9, 153.4, 157.4, 198.9.

<img file="MX9704718A_D0326.tif" />

<img file="MX9704718A_D0327.tif" />

<img file="MX9704718A_D0328.tif" />

<img file="MX9704718A_D0329.tif" />

-207207

To the mimetric (17) (0.422 g, 1.42 mmol) in THF (50 mL) at 0 ° C and in the presence of N-methyl morpholine (0.19 mL), a solution of 1M toluene isopropylchloroformate (1.71 mL) was slowly added. ). The reaction was stirred at 0 ° C for 30 minutes, to then be treated with aminochloromethyl ketone (20) in small portions. After the addition was complete, the reaction was stirred for 15

<img file="MX9704718A_D0330.tif" />

<img file="MX9704718A_D0331.tif" />

waved

<img file="MX9704718A_D0332.tif" />

then minutes in a row

<img file="MX9704718A_D0333.tif" />

temperature

<img file="MX9704718A_D0334.tif" />

reaction

<img file="MX9704718A_D0335.tif" />

I extracted

<img file="MX9704718A_D0336.tif" />

brine wash and extraction

<img file="MX9704718A_D0337.tif" />

solvent

<img file="MX9704718A_D0338.tif" />

<img file="MX9704718A_D0339.tif" />

followed with acetate

<img file="MX9704718A_D0340.tif" />

<img file="MX9704718A_D0341.tif" />

10% aqueous citric acid.

<img file="MX9704718A_D0342.tif" />

later without (21)

<img file="MX9704718A_D0343.tif" />

<img file="MX9704718A_D0344.tif" />

96%) organic

<img file="MX9704718A_D0345.tif" />

purification.

iH NMR (CDC13) d 0.07-0.97 (m, 1H), 1.15-1.41 (m, 7H), 1.62-

1.91 (m, 10H), 2.10-2.16 (m, 1H), 2.43-2.48 (m, 1H), 2.74-2.80 (m, 1H), 3.01-3.07 (m, 1H), 3.87-3.94 (m, 1H) ), 4.11-4.19 (m,

2H), 4.60-4.66 (m, 1H), 4.74-4.86 (m, 2H), 5.09-5.24 (m, 4H),

-208208

7.30-7.39 (τη, 10Η), 7.95 (d, IH, J = 8 Hz), 9.4 (br s, IH),

9.56 (br s, IH).

EXAMPLE 3.

<img file="MX9704718A_D0346.tif" />

(Nt-BOC-N-tosyl) butyrylketoarginine (240 mg, 0.515 mmol) was deprotected, using 30% TFA in dichloromethane.

The unprotected arginine derivative was coupled with mimetic (8) (100mg, 0.343mmol) in DMF under basic conditions (Et.N, pH = 8-9), using BOP reagent (228mg, 0.52mmol) and the agent dehydrating. Typically the reaction was complete within 2-4 hours. Extraction with ethyl acetate, followed by successive washing with brine and 10% aqueous citric acid produced the primary product. The

-209209 primary product was purified by column chromatography, yielding 180 mg (76%) of the pure product. This product was then treated with HF to extract the tosyl group. Purification of the isolated deprotected product by HPLC produced BCH-2737.

EXAMPLE 4

<img file="MX9704718A_D0347.tif" />

<img file="MX9704718A_D0348.tif" />

mercapto acetic acid reaction at temperature extraction treated with NMM g, 0.245 mmol) (0.036 mL) followed by mmol). The reaction mixture was stirred throughout the environment with overnight acetate.

Ethyl, followed by successive washes with brine and citric acid

-210210 10% aqueous and evaporation of the organic solvent produced the primary product which was purified by column chromatography to produce foamy solid as product (0.125g, 62%).

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

The products of the reactions described above can be isolated in free form or in the form of salts. Furthermore, the products can be obtained as pharmaceutically acceptable acid addition salts by producing a reaction of

-211211 one of the free bases with an acid. Similarly, the product can be obtained as pharmaceutically acceptable salts by reacting the free carboxylic acids with a base. In the same way, treatment of the salts with a base or acid results in the regeneration of the free amide.

EXAMPLE — 5

A general method of synthesizing the compound of the formula

II or III:

-212212

EXAMPLE 5

Br

<img file="MX9704718A_D0349.tif" />

ch₃cn

NaCNBH₃

ΠΤ 1Θ tirs

<img file="MX9704718A_D0350.tif" />

fosyl Ng

KHMDS / THF

<img file="MX9704718A_D0351.tif" />

cocm.ch.co.m ·

<img file="MX9704718A_D0352.tif" />

<img file="MX9704718A_D0353.tif" />

-213213

EXAMPLE 6

Synthesis of:

STEP 1

<img file="MX9704718A_D0354.tif" />

Ester synthesis of

4-tert-butyl hydroxybutyl acid.

