Derivatives cyclopeptides as inhibitors of adhesion

 

(57) Abstract:

The invention relates to new compounds of General formula I

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where a is Gly; the remainder of the formula II

< / BR>
m= 0 or 1; n= 2 or 3; R1and R2each independently of the other represents H, R1and R2both together represent also

< / BR>
or

< / BR>
where IS -(CO)-(CH2)q-(CO)rwhere q=1, 2, or 3, r=0 or 1, or-CO-CH=CH-CO-; X IS H, Cl or1-C6alkyl; and if the mean remains optically active amino acids and derivatives of amino acids, are included as D-and L-forms, and their salts, process for the preparation of compounds of formula I and their salts; pharmaceutical composition having the ability to inhibit integrin containing in its structure at least one compound of the formula I and/or one of its physiologically acceptable salts. 4 S. and 2 C.p. f-crystals, 2 tab.

The invention relates to a compound of formula I

< / BR>
where And denotes Gly, Ala or NH-NH-CO, and these amino acids can also be derivatization,

In denotes a residue of formula II

< / BR>
With means -(CO)p-(CH2)q-(CO)r- or -(CO)p-CH= CH-(CO)r-, m, p, r each independently ody independently of each other denotes H or alkyl,

R1and R2both together represent also

< / BR>
R7, R8, R9, R10each independently of the other represents H, alkyl, AG, OR6Hal, NO2, NR6R6, NHCOR6CN, NHSO2R6, COOR6or COR6,

X denotes H, Hal, alkyl or AG,

Ar denotes unsubstituted or one-, two - or three-fold substituted by the radicals R3, R4or R5phenyl or unsubstituted naphthyl,

R3, R4, R5each independently of one another denotes R6, OR6Hal, NO2, NR6R6, NHCOR6CN, NHSO2R6, COOR6or COR6,

R6, R6'each independently of the other represents H, alkyl, phenyl or benzyl and

Hal denotes F, Cl, Br or I,

moreover, if the mean remains optically active amino acids and derivatives of amino acids, are included as D-and L-forms and their salts.

Such compounds of cyclic peptides are known, for example, application DE 4310643 or EP 0683173.

The basis of the invention was based on the task to obtain new compounds with valuable properties, especially those that could be used to manufacture medicines.

v-,3or5-integrin with a ligand, such as, for example, the binding of fibrinogen to the receptor 3-integrin. Particularly effective compounds exhibit in the case of integrinsv1,v3,v5,IIb3andv6andv8. The effectiveness of this action can be confirmed using the method described by J. W. Smith and others in Journ. Biol Chem. 265, 12267-12271 (1990). The dependence of the occurrence of angiogenesis from the interaction between vascular integrins and extracellular matrix proteins described in the P. C. Brooks, R. A. Clark, and D. A. Cheresh in Science 264, 569-571 (1994).

The possibility of suppression of this interaction and thereby the opportunity for early apoptosis (programmed cell death) of angiogenic vascular cells via cyclic peptide described by P. C. Brooks, A. M. Montgomery, M. Rosenfeld, R. A. Reisfeld, T.-No, Acting Klier and D. A. Cheresh in Cell 79, 1157-1164 (1994).

The compounds of formula I, blocking the interaction of integrin receptors with ligands, for example, the binding of fibrinogen with fibrinogen receptor (glycoprotein IIb/IIIa), prevent as the wound of cancer cells from the local tumor in the vascular system occurs due to the formation of microaggregates (microthrombi), due to the interaction of cancer cells with platelets. Cancer cells are blocked by protecting microaggregate and are not recognized by cells of the immune system. Microaggregate can be fastened in the walls of blood vessels, which facilitates the further penetration of cancer cells into the tissue. Due to the fact that the formation of microthrombi occurs indirectly by binding of fibrinogen with fibrinogen receptors on activated platelets, GPa/IIIb antagonists can be considered as effective inhibitors of metastasis.

The compounds of formula I can be used as active substances in medicinal products intended for use in medicine and veterinary medicine, primarily for the prevention and/or therapy of diseases of the cardiovascular system, thrombosis, myocardial infarction, arteriosclerosis, inflammation, apoplexy, angina, cancer, osteolytic diseases such as osteoporosis, pathological angiogenic diseases, such as inflammation, eye diseases, diabetic retinopathy, molekulyarnoi degeneration, myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, robotically glaucousness infection, bacterial infection, fungal infection, acute renal failure and in healing wounds as a means of facilitating the healing process.

The compounds of formula I can be used as possessing antimicrobial activity of substances in operations where used biomaterials, implants, catheters or pacemakers. They are antiseptic effect. The effect of antimicrobial activity can be confirmed using the method described in the P. Valentin-Weigund and others in Infection and Immunity, 2851-2855 (1988).

