SK2132000A3 - Cyclopeptide derivatives as adhesion inhibitors - Google Patents

Abstract

Compounds of formula (I), in which X, A, B and C have the meanings given in Claim 1, as well as their salts, can be used as integrin inhibitors, in particular for the prevention and treatment of circulatory diseases, thrombosis, heart infarct, coronary heart diseases, arteriosclerosis, in pathological conditions maintained and propagated by angiogenesis, and for tumour therapy.

Description

Cyclic peptide derivatives as inhibitors of adhesion

Technical field

The invention relates to cyclic adhesion inhibiting peptide derivatives, to a process for its preparation, to its use and to pharmaceutical compositions containing it.

The invention relates to compounds of formula I

where it means

A Gly, Ala or NH-NH-CO, said amino acids optionally being in the form of their derivatives,

B is a group of formula II

I H

(II)

C group - (CO) _p - (CH ₂ ) _q - (CO) _r - or - (CO) _p -CH = CH- (CO) _r -, m, p, r independently of one another 0 or 1, n, q independently of 1, 2, 3 or 4,

R ¹ and R ² each independently H or alkyl,

R ^{@ 1} and R ^{@ 2 taken} together are also C1 / C

II c / \

or

R ⁷ , R ⁸ , R ⁹ and R ¹⁰ independently of one another are hydrogen, alkyl, Ar, OR ⁶ , Hal, NO 2, NR ⁶ R ⁶ ', NHCOR ⁶ , CN, NHSO ² R ⁶ , COOR ⁶ or COR ⁶ ,

X is hydrogen, Hal, alkyl or Ar,

Ar is a phenyl group optionally substituted with one, two or three R ⁸ , R ⁴ or R ⁵ groups or an unsubstituted naphthyl group,

R ³ , R ⁴ , R ⁵ independently of one another R ⁶ , OR ⁶ , Hal, NO 2, NR ⁶ R ⁶¹ , NHCOR ⁶ , CN, NHSO ² R ⁶ , COOR ⁶ or COR ⁶ ,

R ⁶ , R ⁶ 'independently of one another are hydrogen, alkyl, phenyl or benzyl; and

Hal atom of fluorine, chlorine, bromine or iodine with respect to residues of optically active amino acids and amino acid derivatives, both D and L forms and their salts are included

BACKGROUND OF THE INVENTION

Similar cyclic peptide derivatives according to the invention are described, for example, in DE 43 10 643 or EP 0 683173.

SUMMARY OF THE INVENTION It is an object of the present invention to provide novel compounds having valuable properties which are particularly suitable for the manufacture of medicaments.

SUMMARY OF THE INVENTION

The present invention provides a cyclic peptide derivative as an adhesion inhibitor.

Surprisingly, it has been found that the compounds of the formula I and their salts have very valuable pharmacological properties with good tolerability. They act especially as integrin inhibitors, in particular inhibiting the interaction of _α ν - ^ 3 ^'and P ₅ l®bo -integrínových receptors with ligands, such as the binding of fibrinogen to the _.beta.3 receptor -integrínový. The compounds of the formula I have particular activity in the case of integrins α _ν β _Σ , α _ν β ₃ , α _ν β ₅ , and _IT ^ ₃ as well as α _ν β ₆ and α _ν βθ.

This action can be demonstrated, for example, by the method of J. W. Smith et al. (J. Biol. Chem. 265: 12267-12271, 1990).

P.C. Brooks, R.A. Clark and D.A. Cheresh, Science 264, p.

569-571 (1994) report that the development of angiogenesis depends on the interaction of vascular integrins and extracellular matrix proteins.

The possibility of using inhibition of this interaction and thereby inducing apoptosis (programmed cell death) of angiogenic vascular cells by the action of a cyclic peptide has been described by P.C. Brooks, A.M. Montgomery, M. Rosenfeld, R.A. Reisfeld, T.-Hu, G. Klier, and D.A. Cheresh (Cell 79, 1157-1164, 1994).

Compounds of formula I that block the interaction of integrin receptors and ligands, such as fibrinogen to the fibrinogen receptor (glycoprotein IIb / IIIa) prevent, as GPIIb / IIIa antagonists, the spread of tumor cells by metastasis. This is illustrated by the following observations.

The spread of tumor cells from a localized tumor to the vascular system occurs by the formation of micro-registers (microthrombi) as a result of the interaction of tumor cells and platelets. Tumor cells are shielded by protection in the microaggregate and are not recognized from cells of the immune system.

The microaggregates can settle on the vessel walls, thereby facilitating further penetration of tumor cells into the tissue. Since microtromb formation is mediated by the binding of fibrinogen to fibrinogen receptors on activated platelets, GPIIa / IIIb antagonists may be considered as potent metastasis inhibitors.