NHZ (Cyclohexyl)₂NH.HOOC ^ U_{co t} (16)

<img file="MX9704718A_D0355.tif" />

2-benzyloxycarbonylaminoD} -ο^Λα

NMM, THF

2) NaBH₄, MeOH

NHZ

H0 '^X^^x^ CO₂t-bu

-214214

To a solution of the protected aspartic acid (1) (Bachera 2.50 g, 4.95 mmol) in 50 mL of dry tetrahydro furan (THF), at -10 ° C, under N₂, N-methylmorpholine (109 µΒ, 0.2 eq) and isopropyl chloroformate (1.0 M / toluene: 384 µβ, 1.1 eq) were added. The solution was stirred at -10 ° C for 60 minutes. In another flask, NaBH was suspended₄ (3 75 mg, 2 eq, in a dry 5: 1 mixture of THF / MeOH (50 mL), at -78 ° C under N₂. This suspension was stirred at -78 ° C for 30 minutes. The mixed anhydride solution was then added to the NaBH suspension₄dropwise via cannula, and the final solution was stirred at -78 ° C for 3 hours. Then acetic acid (2.8 mL, 10 eq) was added and the solution was warmed to room temperature (30 minutes). Solvents were evaporated, the residue was taken in EtOAc and washed with NaHCO₃ saturated and aqueous (2x) and brine. The organic layer was dried over MgSO₄, the solids were filtered and the solvent was evaporated to produce 1.53 g (4.95 mmol, 100%) of the alcohol (2) as a transparent oil.

iH NMR (CDC1₃, 400 MHz): at 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,

-215215

1Η, H-2), 3.69 (m, 2H, H-4), 2.17 (m, 1H, H-3), 1.63 (m, 1H,

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

STEP 2.

Tert-butyl ester of

2-benzyloxycarbonylamino-4-iodobutyric acid

ΝΉΖ

HO

<img file="MX9704718A_D0356.tif" />

<img file="MX9704718A_D0357.tif" />

Imidazole, -10 ° C

C ^ CN / E ^ O

NHZ, t-Bu

To a solution of the alcohol (2) (1.53 g,

4.95 mmol) in a 1: 1 mixture of CH₃CN / Et₂O (50 mL), at -10 ° C, under N₂Imidazole (607 mg, 1.8 eq) and Ph were successively added₃P (2.21 g, 1.7 eq). Then iodine (2.14 g, 1.7 eq) was added in 15 small portions over a period of 15 minutes. After the addition was complete, a white precipitate formed and the solution was brown. It was stirred at -10 ° C for 45 minutes. Then it was poured into Et₂O and the organic phase was washed with Na₂SW₃ aqueous and saturated, CuSO₄ aqueous and saturated, H₂O and dried over MgSO₄.

-216216

Solids were filtered and the solvent was evaporated to produce a yellow oil which was purified by evaporation chromatography (silica gel, 5% a

20%

EtOAc / Hex). Iodide (3) was obtained in a yield of

83% (1.71 g) as a clear oil.

<td>^{X &lt;}&gt; H NMR (CDC1₃, 400</td><td>MHz)</td><td>: to</td><td>7.41-7.31 (m,</td><td>5H,</td><td>ArH), 5.3 5 (bd,</td>

<td>1H, J = 7.3, NH), 5.</td><td>13 (s,</td><td>2H,</td><td>CH₂Ph), 4.30</td><td>(m,</td><td>1H, H-2), 3.22-</td>

<td>3.12 (m, 2H, H-4),</td><td> 2.42</td><td>(m,</td><td>1H, H-3), 2.20</td><td>(m,</td><td>1H, H-3), 1.48</td>

(s, 9H, t-Bu).

STEP 3

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

NHZ

I ^^^ CO₂t-Bu, Cul

IHF, -78 ° C

NHZ

To a suspension of Cul (2.27 g, 5 eq) in dry THF (20 mL) at -78 ° C under N₂slowly added a

-217217 1. OM solution in THF of vinyl magnesium bromide (23.4 mL, 9.8 eq). The solution was then heated to -10 ° C for minutes (its color was then black) and re-cooled to -78 ° C. A solution of iodide (3) (1.00 g, 2.39 mmol) in dry THF (3.5 mL) was slowly added to the cuprate solution.

The reaction mixture was stirred at -78 ° C for 2.5 hours. NH added₄C1 aqueous and saturated (50 mL) and the mixture was returned to room temperature with vigorous stirring. Then it was poured into Et₂0 and stirred for 5 minutes. The dark suspension was filtered through a funnel and the phases were separated. The aqueous phase was extracted with Et₂O (2x) and the combined organic extracts were dried over MgSO₄. The solids were filtered, the solvents were evaporated and the primary oil was purified by evaporation chromatography (silica gel, 15 5% AcOEt / Hex) to produce 0.51 g (67%) of pure alkene (4).

^XH NMR (CDC1₃, 400 MHz): at 7.37-7.31 (m, 5H, ArH), 5.80 (m,

1H, H-5), 5.3-3 (d, 1H, J-7.8 NH), 5.12 (s, 2H, CH₂Ph), 5.05 (d, 1H, J = 17.2, H-6), 5.01 (d, 1H, J-10.4, H-6), 4.30 (q, 1H,

-218218

J = 7.4, H-2), 2.16-2.08 (m, 2H, H-4), 1.92 (m, IH, H-3), 1.74 (m, IH, H3), 1.48 (s. 9H, t- Bu).