Due to the fact that the compounds of formula I are inhibitors of the binding of fibrinogen and thereby ligands fibrinogen receptors on the platelets, they can be used in vivo as a diagnostic tools for the detection and localization of thrombi in the vascular system, as they may be replaced, for example, radioactive or determined by UV radiation balance.

The compounds of formula I as inhibitors of the binding of fibrinogen can also be used as tools to study the metabolism of platelets at different stages of activation or intracellular mechanisms penuh3N-isotopes, allows, after binding to the receptor, to study these mechanisms.

Above and beyond the abbreviations for amino acid residues indicate residues of the following amino acids:

Ala alanine

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Arg arginine

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Gly glycine

H2N-CH-COOH

Next adopted the following abbreviations:

AC acetyl

SIDE tert-butoxycarbonyl

Bsoc benzyloxycarbonyl

DCCI dicyclohexylcarbodiimide

DMAP 4-dimethylaminopyridine

DMF dimethylformamide

EDCI N-ethyl-N,N'-(dimethylaminopropyl)carbodiimide

Et ethyl

The FCC fluoresceinlabeled acid

Fmoc 9-fluorenylmethoxycarbonyl

HOBT 1-hydroxybenzotriazole

MBGA 4-methylbenzhydrylamine

Me methyl

Pri 4-methoxy-2,3,6-trimethylphenylsulfonyl

N-MM N-methylmorpholin

OBzl benzyl ether

Okt octanoyl

OEt ethyl ester

OMe methyl ester

OtBu tert-butyl ether

FOA phenoxyacetyl

Sal salicyloyl

TFK triperoxonane acid

TRT, trityl (triphenylmethyl).

Since the above-mentioned amino acids can be represented in several enantiomeric forms, for example, as the components of the Roma in amino acids, for example, as components of compounds of formula I, can be provided with the relevant known protective groups.

Proposed according to the invention compounds also include the so-called proletarienne derivatives, i.e., modified with, for example, alkyl or acyl groups, sugars or oligopeptides of the compounds of formula I, in which the body quickly split up to the effective compounds according to the invention.

To this group belong also biodegradable polymer derivatives of the compounds according to the invention, as described, in particular, in Int. Journ. Pharm. 115, 61-67 (1995).

Amino acids, the configuration of which is not otherwise specified, have the (S)- or (L)-configuration.

The object of the invention is further a method of producing compounds of the formula I according to paragraph 1 of the formula and their salts. The method differs in that

a) compound of formula III

H-Z-OH (III)

in which Z represents a

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< / BR>
< / BR>
or

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and X, a, b and C have the meanings specified in paragraph 1 of the formula or of a reactive derivative of the compound of formula III is treated with a cyclization agent, or

b) the compound of formula I release from real is either acidic compound of formula I is transferred by treatment with acid or base in one of its salts.

Unless otherwise stated, the above and subsequent residues X, a, b, C, R1, R2, m, n, p, q and Z have the values listed in formulas I, II and III.

In the above formulas, X denotes H, Hal or alkyl, preferably H, C1 or CH3.

In the above formulas, the alkyl has 1-6 C-atoms and represents a methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, and, in addition, also pentyl, 1-, 2 - or 3-methylbutyl, 1,1-, 1,2 - or 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3 - or 4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1 - or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2 - or 1,2,2-trimethylpropyl. Particularly preferably alkyl means methyl.

R7, R8, R9and R10denote preferably H.

These amino acids and amino acid residues can also be derivatization, preferably N-methyl, N-ethyl, N-sawn, N-benzyl or C-methyl derivatives.

To a preferred further include primarily methyl, ethyl, propyl, butyl, tert-butyl, neopentylene or benzyl esters of the side-chain carboxyl group, in addition, also derivatives of AIBO ethoxycarbonylphenyl balance.

R6and R6'denote preferably, for example, H, methyl or ethyl, and benzyl or phenyl.

OR6means is preferably, for example, hydroxy - or methoxy group.

COR6is alkanoyl and preferably denotes formyl, acetyl, propionyl, butyryl, pentanoyl or hexanoyl.