The compounds of the formula I can be used as active medicaments in human and veterinary medicine, in particular for the prophylaxis and treatment of circulatory diseases, thrombosis, myocardial infarction, arteriosclerosis, inflammation, apoplexy, angina pectoris, tumor diseases, osteolytic diseases such as osteoporosis. pathologically angiogenic diseases such as inflammation, ophthalmological diseases, diabetic retinopathy, corneal degeneration, myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, ulcerative glaucoma, ulcerative colitis, angioplasty, angioplasty, psoriasis, psoriasis, atherosclerosis, atherosiosis, atherosclerosis bacterial infections, fungal infections, acute renal failure, and in support of wound healing and healing processes.

The compounds of the formula I can be used as antimicrobial agents in operations using biological material, implants, catheters or cardiac stimulators. These compounds have an antiseptic effect. The efficacy of antimicrobial activity can be demonstrated by the method of P. Valentin-Weigund et al. (Infection and Immunity, 1988, 2851-2855).

Since the compounds of formula (I) are inhibitors of fibrinogen binding and thus fibrinogen receptor ligands to platelets, they are useful as diagnostics for the detection and localization of thrombi in the vascular system in vivo when, for example, substituted with a radioactive or UV detectable residue.

The compounds of formula I as inhibitors of fibrinogen binding can also be used as effective adjuvants for the study of platelet metabolism at various activation stages or in the intracellular signaling mechanisms of fibrinogen receptors. The detectable unit of the embedded label, for example the ³ H isotopic label, allows the mechanisms to be investigated after binding to the receptor.

Amino acid residue abbreviations are the following amino acid residues:

Ala alanine NH, 1 H ₃ C-CH-COOH Arg Arginine NH, 1'NH-CH, CH, CH, -CH-COOH HN <2 2 2 nh ₂ Gly glycine H ₂ N - CH - COOH

In addition, abbreviations have the following meanings:

Ac acetyl

BOC tert-butoxycarbonyl

CBz or Z benzyloxycarbonyl

DCC1 dicyclohexylcarbodiimide DMAP 4-dimethylaminopyridine DMF dimethylformamide EDC1 N-ethyl-N, N '- (dimethylaminopropyl) carbodiimide et ethyl FCA Fluoresceinecarboxylic acid Fmoc 9-fluórenylmetoxykarbony1 HOBt 1-hydroxybenzotriazole HONSu N-hydroxysuccinimide MBHA 4-Methylbenzhydrylamine Me methyl Mtr 4-methoxy-2,3,6-trimethylphenylsulfonyl NMM N-methylmorpholine OBzl benzyl Oct octanoyl OEt ethyl OMe methyl ester OtBu tert-butyl POA phenoxyacetyl

Sal saliciloyl TFA trifluoroacetic acid Trt trityl (triphenylmethyl)

Where the above amino acids may be in several enantiomeric forms, for example as a proportion of the compounds of formula (I) according to the invention, all these forms as well as mixtures thereof (e.g. DL forms) are included in the invention. In addition, amino acids, for example as a proportion of compounds of formula I, may be protected by known protecting groups.

The compounds according to the invention also include so-called prodrug derivatives, i.e., alkyl or acyl groups, sugars or oligopeptides-altered compounds of the formula I which are rapidly cleaved in the body into active compounds according to the invention. Also included in this group are biodegradable polymer derivatives of the compounds of the invention described in the literature (e.g. Int. J. Pharm. 115, 61-67 (1995)).

Amino acids not specifically listed have the (S) - or (L) -configuration.

A process for the preparation of a compound of formula (I), wherein the individual symbols are as defined above, or salts thereof, according to the invention is characterized in that:

a) the compound of formula III

H-Z-OH (III) wherein is

Z is of the formula ## STR6 ## _in t1-NH

NH-C-B-A-

or

OH

HN-A-B-C- and X, A, B and C are as defined above, or a reactive derivative of a compound of formula III is treated with a cyclizing agent, or

b) releasing a compound of formula I from its functional derivative by treatment with a solvolysis or hydrogenolysis agent and / or converting a basic compound or acidic compound of formula I by treatment with an acid or base into its salt.

The above symbols X, A, B, C, R ¹ , R ² , m, n, p, q and Z have the meanings given in formulas I, II and III unless otherwise expressly indicated.

In the above formulas, X is preferably hydrogen, Hal or alkyl, especially hydrogen, chlorine or methyl.

In the above formulas, alkyl is understood to mean alkyl of 1 to 6 carbon atoms, such as, in particular, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, and 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 and 1,1 , 2- or 1,2,2-trimethylpropyl. Particularly preferably, alkyl is methyl.

R @ ⁷ , R @ ⁸ , R @ ⁹ and R @ ¹⁰ are preferably hydrogen.

