Inhibitors of factor viia

 

(57) Abstract:

The invention relates to new compounds of General formula I

R1-A-B-D-En-R2 (I)

in which R1 represents R12C(O), and R12 is selected from the group consisting of alkenyl, alkenylacyl or alkenylamine; And is a group A1-A2-A3, where A1 represents NH, A2 is a CHR93 in which R93 is 4-amidinophenoxy; A3 represents C(O); is a group B1-B2-B3, where B1 represents NH; B2 is a CHR97 where R97 represents ethyl, which is substituted in position 2 by hydroxycarbonyl or allyloxycarbonyl; B3 represents C(O); D represents a group D1-D2-D3, where D1 represents NH, D2 represents CR81R82 where R81 and R82 are independently selected from the group consisting of hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroallyl; D3 represents C(O); En is a (E1-E2-E3)nin which n is 0 or 1; E1 represents NR70, where R70 is H; E2 represents CR71R72, where R71 and R72 include independently selected from the group consisting of hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroaryl the x or substituted alkyl residues, aryl, arylalkyl, heteroallyl and geterotsiklicheskikh, and the alkyl contains from 1 to 13 carbon atoms, alkenyl contains from 2 to 13 carbon atoms, aryl and heteroaryl contain from 5 to 13 ring carbon atoms, where in the rest of heteroaryl one or more carbon atoms are replaced by heteroatoms selected from the group consisting of N, O and S; heteroseksualci contains from 3 to 8 ring carbon atoms, of which from one to three carbon atoms are replaced by heteroatoms selected from the group consisting of N, O and S; in any stereoisomeric forms or their mixtures in any ratio, and their pharmaceutically acceptable salts; the method of obtaining compounds of General formula I, including linking protected amino acids; to pharmaceutical compositions which are able to exert an antithrombotic effect by activated factor VII(FVIIa) blood coagulation. 3 C. and 16 h.p. f-crystals, 4 PL.

The present invention relates to new compounds, their reception, their use and to pharmaceutical compositions containing these compounds, which have a strong antithrombotic effect through reversible inhibition of activated factor VIIa (FVIIa) blood clotting.

Blood clotting is a complex process involving progressive progressive cascade of reactions the activation of enzymes, which by limited proteolysis sequentially activated Imogene plasma. The cascade of blood coagulation was mechanically divided into internal and external paths that converge at the activation of factor X; and the subsequent generation of thrombin continues a single common path (see diagram 1).

Figure 1: the Cascade of blood coagulation

Currently available evidence suggests that the internal path plays an important role in maintaining and growing the formation of fibrin, while the external path is crucial in the initiation phase of blood coagulation (H. Cole, Aust.J.Med.Sci. 16 (1995) 87; G. J. Broze, Blood Coagulation and Fibrinolysis 6, Suppl. l (1995) S7-S13). It is generally recognized that the blood clotting physically triggered by the formation of the complex of tissue factor (TF) / factor VIIa. Once formed, this complex rapidly initiates coagulation by activating FMU with factor Va and phospholipids to form complex prothrombinase, who is responsible for the conversion of soluble fibrinogen into insoluble fibrin by activated thrombin from its precursor prothrombin. Over time the activity of the complex of factor VIIa/tissue factor (outer path) is suppressed protein-protease inhibitor Kunitz type, TFPI, which in the formation of a complex with factor XA can directly inhibit the proteolytic activity of factor VIIa/tissue factor. To maintain the clotting process in the presence of inhibited exogenous system under the influence of mediated thrombin activity internal path produces an additional factor XA. Thus, thrombin plays a dual autocatalytic role Poreba its own production and the conversion of fibrinogen to fibrin.

Autocatalytic nature of the generation of thrombin is an important protective mechanism against uncontrolled bleeding, and it ensures that if this threshold level prothrombinase coagulation will continue until completion. The ability to form blood clots is vital for survival. However, under certain pathological conditions about which the Eney, under such pathological conditions desirable to completely inhibit the clotting system, because it would have caused life-threatening bleeding. Thus, it is most desirable to develop tools that inhibit coagulation by inhibiting factor VIIa without direct inhibition of thrombin.

Many clinical applications there is a great need in the prevention of intravascular blood clots, or in some anticoagulant treatment. Many specific types of clinical application of currently available drugs are unsatisfactory. For example, almost 50% of patients who underwent total hip arthroplasty, developing deep vein thrombosis (DVT). Approved in present methods of treatment are the use of fixed-dose low molecular weight heparin (NMG) and different doses of heparin. Even with these schemes of drug therapy in 10-20% of patients develop DVT, and 5-10% develop complications related to bleeding.

Another clinical situation in which required more sophisticated anticoagulants, for persons exposed to angina. The current conventional therapy, which consists in the introduction of heparin and aspirin, is associated with the frequency of sudden occlusion of vessels within 24 h after the procedure, component from 6% to 8%. The frequency associated with bleeding complications requiring transfusion therapy due to the use of heparin, is approximately 7%. Moreover, even though significant remote occlusion of vessels, the introduction of heparin after the intervention is ineffective and can have harmful effects.

The most widely used inhibitors of blood coagulation are heparin and related sulfated polysaccharides, NMG and heparansulfate. These molecules exert their anticoagulant effects, contributing to the binding of the natural regulator of the coagulation process, anti-thrombin III, thrombin and factor XA. Inhibiting activity of heparin primarily focused on thrombin, which is inactivated in approximately 100 times faster than a factor of CA. Hirudin and hirulog are two more specific in relation to anticoagulant thrombin, which are currently undergoing clinical trials. However, e is the cue studies in baboons and dogs showed that sighting guides enzymes involved in the early stages of the coagulation cascade, such as factor XA or factor VIIa, prevent the formation of a clot, without causing side effects such as bleeding observed in the use of direct thrombin inhibitors (T. Yokoyama, A. B. Kelly, U. M. Marzec, S. R. Hanson, S. Kunitada, L. A. Harker, Ciculation 92 (1995) 485-491; L. A. Harker, S. R. Hanson, A. B. Kelly, Thromb. Hemostas 74 (1995) 464-472; C. R. Benedict, J. Ryan, J. Todd, K. Kuwabara, P. Tyburg, Jr., J. Cartwright, D. Stern, Blood 81 (1993) 2059-2066).

Specific inhibition of the catalytic complex of factor VIIa/TF using monoclonal antibody (international patent application No. W92/06711) and protein, such as an inactivated by chloromethylketone factor VIIa (international patent application No. W96/12800 and W97/47651) is a highly effective means of controlling formation of a blood clot, caused by acute damage to the arteries, or thrombotic complications associated with bacterial septicemia. There is also experimental evidence, indicating that the inhibition of the activity of factor VIIa/TF inhibits restenosis after balloon angioplasty (L. A. Harker, S. R. Hanson, J. N. Wilcox, A. B. Kelly, Haemostasis 26 (1996) S1:76-82). Of bleeding studies were conducted on baboons, and they show that the inhibition of koska bleeding from all the tested approaches to anticoagulation therapy, including the inhibition of thrombin, platelets and factor XA (L. A. Harker, S. R. Hanson, A. B. Kelly, Thromb. Hemostas 74 (1995) 464-472).

Specific inhibitor of factor VIIa would have significant practical value in clinical practice. In particular, the inhibitor of factor VIIa would be effective in the conditions under which the currently used drugs of choice, heparin and related sulfated polysaccharides, ineffective or have only a borderline effect. Thus, there is a need for low-molecular-specific factor VIIa the coagulation inhibitor, which is effective but does not cause unwanted side effects. The present invention meets this requirement by providing derivatives of the formula I, inhibiting the activity of factor VIIa, and also by providing related advantages.

The compounds of formula I are inhibitors of the enzyme factor VIIa coagulation. The invention also relates to methods of preparing compounds of the formula I, to a method of inhibiting the activity of factor VIIa and inhibition of blood clotting, to the use of compounds of formula I for the treatment and prevention of diseases that can be cured or to prevent the stenosis, heart attack and angina, and to the use of compounds of formula I for the preparation of drugs, which are expected to apply in such cases. The invention also relates to compositions containing compounds of formula I, mixed or connected in other ways with an inert carrier, in particular, to pharmaceutical compositions containing a compound of formula I together with pharmaceutically acceptable substances-carriers or fillers and/or additional substances or additives.

The invention

The present invention provides compounds that specifically inhibit the activity of factor VIIa. In particular, the object of the present invention are the compounds of formula I

R1-A-B-D-En-R2 (I)

in which R1 represents R13, R12C(O) or 1 to 3 amino acids, N-end which can be substituted by the Deputy selected from the group consisting of R14C(O), R15S(O)2and group protecting the amino group in which

R12 is selected from the series consisting of alkyl, alkenyl, quinil, alkyloxy, alkylamino, alkynylamino, alkynylamino, alkenylacyl, alkyloxy, aryl, heteroaryl, geterotsiklicheskie, arylalkyl, heterokedasticity,

R13 is selected from a range consisting of a group protecting the amino group, hydrogen, alkyl, aryl, arylalkyl, heteroaryl, heteroaromatic, geterotsiklicheskie and geterotsiklicheskikh,

R14 and R15 are independently selected from the series consisting of alkyl, aryl, arylalkyl, heteroaryl, heteroaromatic, geterotsiklicheskie and geterotsiklicheskikh,

And is a group A1-A2-A3, in which

A1 represents NH,

A2 represents CHR93 in which R93 is a 4-medinformatix,

A3 represents C(O)

Represents a group B1-B2-B3, in which

B1 represents NR95, in which R95 is selected from the series consisting of hydrogen and alkyl,

B2 represents CHR97 in which R97 represents ethyl, which is substituted in position 2 by a Deputy chosen from the series consisting of hydroxycarbonyl, allyloxycarbonyl and arylalkylamine,

B3 represents S(O)

D is a group D1-D2-D3, in which

D1 represents NH,

D2 represents CR81R82 where R81 and R82 are independently selected from the series consisting of hydrogen and unsubstituted or substituted stadelschule WITH a(O)

En is a (E1-E2-E3)nin which n is 0, 1, 2 or 3,

E1 represents the NR70, where R70 is selected from the series consisting of hydrogen, alkyl, aryl, arylalkyl, heteroaryl, heteroaromatic, geterotsiklicheskie and geterotsiklicheskikh,

E2 represents CR71R72 where R71 and R72 include independently selected from the series consisting of hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroaryl, heteroaromatic, geterotsiklicheskie and geterotsiklicheskikh,

E3 represents S(O)

R2 is selected from the series consisting of NR21R22, OR23 and R24, in which R21, R22, R23 and R24 are independently selected from the series consisting of hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroaryl, heteroaromatic, geterotsiklicheskie and geterotsiklicheskikh,

alkyl and heteroalkyl contain from 1 to 13 carbon atoms, where in the rest of heteroalkyl one or more carbon atoms are replaced by heteroatoms selected from the series consisting of N, O and S;

alkenyl, quinil, heteroalkyl and heteroalkyl contain from 2 to 13 carbon atoms, where the remainder heteroalkyl and heteroalkyl one or more carbon atoms substituted gets the carbon atoms, where in the rest of heteroaryl one or more carbon atoms are replaced by heteroatoms selected from the series consisting of N, O and S;

heteroseksualci contains from 3 to 8 ring carbon atoms, of which from one to three carbon atoms are replaced by heteroatoms selected from the series consisting of N, O and S;

in any stereoisomeric forms and mixtures in any ratio, and their pharmaceutically acceptable salts.