STEP-l

Synthesis of 1-benzyloxycarbonyl-5-hydroxymethyl-2-pyrrolidinocarboxylic acid tertobutyl ester

NHZ

<img file="MX9704718A_D0358.tif" />

CO₂t-bu

1) Hg (OAc)₂THF

2) NaHCO₃, KBr

3) NaBH₄, OR₂

<img file="MX9704718A_D0359.tif" />

To a solution of alkene (4) (50 mg, 0.157 mmol) in dry THF (3.1 mL) at room temperature, under N₂, mercuric acetate (75 mg, 1.5 eq) was added. The solution was stirred at room temperature for 18 hours, after which it was cooled to 0 ° C. Then NaHCO was added₃ aqueous and saturated (2 mL) and the mixture was stirred at 0 ° C for 30 minutes. KBr (0.11 added

-219219 g, and eq) and the mixture was stirred at room temperature for 2 hours. Then it was poured into H₂O / Et₂Or and the phases were separated. The aqueous phase was extracted with Et₂O (2x), and the combined organic extracts were dried over MgSO₄. Solids were filtered and solvents were evaporated. Oxygen bubbles were passed (0₂) in a suspension of NaBH₄ (3.3 mg,

0.55 eq) in dry DMF (0.4 mL) for 1 hour, and to this was added dropwise (syringe pumps, 3 mL / hr) a solution of organomeruric bromide in DMF (3.1 mL) with continuous introduction of O₂. Bubbling was continued for 1 hour and Et was added₂O (5 mL). The gray suspension was filtered through celite and the filtrate was evaporated. The residue was chromatographed (silica gel, 6: 4 Hex / EtOAc) to produce pyrrolidinol (5) (30mg, 57%) as a clear oil.

iH NMR (CDC1₃, 400 MHz): at 7.37-7.28 (m, 5H, ArH), 5.22 5.09 (m, 2H, CH₂Ph), 4.30 (dd, 1H, J = 1.4, 8.3, H-2), 4.24 (m, 1H, H-5), 3.70-3.57 (m, 3H, CH₂-OH), 2.25 (m, 1H), 2.13 (m, 1H), 1.92 (m, 1H), 1.70 (m, 1H), 1.34 (s, 9H, t-Bu).

-220220

STEP 5

Synthesis of 1-benzyloxycarbonyl-5-carboxy ”2-pyrrolidinocarboxylic acid tertobutyl ester

<img file="MX9704718A_D0360.tif" />

<img file="MX9704718A_D0361.tif" />

Ot-Bu

<td></td><td></td><td colspan="2">I know</td><td>added</td><td>to</td><td colspan="2">the alcohol solution</td><td> (5)</td><td> (50</td><td>mg,</td>

<td></td><td> 0.149</td><td>mmol)</td><td>and</td><td colspan="3">Et₂N (62μΣ, 3 eq) in CH₂C1₂ dry</td><td> (0.</td><td>8 mL)</td><td>low</td><td>n_2/</td>

<td> 10</td><td>at 0 ° C</td><td>, a</td><td colspan="2">solution</td><td>of</td><td>SO complex₃-pyridine</td><td> (71</td><td>mg,</td><td>3 eq)</td><td>in</td>

<td></td><td>DMSO</td><td>dry.</td><td>I know</td><td>waved</td><td>the</td><td>solution at 0 ° C for</td><td> 30</td><td colspan="2">minutes and</td><td>I know</td>

added 10% citric acid (2 mL). The pH was adjusted to 4 with 1M NaOH and the aqueous phase was extracted with Et₂Or (3x). The combined organic extracts were dried over MgSO₄. Solids were filtered and solvents were evaporated to produce a primary oil which was purified by chromatography of

-221221 evaporation (silica gel, 7: 3 Hex / EtOAc). Pure aldehyde (6) was obtained as a clear oil (45 mg, 90%).

^XH NMR (CDC1₃, 400 MHz): at 9.68 + 9.56 (ds, 1H, CHO), 7.365 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).

PASQ.-Á (7)

<img file="MX9704718A_D0362.tif" />

(6)

BOCHN

NH

Eat

1) 2

MgSO₄, CH₂C1₂

2) NaBH (OAC)₃AcOH, THF

<img file="MX9704718A_D0363.tif" />

-222222

The pyrrolidine-aldehyde (6) was coupled with the protected diamino propionic acid (7), first forming the imine (8) (MgSO₄, CH₂C1₂). The imine ¢ 8) was isolated by filtration of MgSO₄ and the evaporation of the solvent. The primary imine 5 was then treated with NaBH (OAc) and with acidic acid (AcOH) in THF for 15 hours to obtain the amine (8) after extraction maneuvers.

STEP 7

<img file="MX9704718A_D0364.tif" />

1 HOUR₂, Pd / C / MeOH (8)

<img file="MX9704718A_D0365.tif" />

<img file="MX9704718A_D0366.tif" />

(9)

The CB7 protecting group (7) from the amine (8) was removed by hydrogenation with 10% palladium on carbon as catalyst in methanol (MeOH). The catalyst was filtered and

-223223 evaporated the MeOH to produce the primary diamine (9) which can be used without any purification.