Said amino acids or amino acid residues may also be derivatized, with N-methyl, N-ethyl, N-propyl, N-benzyl or Ca-methyl derivatives being preferred.

Particularly preferred are also methyl, ethyl, propyl, butyl, tert-butyl, neopentyl or benzyl esters of carboxyl side chains, furthermore also arginine derivatives which are optionally substituted on the group of the formula -NH-C (= NH) -NH ₂ acetyl, benzoyl, methoxycarbonyl or ethoxycarbonyl radical.

R ⁶ and R ⁶ 'are preferably, for example, methyl, ethyl, also benzyl or phenyl, OR ⁸ is preferably hydroxy or methoxy and COR ^{6 is} alkanoyl, and especially formyl, acetyl, propionyl, butyryl, pentanoyl or hexanoyl.

Ar is unsubstituted preferably as mentioned above monosubstituted phenyl group, preferably phenyl, o-, m- or p-tolyl, o-, m- or p-ethylphenyl, o-, male- or p-propylphenyl, o-, m- or p-isopropylphenyl, o-, m- or p-tert-butylphenyl, o-, m- or p-trifluoromethylphenyl, o-, m- or p-hydroxyphenyl, o-, m- or p-nitrophenyl, ο-, m- or p-aminophenyl, o-, m- or p (N-methylamino) phenyl, o-, m- or p-acetamidophenyl, o-, m- or p- (trifluoromethoxy) phenyl, o-, m- or p-cyanophenyl, o-, m- or p-methoxyphenyl, o-, m- or p-ethoxyphenyl, o- m- or p-carboxyphenyl, o-, m- or p-methoxycarbonylphenyl, o-, m - or p-ethoxycarbonylphenyl, o-, male- or p-benzyloxycarbonylphenyl, o-, m- or p- (carboxymethyloxy) phenyl, o-, m- or p- (methoxycarbonylmethyloxy) phenyl, o-, m- or p- (methoxycarbonylethyloxy) ) phenyl, o-, male- or p- (N, N-dimethylamino) phenyl, o-, m- or p- (N-ethylamino) phenyl, o-, m- or p- (N, N-diethylamino) phenyl, o-, m- or p-fluorophenyl, o-, m- or p-bromophenyl, o-, p-chlorophenyl, o-, m- or p- (difluoromethoxy) phenyl, o ^-, m- or p- (fluoromethoxy) phenyl, o-, m- or p-formylphenyl, o-, m- or p-acetyl-phenyl, o, m or ppropionylphenyl, o-, m- or p-butyrylphenyl, o-, male- or p-pentanoylphenyl, o-, m- or p- (methylsulfonamido) phenyl, o-, m- or p-phenoxyphenyl, o-, m- or p-methylthiophenyl, o-, m- or p-methylsulfinylphenyl, o-, p- or p-methylsulfonylphenyl or naphthyl groups.

Amino-protecting group is preferably acetyl, propionyl, butyryl, phenylacetyl, benzoyl, toluyl, POA, methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOC, 2-iodoethoxycarbonyl, CBZ (carbobenzoyl) -methoxybenzyloxycarbonyl, FMOC, Mtr or benzyl.

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

Accordingly, the invention relates in particular to compounds of the formula I in which at least one of the symbols has the above-mentioned preferred meaning. Some preferred groups of compounds of formula (I) are the following compounds of formulas (Ia), (Ib) and (Ic), wherein not particularly indicated symbols have the meanings given in formula (I) in the formula

la) X hydrogen, alkyl R ^1, R ² a hydrogen atom, m 0 n 3 P, r 1 a q 2 or 3;

a halogen atom or group,

Ib) X is hydrogen, alkyl

R ^{@ 1} , R ^{@ 2} is hydrogen, a group or a halogen atom,

m about, n 3 P I r 0 a q i; Ic) x hydrogen, alkyl R ¹ , R ² together

a halogen atom or group,

H \ / C

II c / \

H

m1, n2, p, r and q2.

The compounds of formula (I) and the starting materials for their preparation are prepared by known methods described in the literature (for example, in standard publications such as Houben-Weyl, Methoden der Organischen Chemie, Georg-Thieme Verlag, Stuttgart) under reaction conditions, which are known and suitable for the above reactions. It is also possible to use known variants which are not described here in greater detail.

The starting materials may optionally be formed in situ, i.e. they are not isolated from the reaction mixture, but the reaction mixture is immediately used to prepare compounds of formula I.

For example, the compounds of formula I can be obtained by the method of Schemes 1 and 2 below.

The important 3-amino-3- (3-nitrophenyl) propionic acid as the building block of the process of Scheme 1 is prepared as described in J. Org. Chem. 25, p. 1758 (1960), from 3-nitrobenzaldehyde from malonic acid and from ammonium acetate. The corresponding nitrobenzaldehyde derivatives are used in the preparation of analogous compounds.