The present invention provides peptides of formula I

R1-A-B-D-En-R2 (I)

in which R1, R2, a, b, D, E, and n are defined as above, and which are compounds that inhibit the activity of factor VIIa, but does not substantially inhibit the activity of other proteases involved in the pathway of blood coagulation. In the compounds of the formula I, for example, in groups a, B, D or E, or a group R1 if R1 represents 1, 2 or 3 amino acids, contains structural elements that represent amino acids or their derivatives, or analogs of amino acids, or mimetic patterns, and which type of peptides linked to the adjacent groups via amide linkages(O)-N formed between the carboxyl group of one such amino acids and t the or group, like a, b, D or E is present in formula I, is obtained from the corresponding amino acids in the formal removal of a hydrogen atom from an amino group and a hydroxyl group from the carboxyl group.

Used herein, the term “amino acid” is used in its broadest sense to refer to the 20 naturally occurring amino acids, which are translated from the genetic code and contain the building blocks of proteins, including, unless specifically indicated otherwise, L-amino acids and D-amino acids, chemically modified amino acids, such as analogs of amino acids, naturally occurring amino acids that are not usually incorporated into proteins, such as norleucine, and chemically synthesized compounds having properties known in this field, characteristic of amino acids. For example, analogs or mimetics of phenylalanine or Proline, which provide the same conformational restriction of the peptide compounds as natural Phe or Pro included in the definition of “amino acids” and well-known experts in this field. Such analogs and mimetics are referred to here as the “functional equivalent” amino acids. Other examples of amino acids and analogs is c. New York, 1983, which is incorporated here by reference). Abbreviations of amino acids, amino acid analogues and mimetic structures, and other abbreviations used in the application are listed in the table.1.

Table 1

Abbreviations used in the application

Connection/residue Abbreviation

Acetic acid Asón

Acetamidomethyl AFM

Alanine l

Allyloxycarbonyl Alloc

para-Amitiptyline pAph

2-Aminobutyric acid 2-Abu

Arginine Arg

Asparagine is Asn

Aspartic acid Asp

Benzyl Bzl

tert-Butyloxycarbonyl Boc

tert-Butyl tBu

Cyclohexylglycine Chg

Cyclohexyl Chx

Cyclohexylamin Cha

Cysteine Cys

2,4-Deaninmesa acid Dab

2,3-Diaminopropionic acid Dap

Dichloromethane DCM

Diisopropylcarbodiimide DIC

Diisopropylethylamine DIEA

N,N-Dimethylformamide DMF

Dimethyl sulfoxide DMSO

9-Fertilityscore Fmoc

Glutamic acid Glu

Glutamine Gln

Glycine Gly

Histidine His

N-Hydroxyl is in Leu

Lysine Lys

Methyl Me

N-Mei NMI

N-Methylmorpholine NMM

2,2,5,7,8-Pentamethylchroman-6-sulfonyl Pmc

The Orn ornithine

Phenyl Ph

Phenylalanine Phe

Phenylglycine Phg

Proline Pro

Serine Ser

Tetrahydrofuran THF

Tetramethylpiperidinyloxy TFFH

Threonine is Thr

Triperoxonane acid TFA

Trail Trt

Tryptophan is Trp

Valine Val

In the absence of other refinements of the above amino acids in the form of the abbreviations have the L-configuration. Amino acids are the D-configuration indicated by the prefix D using the three-letter code (for example, D-Ala, D-Cys, D-Asp, D-Trp, D-pAph). Abbreviations, such, for example, Phe(4-CN) and Phe[4-C(-S-CH2-CH2-S-)-Ph] mean the rest of the amino acids Fe-nylalanine, which is in position 4 of the phenyl group bears, respectively, cyanogaster or Deputy 2-phenyl-1,3-ditiolan-2-yl. Abbreviation, like, for example. Dap[-C(=NH)-NH2] means the balance of amino acids 2,3-diaminopropionic acid in which the amino group in the side chain, i.e., the amino group at position 3 substituted by amidinopropane-C(=NH)-NH2(carbamimidoyl group), the Yu 3 propionic acid. Abbreviations, such, for example, Orn[-C(=NH)-NH2] or Cys(Me), mean residue of the amino acid ornithine, in which, respectively, the amino group in the side chain carries amidinopropane or the remainder of the amino acid cysteine, which mercaptopropyl carries a methyl group.

Terms TOTU, HATU and a THIEF means, respectively, O-[cyan(etoxycarbonyl)methylamino]-1,1,3,3-tetramethylpropylenediamine, O-(7-asobancaria-1-yl)-1,1,3,3-tetramethyleneglutaric and 1 benzothiazolinone-Tris-(dimethylamino)-fosfodiesterasa.

Used herein, the term “specific” when used with reference to the inhibition of the activity of factor VIIa means that the compound of the formula I can inhibit the activity of factor VIIa without significantly inhibiting the activity of other specific proteases, including plasmin and thrombin (using the same concentration of the inhibitor). Such proteases are involved in the cascade of blood coagulation and fibrinolysis.

Used herein, the term “Deputy” refers to any of various chemical groups that substituted for those disclosed here, the frame of the peptide or the side chain of a peptide analogue of a peptide mimetic or organic compound. Zamestitelyami, publication Giannis and Kolter, Angew. Chem. Int. Ed. Engl. 32 (1993) 1244-1267, which is incorporated here by reference).

Used herein, the term “alkyl” is used in its broadest sense to denote a saturated or unsaturated, linear, branched or cyclic chains of about 1-13 carbon atoms, in which unsaturated alkyl group, of course, contains at least 2 carbon atoms, and cyclic alkyl group of at least 3 carbon atoms. Unsaturated group may contain one or more double bonds and/or triple bonds. Thus, the term “alkyl” includes, for example, methyl, ethyl, n-sawn, ISO-propyl, n-boutelou, isobutylene, second-boutelou, tert-boutelou, 1-methylbutyl, 2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropylene, n-pentelow and n-hexoloy group, alkylene group, a cyclic chain of carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentamine, tsiklogeksilnogo and cycloheptyl groups, and combinations of linear or branched chains and cyclic chains of carbon atoms, such as methylcyclohexyl, cyclohexylmethyl, 1-cyclohexylethyl, 2-cyclohexylethyl, cyclopentylmethyl or cyclopropylmethanol group. Thus, alkyl also includes cyclic alkyl groups, which carry one or more alkyl substituents. Further examples of alkyl are mentioned below specific unsaturated group. In addition, it should be understood that the particular here, the alkyl may be substituted by one or more identical or different substituents, for example one, two, three or four substituents which may be present in any desired suitable position.

The term “alkyl” preferably means a saturated, linear or branched chain of 1 to 6 carbon atoms, unsaturated linear or branched chain of 2 to 6 carbon atoms or a cyclic alkyl group of 3-8 carbon atoms, in particular from 3 to 6 or 4 to 6 ring carbon atoms. From the point of view of unsaturated alkyl chains are preferred (C2-C6)-alkenyl and (C2-C6)-quinil. Examples of unsaturated alkyl groups are alkenylphenol and alkylamino groups such as vinyl, prop-1-enyl, prop-2-enyl (=allyl), but-2-enyl, butene-3-yl, 3-methylbut-2-enyl, ethinyl, prop-2-inyl, but-2-inyl and the like.

Similarly, the term “acyl” ispolzuetsia chains of about 1-13 carbon atoms, or aryl groups, having from 5 to 13 ring carbon atoms, which is attached to a carbonyl group-C(O)- and it is connected through the specified carbonyl group. The acyl group can be considered as obtained from the corresponding compounds containing carboxyl group(ABOUT)-HE by formal removal of the hydroxyl group. Thus, the term “acyl” includes, for example, such groups as formyl, acetyl, benzoyl and the like. A preferred group of the acyl group covers mentioned here above saturated or unsaturated, linear, branched or cyclic chain, with a preferred range of carbon atoms, which optionally contain a carbonyl group, through which they are associated.

The term “aryl” refers to aromatic groups containing from about 5 to 13 ring carbon atoms and at least one ring group having a conjugate system of pi electrons. Preferably, the term “aryl” refers to aromatic groups having from 6 to 10 ring carbon atoms. Examples of aryl include, for example, phenyl, naftalina, such as 1-naftalina and 2-naftalina, tortilla, biphenylene groups and their analogues and proizvodnymi, identical or different substituents which may be present in any desired suitable position. For example, one-deputizing phenyl group may be substituted in position 2-, 3 - or 4-, disubstituted phenyl group in position 2,3-, 2,4-, 2,5-, 2,6-, 3,4- or 3.5.

The term “arylalkyl” refers to alkyl as defined above, substituted by one or more, for example one or two, identical or different groups. Suitable arylalkyl groups include benzyl, phenylethyl, such as 1-phenylethyl and 2-phenylethyl, diphenylmethyl, diphenylether, such as 1,2-diphenylether and 2,2-diphenylether, phenylpropyl, such as 1-phenylpropyl, 2-phenylpropyl and 3-phenylpropyl, diphenylpropyl, such as 2,3-diphenylpropyl and 3.3-diphenylpropyl, naphthylmethyl, naphtalate, such as 1-naphtalate and 2-naphtalate, naftilamin, such as 1-naftilamin, 2-naftilamin and 3-naftilamin, 1,2,3,4-tetrahydro-1-naphthyl, 1,2,3,4-tetrahydro-2-naphthyl and the like, all of which may be optionally substituted.

Used herein, the terms “heteroalkyl”, “heteroalkyl”, “heteroalkyl”, “heteroaromatic and heteroaryl” refer respectively to alkyl, arylalkyl and aryl groups in which one or the AMI, such as N, O or S. in Addition, the term “heteroseksualci” is used when referring to a cyclic alkyl group in which one or more ring carbon atoms are replaced by heteroatoms. Preferably, the term “heteroseksualci” means a cyclic alkyl group having 3 to 8 ring carbon atoms, 1, 2 or 3 of which are substituted by identical or different heteroatoms, such as N, O or S.

All of these groups can be linked via any desired position, including a suitable ring nitrogen atoms in the case of nitrogen heterocycles. Suitable heteroaryl groups, heteroallyl group, heteroalkyl group and heterocytolysine groups include, for example, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-thienyl, 3-thienyl, indolyl, imidazolyl, furyl, piperonyl, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, 1-(2-pyridyl)ethyl, 1-(3-pyridyl)ethyl, 1-(4-pyridyl)ethyl, 2-(2-pyridyl)ethyl, 2-(3-pyridyl)ethyl, 2-(4-pyridyl)ethyl, picolyl, pyrrolidinyl, piperidinyl, tetrahydrofuryl, tetrahydrofuran-2-ylmethyl, morpholinyl, 4-morpholinyl, 2-(4-morpholinyl)ethyl, piperazinil, 2-(4-methylpiperazin-1-yl)ethyl and the like, all of which may be optionally substituted by one or more, the ACLs to the invention can be modified at the N-end and/or the end through reaction with suitable reagents or by introducing (or presence), accordingly, the group protecting the amino group, or a group protecting the carboxyl group. N-end of the peptide or analogue of the peptide may be chemically modified so that the N-terminal amino group is substituted, for example, acyl group (e.g. acetyl, cyclopentanecarbonyl, skinlightening, frailey, toiley, bentilee, pyrazinecarboxamide or other such groups) through reaction with isocyanate, chloroformate, alkylating agent or by introducing another such group, all of which can be substituted by the Deputy, as described above. It should be understood that the term “amino group” is used here broadly to refer to any free amino group, including primary, secondary, or tertiary amino group present in the peptide. For comparison, the term “N-end” refers to the-amino group of the first amino acids present in the peptide, written in the usual way.