STEP 8

<img file="MX9704718A_D0367.tif" />

(9)

Heat

<img file="MX9704718A_D0368.tif" />

(10)

The cyclization is carried out by heating the primary oil (9) in step 7, slightly above the boiling point of the methanol. Bicyclic lactane (10) was purified by evaporative chromatography

STEP 9

-224224

<img file="MX9704718A_D0369.tif" />

(10) ((--COCI

----------- . ;&gt;

Pyridine

<img file="MX9704718A_D0370.tif" />

(11)

The secondary amine of the bicyclic lactam (10) was protected as an amide using benzoyl chloride in pyridine. Pyridine evaporation and extraction maneuvers produced cyclic lactam-amide (11).

STEP 10

<img file="MX9704718A_D0371.tif" />

(11)

HCl / Et₂OR

-------&gt;

<img file="MX9704718A_D0372.tif" />

(12)

-225225

<td colspan="2"></td><td>The</td><td>groups</td><td colspan="2">BOC protectors</td><td>and</td><td>of</td><td>this</td><td>'t-</td>

<td>butyl</td><td>of</td><td>lactam</td><td colspan="2">bicyclic amide</td><td colspan="2">(11) were</td><td colspan="2">extracted</td><td>low</td>

<td colspan="2">terms</td><td>acidic</td><td>(HCl)</td><td>in ether</td><td>ethyl</td><td>(Et</td><td> - 0):</td><td>). The</td><td>Salt</td>

<td>amine</td><td> ¢12:</td><td colspan="2">) precipitated</td><td>of the</td><td>solution</td><td>and</td><td colspan="2">is collected</td><td>by</td>

filtration.

<img file="MX9704718A_D0373.tif" />

The primary amine of compound (12) is protected with the CBZ group by reacting it with benzyl chloroformate in acetonitrile (CH₃CN) with I ^ CO as base. The 15 extraction maneuvers provided a fully protected carboxylic acid (13) that can be used for step 12 without purification.

-226226

STEP 12

<img file="MX9704718A_D0374.tif" />

(15)

Carboxylic acid (13) is coupled with benzothiazole (14) ketoarginine in DMF using BOP as a coupling agent in the presence of diisopropylethylamine 10 (EtNiPr₂). Extraction with ethyl acetate (EtOAC) produces compound (15) as a solid which is purified by chromatography.

STEP 13

-227227

<img file="MX9704718A_D0375.tif" />

(16)

The two CBZ (Z) protecting groups of compound (15) are removed by catalytic hydrogenation with 10% Pd / C as catalyst. The catalyst is filtered and the solvent is evaporated to produce aminoguanidine (16).

EXAMPLE 7

SYNTHESIS OF THE COMPOUND (10)

-228228

<img file="MX9704718A_D0376.tif" />

-229229

4-Methylmorpholine (NMM) was added to a solution of the carboxylic acid (2) (1.7 g, 4.9 mmol, 1.0 eq), 4-hydroxyproline (3) (5.39 mmol, 1.1 eq), and BOP reagent (2.17 g, 4.9 mmol, 1.0 eq) in anhydrous DMF (10 mL) at room temperature. The reaction mixture was stirred at room temperature overnight, quenched with brine

<td></td><td>(50 mL) and ethyl acetate</td><td>(100 mL)</td><td>. I know</td><td>washed organic coat</td>

<td></td><td>with aqueous citric acid</td><td> (10%, 2</td><td>x 50</td><td>mL), bicarbonate of</td>

<td></td><td>sodium (10%, 2 x 50 mL) and</td><td>brine</td><td> (50</td><td>mL). Layer dried</td>

<td> 10</td><td>resulting organic about</td><td>sulfate</td><td>of</td><td>anhydrous magnesium,</td>

<td></td><td>filtered and evaporated the</td><td>solvent.</td><td>The</td><td>primary waste is</td>

purified by evaporation chromatography (5: 4: 1, ethyl acetate-hexane-methanol). 1.1 g of the pure product (4) was recovered in a 48% yield.

STEP 2

-230230

<img file="MX9704718A_D0377.tif" />

(4) o ° c

1) trielhylamine

2) MsCI

------------&gt;

<img file="MX9704718A_D0378.tif" />

(5)

<td></td><td colspan="7">To a 4-hydroxyproline derivative solution</td>

<td> 5</td><td> (4)</td><td>(115 mg, 240</td><td>umol,</td><td> 1.0</td><td>eq) in</td><td>dichloromethane (10</td><td>mL,</td>

<td></td><td colspan="2">anhydrous) at 0 ° C</td><td>added</td><td colspan="2">triethylamine</td><td>(72 mg, 72 0 umol,</td><td> 3.0</td>

eq) and metasulfonyl chloride (28 mg, 240 umol, 1.0 eq) and the reaction mixture was stirred at room temperature. The mixture was then quenched with an aqueous 10 ammonia chloride solution and extracted with ethyl acetate. The organic layer was washed with 10% citric acid and brine, dried, filtered, and the solvent was evaporated to dryness, yielding compound (5).

STEP 3

-231231

<img file="MX9704718A_D0379.tif" />

(5)

1) Hg (0Ac)₂CF3C00H / 0 ° C

------------►

2) NaBH₄

<img file="MX9704718A_D0380.tif" />

(6)

Enamine (5) (1.0 eq) is treated with mercuric acetate (1.1 eq) in THF. The solvent is evaporated until dry and the residue is dissolved in methanol. The resulting organomercuric is reduced with sodium borohydride (1.3 eq). The resulting primary lactam thioether is purified by evaporation chromatography on silica gel to produce compound (6).