Scheme 1

O o hoAA

· »

SOCI ^ MeOH

1. Bcc-Gly-QH MeO

2. HCl / Dioxane

----------------- ►

3. Boc-Arc (Mtr) -OH

NH

Gly — Arg (Mtr) —Hoc

IN"

1. HCl / Dioxane

Second

or

Arg (Mtr) I.RaNi

2. cyclization

3. saponification

4. cleavage Mtr = 2 or 3

VIII

! A

Scheme 2

IX

X

cleavage of the group

In particular, the compounds of formula I can be obtained by cyclization of compounds of formula III under peptide synthesis conditions. In this connection, the known peptide synthesis methods described in the literature are expediently carried out (for example, HoubenWeyl, I..c., Vol. 15/11, pp. 1-806, 1974).

Preferably, the reaction proceeds in the presence of a dehydrating agent selected from, for example, a carbodiimide such as DCCl or EDCl, propanephosphonic anhydride (Angew. Chem. 92, 129, 1980), diphenylphosphoryl azide and 2-ethoxy-N-ethoxycarbonyl- 1,2-dihydroquinoline, in an inert solvent such as a halogenated hydrocarbon such as dichloromethane; an ether such as tetrahydrofuran or dioxane; an amide such as dimethylformamide or dimethylacetamide; a nitrile such as acetonitrile; dimethyl sulfoxide or a mixed solvent thereof, at a temperature of about -10 to 40C, preferably 0 to 30 C ^e to promote intramolecular cyclization over intermolecular peptide bonding, it is expedient to work in dilute solutions. The reaction time is between a few minutes and 14 days, depending on the reaction conditions used.

Instead of the compounds of the formula III, suitable reactive derivatives of these compounds can also be used for the reaction, for example a pre-activated carboxylic acid or carboxylic acid halide, symmetrical or mixed anhydride or active ester. Such groups for activating the carboxyl group in typical acylation reactions are described in the literature (for example, in standard publications such as Houben-Weyl, Methoden der Organischen Chemie, Georg-Thieme Verlag, Stuttgart). The activated esters are conveniently formed in situ, for example by the addition of HOBt or N-hydroxysuccinimide.

The reaction is generally carried out in an inert organic solvent, using a carboxylic acid halide in the presence of an acid binding agent, preferably an organic base such as triethylamine, dimethylaniline, pyridine or quinoline.

It may also be beneficial to add an alkali metal or alkaline earth metal hydroxide, carbonate or bicarbonate or other weak alkali metal or alkaline earth metal salt, preferably potassium, sodium, calcium or cesium.

The starting materials of the formula III are generally novel. They can be prepared by known methods of peptide synthesis.

The compounds of the formula I can also be obtained by liberating from their functional derivatives by solvolysis, in particular by hydrolysis or hydrogenolysis.

Suitable starting materials for solvolysis or hydrogenolysis are compounds having, instead of one or more free amino and / or hydroxyl groups, corresponding protected amino and / or hydroxyl groups, preferably compounds which, instead of one H-atom which is linked to N the atoms have an amino protecting group, for example compounds which correspond to the general formula I, but instead of the NH ₂ group they have an NHR 'group (where R' represents an amino protecting group such as BOC or CBZ).

Further suitable as starting materials are compounds which have a hydroxyl protecting group in place of the H-atoms of the hydroxyl group, for example compounds which correspond to the general formula I, but instead of the hydroxyphenyl group have an R-phenyl group (where R is a hydroxyl protecting group).

There may also be several identical or different amino and / or hydroxy protecting groups in the molecule of the starting material. If the molecule contains from each other from 17

Ear protecting groups, in many cases these protecting groups can be selectively removed.

The term amino protecting group is well known and refers to groups which are suitable for protecting (blocking) the amino group from chemical reactions, but which are readily removable when the desired reaction is carried out elsewhere in the molecule. Typically, such groups are unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkyl groups. Since the amino-protecting groups are removed after the desired reaction (or sequence of reactions), their type and size are not critical. However, groups having 1 to 20, and in particular 1 to 8, carbon atoms are preferred. The term acyl is always understood in the broadest sense. It includes acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids, in particular alkoxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonyl groups. Examples of such acyl groups include alkanoyl groups such as acetyl, propionyl, butyryl; aralkanoyl such as phenylacetyl; aroyl such as benzoyl or toluyl; aryloxyalkanoyl such as phenoxyacetyl (POA); alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, tert-butoxycarbonyl (BOC), 2-iodoethoxycarbonyl; aralkoxycarbonyl as a benzyloxycarbonyl (carbobenzoxy-CBZ) group,

A 4-methoxybenzyloxycarbonyl or 9-fluorenylmethoxycarbonyl (FMOC) group and an arylsulfonyl group such as a 4-methoxy-2,3,6-trimethylphenylsulfonyl (Mtr) group. Preferred amino protecting groups are BOC and Mtr as well as CBZ, Fmoc, benzyl and acetyl.