N-end of the peptide according to the invention can be protected by attaching to it the group protecting the amino group. The term “group protecting the amino group” is used here broadly to refer to a chemical group that can react with a free amino group, including, is based amino group, protects reactive otherwise, the amino group against undesirable reactions which can occur, for example, during the procedure of synthesis or due to the activity of ectopeptidases with respect to the target connection. Modification of the amino group can also provide additional benefits, including, for example, increasing the solubility or activity of the compounds. Here disclosed or otherwise known in the field of various group protecting the amino group, and they include, for example, acyl groups such as acetyl, tert-butyloxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl group or benzoline group, and aminoallyl a residue that can be altered with the help of a group protecting the amino group. Other group protecting the amino group described, for example, in the book " the Peptides, eds. Gross and Meienhofer, vol.3 (Academic Press, Inc., New York, 1981); and in the publication of Greene and Wuts Protective Groups in Organic Synthesis, 2nd ed., pages 309-405 (John Wiley&Sons, New York, 1991), each of which is included here as a reference. The product of any such modification, N-terminal amino group of the peptide or analogue peptide of the invention referred to here as the “N-terminal derivative”.

Similarly, a carboxyl group, such as Kauppi, protecting the carboxyl group. The terms “carboxyl group” and “the end” are used in a consistent manner with above defined terms “amino group” and “N-end.” Carboxyl group, such a group, which is present at the C-end of the peptide can be modified by restoring the C-terminal carboxyl group to the alcohol or aldehyde or through the formation of ester for oral use, or by substitution of the carboxyl group such as Deputy thiazolyl, cyclohexyl or another group. Esters for oral use are well known in this field and include, for example, acyloxymethyl groups, such as methoxymethyl, ethoxymethyl, supraoccipital and the like; 1-((C1-C4)-alkyloxy)ethyl groups such as methoxyethyl, ethoxyethyl, propoxyethyl, isopropoxide and the like; 2-oxo-1,3-dioxolan-4-ylmethylene groups such as 5-methyl-2-oxo-1,3-dioxolan-4-ylmethyl, 5-phenyl-2-oxo-1,3-dioxolan-4-ylmethyl and the like; ((C1-C3)-alkylthio)methyl group, such as methylthiomethyl, ethylthiomethyl, isopropylaminomethyl and the like; acyloxymethyl groups, such as pivaloyloxymethyl, acetoxymethyl the Thiel; 3-phthalidyl or 5,6-dimethylpyridine group; 1-(((C1-C4)-alkyloxy)carbonyloxy)ethyl groups such as 1-(ethoxycarbonyl)ethyl group; and 1-(((C1-C4)-alkylamino)carbonyloxy)ethyl group, such as 1-(methylaminorex)ethyl group.

The peptide according to the invention can be modified by linking with it group protecting the carboxyl group. Group, a carboxyl protecting group well known in this area, and by binding with the peptide they protect the carboxyl group against undesirable reactions (see, for example, Greene and Wuts, supra, pages 224-276 (1991), which is incorporated here by reference). The specialist will be clear that the above-described modifications that may be made against the N-terminal amino group or the carboxyl end group of a peptide in a similar manner can be carried out in respect of any reactive amino groups or carboxyl groups present, for example, on the side chain of the amino acid or analogue of the amino acids in the peptide according to the invention. Methods of performing such modifications are disclosed here or known in this area from other sources.

The choice of inclusion of L - or D-and is. what, for example, the inclusion of one or more D-amino acids can make the connection enhanced stability in vitro or in vivo. The inclusion of one or more D-amino acids can also increase or decrease the pharmacological activity of the compounds. In some cases it may be desirable to provide the connection to remain active for only a short period of time. In such cases, the inclusion of one or more L-amino acids in the connection can allow endogenous peptidases from the individual to digest the compound in vivo, thereby limiting the impact on the individual active compounds. An experienced specialist can determine the desirable characteristics required of the compounds according to the invention, taking into account, for example, age and General health of the individual. In General, the present invention relates to compounds of the formula I, in all their stereoisomeric forms and to mixtures of two or more stereoisomers in all ratios, for example, to the pure enantiomers, pure diastereomers, mixtures of the two enantiomers in all ratios, including the racemates, mixtures of diastereomers, cisisomer, TRANS-isomers, E-isomer or Z-isomer. The invention also relates to the s of the formula I, such as esters, Amida, aldehydes or alcohols, which, as mentioned, can be obtained from carboxyl groups, or acyl derivative, such (C1-C6)-alkylcarboxylic, (C1-C6)-allyloxycarbonyl or aryl-(C1-C4)-allyloxycarbonyl derivatives, which can be obtained from alleluiah groups including amino groups, aminogroup, guanidinium and amidinopropane, and the invention also relates to the active metabolites of compounds of formula I.

In the compounds of formula I, the group R1 preferably represents R12C(O). A special group definitions R12 formed by a group of alkyl, alkenyl, quinil, alkyloxy, alkylamino, alkynylamino, alkynylamino, alkenylacyl, alkyloxy, aryl, heteroaryl, geterotsiklicheskie, heteroallyl, geterotsiklicheskikh, heteroalkyl, heteroalkyl and heteroalkyl, all of these residues can be unsubstituted or substituted. R12 preferably represents alkyl, alkenyl, quinil, alkyloxy, alkenylacyl, alkyloxy, alkylamino, alkynylamino, alkylamino, aryl, heteroaryl, arylalkyl or heteroaromatic, more preferably, alkenylacyl, alkenylamine or aryl, all of these ostly alkenylamine, like (C2-C6)-alkenylamine or (C2-C6)-alkynylamino, each of which contains one double bond, for example, allyloxy or allylamine. In addition, preferably R12 is a (C2-C6)-alkenylamine. The remains representing R12 may be unsubstituted or substituted. In the substituted residues residues R12 is preferably substituted by one or more identical or different substituents chosen from the series consisting of halogen, i.e. fluorine, chlorine, bromine or iodine, trifloromethyl, hydroxy, nitro, amino, cyano, carboxy, aminocarbonyl, alkylsulfonyl, aminosulfonyl, alkyloxy, alkylcarboxylic and mono - or dialkylamino. Similarly remains representing R13, R14 and R15 may be unsubstituted or substituted, for example, substituents which may be present in R12, where R14 and R15 independently of each other and can be identical or different.

Group a in the compounds of formula I, which represents the bivalent residue of 4-amitiptyline-NH-CH[-CH2-C6H4TO(4-C(=NH)-NH2)]-C(O)-, preferably represents the residue of (L)-4-amitiptyline (=residue (S)-4-amitiptyline). The group, which Pavlichenko acceptable salt or ester, preferably represents the residue of (L)-glutamic acid (=residue (S)-glutamic acid) or its pharmaceutically acceptable salt or ester.

R95 preferably represents hydrogen or (C1-C4)-alkyl, more preferably hydrogen or methyl, particularly preferably hydrogen.

Substituted residues R81 and R82 are independently bear one or more, for example 1, 2, 3 or 4 identical or different residues, which are preferably chosen from the series consisting of amino, aminocarbonyl, amidine, guanidine, aminoalkyl, hydroxy, mercapto, all of which can be substituted by a protective group, and acetamido (-C(=NH)-CH3), nitro and cyano. As for the nitro group in compounds of formula I in accordance with the invention preferably are present only up to two nitro groups. Suitable protective groups for the listed groups are known to the expert in this area and can be found in the above references, such publications Greene and Wuts, Protective Groups in Organic Synthesis, 2nd ed., (John Wiley & Sons, New York, 1991), which is incorporated here by reference. Examples of protective groups are the above-mentioned group protecting the amino group, such tertbutyloxycarbonyl, Gruppo and guanidinopropionic, the nitrogroup, which can be used to protect guanidinium or groups such as benzyl, stands, tritely or acetamidomethyl that can be used to protect groups such as hydroxy, mercapto, and other. Preferably R81 and R82 is selected from the series consisting of hydrogen, alkyl, such (C1-C6)-alkyl, aryl such as phenyl, arylalkyl, such phenyl-(C2-C2)-alkyl, and heteroalkyl, such heteroaryl-(C1-C2)-alkyl, all of which can be unsubstituted or substituted and in which heteroaryl preferably represents the balance monocyclic or bicyclic aromatic ring system containing one or two identical or different ring heteroatoms such as N, O or S. More preferably R81 represents a hydrogen, and R82 represents a higher unsubstituted or substituted residue.

Particularly preferably the group D represents a residue selected from the series consisting of Arg, Dap, Dab, Orn, Lys, Dap[-C(=NH)-NH2], Dab[-C(=NH)-NH2], Lys[-C(=NH)-NH2], Lys[-C(=NH)-CH3], Orn[-C(=NH)-CH3], Dab[-C(=NH)-CH3], Dap[-C(=NH)-CH3], Dab(Alloc), Asn, Gln, Met, Ser, Thr, Ser(Bzl), Thr(Bzl), Cys(Me), Cys(Bzl), Cys(Acm), Arg (NO2), His, Sub-group balances, are selected especially preferred residues D, is formed next, consisting of Arg, Dap, Dab, Orn, Lys, Dap[-C(=NH)-NH2], Dab[-C(=NH)-NH2], Lys[-C=NH)-NH2], Asn, Ser, Thr, Ser(Bzl), Arg (NO2), Trp, Phg, Ala, Val, Ile, Leu, Phe, 2-Abu, Ala(3-CN), Ala(3C(=NH)-NH2), 2-Abu(4-CN) and 2-Abu(4-C(=NH)-NH2).

The number n is preferably 0, 1 or 2, more preferably 0 or 1. If n is 0, the group R2 is directly linked to a carbonyl group, representing the D3. If n is non-zero, the group R2 is linked to the carbonyl group, represent an end group E3. If n is 2 or 3, all group E may be identical or different.