AG represents an unsubstituted, preferably, as indicated above, one-deputizing phenyl, particularly preferably phenyl, o-, m - or p-tolyl, o-, m - or p-ethylphenyl, o-, m - or p-propylphenyl, o-, m - or p-isopropylphenyl, o-, m - or p-tert-butylphenyl, o-, m - or p-triptoreline, o-, m - or p-hydroxyphenyl, o-, m - or p-nitrophenyl, o-, m - or p-AMINOPHENYL, o-, m - or p-(N-methylamino)phenyl, o-, m - or p-acetamidophenyl, o-, m - or p-(triptoreline)phenyl, o-, m - or p-tianfeng, o-, m - or p-methoxyphenyl, o-, m - or p-ethoxyphenyl, o-, m - or p-carboxyphenyl, o-, m - or p-ethoxycarbonylphenyl, o-, m - or p-ethoxycarbonylphenyl, o-, m - or p-benzyloxycarbonyl, o-, m - or p-(carboxymethoxy)phenyl, o-, m - or p-(ethoxycarbonylmethoxy)phenyl, o-, m - or p-(ethoxycarbonylmethoxy)phenyl, o-, m - or p-(N,N-dimethylamino)phenyl, o-, m - or n-(N-ethylamino)phenyl, o-, m - or p-(N is hydroxy)phenyl, o-, m - or p-(formatosi)phenyl, o-, m - or p-formylphenyl, o-, m - or p-acetylphenyl, o-, m - or p-propionitrile, o-, m - or p-boutillier, o-, m - or p-pentanolide, o-, m - or p-(phenylsulfonyl)phenyl, o-, m - or p-phenoxyphenyl, o-, m - or p-methylthiophenyl, -, m - or p-methylsulfinylphenyl, o-, m - or p-methylsulfinylphenyl or naphthyl.

The concept of "aminosidine group" means preferably acetyl, propionyl, butyryl, phenylacetyl, benzoyl, toluyl, FOA, methoxycarbonyl, etoxycarbonyl, 2,2,2-trichlorocyanuric, SIDE, 2-iodoxybenzoic, Bsoc (carbobenzoxy"), 4-methoxybenzeneboronic, Fmoc, MTP or benzyl.

The compounds of formula I have at least two Cherednik center and therefore can be represented in different stereoisomeric forms. Formula I includes all these forms.

In accordance with this object of the invention are such compounds of formula I in which at least one of these residues has one of the abovementioned preferred meanings. Some preferred groups of compounds can be represented by the following fall under the formula I subformulae Ia, Ib and IC, where

in Ia X Is H, alkyl or Hal,

R1, R2- N,= 0,

n = 3,

p = 1,

r = 0 and

q = 1 and

in the IC X - H, alkyl, or Hal,

R1and R2both together constitute the

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m = 1,

n = 2,

R, r = 1 and

q = 2.

The compounds of formula I, as well as source materials for their production in the rest get by known methods described in the literature (for example, in such fundamental publications, as Houben-Weyl, Methods der organischen Chemie, published by Georg-Thieme-Verlag, Stuttgart), namely under conditions known and suitable for the implementation of these reactions. This may be also known, is not illustrated in the present description more options.

The source of the substance, if necessary, can also be formed in situ, eliminating their compulsory isolation from the reaction mixture and allows their subsequent conversion into the compounds of formula I.

The compounds of formula I can be obtained, for example, according to schemes 1 and 2 (see the end of the description).

An important structural element, 3-amino-3-(3-nitrophenyl)propionic acid, from scheme 1 are as under Journ. Org. Chem. 25, page 1758 (1960) of 3-nitrobenzaldehyde, malonic acid and ammonium acetate. During the synthesis of similar compounds used from box by cyclization of compounds of formula III under the conditions of peptide synthesis. It is helpful to work through conventional methods such synthesis are described, for example, in Houben-Weyl, 1.S., volume 15/II, pages 1 to 806 (1974).

The reaction is carried out preferably in the presence of a dehydration agent, such as carbodiimide, such as DCCI or EDCI, can also be used, for example, anhydride papapostolou acid (compare Angew. Chem. 92. 129 (1980)), diphenylphosphoryl or 2-ethoxy-N-etoxycarbonyl-1,2-dihydroquinoline, in an inert solvent, for example, in a halogenated hydrocarbon, such as dichloromethane, in a simple ether, such as tetrahydrofuran or dioxane, in amide, such as DMF or dimethylacetamide, a nitrile such as acetonitrile, dimethylsulfoxide, or in the presence of these solvents, at temperatures in the range of from about -10 to 40, preferably from 0 to 30oC. For more efficient intramolecular cyclization in front of intramolecular binding peptides suitable to work in dilute solutions. Duration of response depending on the specific conditions of its carrying out is from a few minutes up to 14 days.