The term hydroxy protecting group is also generally known and refers to groups which are suitable for protecting (blocking) the hydroxy group from chemical reactions but which are readily removable when the desired reaction elsewhere in the molecule is carried out. Typical of such groups are the above-mentioned unsubstituted or substituted aryl, aralkyl or acyl groups as well as alkyl groups. Since the hydroxyl protecting groups are removed after the desired reaction (or reaction sequence), their type and size are not critical. However, groups having 1 to 20 and in particular 1 to 10 carbon atoms are preferred. Examples of hydroxyl protecting groups are benzyl, p-nitrobenzyl, p-toluenesulfonyl, tert-butyl and acetyl, with benzyl and tert-butyl being particularly preferred. The COOH groups are primarily protected in the form of their tert-butyl esters.

Depending on the protecting group used, the liberation of the compounds of the formula I from their functional derivatives is successful, for example with strong acids such as, for example, trifluoroacetic acid and perchloric acid, but also other strong inorganic acids such as hydrochloric or sulfuric acid, strong organic carboxylic acids such as trichloroacetic acid. an acid or sulfonic acids such as benzenesulfonic acid or p-toluenesulfonic acid. It is possible, but not always necessary to carry out the reaction in the presence of an additional inert solvent. Suitable inert solvents are organic, for example, carboxylic acids such as acetic acid, ethers such as tetrahydrofuran (THF) or dioxane, amides such as dimethylformamide (DMF), halogenated hydrocarbons such as dichloromethane, and also alcohols such as methanol , ethanol or isopropanol as well as water. Mixtures of these solvents may also be suitable. Trifluoroacetic acid is preferably used in excess without addition of additional solvents, perchloric acid in the form of a 9: 1 mixture of acetic acid and 70% perchloric acid. The reaction temperature for the cleavage is expediently about 0 to about 50 ° C, preferably 15 to 30 ° C (room temperature).

For example, the BOC, OBut, and Mtr groups may be conveniently cleaved with trifluoroacetic acid in dichloromethane or about 3 to 5N hydrochloric acid in dioxane at 15 to 30 ° C, while the FMOC group is about 5 to 50% dimethylamine, diethylamine or piperidine in dimethylformamide. at 15 to 30 ° C.

The trityl group is used to protect the amino acids of histidine, asparagine, glutamine and cysteine. The deprotection is carried out according to the desired end product with trifluoroacetic acid / 10% thiophenol, whereby the trityl group is cleaved from said amino acids. When using the trifluoroacetic acid / anisole or trifluoroacetic acid / thioanisole system, the trityl group is cleaved from histidine, aparagine and glutamine, but remains on the Cys side chain.

Hydrogenolytically removable protecting groups (e.g. CBZ or benzyl groups) can be removed, for example, by treatment with hydrogen in the presence of a catalyst (e.g., a noble metal catalyst such as palladium, preferably on a support such as coal). Suitable solvents are the abovementioned solvents, in particular, for example, alcohols such as methanol or ethanol or amides such as dimethylformamide. The hydrogenolysis is generally carried out at a temperature of about 0 to 100'C, at a pressure of about 0.1 to 20 MPa, preferably at a temperature of 20 to 30'C, at a pressure of about 0.1 to 1 MPa. Hydrogenolysis of the CBZ group, for example, performs well on 5 to 10% palladium on carbon in methanol, or when using ammonium formate (instead of hydrogen) on palladium on carbon in a methanol / dimethylformamide system at 20-30 ° C.

The base of formula (I) can be converted by acid into the appropriate acid addition salt, for example by reacting an equivalent amount of base and acid in an inert solvent such as ethanol and then evaporating the solvent. Suitable acids for this reaction are, in particular, those which give physiologically acceptable salts. Inorganic acids such as sulfuric, nitric, hydrohalic acids such as hydrochloric or hydrobromic acids, phosphoric acids such as orthophosphoric acid, sulfamic acid and organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic monohydric or non-polybasic carboxylic acids, sulfonic acids may be used. or sulfuric acids such as formic, acetic, propionic, pivalic, diethyl acetic, malonic, succinic, pimelic, fumaric, maleic, lactic, tartaric, malic, citric, gluconic, ascorbic, nicotinic, isonicotinic, ethanesulfonic, methanesulfonic, methanesulfonic

2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalene monosulfonic acid, naphthalenedisulfonic acid and lauryl sulfuric acid. Salts with physiologically unacceptable acids, for example picrates, can be used for the isolation and / or purification of the compounds of formula I.