Substituted residues R71 and R72 include can independently bear one or more, for example 1, 2, 3 or 4 identical or different residues, which are preferably chosen from the series consisting of alkyl, alkyloxy, halogen, trifloromethyl, nitro, cyano, alkylsulfonyl, alkylcarboxylic, vinylcarbazole and 2-phenyl-1,3-ditiolan-2-yl, which may optionally be substituted. A subgroup of the substituents that can be present in R71 and R72 include, formed adjacent consisting of alkyl, alkyloxy, halogen, trifloromethyl, nitro, cyano, alkylsulfonyl, alkylcarboxylic, which may be optionally substituted. Preferably R7172 preferably represents alkyl, in particular, (C3-C8)-alkyl, including cyclic alkyl, such cycloalkenyl, such as cycloalkyl-(C1-C2)-alkyl, or aryl, in particular phenyl or arylalkyl, in particular, phenyl-(C1-C2)-alkyl, or heteroaromatic, in particular heteroaryl-(C1-C2)-alkyl, where these residues can be unsubstituted or substituted, and where heteroaryl preferably represents a monocyclic five-membered or six-membered aromatic ring containing 1 or 2 identical or different ring heteroatoms such as N, O and S. the Group or groups E, in particular, if the number n is equal to 1, preferably chosen from the series consisting of Phe, which is not substituted or substituted in the phenyl group, Cha and Chg. Especially preferred group E is selected from the series consisting of Cha, Chg, and Phe[4-C(S-CH2-CH2-S-)-Ph]. The group of R70, present in group E, represents preferably hydrogen, alkyl, in particular (C1-C4)-alkyl, including methyl, or arylalkyl, in particular, phenyl-(C1-C4)-alkyl, including benzyl and 2-phenylethyl, which may be unsubstituted or substituted in the phenyl group. Particularly preferably, R70 represents hydrogen.

For the personal residue, which are preferably chosen from the series consisting of halogen, in particular F, Cl, Br, hydroxy, trifloromethyl, nitro, cyano, dialkylamino, alkyloxy, such metiloksi, alkylenedioxy, alkylsulfonyl, aminosulfonyl and oxo(=O), which may be optionally substituted. Examples of alkylenedioxy are methylenedioxy (O-CH2-Oh) or 1,2-Ethylenedioxy. Examples of dialkylamino are dimethylamino, diethylamino or dibutylamino, examples alkylsulfonyl are methylsulphonyl, ethylsulfonyl or butylsulfonyl. R2 preferably represents NR21R22 where R21 and R22 are as defined above groups. R21 preferably represents hydrogen, (C1-C4)-alkyl or phenyl-(C1-C4)-alkyl, which is not substituted or substituted in the phenyl group. Particularly preferably, R21 represents hydrogen, i.e., particularly preferably NR21R22 is a NHR22, and, thus, particularly preferably R2 represents NHR22. R22 preferably represents a residue selected from the series consisting of hydrogen, alkyl, in particular (C1-C12)-alkyl, including cyclic alkyl, such cycloalkenyl, such as cycloalkyl-(C1-C4)-album or two residues (C6-C12)-aryl, heteroaromatic, in particular (C1-C4)-alkyl substituted monocyclic or bicyclic residue of heteroaryl containing 1 or 2 identical or different heteroatoms, such as N, O or S, and geterotsiklicheskikh, in particular (C1-C4)-alkyl substituted monocyclic 4-, 5-, 6 - or 7-membered heterocyclizations group containing 1 or 2 identical or different heteroatoms, such as N, O or S, all of these residues, as described above, can be unsubstituted or substituted.

Particularly preferably, R22 represents a residue selected from the series consisting of hydrogen, benzyl, naphthylmethyl, pyridylmethyl, phenylethyl, naphtylamine, pyridylethyl, phenylpropyl, naphtylamine, pyridylamino, fertile, diphenylmethyl, diphenylamine and diphenylpropyl, and these balances are not substituted or substituted by one or more, for example one, two, three or four identical or different substituents, which are preferably chosen from the series consisting of F, Cl, Br, hydroxy, methoxy, methylenedioxy, nitro, cyano, dialkylamino, alkylsulfonyl, aminosulfonyl and trifloromethyl, which may be optionally substituted. A number of particularly prepulse a NHR22, and R22 represents hydrogen. Another series of especially preferred compounds is formed by compounds in which at the same time n is 0, R2 is a NHR22, and R22 is other than hydrogen, and in this series of compounds, a preferred group designation D represents Asn.

Preferred compounds of formula I are those compounds in which one or more groups or residues are preferred designations, and all combinations of the preferred notation is the subject of the present invention.

A group of preferred compounds of the invention is formed by compounds of the formula I, in which

R1 represents R12C(O), in which R12 represents a certain above the rest,

But a certain above the rest,

Represents a certain above the rest, and preferably represents NH-CHR97-C(O), where

R97 represents ethyl, which is substituted in position 2 by hydroxycarbonyl or its salt or allyloxycarbonyl like (C1-C4allyloxycarbonyl,

D represents NH-CHR82-C(O), in which R82 represents a higher OCTA which represents NH,

E2 represents CHR72, which remains R72 include, which are independent from each other and are identical or different, are as defined above groups

E3 represents C (O), and

R2 represents a certain above the rest,

in any of their stereoisomeric forms or their mixtures in any ratio, and their pharmaceutically acceptable salts, amides and esters.

A group of especially preferred compounds is formed by compounds of the formula I, in which

R1 represents allyloxycarbonyl or arylamination,

And represents the balance (L)-4-amitiptyline,

Represents a residue of (L)-glutamic acid or a pharmaceutically acceptable salt or ester (L)-glutamic acid,

D is a residue selected from the series consisting of Arg, Dap, Dab, Orn, Lys, Dap[-C(=NH)-NH2], Dab[-C(=NH)-NH2], Lys[-C(=NH)-NH2], Lys[-C(=NH)-CH3], Orn[-C(=NH)-CH3], Dab[-C(=NH)-CH3], Dap[-C(=NH)-CH3], Dab(Alloc), Asn, Gln, Met, Ser, Thr, Ser(Bzl), Thr(Bzl), Cys(Me), Cys(Bzl), Cys (Acm), Arg(NO2), His, Trp, Phg, Gly, Ala, Val, Ile, Leu, Phe, Phe(4-NO2), Phe(4-NH-C(=NH)-NH2), 2-Abu, Ala(3-CN), Ala[3-C(=NH)-NH2], 2-Abu(4-CN) and 2-Abu[4-C(=NH)-NH2],

n is 0 or Ph],

R2 represents NHR22, R22 represents hydrogen or a residue selected from the series consisting of benzyl, naphthylmethyl, pyridylmethyl, phenylethyl, naphtylamine, pyridylethyl, phenylpropyl, naphtylamine, pyridylamino, fertile, diphenylmethyl, diphenylamine and diphenylpropyl, and these balances are not substituted or substituted by one or more identical or different substituents chosen from the series consisting of F, Cl, Br, hydroxy, methoxy, methylenedioxy, nitro, cyano, dialkylamino, alkylsulfonyl, aminosulfonyl and trifloromethyl,

in any of their stereoisomeric forms or mixtures thereof in any ratio, and their pharmaceutically acceptable salts, amides and esters.

Particular examples of compounds according to the invention include, for example, the compounds listed below in table. 2, and in the examples section, and their pharmaceutically acceptable salts, amides and esters.

The compounds of formula I can be obtained, for example, in accordance with the methods of solid-phase chemistry using a method that includes:

A1) the binding of the compounds of formula Fmoc-En-OH, in which n is 1, 2 or 3, p is sensitive to acid linker that is attached to the resin or svobodnoe amino group and again the removal of the protective Fmoc group, or to obtain the compounds of formula I in which n is 0, the binding compounds of the formula Fmoc-D1-D2-C(O)HE is sensitive to acid linker that is attached to the resin, or in General to a solid support, and the removal of the protective Fmoc group;

A2) the binding of the compounds of formula Fmoc-B1-B2-C(O)HE with a free amino group obtained in step A1), and cleavage of the protective Fmoc group;

A3) the binding of the compounds of formula R1-A1-A2-C(O)HE with a free amino group obtained in step A2);

A4) the separation of compounds obtained in accordance with steps A1) to A3), from the resin by triperoxonane acid.

Resin or the linker used in this way, can relate to this type, in which the carboxyl group in connection associated, respectively, with the resin or with a linker, is transformed into an amide group C(O)-NH2such as Knorr linker or amide resin Rink. The connection is obtained, in which the number n is 2 or 3 can also be accomplished via a stepwise Assembly of element Enin the following way. At step A1) instead of the compound of the formula Fmoc-En-OH, in which n is 2 or 3, first, the compound of the formula Fmoc-En-OH, in which n is 1, is associated with sensitive acid linkarray n is 1, associated with received a free amino group. Then, to obtain compounds in which n is 1, otscheplaut the protective Fmoc group, and the third compound of the formula Fmoc-En-OH, in which n is 3, is associated with received a free amino group. Finally, otscheplaut the protective Fmoc group, and this is followed by the steps A2) - A4).

Another method of preparing compounds of formula I includes:

b1) the binding of the carboxylic acid side chain of the compounds of formula Fmoc-B1-CHR97-C(O)OPG in which R97 represents a 2-hydroxycarbonylmethyl, a PG is a protective group is sensitive to acid linker type benzyl alcohol, attached to the resin with a functional amino group;

b2) cleavage of the protective group PG;

b3) the binding of the compounds of formula H2N-D2-D3-En-R2, in which n is 0, 1, 2 or 3, with the free carboxylic acid obtained in step b2);

b4) the removal of the protective Fmoc group;

b5) the binding of the compounds of formula R1-A1-A2-C(O)HE with a free amino group obtained in step b4);

b6) cleavage of the compounds obtained in accordance with steps bl) - b5) from the resin by triperoxonane acid.

Similar to the modification described is estatica on the resin. In accordance with another method similar to this method, the compounds of formula I can also be obtained by linking first group of carboxylic acid, which is present in the side chain group D2 group D, i.e., which is present in one of the groups R81 and R82, with the linker attached to the resin. Similarly to the above-mentioned compound of the formula Fmoc-B1-CHR97-C(O)OPG, such a connection can, for example, have the formula Fmoc-NH-CR81R82-C(O)OPG in which R82 represents the connection defined above, provided that it contains a group WITH(ABOUT)IT, and R81 represents a certain above the rest. For example, R81 may be a hydrogen, and R82 may be hydroxycarbonylmethyl, and the compound of the formula Fmoc-NH-CR81R82-C(O)OPG may thus represent a protected derivative of aspartic acid. After removing the protective group C(O)OPG, a carbonyl group which is a group D3 in the formula (I received a free carboxylic acid group is associated with a connection that is similar to the H2N-En-R2 H-R2. Then, after removal of the protective Fmoc group the resulting amino group is associated with the compound of the formula Fmoc-B1-B2-C(O)IT, and after removing the protection of the amino group of the product associated with soedinenii connection, which is associated with the resin or with a linker, is transformed into an amide group C(O)-NH2. For example, using amide resin element aspartic acid attached to the resin may be in the final connection is converted into aspartic element.

The connection according to the invention can be chemically synthesized using, for example, automatic synthesizer (see example 1). Selective modification of reactive groups, such as group present on the side chain of an amino acid, or N-terminal or C-terminal reactive group of the peptide, can make the connection according to the invention the desired characteristics such as increased solubility or enhanced inhibitory function. When using solid-phase synthesis methods the chemical composition of the connection can be changed, while the resin formed is attached peptide or after the peptide was derived from the resin to obtain, for example, N-terminal derivative, such as compound acylated on the N-end, for example acetylated compound. Similar modifications can also be made in the carboxyl group of compound including a carboxyl group-end, which mouyh amino acids in accordance with the methods of traditional pharmaceutical chemistry or by using known in the field of standard procedures of liquid-phase organic chemistry and unprotect to the target molecule. Generally, suitable reaction for the synthesis of compounds of formula I using solid-phase or liquid-phase methods, and experimental details, such suitable binding agents, such as carbodiimide, TOTU or HATU, or the solvent and the reaction temperature, is well known to specialists in this field and they can also be found in standard references, including references mentioned here and below as examples.