Instead of the compounds of formula III can also be used derivatives of compounds of formula III, preferably the pre-orangered or activated ester. Remnants of this type is used to activate carboxyl groups in a typical acylation reactions described in the literature (for example, in such fundamental publications, as Houben-Weyl, Methods der organischen Chemie, published by Georg-Thieme-Verlag, Stuttgart). Activated esters suitable to form in situ, for example, by adding HOBT'a or N-hydroxysuccinimide.

The reaction is carried out usually in an inert solvent using gelegenheid carboxylic acid in the presence of an acid binding agent, preferably an organic base such as triethylamine, dimethylaniline, pyridine or quinoline. Appropriate and useful can be also supplements hydroxide, carbonate or bicarbonate of alkali or alkaline earth metal or other salts of weak acids, formed by interaction with alkaline or alkaline earth metals, preferably salts of potassium, sodium, calcium or caesium.

Educt of the formula III are generally new. They can be obtained by known methods of peptide synthesis.

Another possibility of obtaining compounds of formula I consists in the fact that these compounds release from their functional derivatives way is Tami for solvolysis, accordingly hydrogenolysis are those which, instead of one or more free amino and/or hydroxyl groups contain appropriate protective amino and/or hydroxy-group, preferably such that instead of the N-atom with a N-atom, are aminosidine group, for example, those which fall under the formula I, but instead of NH2groups contain other'group (where R' denotes aminosidine group, for example, SIDE or Bsoc).

It preferred are further educt bearing instead of the N-atom of the hydroxy-group hydroxyamino group, for example, those which fall under the formula I, but instead hydroxyproline groups contain R"O-phenyl group (where R' denotes hydroxyamino group).

It is also possible the presence in the molecule of the original substance of several identical or different, amino - and/or hydroxyamine groups. When there are differing from each other protective groups in many cases it is possible by selective to split.

The concept of "aminosidine group" is well known and relates to groups which are capable of protecting (blocking) an amino group from chemical reactions but which can be easily removed by zavershenija just unsubstituted or substituted acyl, aryl, arelaxation or kalkilya group. Because aminosidine group upon completion of the desired reaction (or the stage) is removed, the type and extent otherwise not play a significant role, preferred, however, groups with 1-20, especially with 1-8 C-atoms.

The term "acyl group" in relation to the proposed method is used in the broadest sense. It includes acyl groups derived from aliphatic, alifaticheskih, aromatic or heterocyclic carboxylic acids or sulphonic acids, and above all alkoxycarbonyl, aryloxyalkyl first alcoxycarbenium group. As examples of such apilink groups can be called alkanoyl, such as acetyl, propionyl, butyryl; arkanoid, such as phenylacetyl; aroyl, such as benzoyl or toluyl; aryloxyalkanoic, such as the FOA; alkoxycarbonyl, such as methoxycarbonyl, etoxycarbonyl, 2,2,2-trichlorocyanuric, SIDE, 2-iodoxybenzoic; Uralelectromed, such as BSAC (carbobenzoxy"), 4-methoxybenzeneboronic, Fmoc; arylsulfonyl, such as Inventory, Preferred aminosidine groups are SIDE and Inventory, as well as BSAC, Fm, benzyl and the us to protect the hydroxy-group from chemical reactions, but that can be easily removed upon completion of the desired chemical reactions in other parts of the molecule. Typical representatives of such groups are the abovementioned unsubstituted or substituted aryl, kalkilya or acyl groups, and alkyl groups. The type and extent hydroxyamine groups do not play a significant role, since upon completion of the desired chemical reaction or an appropriate stage again remove; preferred group with 1-20, especially with 1-10 C-atoms. As examples hydroxyamine groups can be named, among others, benzyl, p-nitrobenzoyl, p-toluensulfonyl, tert-butyl and acetyl, and particularly preferred of them benzyl and tert-butyl. To protect COOH-groups, it is preferable to use tert-butyl ethers.

The release of the compounds of the formula I from their functional derivatives is carried out - depending on the protective group - for example, by using a strong acid, preferably TFA or perchloro acid, but also using other strong inorganic acids, such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids, such as trichloroacetic acid, or sulfonic acids such as benzene and the flax. As the inert solvent preferably suitable organic solvents, for example carboxylic acids, such as acetic acid, ethers, such as tetrahydrofuran or dioxane, amides, such as DMF, halogenated hydrocarbons such as dichloromethane, in addition, alcohols such as methanol, ethanol or isopropanol, and also water. Can be used and mixtures of the mentioned solvents. TFK preferably used in excess without the addition of another solvent. Perchloro acid is used in the form of a mixture of acetic acid and 70% perchloro acid in the ratio 9:1. The temperature required for the cleavage is preferably in the range from approximately 0 to approximately the 50oWith, it is advisable to operate in the range of from 15 to 30oC (room temperature).