STEP 4

-232232

<img file="MX9704718A_D0381.tif" />

To a solution of lactam thioether (6) (1.0 eq) in dichloromethane N-chlorosuccinimide (1.0 eq) is added

0 ° C. The reaction mixture is warmed up to room temperature. When the mixture shows no more starting material, the solid is filtered and the solvent is evaporated until it is dry. The primary material (7) is used without any other purification for step 5.

STEP 5

-233233

<img file="MX9704718A_D0382.tif" />

<img file="MX9704718A_D0383.tif" />

(S)

To a solution of the alphachlorothioether (7) (1.0 eq) in THF (anhydrous), a solution of phenylcuprate (1.0 eq) (prepared according to the literature procedure) is added at room temperature. When the reaction mixture no longer has initial chlorothioether, brine and ethyl acetate are added. The organic layer is dried, filtered and evaporated to dryness to produce the desired product (8).

PASQ .. 6.

-234234

<img file="MX9704718A_D0384.tif" />

<img file="MX9704718A_D0385.tif" />

(8) is hydrolyzed with one equivalent of lithium hydroxide in a 1: 1 mixture of

THF and water. The mixture is stirred at room temperature for one hour. The primary mixture is extracted with ether, and the resulting solution is poured

10% and extracted with dichloromethane to produce the corresponding carboxylic acid (9).

PASQ £

-235235

<img file="MX9704718A_D0386.tif" />

<img file="MX9704718A_D0387.tif" />

The primary carboxylic acid (9) is coupled with benzodiazole ketoarginine in DMF using BOP as a coupling reagent in the presence of diisopropylethylamine. Extraction with EtOAc produces a solid which is purified on silica gel to produce the protected amide. The CBZ protecting group 10 is extracted with BBr in dichloromethane at room temperature, which finally produces the bicyclic benzothiazole ketoarginine inhibitors (10).

The following compounds were produced in accordance with the above, except that the appropriate substitution of the products were made in order to obtain the final compounds.

-236236

COMPOUND # 11

<img file="MX9704718A_D0388.tif" />

NH

Π.1) ^ = nh

Η N

COMPOUND # 12

<img file="MX9704718A_D0389.tif" />

NH

J = NH

Η N

0-2)

-237237

EXAMPLE 8

STEP 1

<img file="MX9704718A_D0390.tif" />

Commercially available glutaric acid monomethyl ester chloride (1) (20 ml, 0.144 mol) was dissolved in 40 ml of dry tetrahydrofuran (THF) and cooled to 15 ° C. Freshly prepared excess diazomethane was introduced to

300 ml of ether via cannula at -15 ° C to the solution. The mixture was allowed to warm to room temperature overnight. The excess diazomethane was evacuated from the flask with a

-238238 argon stream. To terminate the reaction, 75 ml of 1N HC1 in ether was added at 0 ° C, and allowed to warm to room temperature for 5 hours. Solvent volume was reduced and then washed with 2x 5% NaHCO₃, dried 5 over Na₂CO₃ and evaporated to produce primary chloromethylketone (20.46 g, 79%) which is used in the next step without further purification.

NMR (CDCL₃, 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).

PASQ. 2

-239239

<img file="MX9704718A_D0391.tif" />

Primary chloromethylketone (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 salt (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 hours 30 minutes. 2 00 ml of methanol (MeOH) was added to dissolve the solids, and the pH was adjusted to 2 with IN of HC1. The mixture was then basified with NaHCO₃ saturated until pH = 8 was obtained. MeOH was evaporated and the remaining aqueous solvent was washed with ethyl acetate and dichloromethane. Solvents were combined, dried over Na₂SW₄ and

-240240 evaporated. The primary residue was purified by silica gel evaporation column chromatography using a gradient of ethyl acetate / hexane eluents in the following proportions (3: 7, 5: 5, 6: 4, 7: 3) yielding the compound cyclical (3).

NMR (CDC1₃, 400 MHz) of compound (3) d 1.21-1.27 (t, 3H,

J = 7.06 Hz), 1.41-1.48 (m, 2H), 1.65-1.73 (τη, 2H), 2.28-2.39 (m, 4H), 2.57-2.63 (t, 1H, J = 10.9), 2.72-2.76 ( dd, 1H, J = 10.7

Hz), 2.8-2.86 (m, 1H), 3.6-3.64 (d, 4H, .J = 2.55 Hz), 3.63 (s,

3H), 4.13-4.2 (m, 2H).

¹³C NMR (CDC1₃, 400 MHz) 13.078, 19.888, 28.326, 31.133,

32.741, 35.277, 50.462, 56.394, 59.149, 60.188, 69.713,

170.182, 172.52.