On the other hand, the compounds of formula I can be converted into their physiologically acceptable metal or ammonium salts by reaction with a base. Suitable salts are, in particular, the sodium, potassium, magnesium, calcium and ammonium salts, further substituted ammonium salts, e.g. .

The compounds of the formula I and their physiologically acceptable salts can be used for the production of pharmaceutical compositions, in particular by a non-chemical route. For this purpose, they can be converted into a suitable dosage form with at least one solid or liquid and / or semi-liquid carrier or excipient and optionally in admixture with one or more other active substances.

The invention therefore also relates to compositions, in particular pharmaceutical compositions, comprising at least one compound of the formula I and / or a physiologically acceptable salt thereof.

The compositions of the invention may be used as medicaments in human and veterinary medicine. Suitable carriers are inorganic or organic substances which are suitable for enteral (e.g. oral) or parenteral or topical administration or for administration in the form of inhalation sprays and which do not react with compounds of the formula I, such as water, vegetable oils, benzyl alcohols , alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starches, magnesium stearate, talc and petrolatum. Especially suitable for oral use are tablets, pills, dragees, capsules, powders, granules, syrups, juices or drops, suppositories for rectal use, solutions for parenteral use, especially oily or aqueous solutions, further suspensions, emulsions or implants, for topical use ointments, creams or powders. The compounds of the invention may also be lyophilized and the resulting lyophilisates used, for example, for the preparation of injectables. The compositions may be sterilized and / or may contain adjuvants such as glidants, preservatives, stabilizing agents and / or wetting agents, emulsifiers, salts for affecting the osmotic pressure, buffers, coloring agents, flavoring agents and / or one or more others. other active ingredients such as vitamins.

For administration in the form of inhalation sprays, the active ingredient is dissolved or suspended in a propellant or propellant mixture (such as, for example, carbon dioxide or fluorocarbon). In this case, the active compounds are used in micronized form, and at least one physiologically compatible solvent, for example ethanol, can be added. Inhalable solutions may be administered using known devices for this purpose.

The compounds of formula I and their physiologically acceptable salts can be used for the treatment of diseases such as, in particular, circulatory diseases, thrombosis, heart attack, coronary heart disease, arteriosclerosis, apoplexia, angina pectoris, cancer, osteoporosis, inflammation, infection and restenosis after angioplasty. .

The compounds of formula I and / or their physiologically acceptable salts are also suitable for use in pathological processes of sustained and spread angiogenesis, in particular tumors, or rheumatoid arthritis.

The compounds of the formula I according to the invention are generally used in doses similar to commercially known peptides, in particular analogously to the compounds of U.S. Pat. No. 4,472,305, preferably at a dose of about 0.05 to 500 mg, in particular 0.5 to 100 mg per dosing. unit. The daily dose is preferably about 0.01 to 2 mg / kg body weight. The dose for each individual patient will depend on a variety of factors, such as the efficacy of the particular compound used, age, body weight, general health, sex, diet, time and route of administration, elimination rate, drug combination and severity of the disease. . Parenteral administration is preferred.

In addition, the compounds of formula I as integrin ligands can be used to produce affinity chromatography columns to produce pure integrin. The ligand, i.e. the compound of the formula I, is coupled to the polymeric carrier by means of an anchoring function, for example the carboxyl group Asp.

Suitable polymeric carriers are those known from the peptide chemistry of known polymeric solid phases, preferably having hydrophilic properties, for example, crosslinked polycarbons, such as cellulose, sepharose or the like.

Sephadex ^R , acrylamides, polymers based on polyethylene glycol or Tentacelpolymer ^R.

The production of affinity chromatography materials for purifying integrin is carried out under conditions which are conventional and known for the condensation of amino acids.

The compounds of formula I may have one or more chiral centers and may therefore be in racemic or optically active form. Alternatively, the racemates obtained can be separated into their enantiomers mechanically or chemically by known methods. Preferably, diastereomers are formed from the racemate by reaction with an optically active resolving agent. Examples of such resolving agents are optically active acids such as the D and L forms of tartaric, diacetyltartaric, dibenzoyltartaric, mandelic, malic or lactic acid or various optically active camphorsulfonic acids, such as β-phosphorsulfonic acid. Also preferred is the resolution of enantiomers by columns packed with optically active resolving agents (e.g. dinitrobenzoylphenylglycine); a suitable eluent is, for example, hexane / isopropanol / acetonitrile, for example in a 82: 15: 3 by volume ratio.

It is of course also possible to obtain the optically active compounds of the formula I by one of the abovementioned processes, starting from optically active substances.