The synthesized compound can be purified using well known methods such as high performance liquid chromatography with reversed phase (RP-HPLC; see example 1) or other methods of separation based on, for example, on the size, charge or hydrophobicity of the compounds. Similarly to characterize the structure of the compounds according to the invention can be used well-known methods, such as analysis of the amino acid sequence or mass spectrometry (MS), see example 1.

Various compounds containing the various locations of the substituents, exhibit various levels of inhibitory activity against factor VIIa. For example, the choice of substituents affects the affinity of binding compounds. These soybean is the dominant activity was carried out using quantitative analysis, described in example 22. Using such methods, the person skilled in the art can synthesize the compound, as disclosed here, including its modification, and to determine the inhibitory activity of compounds against factor VIIa. The composition of the present invention may be presented in the form of a homogeneous composition or a mixture of compounds containing various combinations of substituents. The flexibility afforded by the choice of substituents, provides significant control of biological and physico-chemical properties of the compounds and compositions according to the invention.

The invention provides compounds that specifically inhibit the activity of factor VII. Such compounds preferably have inhibitory activity (Ki)500 nmol, more preferably 50 nmol for the activity of factor VIIa and do not have a significant inhibitory effect on the activity of other proteases involved in the coagulation cascade and fibrinolysis, relative inhibition of the factor VIIa (using the same concentration of the inhibitor). Other proteases include, for example, factor XA, thrombin and plasmin.

In the following table.2 shows the inhibitory active is also provided to illustrate the invention.

Inhibiting thrombin activity of the above compounds can be expressed as Ki values, which in General is much higher than the above inhibitory activity against factor VIIa, for example, approximately 200 times or approximately 500-fold, or about 1000 times higher than the inhibitory activity against factor VIIa. Inhibiting activity of the above compounds of factor XA can also be expressed in terms of Ki, which in General is much higher than the above inhibitory activity against factor VIIa, for example, approximately 100 times higher than inhibiting the activity of factor VIIa.

These data show that the compounds of formula I can be used as inhibitors of factor VIIa, but do not have a significant inhibitory effect on the activity of factor XA or serine proteases, such as thrombin, which are involved in blood coagulation and fibrinolysis.

The connection according to the invention can be used mainly as an anticoagulant, which may come in contact with the blood sample to prevent clotting. For example, effective to the I of the blood sample. Used herein, the term “effective amount” when used with reference to the connection according to the invention means the amount of compound that inhibits the activity of factor VIIa. An experienced specialist should be understood that the effective amount of the compounds according to the invention can be determined using the disclosed here (see example 22) or other methods known in this field. From the point of view of possible applications of the compounds according to the invention, an experienced specialist will be clear that a tool such as heparin may be replaced by a connection according to the invention. This use of the compounds of this invention may, for example, result in cost savings compared with other anticoagulants.

In addition, the connection according to the invention can be administered to individuals to treat various clinical conditions, including, for example, the treatment of cardiovascular disorders or complications associated with, for example, infection or surgery. Examples of cardiovascular disorders include restenosis after angioplasty, respiratory distress syndrome of adults, multiple organ failure, stroke and is licheskimi interventions include, for example, deep vein thrombosis and proximal veins, which can occur after surgery. Thus, the connection according to the invention can be used as a drug for reducing or inhibiting undesirable coagulation of blood in the individual.

Because the connection according to the invention may inhibit the activity of factor VIIa, such a connection can generally be used for reducing or inhibiting blood clotting in an individual. Used herein, the term “individual” means a vertebrate animal, including a mammal, such as man, in which the factor VIIa participates in the clotting cascade.

Blood clotting the individual can be reduced or ingibirovalo by administering to the individual a therapeutically effective amount of the compounds according to the invention. Used herein, the term “therapeutically effective amount” means the dose of a compound, you need to enter the individual for inhibiting the activity of factor VIIa in the individual. More specifically, a therapeutically effective amount of the compounds according to the invention inhibits the catalytic activity of factor VIIa or direct the investments of the Assembly of factor VIIa in complex prothrombinase. The preferred compounds can inhibit the activity of factor VIIa in Ki500 nmol, and more preferred connection - when Ki50 nmol. therapeutically effective amount can be determined using the above methods, for example, in example 22, or using other methods known in this field.

When using the method of treatment according to the invention a specific dosage to obtain a therapeutically effective amount of a pharmaceutical composition, which will enter the individual will depend on many factors that should be considered, including, for example, the nature or severity of the disease, the scheme of administration, the age and physical characteristics of the individual. The appropriate dosage can be set using clinical approaches that are well known in the field of medicine. Thus, the invention represents a specific way of inhibiting the activity of factor VIIa by providing contact of factor VIIa with a compound having the formula R1-A-B-D-En-R2. The invention also provides a method of reducing or inhibiting the formation of blood clot in an individual by administering a therapeutically effective amount of soemu in the form of a composition, containing one or more compounds of the formula I, and pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” refers to a medium or composition that is non-toxic for the individual or has acceptable toxicity for determining the relevant regulatory agencies. Used herein, the term “pharmaceutically acceptable carrier” encompasses any of the standard pharmaceutical carriers include solid carriers, such corn starch, lactose, fats, waxes, etc. or liquid, such as saline solution with phosphate buffer, water, emulsions such as emulsions oil-in-water or water-in-oil, and/or conventional additives, for example, any of various types of wetting means. Suitable pharmaceutical carriers and their compositions described Martin's guide Remington''s Pharmaceutical Sciences, 15th ed. (Mack Publishing Co., Easton, 1975), which is included here by reference. Such compositions will generally contain a therapeutically effective amount of the compounds according to the invention together with a suitable amount of carrier so as to contain the appropriate dosage for administration to the individual. Thus, the claimed compounds can ndividual.

Pharmaceutical compositions or pharmaceutical preparations according to the invention can be administered orally, for example in the form of pills, tablets, glossy tablets, coated tablets, granules, hard and soft gelatine capsules, solutions, syrups, emulsions, suspensions or aerosol mixtures. However, administration can also be carried out rectally, for example in the form of suppositories, or parenterally, for example intravenously, intramuscularly or subcutaneously, in the form of solutions for injection or solution for infusion, in the form of microcapsules, implants or rods, or transdermally, or topically, e.g. in the form of ointments, solutions or tinctures, or in other ways, for example in the form of aerosols or aerosol compositions for intranasal spray. The amount of active ingredient of formula I or its pharmaceutically acceptable salt or derivative in the standard dose of the pharmaceutical composition is typically from about 0.5 mg to 1000 mg, preferably from about 1 mg to 500 mg, but, depending on the type of pharmaceutical composition, may be higher. Daily dose of the compounds of formula I, which are going to enter, can be a single daily dose Ile media may also include, for example, other environment, connection or modification in addition to the inhibitory factor VIIa to the compound of formula I, which maximizes its pharmacological function. Pharmaceutically acceptable medium can include, for example, pharmaceutically acceptable salt. Salt accession acids of compounds of formula I may be formed, for example, inorganic acid, such as hydrochloric acid, Hydrobromic acid, phosphoric acid, sulfuric acid or Perlina acid, or with organic carboxylic acid, such as acetic acid, oxalic acid, maleic acid, axentra acid, formic acid, lactic acid, tartaric acid, citric acid, succinic acid or malonic acid, with an organic acid, such as methanesulfonate acid or para-toluensulfonate acid. The acid group in the compound of formula I, for example, the carboxyl group may be present in the form of a metal salt, the cation of which is based on the alkali and alkaline earth metals such as sodium, lithium, potassium, calcium or magnesium, or also non-toxic ammonium salts, including salts of Quaternary ammonium salt and attach acids with amines, for example ammatillinen or tetraethylammonium salt.

Examples of modifications that enhance the pharmacological function compounds include, for example, esterification, such as education (C1-C6)-alilovic esters, preferably (C1-C4)-alilovic esters in which the alkyl group is a straight or branched chain. Other acceptable esters include, for example, (C5-C7)-cycloalkyl esters and arylalkylamine esters, such as benzyl ethers. Such esters can be obtained from the compounds described herein using conventional methods well known in the field of peptide chemistry.

Pharmaceutically acceptable modification can also include, for example, the formation of peptide amides. Such amide modifications that may be made in respect of the compounds according to the invention include, for example, modification, obtained from ammonia, primary (C1-C6)-alkylamines followed and secondary di(C1-C6)-bonds alkylamines, where the alkyl groups are straight or branched chain, or arylamine having different substitution. In the case of secondary amines amine may also be in the form of a 5 - or 6-membered heterocycle, which, in the and the sulfur atom. Methods of obtaining such amides are well known in this field.

In another embodiment of the invention the compound according to the invention can be used in quantitative analysis to identify the presence of factor VIIa or to highlight factor VIIa essentially in purified form. Preferably the connection according to the invention have been labelled, for example, a radioisotope, and labeled compound identified using the usual method used to identify a particular label. In addition, the connection according to the invention can be used mainly as a probe to identify the type or amount of activity of factor VIIa in vivo, in vitro or ex vivo.

It is clear that in the open here, the invention includes modifications that do not significantly affect the activity of different implementations of this invention. Accordingly, the following examples are intended to illustrate and not to limit the present invention.

Example 1. Procedures peptide synthesis and General procedures synthesis

The source materials used in the synthesis are companies that sell chemical products, such as Aldrich, Sigma, Fluka, Nova Biochem and Advanced Chemtech. During sinestrari adverse reactions during linking. Examples of suitable protective groups and their use are described in the publication of The Peptides, supra, 1981, and vol.9, Udenfriend and Meienhofer (eds.), 1987, which are incorporated here by reference.

General solid-phase peptide synthesis is used to produce compounds according to the invention. Such methods are described, for example, in the leadership of Stewart and Young, Solid Phase Peptide Syntesis (Freeman&Co., San Francisco, 1969, which is incorporated here by reference.

In the absence of other indications, the peptides are synthesized on the resin TentaGel S NH2(Rapp Polymere, Tubingen, Germany). Sensitive to acid binder pair-[(R,S)--[1-(N-foren-9-yl)metoclopramide]-2,4-dimethoxybenzyl]Phenoxyethanol acid (binder Knorr) associated with solid media (publication Bernatowicz et al., Tetr. Lett. 30 (1989) 4645, which is incorporated here by reference). Alternatively, the peptides are synthesized on polystyrene resin, cross crosslinked with 1% divinylbenzene, modified sensitive to acid linker (Rink resin) (Rink, Tetr. Lett. 28(1987)3787; Sieber, Tetr. Lett. 28(1987)2107, each of which is included here for reference). When the peptides are synthesized first by linking the carboxylic acid side chain of the compounds of formula Fmoc-B1-CHR97-C(O)OPG resin used resin TentaGel S NH2modified prereplicative equivalent amount of HOBt, except Alloc-pAph-OH where the use of 2 equivalents (EQ.) HOBt. All binding spend or N,N-dimethylformamide (DMF), or DMF/DMSO (1/1 mixture) at room temperature (RT). The end link control using ninhydrin test. The second (double) linking execute when in the first case is incomplete binding.