STEP 4

-241241

<img file="MX9704718A_D0392.tif" />

Cyclic compound (3) (913 mg, 3.32 mmol) was dissolved in 50 ml of dry toluene. (IS) - (+) - 10-camphor sulfonic acid (92 mg, 0.39 mmol) was added and the mixture was allowed to reflux for 4 days. When the TLC showed that all the starting material was consumed, the mixture was maneuvered by evaporation of the solvent, dissolving residues in ethyl acetate and washing it with 2x5% NaHCO₃. The ethyl acetate layer was dried over Na₂SW₄ and evaporated. The primary residue was purified by silica gel evaporation column chromatography using 60% ETOAC / 40% hexane followed by

-242242

70% ETOAC / 30% hexane yielding 62.5% of the bicyclic compound (4).

NMR (CDC1₃, 400 MHz) of compound (4) d 1.27-1.31 (t, 3H,

J = 7 Hz), 1.5-1.6 (m, 1H), 1.72-1.87 (m, 2H), 2.02-2.1 (m, 1H),

2.33-2.46 (m, 2H), 2.83-2.88 (dd, 1H, J = 14,

STEP 5 (d,

1H), 3.78-3.85 (m,

1H), 4.2-4.27 (q, 2H,

<img file="MX9704718A_D0393.tif" />

<td> 15</td><td>Bicyclic dissolved</td><td>(4) (36 mg,</td><td>1.5 mmol)</td>

<td></td><td>in 25 mL of THF and 5 mL of H₂Or hydroxide</td><td colspan="2">lithium monohydrate</td>

<td></td><td>(1.1 eq, 7.05 mg, 1.68 mmol) was added</td><td>in 2.3 mol</td><td>from H₂Or, to</td>

-243243

0 ° C, and the mixture was allowed to stir at 0 ° C for 1 hour and at room temperature for 3 hours. THF was then evaporated and the remaining aqueous mixture was acidified by adding citric acid until obtaining a pH equal to 2. Extraction of the aqueous mixture with 2 x CH₂CL₂ and 2 X ETOAC, drying the organic layers combined with NA₂SW₄ and evaporation produced a primary residue which was purified by a silica gel evaporation chromatography column using 70% ETOAC / 30% hexane followed by 4.7% HOAC / ethyl acetate, yielding 10 pure acid (5) in yield 54%. 16% of the initial material was recovered (4).

^XH NMR (MeOD, 400 MHz) of compound (5) d 1.57-1.69 (m, 1H),

1.70-1.80 (m, 1H), 1.81-1.89 (m, 1H), 2.05-2.12 (m, 1H), 2.3515 2.5 (m, 2H), 2.51-2.66 (m, 2H), 2.86-2.91 (dd, 1H, J = 13.8, 4

Hz), 3.12-3.17 (d, 1H), 3.3-3.32 (m, 1H), 3.78-3.84 (m, 1H),

5.76-5.78 (t, IN, J = 3.53 Hz).

^{1 3}C NMR (MeOD, 400 MHz) d 17,052, 07/27, 28,928, 31,382,

32.096, 51.016, 55.138, 170.088, 171.24

-244244

<img file="MX9704718A_D0394.tif" />

(trimethylsiril) (5

<img file="MX9704718A_D0395.tif" />

A solution of lithium amide bis ml of 1M THF solution, mmol) in THF (10 ml) was added at -78 ° C a solution of carboxylic acid (5) (500 mg, 2.32 mmol). The resulting solution was stirred at 10 78 ° C for 1 hour. Benzyl bromide (0.26 ml, 2.22 mmol) was then added and the mixture was allowed to come to room temperature and stirred for 15 hours. The mixture was then poured into 10% HC1 (50 ml) and extracted with dichloromethane (4x 60ml). The combined organic phases were dried over MgSO₄ and the solvent was removed by evaporation to produce the primary alkylated amide (6).

-245245

PASQ 7

<img file="MX9704718A_D0396.tif" />

<img file="MX9704718A_D0397.tif" />

The primary alkylated amide (6) is coupled with benzithiazole ketoarginine in DMF using BOP as a coupling reagent in the presence of isopropylethylamine.

Extraction with EtOAc produces a solid which is purified on silica gel to produce the protected amide. The CBZ protected group extracts with BBr in dichloromethane at temperature

-246246 environment, which eventually produces the bicyclic benzothiazole ketoarginine inhibitors (7),

<td></td><td>According to</td><td>previous</td><td>I know</td><td>produced</td><td>the compound</td>

<td> 5</td><td>next except</td><td>That the</td><td colspan="2">substitution</td><td>appropriate of</td>

<td></td><td colspan="2">the products were made with</td><td>the</td><td>end of</td><td>get the</td>

<td></td><td>final compounds.</td><td></td><td></td><td></td><td></td>

<img file="MX9704718A_D0398.tif" />

EXAMPLE 9

-247247

Determination of K Values ​​of heterocyclics.

The affinity of the inhibitors for thrombin were measured according to the procedures described in (DiMaio et al., J. Bio. Chem., 1990, 265: 21698). Inhibition of amidolytic activity of human thrombin was measured fluorometrically using Tos-Gly-Pro-Arg-AMC as a fluorogenic substrate in 50 nM Tris-HCl buffer (pH

7.52 at 37 ° C) containing 0.1 M NaCl and 0.1% poly (ethylene glycol) 8000 at room temperature, and (Szewczuk et al., Biochemistry, 1992 31: 9132).