Cleavage of SIDE groups, OtBu and Inventory can be carried out, for example, preferably using TFA in dichloromethane or using approximately 3 BC-5 BC Hcl in dioxane at 15-30oWith, Fmoc-group can be split with approximately 5-50% solution of dimethylamine, diethylamine or piperidine in DMF at 15-30oC.

Trityloxy group used to protect amino acids such as get using TFA/10% thiophenol, and trailing group otscheplaut from all the above-mentioned amino acids; with TFA/anisole or TFA/thioanisole trityloxy group otscheplaut only from histidine, asparagine and glutamine, whereas its presence in the side chain of cysteine is saved.

Remove gidrodinamicheskim by a protective group (for example, BSAC or benzyl) can be chipped off, for example, by treatment with hydrogen in the presence of a catalyst (e.g. catalyst based on a noble metal such as palladium, preferably on a medium such as coal). As solvents it is possible to use any of the above, first of all, for example, alcohols, such as methanol or ethanol, or amides, such as DMF. The hydrogenolysis is carried out, as a rule, at temperatures in the range from approximately 0 to 100oC and a pressure ranging from about 1 to 200 bar, preferably at 20-30oC and 1-10 bar. Hydrogenolysis of Bsoc group is expedient to carry out, for example, in the presence of 5-10% Pd/C in methanol or using ammonium formate (instead of hydrogen) in the presence of Pd/C in methanol/DMF at 20-30oC.

The basis of the formula I can be translated using acid sootvetstvuem solvent, such as ethanol and subsequent evaporation. For such a reaction is primarily suitable acid, which form physiologically acceptable salts. Thus, in particular, it is possible to use such inorganic acids as sulfuric acid, nitric acid, halogenation acid, such as hydrochloric acid or Hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfanilate, and also organic acids, especially aliphatic, alicyclic, analiticheskie, aromatic or heterocyclic one - or polybasic carboxylic, sulfonic or sulfuric acids, such as formic acid, acetic acid, propionic acid, pavlikova acid, diethyloxalate acid, malonic acid, succinic acid, Emelyanova acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinamide acid, methane - or econsultation, ethicalfashion, 2-hydroxyethanesulfonic, benzosulfimide, p-toluensulfonate, naphthalenamine and dissolvability, louisanna acid. About salts palenia and/or purifying compounds of formula I.

On the other hand, the acid of formula I by the interaction with the base can be translated into one of its physiologically acceptable metal or ammonium salts. As such salts may be treated primarily salts of sodium, potassium, magnesium, calcium and ammonium, and substituted ammonium salts, e.g. salts of dimethyl-, diethyl - or Diisopropylamine, salt monoethanol-, diethanol - or Diisopropylamine, salt cyclohexyl-, dicyclohexylamine, salts of dibenzylethylenediamine, in addition, for example, salts with arginine or lysine.

Another object of the invention is the use of compounds of the formula I and/or their physiologically acceptable salts for pharmaceutical compositions, primarily non-chemical way. Thus they can be used in conjunction with at least one solid, liquid and/or semi-liquid carrier or auxiliary substance, and optionally in combination with one or more other active substances for the manufacture of appropriate dosage forms.

The object of the invention is further pharmaceutical composition, containing in its composition at least one compound of the formula I and/or one of its physiologically III. As carriers for these compositions can be considered organic or inorganic substances suitable for enteral (for example oral), parenteral, local injection or for use in the form of an inhalation spray and do not react with the new compounds, for example water, vegetable oils, borzilova alcohols, alkalophile, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, vaseline. For oral assignments are designed primarily for tablets, pills, coated tablets, capsules, powders, granules, syrups, medicine or drops for rectal use - suppositories, for parenteral use, solutions, preferably oily or aqueous solutions, furthermore suspensions, emulsions or implants, for topical application suitable ointments, creams or powders. The new compounds can also be lyophilized and the resulting lyophilizate be used, for example, for the preparation of injection preparations. These compositions can be sterilized and/or they may contain auxiliary substances, such as oil, preservatives, stabilizers and/or wetting, emulsifying agents,several other active substances, for example, one or more vitamins.

As of spray for inhalation can be used sprays, which contain the active substance in dissolved or suspended form in the same propellant or propellant mixture of (for example, in CO2or in perchloroethane). It is advisable to apply the active ingredient in micronized form, and you can add one or more additional physiologically acceptable solvents such as ethanol. Inhalation solutions can be performed using a conventional inhalers.

The compounds of formula I and their physiologically acceptable salts can be used as integrin inhibitors primarily in the fight against diseases such as diseases of the cardiovascular system, thrombosis, myocardial infarction, ischemic heart disease, arteriosclerosis, apoplexy, angina, cancer, osteoporosis, inflammation, infection and restenosis after angioplasty.