Deleting a group protecting group Fmoc, carried out using 50% piperidine in DMF for 2-10 minutes Number of released Fmoc determined by absorbance at 300 nm solution after removal of the protective group, in terms of washing and weight of the resin used in the synthesis.

The cycle of each link was as follows:

After Assembly of the peptide on the resin, if necessary, perform the final destruction of the group, Fmoc protecting. Then the peptide resin was sequentially washed with DMF and DCM, and then, in the absence of other indications, the peptide decompose and remove the protective group using a mixture of TFA/thioanisole (95/5) for 1.5 hours the Resin was washed with DCM, and DCM wash combined with backordered when washing TFA. The solution is evaporated, the product is precipitated using simple anhydrous diethyl ether, and the solid odoe substance re-dissolved in a mixture of water and acetonitrile and lyophilizers.

The dried peptide was subjected to purification using HPLC using a gradient of 0.1% TFA in water and acetonitrile (ACN). After collecting the peak containing the proposed synthetic product, a solution of peptide lyophilizer and the peptide is subjected to the process of identification, which includes electroaerosol mass spectrum (MS) and/or NMR, and/or amino acid analysis to confirm that the synthesized proper connection.

For HPLC analysis of the product sample analyzed using device HPLC Beckman (consisting of feeder solvent 126, the device for automatic sampling e with programmable detector module 166 controlled by the control unit Data Station software Gold Nouveau) and column YMC ODS-AM 4,h mm at 230 nm, flow rate 1 ml/min

Cleaning product sample lyophilized crude peptide was dissolved in a mixture of 0.1% aqueous TFA containing from 10% to 50% ACN. The peptide solution is typically filtered through a syringe connected to the filter ACRODISC”13CR PTFE (Gelman Sciences; Ann Arbor MI) with a pore size of 0.45 μm. An appropriate volume of the filtered solution of the peptide Inuktitut in prepreparation C18 column (Vydac Protein and Peptide C18, TR (22250 mm); Th of 0.1% TFA buffer and ACN (HPLC grade) as eluent support using programmable solvent module 126 and programmable detector module 166, managed using the software “SYSTEM GOLD” devices Beckman SYSTEM GOLD HPLC (Beckman System Gold). Elution peptide control using UV detection at 230 nm. After identification using MS peak corresponding synthesized the connection, the connection is collected, lyophilizers and carry out its biological testing. MS carried out using the device VG Piatforn (Fisons Instruments) ES+. For NMR samples typically measured in DMSO-d6(Aldrich) using the instrument Bruker Avance DPX300.

Example 2. Synthesis of Alloc-pAph-OH

The same procedure is applicable for Alloc-pAph-OH.

Alloc-Phe(4-CN)-HE

5.7 g (30 mmol) H-Phe (4-CN)-HE dissolved in 100 ml of 1M NaOH with the addition of 2M NaOH to pH 10 with ice cooling. With vigorous stirring, slowly add allylchloroformate (7.5 ml) (pH is maintained at level 10 with 2M NaOH). The reaction mixture was stirred at 0°C for 15 min and at RT for 30 min, acidified with Hcl to pH 2, extracted with ethyl acetate (3 times), dried with MgSO4and evaporated. The residue is subjected to recrystallization from ethyl acetate/hexane to obtain a white solid. Yield: 7.0 g (85%).

Alloc-Phe[4-C(=S)-NH2]-

2,74 g Alloc-Phe(4-CN)-HE dissolved in a mixture of pyridine (50 m is evaporated. Drying in a high vacuum gives 3,21 g solid foam untreated thioamide, which is directly transformed into metaltometal.

Alloc-Phe[4-C(=NH)-SCH3]-OH·HI

1 g Alloc-Phe[4-C(=S)-NH2]-IT is dissolved in acetone (50 ml) and add methyliodide (5 ml). The reaction mixture was kept overnight at RT, the volatile solvent is evaporated rapidly, a maximum of 35°C), the residue is treated simple diethyl ether. After 1 h at 0°C. the ether is decanted, the product is washed with simple diethyl ether and dried in vacuum. Receives a yellow solid foam, which is directly transformed into amidin.

Alloc-pAph-OH

All the above Alloc-Phe(4-C(=NH)-S3)-OH·HI dissolved in 50 ml of methanol with 300 μl of acetic acid and add 0.5 g of ammonium acetate. The mixture is heated for 3 h at 55°C, evaporated and add 10 ml of acetone. After 2 h at 0°C. the solid product was filtered, washed with a small amount of cold acetone, a small amount of cold methanol and simple diethyl ether and dried in vacuum to obtain a yellow solid. Output: 0,53 g

Example 3. Synthesis of Alloc-pAph-Glu-Arg-Cha-NH2

To 1 g of resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. the ha-OH, Fmoc-Arg(Pmc) -, Fmoc-Glu(OtBu)-HE Alloc-pAph. The peptide digest and use the exposure for 3 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 729,1, according to the calculations 729,4.

Example 4. Synthesis of allyl-NH-C(O)-pAph-Glu-Arg-Cha-NH2

To 0.5 g of the resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Arg(Pmc) -, Fmoc-Glu(OtBu)-HE and Fmoc-Phe(4-CN). After removing the protection of the N-terminal Fmoc resin for 2 h, treated with a solution of 1 mmol arylisocyanate in 3 ml of DMF. Then the resin was washed with DMF and triethylamine/pyridine (1/2) and during the night treated with a saturated solution of H2S in pyridine/triethylamine. The resin is washed with acetone for 6 h conduct the reaction thioamides resin with methyliodide (3 ml of 10% solution under the conditions in acetone). Methylthiomethyl the resin is washed with acetone, methanol and treated with a solution of 0.2 g of ammonium acetate, 100 µl of acetic acid in 3 ml of methanol at 55°C for 3 h, the Resin was washed with methanol, DMF and DCM and the peptide cleaved and removed the th material purified using HPLC, as described in example 1 and characterized with MS. (M+H)+: detected 728,3, according to the calculations 728,4.

Example 5. Synthesis of Alloc-pAph-Glu-Arff-Chg-NH2

To 1 g of resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Chg-OH, Fmoc-Arg(Pmc) -, Fmoc-Glu(OtBu)-HE Alloc-pAph. The peptide digest and use the exposure for 3 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: found 715,8, according to the calculations 715,4.

Example 6. Synthesis of Alloc-D-pAph-Glu-Arg-Cha-NH2

To 1 g of resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Arg (Pmc) -, Fmoc-Glu(OtBu)-HE Alloc-D-pAph-HE (synthesized in accordance with the same procedure as Alloc-pAph-HE in example 2). The peptide digest and use the exposure for 3 h TFA/thioanisole (95/5) remove the protective group and treated as described in example 1. The crude compound purified using HPL 7. Synthesis of Alloc-pAph-Glu-Phe(4-guanidino)-Cha-NH2

To 0.25 g of the resin TentaGel S NH2(substitution of 0.23 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Phe(4-NH-C(=NBoc)-NH-Boc)-OH, Fmoc-Glu(OtBu)-HE Alloc-pAph-OH. The peptide digest and use the exposure for 1 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 777,1, according to the calculations 777,4.

Example 8. Synthesis of Alloc-pAph-Glu-Dap[-C(=NH)-NH2]-Cha-NH2

To 0.25 g of the resin TentaGel S NH2(substitution of 0.23 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Dap[-C(=NBoc)-NH-Boc]-OH, Fmoc-Glu(OtBu)-HE Alloc-pAph-OH. The peptide is cleaved and using impact during 1H TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 729,1, according to the calculations 729,4.

Example 9. Synthesis of Alloc-pAph-Glu-Dap[-C(=NH-CH3]-Cha-NH3)4within 3 hours the Resin was washed with DMF and treated with a solution of 150 mg of 2-methylnaphtaleneamine in 4 ml of ethanol/DMSO (3/1) within 1 h After washing DMF remove the protecting group Fmoc (1-5min) and bind the N-terminal Alloc-pAph-OH. The peptide digest and use the exposure for 1 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 700,1, according to the calculations 700,4.

Example 10. Synthesis of Alloc-pAph-Glu-Ala[3-C(=NH)-NH2]-Cha-NH2

To 0.25 g of the resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Ala(3-CN) -, Fmoc-Glu(OtBu)-OH and Alloc-Phe(4-CN)-HE. A mixture of pyridine and triethylamine (2/1) saturated with N2S (CT, 15-30 min), and this solution is added to the resin, predvaritel treated with 20% methyliodide in acetone. Then the resin is washed with acetone and methanol. Associated with resin metaltometal then turn in amidin by using heat (water bath to 55°C, 3 h) resin with a solution of 10 EQ. ammonium acetate in methanol containing 5% acetic acid. After this final conversion of the resin washed with methanol, DMF, and DCM. The peptide is cleaved and using impact during 1H TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 685,9, according to the calculations 686,4.

Example 11. Synthesis of Alloc-pAph-Glu-Asn-Cha-NH2

With 0.125 g of Rink resin (substitution of 0.78 mmol/g) after removal of the Fmoc protection in accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Asn-OH, Fmoc-Glu(OtBu)-HE Alloc-pAph-HE. The peptide is cleaved and using impact during 1H TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 686,9, according to the calculations 687,3.

Example 12. Synthesis of Alloc-pAph-Glu-Dab-Cha-NH2

To 0.25 g of the resin TentaGel S NHwhat the example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Dab(BOC) -, Fmoc-Glu(OtBu)-HE Alloc-pAph. The peptide digest and use the exposure for 1 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 673,2, according to the calculations 673,4.

Example 13. Synthesis of Alloc-pAph-Glu-Ala[3-C(=NH)-NH2]-NH2

To 0.25 g of the resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Ala(3-CN) -, Fmoc-Glu(OtBu)-HE Alloc-Phe(4-CN)-HE. A mixture of pyridine and triethylamine (2/1) is saturated with H2S (CT, 15-30 min), and this solution is added to the resin, pre-washed with pyridine/triethylamine (2/1). After storage overnight, the resin was washed with acetone and during the night is treated with a solution of 20% under the conditions in acetone. Then the resin is washed with acetone and methanol. Associated with resin metaltometal then turn in amidin by using heat (water bath to 55°C, 3 h) resin with a solution of 10 EQ. ammonium acetate in methanol containing 5% acetic acid. After this final prevrasheniya protection and process, as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+N)+: detected 533,3, according to the calculations 533,2.

Example 14. Synthesis of Alloc-pAph-Glu-Gly-Cha-NH2

With 0,150 g of Rink resin (substitution of 0.78 mmol/g) after removal of the Fmoc protection in accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Cha-OH, Fmoc-Gly-OH, Fmoc-Glu(OtBu)-HE Alloc-pAph-OH. The peptide digest and use the exposure for 1 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 630,1, according to the calculations 630,3.