The invention is illustrated by the following non-limiting examples. Temperatures are always given in degrees Celsius, room temperature refers to 22C. In the following examples, the expression conventional processing means:

Optionally, water is added or the pH is adjusted to 2-10 depending on the constitution of the final product, the reaction mixture is extracted with ethyl acetate or dichloromethane, the organic phase is separated and dried over sodium sulfate, filtered by evaporation and purified by silica gel chromatography and / or crystallization.

time (in minutes) for HPLC in the following

Lichrosorb ^R RP 18 (250 x 4; 5 μπι);

A: 0.1% TFA in water

B: 0.1% TFA in 90% acetonitrile,% water mL / min to 95% B / 50 min at 215 nm.

RT is retention systems: column: eluent eluent flow: gradient: detection:

The separation of the diastereomers is preferably carried out under the above conditions.

Mass spectrometry (MS): FAB (fast atom bombardment) (M + H) ⁺

DETAILED DESCRIPTION OF THE INVENTION

Example 1

8S.14S-IB

Preparation of the cyclizing compound

By the method of Scheme 1, (3R, S) -

3-Amino-3- (4-methyl-3-nitrophenyl) propionic acid (3R, S-IX). This ester is cleaved into the enantiomers by known methods and (3S) -3-amino-3- (4-methyl-3-nitrophenyl) propionic acid methyl ester (3S-IX) is reacted subsequently according to Scheme 2 to give the Mtr protected compound, which is 3S-amino-3- (3- {3- [1- (carboxymethylcarbamoyl) -4-guanidino-1S-butylcarbamoyl] propionylamino} -4-methylphenyl] propionic acid methyl ester (S, S-XIV).

cyclization

Dissolve 616 mg of the Mtr-protected compound 3S-amino-3- (3- {3- [1- (carboxymethylcarbamoyl) -4-guanidino-1S-butylcarbamoyl] -propionylamino) -4-methylphenyl] propionic acid methyl ester (S, S-XIV) in 80 ml of dimethylformamide and diluted with 800 ml of dichloromethane. The mixture was cooled to -20 C and ^E are added successively 300 mg of EDC1, 98 mg of DMAP and 0.176 ml of NMM. The mixture was allowed to warm to room temperature and stirred overnight. The solution is concentrated and the residue is taken up in 200 ml of semi-saturated sodium bicarbonate solution. The precipitate is filtered off with suction and washed. 400 mg of product are obtained, which is purified by preparative HPLC. Chromatography gave 44 mg of the Mtr protected cyclic compound (8S, 14S) - [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo-2,7,10,13] methyl ester -tetraazabicyclo [13.3.1] nonadeca-1 (18), 15 (19), 16-trien-14-yl] acetic acid. RT 26.1; FAB 716.

Saponification and cleavage of the protecting group Mtr

Saponification in potassium hydroxide / methanol provides the Mtr-protected cyclic compound which is (8S, 14S) [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo-2,7,10] 13 tetraazabicyclo [13.3.1] nonadeca-1 (18), 15 (19), 16-trien-14-yl] 26 acetic acid. Dissolve 25 mg of this compound in 4.3 mL of 98% trifluoroacetic acid and stir overnight at room temperature. The solution was centrifuged and purified by preparative HPLC. 9.5 mg of (8S, 14S) - [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo-2,7,10,13-tetraazabicyclo [13.3.1] are obtained. nonadeca-1 (18), 15 (19), 16-trien-14-yl] acetic acid (8S, 14S-IB). RT 20.2;

FAB 490.

Examples 2 to 3

Example 2 (q = 2):

Of the 85 mg Mtr-protected compound which is (8S, 14S) -2- (8- (3-guanidinopropyl) -3,6,9,12-tetraoxo-2,7,10,13-tetraazabicyclo [13.3.1] nonadeca) (16,18,19-trien-14-yl) acetic acid is obtained analogously to Example 1 by reaction with

14.7 ml 98% trifluoroacetic acid 13 mg (8S, 14S) -2- (8- (3guanidinopropyl) -3,6,9,12-tetraoxo-2,7,10,13-tetraazabicyclo [13.3.1] nonadeca-16,18,19-trien-14-yl) acetic acid.

RT 17.2; FAB 476.

Example 3 (q = 3):

Of the 300 mg Mtr protected compound which is (9S, 15S) -2- (9- (3-guanidinopropyl) -3,7,10,13-tetraoxo-2,8,1114-tetraazabicyclo [14.3.1] eicosan -17,19,20-trien-15-yl) acetic acid similar to Example 1 yielded 74 mg of (9S15S) -2- (9- (3-guanidinopropyl) -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

Of the Mtr protected compound which is (6S, 12S) [6- (3-guanidinopropyl) -4,7,10-trioxo-2,5,8,11-tetraazabicyclo [11.3.1] heptadeca-1 (17 (13,15-trien-12-yl) -acetic acid analogously to Example 1 gives (6S, 12S) - (6- (3-guanidinopropyl) -4,7,10-trioxo-2,5,8), 11-tetraazabicyclo [11.3.1] heptadeca-1 (17), 13,15-trien-12-yl) acetic acid.