The compounds of formula I with distinctive features in claim 1 of the formula and/or their physiologically acceptable salts can find application also in pathological processes associated with or caused by angiogenesis, primarily cancer, z is Oh, can be entered similarly to other known, commercially available peptides, but especially similar to the compounds described in US-A 4472305, preferably in dosages from about 0.05 to 500 mg, especially from 0.5 to 100 mg per uniform dose. The daily dose is preferably from about 0.01 to 2 mg/kg of body weight of the patient. However, when you assign specifically to one or other patient specific dose of the latter depends on various factors such as the effectiveness of the compounds of the age and the patient's weight, General health, sex, diet, time and method of administration, rate of excretion, combination of drugs and the severity of the respective disease, which requires this treatment. Preferably parenteral application.

The compounds of formula I can be used also as integrin ligands for the manufacture of columns for affinity chromatography, designed to produce a clean integrins.

The ligand, i.e. the compound of formula I, attached through the anchoring functional group, for example, carboxyl group, by type of covalent bond to the polymer carrier. In hydrophilic properties, for example, sewn, polisher, such as cellulose, sepharose or Sephadexacrylamide, the polymer is polyethyleneglycol or Tentakelpolymere.

Obtaining materials for affinity chromatography to clean integrin carried out under the conditions customary and known, which is used by condensation of amino acids.

As mentioned above, the compounds of formula I contain at least two chiral center and can therefore be represented in racemic or optically active form. Resulting racemates can by known methods of mechanical or chemical means to separate the enantiomers. Preferably the racemic mixture by reacting with an optically active separating agent form diastereoisomers. As separating agents can be used, for example, optically active acids, such as D - and L-forms of tartaric acid, diatsetilvinny acid, dibenzoyltartaric acid, almond acid, malic acid, lactic acid or the various optically active camphor sulphonic acids, such as camphor acid. Preferably also carry out the separation of the enantiomers using columns filled with optically active Xan/isopropanol/acetonitrile, for example, in a volume ratio 82:15:3.

Optically active compounds of formula I can, of course, also be obtained by the above methods due to the use of starting materials which are already optically active.

Above and beyond all temperatures are in degrees Celsius. Room temperature refers to a temperature equal to the 22oC. Under used in the following examples, the term "normal processing" we mean the following: if necessary, water is added, depending on the structural characteristics of the final product establish, if necessary, for pH values from 2 to 10, extracted with simple ether or dichloromethane, the organic phase is separated, dried over sodium sulfate, filtered, evaporated and purified by chromatography on silica gel and/or by crystallization.

RT = retention time (in minutes) when GHUR in the following systems;

column: LichrosorbRP 18 (2504; 5 μm);

eluent A: 0.1% of TFA in water;

eluent B: 0.1% of TFA in 90% acetonitrile, 10% water;

flow rate: 1 ml/min;

gradient: 20-95% B/50 min;

the detection at 215 nm.

Separation of the diastereomers preferably carried out under these conditions.

BR>
Synthesis ciclismo connection

Similarly to scheme 1 synthesize methyl ester (3R,S)-3-amino-3-(4-methyl-3-nitrophenyl)propionic acid (3R, S-IX). This broadcast by a known method splits the enantiomers and then the methyl ester of (3S)-3-amino-3-(4-methyl-3-nitrophenyl)propionic acid (3S-(IX) similar to the scheme 2 turn in MTP-secure connection - methyl ether 3S-amino-3-(3-{3-[1-(carboximetilkrahmal)-4 - guanidino-1S-butylcarbamoyl]propionamido}-4-were)propofol acid (S,S-XIV).

Cyclization of

616 mg MTP-secure connection - methyl ester 3S-amino-3-(3-{3-[1-(carboximetilkrahmal)-4-guanidino-1S-butylcarbamoyl] propionamido}-4-were)propionic acid (S,S-XIV) is dissolved in 80 ml of DMF and diluted with 800 ml of dichloromethane. Then cooled to -20oWith and consistently add 300 mg EDCI, 98 mg DMAP and 0,176 ml N-MM. Then warmed to room temperature and stirred over night. Then, the solution concentrated and the residue was stirred in 200 ml Polynesians solution Panso3. The precipitate is removed by using a suction filter and washed. The result is 400 mg of the substance, which is purified by preparative GHUR. After chromatography receive 44 mg MTP-protected cyclo[13.3.1] endeca-1(18), 15(19), 16-trien-14-yl]acetic acid, RT 26,1; FAB 716.