Example 15. Synthesis of Alloc-pAph-Glu-Asn-(Ph-CH2-CH2-)Gly-NH2

For N-substituted glycinol use the technique Zuckermann et al. (J. Am. Chem. Soc.ll4 (1992) 10646, which is incorporated here by reference). With 0.1 g of Rink resin (substitution of 0.78 mmol/g) after removal of the Fmoc protection associated bromoxynil acid by symmetrical anhydride in DCM/DMF. After 10 min the resin was washed with DCM and repeat binding. After washing DCM and DMF the resin during the night treated with 1M solution of 2-phenethylamine in DMSO. After p is actwu and conduct the reaction with the symmetrical anhydride of Fmoc-Asn(Trt)-HE's in DCM/DMF. After removal of the Fmoc protection in accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Glu(OtBu)-HE Alloc-pAph-OH. The peptide digest and use the exposure for 1 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 694,9, according to the calculations 695,3.

Example 16. Synthesis of Alloc-pAph-Glu-Thr(Bzl)-NH-CH2-CH2-CH(Ph)2

H-Thr(Bzl)-NH-CH2-CH2-CH(Ph)2·HCl

of 0.62 g (2 mmol) of the BOC-Thr (Bzl)-HE dissolved in 10 ml of DCM is added 2 mmol of triethylamine, and the solution is cooled to 0°C. While mixing, slowly add 2 mmol of isobutylacetate. The cooling bath is removed, the solution is stirred for 15 min and add 2.5 mmol of 3,3-diphenylpropylamine in 2 ml of DMF and stirred at room temperature for 1 h, the Solution is evaporated, dissolved in ethyl acetate and extracted with a solution of 0.5 M KHSO4, a saturated solution NaHC3and with brine, dried with gS4and evaporated. The oily product is dissolved in 10 ml DCM and added dropwise to 10 ml of 4M hydrochloric acid in dioxane. After 10 min rastvornym ether and dried in vacuum to obtain a white solid. MS analysis: (M+N)+: detected 403,1, according to the calculations 403,2.

Alloc-pAph-Glu-Thr(Bzl)-NH-CH2-CH2-CH(Ph)2

To 0.5 g of the resin TentaGel S NH2(substitution 0.26 mmol/g) attach the 4-gidroksietilirovanny acid (3 EQ., activated for 1.5 h DIC/HOBt). To the resin through the side chain attached Fmoc-Glu(OH)-O-allyl using DIC/HOBt/NMI in DMF overnight. Group, allyl protecting group is removed by shaking the resin with PD(h3)4in DMF/AcOH/NMM (10/2/1) for 4 h under argon. Lacking the protection of the carboxyl group is activated with a solution of 0.5 mmol THIEF, 0.5 mmol HOBt, 1.5 mmol DIEA and 0.5 mmol of H-Thr(Bzl)-NH-CH2-CH2-CH (Ph)2·HCl in 1.5 ml DMF for 2 hours After removal of the Fmoc protection Alloc-pAph-OH link in accordance with the General procedure described in example 1. The peptide digest and use the impact for 1.5 h TFA/thioanisole (95/5) remove the protection and processed as described in example 1. The crude compound purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 805,0, according to the calculations 805,4.

Example 17. Synthesis of Alloc-pAph-Glu-Dab-NH-CH2-CH2-Ph

To 0.2 g of resin TentaGel S NH2(substitution 0.26 mmol/g) attach the 4-hydroxymethylene processing resin GVS4(5 EQ.)/h3(5 EQ.) in DCM for 4 hours resulting from the substitution of bromine resin during the night treated with a 2M solution of phenethylamine in DCM. Fmoc-Dab(BOC)-IT is associated with resin using TFFH/DIEA (allford generated in situ). In accordance with the General procedures described in example 1, connect the following protected amino acids: Fmoc-Glu(OtBu)-HE Alloc-pAph-OH. The peptide digest and use the exposure for 2 h TFA/triisopropylsilane (99/1) remove the protection. TFA is evaporated, the peptide is dissolved in H2O/ACN and lyophilizers. The crude material is purified using HPLC as described in example 1 and characterized with MS. (M+N)+: detected 624,2, according to the calculations 624,3.

Example 18. Synthesis of Alloc-pAph-Glu-NH-CH2-CH2-CH

To 0.2 g of resin TentaGel S NH2(substitution 0.26 mmol/g) attach the 4-gidroksietilirovanny acid (3 EQ., activated for 1.5 h DIC/HOBt). To the resin through the side chain attached Fmoc-Glu(OH)-O-allyl using DIC/HOBt/NMI in DMF overnight. Group, allyl protecting group is removed by shaking the resin with PD(h3)4in DMF/AcOH/NMM (10/2/1) for 4 h under argon. Lacking the protection of the carboxyl group activated DIC (3 EQ.)/HOBt (3 EQ.) in Out in accordance with the General procedure, described in example 1. The peptide is cleaved by exposure for 2 h TFA/triisopropylsilane (99/1) remove the protection. TFA is evaporated, the peptide is dissolved in2O/ACN and lyophilizers. The crude material is purified using HPLC as described in example 1 and characterized with MS. (M+N)+: detected 473,1, according to the calculations 473,2.

Example 19. Synthesis of Alloc-pAph-Glu-Asn-NH-CH2-Chx

To 0.1 g of the resin TentaGel S NH2(substitution 0.26 mmol/g) attach the amide linker Knorr. To the linker via the side chain attached Fmoc-Asp(OH)-O-allyl, the group, allyl protecting group is removed as described in example 18. Lacking the protection of the carboxyl group activated DIC (5 EQ.)/ HOBt (5 EQ.) and for 2.5 h add cyclohexylethylamine (5 EQ.) in DMF. After removal of the protecting Fmoc Fmoc-Glu(OtBu)-HE Alloc-pAph-OH link in accordance with the General procedure described in example 1. The peptide is cleaved by exposure during 2H TFA/triisopropylsilane (99/1) remove the protection. TFA is evaporated, the peptide is dissolved in H2O/ACN and lyophilizers. The crude material is purified using HPLC as described in example 1 and characterized with MS. (M+H)+: detected 629,9, according to the calculations 630,3.

Example 20. Synthesis of Alloc-pAph-Glu-Asn-NH-CH2-CH2-Ph

The hydrochloride of 2-(S)-allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionic acid (3,48 g, 10.6 mmol) and the hydrochloride of 5-tert-butyl-1-methyl ester 2-(S)-aminopentanedioic acid (2.7 g, 10.6 mmol) in 20 ml DMF at -15° C. add TOTU (3,83 g, 11,67 mmol) and N-Ethylmorphine (2,7 ml and 21.2 mmol). The mixture is stirred for 1H, and then allowed to warm to room temperature. After evaporation the residue is added ethyl acetate and the organic layer is extracted with aqueous sodium hydrogen carbonate solution, a solution of potassium hydrosulfate and water. The organic layer is evaporated. Yield: 2.8 g (50%). MS: m/z = 491,3 (M+N)+.

5-tert-butyl ester 2-(S)-[2-(S)-Allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionamido]-pentandiol acid

The hydrochloride of 5-tert-butyl-1-methyl ester 2-(S)-[2-(S)-allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionamido]-pentandiol acid (a 3.06 g, 5.8 mmol) in 100 ml of water and 30 add THF hydrate of lithium hydroxide (0,49 g, 11.6 mmol). The solution is stirred for 12 h at room temperature, evaporated and lyophilizers. The residue is purified by chromatography on a column of Sephadex LH20 using as eluent n-butanol /glacial acetic acid/ water (17/1/2). Pure fractions are combined. Races/SUP>.

The hydrochloride of 4-(S)-[2-(S)-Allyloxycarbonyl-3-(4-carbamimidoyl)-propionamido]-4-(2-carbarnoyl-1(S)-(2-phenylethanol)-ethylcarbamate)-butyric acid (Alloc-pAph-Glu-Asn-NH-CH2-CH2-Ph)

5-tert-butyl ether 2-(S)-[2-(S)-allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionamido]-pentandiol acid (48 mg, 0.1 mmol) and the hydrochloride of 2-(S)-amino-N1-phenylethyl-succinamide (27 mg, 0.1 mmol) in 5 ml of DMF at 0°C is added HATU (39 mg, 0.1 mmol) and kallidin (or 24.2 mg, 0.2 mmol). The mixture is stirred for 1 h and allowed to warm to room temperature. After evaporation the residue is purified by chromatography on a column of Sephadex LH20 using as eluent n-butanol /glacial acetic acid/ water (17/1/2). Pure fractions are combined. The solvent is evaporated, the residue is collected in water and aqueous solution lyophilizer. Yield: 45 mg (66%). MS: m/z = 638,4 (M+N)+.

Example 21. Synthesis of Alloc-pAph-Glu-Asn-NH-(3-Chlorobenzyl)

5-tert-butyl ether 2-(S)-[2-(S)-Allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionamido]-pentandiol acid (50 mg, 0,105 mmol) and 2-(S)-amino-N1-(3-Chlorobenzyl)succinamic trifenatate (61 mg, 0.16 mmol) in 5 ml of DMF at 0°C. add TOTU (36 mg, 0.11 mmol) and N-ethylmorpholine (57 μl, 0.4 mmol). Slut using chromatography on a column. Sephadex LH20 using as eluent n-butanol /glacial acetic acid/ water (17/1/2). Pure fractions are combined. The solvent is evaporated, the residue is collected in water, and an aqueous solution lyophilizer. Output 4-(S)-[2-(S)-allyloxycarbonyl-3-(4-carbamimidoyl-phenyl)-propionamido]-4-(2-carbarnoyl-1(S)-(3-chlorobenzylamino)-ethylcarbamate)-butyric acid (Alloc-pAph-Glu-Asn-NH-(3-Chlorobenzyl) or Alloc-pAph-Glu-Asn-3-chlorobenzylamino): 28 mg (41%). MS: m/z = 658,3 (M+N)+.

Other illustrative compounds are obtained analogously to the preceding examples, are listed in table.2.

Example 22. Determination of Ki for the inhibition of factor VIIa

Inhibitory activity (Ki) of each compound relative to the activity of factor VIIa/tissue factor determined using the chromogenic quantitative analysis, essentially in accordance with the previously described procedure (J. A. Ostrem, F. Al-Obeidi, P. Safar, A. Safarova, S. K., Stringer, M. Patek, M. T. Cross, J. Spoonamore, J. C. LoCascio, P. Kasireddy, D. S. Thorpe, N. Sepetov, M. Lebi, P. Wildgoose, P. Strop, Discovery of a novel, potent, and specific family of factor Xa inhibitors via combinatorial chemistry. Biochemistry 37(1998)1053-1059). Kinetic quantitative tests conducted at 25°C in half microtitration plates (Costar Corp., Cambridge, MA), using the kinetic tablet reader (Molecular Devices Spect is the super final concentration), in combination with 40 μl of dilutions of inhibitor in 10% buffered DMSO/TBS-PEG (50 mmol Tris, 15 mmol NaCl, 5 mmol l2; 0,05% PEG 8k, pH 8,15). After preincubation period of 15 min quantitative analysis begin by adding 35 ál of chromogenic substrate S-2288 (D-Ile-Pro-Arg-pNa, Pharmacia Hepar Inc., the final concentration of 500 Microm). Constants visible inhibition calculated by the slope of the curves of the dynamics in their linear parts, usually from 1 to 5 min after addition of the substrate. The actual Ki subsequently determined for each connection using the correction on the substrate concentration (S) and Km using the formula Ki=Ki arr/(1+(S)/Km) (I. H. Segal, Enzyme Kinetics, pp 100-125(John Wiley & Sons, New York, 1975)).