Hydrolysis of the substrate by thrombin was monitored on a Varian-Cary 2000 ™ spectrophotometer in fluorescent mode (XeX = 383 nm, Xem = 455 nm) or on a Hitachi F2000 ™ fluorescence spectrophotometer (λ = 383 nm, λ = 455 nm), and fluorescent intensity was calibrated using AMC. The reaction reached a stable state after three minutes after mixing thrombin with substrate and an inhibitor. Steady state velocity was then measured

-248248 for a few minutes. The compounds of the present invention were also preincubated with thrombin for 20 minutes at room temperature before adding the substrate. Steady state was achieved after 3 minutes and measured for a few minutes. Synthetic data (the steady state rate at various concentrations of the substrate and inhibitors) of competitive inhibition was analyzed using the methods described by Segel (1975). A nonlinear regression program was used, RNLIN in the IMSL library (IMSL, 1987), 10 LMDER in the MINPACK library (More et al., 1980) or

Microsoft ™ Excell ™, to estimate the kinetic parameters (K ,, V and K).

DTT test

Fibrin agglutination test was performed in 50 nM Tris HC1 buffer (pH 7.52 at 37 ° C) containing 0.1 M NaCl and 0.1% polyethylene glycol 8000 with 9.0 x 10-10 M (0.1 NIH unit / mL) and 0.03% (w / v) of the final concentrations of human thrombin and fibrinogen

-249249 bovine, respectively, as reported elsewhere (Szewczuk et al., Supra). Agglutination time was plotted against inhibitor concentrations, and IC was estimated.₅ as the required inhibitor concentration 5 to double the binding time relative to the control. The results are summarized in Tables 1 and 2 below.

Fibrin Clot Assay

The fibrin clot assay was performed essentially as described by Krtenansky et al., FEBS, 1987, 211: 10. The serial dilution of the inhibitor was prepared in 50 mM tris HC1 buffer (pH 7.8 at 23 ° C) containing 0.1 M NaCl and 0.1% (w / v) polyethylene glycol 8000. Human plasma (60 gL, collected in 3.8% sodium citrate, blood / anticoagulant 9: 1) was added to microwells (microwell plate, Falcon) containing 100 pL of various inhibitory dilutions. The solution was mixed after which 50 pL of human thrombin (1 nM concentration) was added.

-250250 final) and mixed for 15 seconds. Clot turbidity was immediately monitored by microplate auto-reader (Dynateck MR 500 0) at 4 05 nm and recorded every 3 minutes. The maximum turbidity in the absence of inhibitors was reached at 60 minutes. IC values ​​at 30 minutes were calculated as the concentration of inhibitors that produced half the optical density of the control.

Aggregation and Secretion of. Platelets

Rat blood was collected in ACD (6/1 v / v) by cardiac puncture. A suspension of washed platelets was prepared as described by Ardlie et al. (Br. J. Haematol. 1970, 19: 7 and Proc. Soc. Exp. Biol, Med.,

1971, 136: 1021). The final suspension medium was a modified thyroid solution (138 mM NaCl, 2.9 nM KC1, 20 mM HEPES, NaH₂PO₄0.42 mM, 12 mM NaHCO, CaCl₂, 1 mM, MgCl, 2 mM, 0.1% glucose, 0.35% albumin, apyrase lpL / mL pH 7.4), Platelet counts were adjusted to 5000,000 / gL.

-251251

To allow measurement of the measure of release of the contents of the dense granules, the platelets were labeled in the first wash solution with ¹⁴C-serotonin (5-HT) (lpCi / 10 mL of wash fluid) and the release of ¹⁴C-serotonin was determined as described in Holmsen et al. (Enzimolgy, 1989, 169: 206). Inipramine (5μΜ final concentration) to date was added for reabsorption of the released serotonin.

Platelet aggregation was recorded at 37 ° C in an aggregometer (BioData PAP-4) at a stirring speed of

1,100 rpm by measuring variations in light transmission. The percentage of the aggregation was determined 3 minutes after adding stimulating agents (human thrombin with a final concentration of 0.1 TU / mL). Inhibitors were preincubated 1 minute at 37 ° C before adding the stimulating agent. IC values₅ they represent the concentration necessary to inhibit platelet aggregation or secretion at 50% of the control.

Arterial Thrombosis Model

-252252

FeCl-Induced Carotid Arterial Damage Model.

FeCl-induced damage₃ in the carotid artery of rats it was induced according to the method described by

Main,

RW, Sandusky, GE,

Thrombosis Research

60; 269-280,

1990 and Schumacher, WA

and collaborators,

J.

Pharmacology and

Experimental

Therapeutics 267; 1237-1242,

1993.

Male rats, breed SpragueDawley (375-410 g) were anesthetized with urethane (1500 mg / kg ip). Animals were placed on a 37 ° C heating pad. A carotid artery was exposed through a mid-cerevic incision.