Saponification and removal of Materials and equipment-protective group

By saponification in KOH/methanol receive MTP-protected cyclic connection (8S, 14S)-[8-(3-guanidinopropionic)-18-methyl-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo [13.3.1] endeca-1(18), 15(19), 16-trien-14-yl] acetic acid. 25 mg of this compound then dissolved in 4.3 ml of 98% triperoxonane acid and stirred overnight at room temperature. In conclusion, the solution is centrifuged and purified by preparative GHUR. The result is 9.5 mg (8S,14S)-[8-(3-guanidinopropionic)-18-methyl-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo [13.3.1] endeca-1(18),15(19),16-trien-14-yl] acetic acid (8S,14S-IB), RT 20,2; FAB 490.

Examples 2-3

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Example 2 (q=2)

On the basis of 85 mg MTP-protected connection (8S,14S)-2-(8-(3-guanidinopropionic)-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo [13.3.1] -endeca-16,18,19-triene-4-yl)acetic acid, analogously to example 1 by interaction with 14.7 ml 98% triperoxonane acid 13 mg of (8S,14S)-2-(8-(3-guanidinopropionic)-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo [13.3.1] endeca-16,18,19-triene-4-yl)acetic acid; RT 17,2; FAB 476.

Example 3 (q=3)

Based on 300 mg MTP-secure connection - (9S,15S)-2-(9-(3-guanidino 1 obtain 74 mg of (9S, 15S)-2-(9-(3-guanidinopropionic)-3,7,10,13-tetraoxo-2,8,11,14-tetraazabicyclo[14.3.1]eicosan-17,19,20-trien-15-yl)acetic acid; RT 18,3; FAB 490.

Example 4

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Based on the MTP-secure connection - (6S,12S)-[6-(3-guanidinopropionic)-4,7,10-trioxo-2,5,8,11-tetraazabicyclo [11.3.1] heptadecan-1(17), 13,15-triene-12-yl] acetic acid, analogously to example 1 receive (6S, 12S)-[6-(3-guanidinopropionic)-4,7,10-trioxo-2,5,8,11-tetraazabicyclo[11.3.1] heptadecan-1(17),13,15-trien-12-yl] acetic acid.

Example 5-8

< / BR>
Analogously to example 1 to synthesize the following compounds (see table 1).

In the following examples, the pharmaceutical composition and the technology of their preparation in the appropriate dosage forms.

Example: Vials for injection solutions

A solution of 100 g of the active substance of the formula I and 5 g of disodium hydrogen phosphate in 3 l of double-distilled water using 2 N. hydrochloric acid set at pH 6.5, sterile filtered, filled flask, lyophilized in sterile sterile conditions and sealed. Each vial contains 5 mg of active substance.

Example B: Suppositories

First prepare a mixture of 20 g of the active substance of the formula I, 100 g of soya lecithin and 1400 is about substance.

Example: Solution

Prepare a solution of 1 g of the active substance of the formula I, 9,38 g NaH2PO42H2O, 28,48 g Na2HPO412H2O and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. Then set to pH 6.8, adjusted to a volume of 1 l and sterilized by irradiation. This solution can be applied in the form of eye drops.

Example D: Ointment

When observing aseptic conditions prepare a mixture of 500 mg of the active substance of the formula I with 99.5 g of petroleum jelly.

Example D: Tablets

Prepare a mixture of 1 kg of active substance of the formula I, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate, which is then tabletirujut by those of ordinary gentle in such a way that each tablet contains 10 mg of active substance.

Example E: Bean

Analogously to example D is pressed tablets, which then according to standard technology is covered by a shell of sucrose, potato starch, talc, tragant and dye.

Example G: Capsules

2 kg of active substance of the formula I according to standard technology to produce capsules with terrorisation coating in such a way that each capsule contains 20 mg of active substance.

Example: Inhalation spray

14 g of the active substance of the formula I are dissolved in 10 l of isotonic NaCl solution and then the solution is poured into a conventional, commercially available cartridges, equipped with a pumping device. The solution can be applied to inhalation mouth and nose. Issued from cans for one pressing portion (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg

The Protocol pharmacological tests

Analysis of inhibition of receptor

The purified integrinv3from human placenta was absorbed in the wells of microtiter tablet and were loaded with biotinylated complementary ligand, vitronectin (VN)v3/in the presence of increasing amounts of the analyzed compounds.

Method: 1 µg-ml-1conjugate Biotin-ligand is incubated with the receptor mucous membranes (1 μg-ml-1in the presence of peptides in serial dilution in 30oC for 3 hours Associated ligand was measured using the conjugate antibiotin-alkaline phosphatase.