Characterization of the compounds of examples

To determine the retention time HPLC (Rt) sample of the product was analyzed using a Beckman HPLC system (including the delivery system solvent 126 (126 Deliver Solvent System) module programmable detector 166 (166 Programmable Programmable Detector Module), autocorrector (dispenser, sampler) e, adjustable item collection and preliminary data processing software Gold Nouveau) and column YMC ODS-AM 4,6250 mm at 230 nm, flow rate 1 ml/min binary gradient formed from water (0.1% of TFU) and is labeled as "DA", were determined using the same system, except that the detector used detector with diode array Beckman 168 Diode Array. Retention time marked VY", was determined under the same conditions, except that the used analytical column Vydac 218TP54 (C18, 5 um, 4.6 to 250 mm).

Mass spectrometry (MS) was performed ES+ using device VG Platform (Fisons Instruments).

The retention time of Rt connections sample descriptions are presented in table 3.

Characteristics of the compounds of Table 2 are presented in Table 4.

1. The compound of the formula I

R1-A-B-D-En-R2 (I)

in which R1 represents R12C(O), and R12 is selected from the group consisting of alkenyl, alkenylacyl or alkenylamine; And is a group A1-A2-AZ, in which A1 represents NH; A2 represents CHR93 in which R93 is 4-amidinophenoxy; A3 represents C(O); is a group B1-B2-B3, where B1 represents NH; B2 is a CHR97 where R97 represents ethyl, which is substituted in position 2 by hydroxycarbonyl or allyloxycarbonyl, B3 represents C(O); D D1 represents a group consisting of hydrogen and unsubstituted or substituted alkyl residues, aryl, arylalkyl, heteroallyl, D3 represents C(O); En is a (E1-E2-OH)nin which n is 0 or 1; E1 represents NR70, where R70 is H, E2 represents CR71R72, where R71 and R72 include independently selected from the group consisting of hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroallyl, E3 represents C(O);

R2 represents NR21R22 where R21 and R22 are independently selected from hydrogen and unsubstituted or substituted residues of alkyl, aryl, arylalkyl, heteroallyl and geterotsiklicheskikh, and the alkyl contains from 1 to 13 carbon atoms, alkenyl contains from 2 to 13 carbon atoms, aryl and heteroaryl contain from 5 to 13 ring carbon atoms, where in the rest of heteroaryl one or more carbon atoms are replaced by heteroatoms selected from the group consisting of N, O and S, heteroseksualci contains from 3 to 8 ring carbon atoms, of which from one to three carbon atoms are replaced by heteroatoms selected from the group consisting of N, O and S, in any stereoisomeric forms or their mixtures in any ratio, and their pharmaceutically acceptable salts.

2. Connection on p. 1, in which the remains representing R81 and R82 may be nezavisiamya, hydroxy, mercapto, all of which can be substituted by a protective group, acetamido, nitro and cyano.

3. Connection on p. 1 and/or 2, in which the remains representing R71 and R72 include, may be independently substituted by substituents selected from the series consisting of alkyl, alkyloxy, halogen, trifloromethyl, nitro, cyano, alkylsulfonyl, alkylcarboxylic, vinylcarbazole and 2-phenyl-1,3-ditiolan-2-yl.

4. The connection of one or more of paragraphs. 1-3, in which the remains representing R21 and R22 may be independently substituted by substituents selected from the series consisting of halogen, trifloromethyl, hydroxy, nitro, cyano, alkyloxy, alkylenedioxy, dialkylamino, alkylsulfonyl, aminosulfonyl, and

5. The connection of one or more of paragraphs.1-4, in which saturated linear or branched alkyl chains have from 1 to 6 carbon atoms, unsaturated linear or branched alkeneamine chains have from 2 to 6 carbon atoms and cyclic alkyl groups have from 3 to 8 carbon atoms.

6. The connection of one or more of paragraphs.1-5, in which R1 represents R12C(O), where R12 is as defined above, D represents NH-CHR82-C(O), where R82 - such as defined above, Enbattle CHR72, where R72 include such as defined above, E3 represents C(O).

7. The connection of one or more of paragraphs.1-6, in which n is as defined above, R2 represents NHR22, where R22, such as defined above.

8. The connection of one or more of paragraphs.1-7, in which R1 represents allyloxycarbonyl or arylamination.

9. The connection of one or more of paragraphs.1-8, in which a represents a residue of (L)-4-amidinopropane.

10. The connection of one or more of paragraphs.1-9, in which a represents the residue of (L)-glutamic acid, or its pharmaceutically acceptable salt, or ester.

11. The connection of one or more of paragraphs.1-10, in which D represents a residue selected from the series consisting of Arg, Dap, Dab, Orn, Lys, Dap[-C (=NH)-NH2], Dab[-C(=NH)-NH2], Lys[-C(=NH)-NH2], Lys[-C(=NH)-CH3], Orn[-C(=NH)-CH3], Dab[-C(=NH)-CH3], Dap[-C(=NH)-CH3], Dab(Alloc), Asn, Gln, Met, Ser, Thr, Ser(Bzl), Thr(Bzl), Cys(Me), Cys(Bzl), Cys(Acm), Arg(NO2), His, Trp, Phg, GIy, Ala, Val, Ile, Leu, Phe, Phe(4-NH-C(=NH)-NH2), Phe(4-NO2), 2-Abu, Ala(3-CN), Ala[3-C(=NH)-NH2], 2-Abu(4-CN) and 2-Abu[4-C(=NH)-NH2].

12. The connection of one or more of paragraphs.1-11, in which E represents a residue selected from the series the which R22 represents a residue selected from the series consisting of hydrogen, alkyl, aryl, arylalkyl, heteroallyl and geterotsiklicheskikh, all of these residues may be substituted by substituents selected from the series consisting of halogen, hydroxy, alkyloxy, alkylenedioxy, nitro, cyano, dialkylamino, alkylsulfonyl, aminosulfonyl and trifloromethyl, which may be optionally substituted.

14. The compound of formula I according to one or more of the paragraphs.1-13, in which R1 represents allyloxycarbonyl or arylamination, And represents the balance (L)-4-amidinopropane, represents the balance (L)-glutamic acid, or a pharmaceutically acceptable salt, or ester (L)-glutamic acid, D represents a residue selected from the series consisting of Arg, Dap, Dab, Orn, Lys, Dap[-C(=NH)-NH2], Dab[-C(=NH)-NH2], Lys[-C(=NH)-NH2], Lys[-C(=NH)-CH3], Orn[-C(=NH)-CH3], Dab[-C(=NH)-CH3], Dap[-C(=NH)-CH3], Dab(Alloc), Asn, Gln, Met, Ser, Thr, Ser(Bzl), Thr(Bzl), Cys(Me), Cys(Bzl), Cys(Acm), Arg(NO2), His, Trp, Phg, GIy, Ala, Val, Ile, Leu, Phe, Phe(4-NR2), Phe(4-NH-C(=NH)-NH2), 2-Abu, Ala(3-CN), AIa[3-C(=NH)-NH2], 2-Abu(4-CN) and 2-Abu[4-C(=NH)-NH2], n = 0 or 1, E represents a residue selected from the series consisting of Cha, Chg, and Phe[4-C(-S-CH2

15. The compound of formula I according to one or more of the paragraphs.1-13, which is a

Alloc-pAph-Glu-Arg-Cha-NH2,

Arylamination-pAph-Glu-Arg-Cha-NH2,

Alloc-pAph-Glu-Arg-Chg-NH2,

Alloc-pAph-Glu-Dap[-C(=NH)-NH2]-Cha-NH2,

Alloc-pAph-Glu-Ala[3-C(=NH)-NH2]-Cha-NH2,

Alloc-pAph-Glu-Asn-Cha-NH2,

Alloc-pAph-Glu-Dab-Cha-NH2,

Alloc-pAph-Glu-Dap[-C(=NH)-NH2]-NH2,

Alloc-pAph-Glu-Gly-Cha-NH2,

Alloc-pAph-Glu-Thr(Bzl)-NH-(CH2)2-CH(Ph)2,

Alloc-pAph-Glu-Dab-NH-(CH2)2-Ph,

Alloc-pAph-Glu-Asn-NH-CH2-Chx,

Alloc-pAph-Glu-Dap[-C(=NH)-CH3]-Cha-NH2,

Alloc-pAph-Glu-Dab[-C(=NH)-NH2]-Cha-NH2,

Alloc-pApP>Alloc-pAph-Glu-Asn-1-(1-naphthyl)ethylamide,

Alloc-pAph-Glu-Asn-2-naphtylamine,

Alloc-pAph-Glu-Asn-3,4-dichloraniline,

Alloc-pAph-Glu-Asn-2-(3-chlorophenyl)ethylamide,

Alloc-pAph-Glu-Arg(NO2)-Cha-NH2,

Alloc-pAph-Glu-Cys(Bzl)-Cha-NH2,

Alloc-pAph-Glu-Trp-Cha-NH2,

Alloc-pAph-Glu-Phg-Cha-NH2,

Alloc-pAph-Glu-Asn-9-fluoroaniline,

Alloc-pAph-Glu-Asn-3,5-bistrifluormethylbenzene,

Alloc-pAph-Glu-Dap[-C (=NH)-NH2]-Phe[4-C(-S-(CH2)2-S-)-Phenyl]-NH2,

Alloc-pAph-Glu-Cys(Bzl)-Cha-NH2,

Alloc-pAph-Glu-Thr(Bzl)-Cha-NH2,

Alloc-pAph-Glu-Phe(4-NO2)-Cha-NH2,

Alloc-pAph-Glu-Asn-3,4-methylenedioxybenzene,

Alloc-pAph-Glu-Asn-2-(2-naphthyl)ethylamide,

Alloc-pAph-Glu-Asn-2-(1-naphthyl)ethylamide,

Alloc-pAph-Glu-Asn-2-(2-pyridyl)ethylamide,

Alloc-pAph-Glu-Asn-2,2-diphenylethylamine,

Alloc-pAph-Glu-Asn-2,4-differentiated or

Alloc-pAph-Glu-Asn-4-dimethylaminobenzene

or their pharmaceutically acceptable salt, amide or ester.

16. The method of obtaining compounds of one or more of the paragraphs.1-15, which includes linking protected amino acids using the methods of traditional medicinal chemistry and removing the protection from the target molecule with SIP is the ability to provide antithrombotic effect through reversible inhibition of activated factor VIIa (FVIIa) blood coagulation, containing an effective amount of a compound according to one of paragraphs.1-15 or its pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

18. The connection of one or more of paragraphs.1-15 or its pharmaceutically acceptable salt for use as an inhibitor of factor VIIa.

19. The connection of one or more of paragraphs. 1-15 or its pharmaceutically acceptable salt for use in the inhibition or reduction of blood clotting, inflammatory reactions, thromboembolic disease or restenosis of blood vessels.

 

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