Careful blunt dissection was used to isolate the vessel from the carotid layer. Using forceps, the artery was raised to provide enough space to insert the small pieces of polyethylene tubing (PE-205¹) under these. A temperature probe (Physitemp MT23 / 3) was placed between one of the pieces of the tubes and the artery. Damage was induced by topical application to the carotid artery on the temperature probe of a small disc (3 millimeters of

-253253 diameter) of Whatman No. 1 filter paper previously dipped in a 35% FeCl solution₃. The incision area was covered with aluminum foil in order to protect the FeCl₃of light degradation. The temperature of the blood vessel 5 was controlled for 60 minutes after the application of

FeCl₃ as an indication of blood flow. Changes of. vessel temperature in a thermistor (ColePalmer Model 08533-41).

The time elapsed between the application of the

FeCl₃ and the time when the temperature of the vessel abruptly decreased (&gt; 2.4 ° C) the time elapsed to occlude the vessel was recorded as time.

Inhibitory compounds were given as iv bolus (mg / kg) followed immediately by iv ^ g / kg / min infusion. through the femoral vein). The dose of inhibitor necessary to double the occlusion time was determined, compared to control animals where the damage was induced in the absence of the inhibitor.

-254254

Table 1

<td rowspan="2">cmpd</td><td colspan="3">Antiplatelet activity μΜ</td><td rowspan="2">dTTIC μΜ</td><td rowspan="2">IC test_Gof plasma fibrin clots IC</td>

<td>ΚΙ μΜ</td><td>Aggregation</td><td>5-HT Secretion</td>

<td> 0005</td><td> 4</td><td>ND</td><td>ND</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></td><td> 162</td><td> &gt;450</td>

<td> 0020</td><td> 2.2</td><td> 18</td><td> &gt;100</td><td> 22</td><td> &gt;450</td>

<td> 0025</td><td> 53</td><td> &gt;100</td><td> 14.2</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>ND</td><td> &gt;100</td><td> 415</td><td> &gt;450</td>

<td> 0050</td><td> 62</td><td>ND</td><td>ND</td><td></td><td> &gt;450</td>

<td> 0065</td><td> 32.7</td><td> 47.5</td><td>ND</td><td> 42</td><td> 200</td>

<td> 0070</td><td> 4.4</td><td> 22</td><td> 52</td><td> 25</td><td> 78</td>

<td> 0080</td><td> 0.048</td><td> 0.4</td><td> 2.1</td><td> 0.375</td><td>ND</td>

<td> 0090</td><td> 0.031</td><td>ND</td><td> 0.38</td><td> 0.33</td><td>ND</td>

<td> 0095</td><td> 26</td><td>ND</td><td>ND</td><td></td><td></td>

<td> 0100</td><td> 19</td><td>ND</td><td>ND</td><td> 165</td><td>ND</td>

* suspension of washed platelets from rats * constant inhibitory cleavage for human α-thrombin

-255255

<td>Compound</td><td>ki (nM)</td><td>dTT (nM)</td><td>Ivb + lnf route</td><td>MOT (min) ± sem.</td>

<td> 0220</td><td> 18</td><td></td><td></td><td></td>

<td> 0225</td><td> 2250 235</td><td></td><td> 0.75-50</td><td> 2317</td>

<td> 0245</td><td> 5 8</td><td></td><td> 0.5-30 0.75-50</td><td> 2713 22.612.6</td>

<td> 0250</td><td> 40</td><td> 350</td><td> 0.25-20 0.75-50</td><td> 2318 2213</td>

<td>0295a</td><td> 1500</td><td></td><td> 0.75-50</td><td> 2011</td>

<td>0295b</td><td></td><td> 5000 520</td><td> 0.75-50</td><td> 1912.7</td>

<td> 0240</td><td> 18</td><td></td><td> 0.75-50</td><td> 1712.6</td>

<td> 0210</td><td> 8</td><td></td><td> 0.75-50</td><td> 14.8310.2</td>

<td> 0225</td><td> 500</td><td></td><td></td><td></td>

<td> 0260</td><td> 16</td><td></td><td> 0.75-50</td><td> 14.8310.2</td>

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

<td>0305b</td><td> 12000</td><td></td><td></td><td></td>

<td>0265a</td><td> 4</td><td></td><td> 0.75-50</td><td> 21.5110</td>

<td>0265b</td><td> 18</td><td></td><td> 0.75-50</td><td> 14.8312.3</td>

<td> 0285</td><td> 10 150</td><td></td><td> 0.75-50</td><td> 11.3311.34</td>

<td>0315a + b</td><td> 45</td><td></td><td> •</td><td></td>

<td>0315b</td><td> 10</td><td></td><td> 0.75-50</td><td> 30.3318.4</td>

<td> 0335</td><td> 25</td><td> 138</td><td> 0.75-50 0.5-30 0.25-20</td><td> 45.8114.2 41.517.27 27.5111.3</td>

<td> 0340</td><td> 0.6</td><td></td><td> 0.25-20 0.75-50</td><td> 3619.6 42.25111.3</td>

<td> 0345</td><td> 2</td><td></td><td> 0.75-50</td><td> 5015.86</td>

<td> 0915</td><td> 1600</td><td></td><td> 0.75-50</td><td> 1511.3</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> 19.610.2</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> 20.313.5</td>

<td>0940a</td><td> 16</td><td></td><td> 0.75-50</td><td> 15.210.82</td>

-256256

<td>0950a</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="5">a = early allusion of RP HPLC single isomer b = late elusion over RP HPLC single isomer a + b = mixture</td>

-257257