Velich.

The tested compounds exhibit inhibitory activity against the bindingv3with vitronectin.

Literature

1. Charo I. F., L. Nannizzi, Smith J. W., Cheresh D. A., Cell. Biol., 111, 2795-2800 (1990).

1. The compounds of formula I

< / BR>
in which A - Gly;

In the remainder of the formula II

< / BR>
where m = 0 or 1, n = 2 or 3;

R1and R2each independently of one another denotes N and R1and R2both together represent also

< / BR>
or

< / BR>
where IS -(CO)-(CH2)q-(CO)r- where q = 1, 2, or 3, and r = 0 or 1, or-CO-CH= CH-CO-;

X is H, CL or1-C6alkyl;

moreover, if the mean remains optically active amino acids and derivatives of amino acids, are included as D-and L-forms,

and also their salts.

2. Enantiomer or diastereoisomer of compounds of formula I under item 1.

3. The compounds of formula I on p. 1:

a) (8S, 14S)-2-(8-(3-guanidinopropionic)-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo[13.3.1] endeca-16,18,19-trien-14-yl)acetic acid;

b) (9S, 15S)-2-(9-(3-guanidinopropionic)-3,7,10,13-tetraoxo-2,8,11,14-tetraazabicyclo[14.3.1] eicosan-17,19,20-trien-15-yl)acetic acid;

in) (8S, 14S)-(8-(3-guanidinopropionic)-18-methyl-3,6,9,12-tetraoxa-2,7,10,13-tetraazabicyclo[13.3.1] endeca-1(18), 15(19), 16-trien-14-yl) who Rijen-12-yl)acetic acid,

and also their salts.

4. The compounds of formula I on p. 1 and their physiologically acceptable salts as inhibitors of integrin to fight with diseases of the cardiovascular system, thrombosis, myocardial infarction, ischemic heart disease, arteriosclerosis, apoplexy, angina pectoris, tumors, osteoporosis, inflammations, infections and restenosis after angioplasty.

5. The method of obtaining compounds of formula I on p. 1 and their salts, characterized in that the compound of formula III

H-Z-OH, (III)

in which Z represents a

< / BR>
In the remainder of the

< / BR>
X, A, B, C, R1, R2n and m have the meanings specified in paragraph 1,

treated with a cyclization agent, after which the compound of formula I release from one of its functional derivatives by treatment with an agent of the solvolysis or hydrogenolysis and/or the resulting compound of formula I is transferred by treatment with acid or base in one of its salts.

6. Pharmaceutical composition having the ability to inhibit integrin, characterized in that it contains at least one compound of formula I under item 1 and/or one of its physiologically acceptable salts.

 

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< / BR>
in which R1selected from the group consisting of hydrogen, unsubstituted or optionally substituted aralkyl, unsubstituted or optionally substituted orelkinoservisa, unsubstituted or optionally substituted allyloxycarbonyl, unsubstituted or optionally substituted alkyl and hydroxyamino group; R2selected from the group consisting of hydrogen, unsubstituted or optionally substituted orelkinoservisa, unsubstituted or optionally substituted allyloxycarbonyl, aminosidine group; R3selected from the group consisting of hydrogen, unsubstituted or optionally substituted alkyl and unsubstituted or optionally substituted aralkyl; R4selected from the group consisting of unsubstituted or optionally substituted alkyl and unsubstituted or optionally substituted aralkyl; R5and R6that may be the same or different, each independently selected from the group consisting of hydrogen, unsubstituted or optionally substituted alkyl, unsubstituted or optionally substituted cycle>and R6taken together with the nitrogen atom to which they are attached, form an unsubstituted or optionally substituted heterocyclic group; R7selected from the group consisting of hydrogen, hydroxy, unsubstituted or optionally substituted alkyl and unsubstituted or optionally substituted aralkyl; R8selected from the group consisting of hydrogen, hydroxy, unsubstituted or optionally substituted alkyl and unsubstituted or optionally substituted aralkyl, and R9selected from the group consisting of hydrogen, hydroxy, amino and a group of the formula-X-Y, in which X is selected from the group consisting of unsubstituted or optionally substituted (C1-C6)-alkylene and unsubstituted or optionally substituted phenylene, and Y denotes a group of formula-a-b or a-B, where a is selected from the group consisting of unsubstituted or optionally substituted (C1-C6)-alkylene, imino and unsubstituted or optionally substituted (C1-C6)-alkylamino, and selected from the group consisting of hydrogen, amino, amidino, acylmethyl, unprotected or optionally protected bis (phosphono)methyl, provided that R7, R8and R are not simultaneously represent
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