Examples 5 to 8

In analogy to Example 1, the following compounds are obtained:

Pr. no. X C m n R ¹ and R ² 5 H -CO-CH = CH-CO- 0 3 H 6 Cl -co-ch ₂ -ch ₂ -co- 0 3 H 7 ch ₃ -co-ch ₂ -ch ₂ -co- 1 2 8 H -co-ch ₂ -ch ₂ -co- 1 2 C ^H M II Ή

The following examples illustrate but do not limit the pharmaceutical compositions of the invention:

Example A

Injection ampoules

A solution of 100 g of an active compound of the formula I and 5 g of disodium hydrogen phosphate in 3 l of double-distilled water is adjusted to pH 6.5 with 2N hydrochloric acid, sterile filtered, filled into injection vials, lyophilized under sterile conditions and sealed. Each ampoule contains 5 mg of active ingredient.

Example B

suppository

A mixture of 20 g of an active compound of the formula I with 100 g of soya lecithin and 1400 g of cocoa butter is melted, poured into molds and allowed to solidify. Each suppository contains 20 mg of active ingredient.

- £

Example C Solution Solution

A solution of 1 g of an active compound of the formula I, 9.38 g of sodium dihydrogen phosphate dihydrate, 28.48 g of dihydrogen phosphate with 12 water molecules and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water is prepared. The pH of the solution was adjusted to 6.8, made up to 1 liter and sterilized by irradiation. This solution can be used, for example, as eye drops.

Example D

ointment

500 mg of an active compound of the formula I are mixed with 99.5 g of petroleum jelly under aseptic conditions.

Example E

tablets

A mixture of 1 kg of active ingredient 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 is compressed in known manner into tablets, each tablet containing 10 mg of the active compound of the formula I .

Example F

dragee

Analogously to Example E, tablets are compressed and coated in a known manner with a coating of sucrose, corn starch, talc, tragacanth and colorant.

Example G

The capsules

2 kg of active ingredient of the formula I are filled into hard gelatin capsules, each capsule containing 20 mg of the active ingredient of the formula I.

Example H

ampoules

A solution of 1 kg of an active compound of the formula I in 60 liters of double-distilled water is sterile filtered, filled into ampoules, lyophilized under sterile conditions and sealed. Each ampoule contains 10 mg of active ingredient.

Example I

Inhalation spray

*> » Dissolve 14 g of an active compound of the formula I in 10 l of isotonic sodium chloride solution and fill in conventional commercial spray cans with a pump mechanism. The solution may be sprayed into the mouth or nose. Each injection (about 0.1 ml) corresponds to a dose of about 0.14 mg. T Industrial usability Cyclopeptide derivative as an integrin inhibitor suitable for the manufacture of pharmaceutical compositions, in particular for the prophylaxis and treatment of diseases such as, in particular, circulatory diseases, thrombosis, heart attack, coronary heart disease, arteriosclerosis, apoplexia, angina pectoris, cancer, osteoporosis, inflammation, infections and restenosis after angioplasty.

j

Claims (10)

PATENT CLAIMS Compounds of general formula I (I) wherein is Gly, Ala or NH-NH-CO, wherein said amino acids are optionally in the form of their derivatives, a group of formula II (II) C - (CO) _p - (CH ₂ ) _q - (CO) _r - or - (CO) _p -CH = CH- (CO) _r -, m, p, r independently of one another 0 or 1, n, q independently of 1, 2, 3 or 4, R ¹ and R ² each independently H or alkyl, R ¹ and R ² together together also form a group \ C II c / or R ⁷ , R ⁸ , R ⁹ and R ¹⁰ independently of one another are hydrogen, alkyl, Ar, OR ⁶ , Hal, NO 2, NR ⁶ R ⁶ ', NHCOR ⁶ , CN, NHSO ² R ⁶ , COOR ⁶ or COR ⁶ , X is hydrogen, Hal, alkyl or Ar, Ar is a phenyl group optionally substituted with one, two or three R ³ , R ⁴ or R ⁵ groups or an unsubstituted naphthyl group, R ³ , R ⁴ , R ⁵ independently of one another R ⁶ , OR ⁶ , Hal, NO 2, NR ⁶ R ⁶ ', NHCOR ⁶ , CN, NHSO ² R ⁶ , COOR ⁶ or COR ⁶ , R ⁶ , R ⁶ 'independently of one another are hydrogen, alkyl, phenyl or benzyl; and Hal is a fluorine, chlorine, bromine or iodine atom, with respect to the residues of optically active amino acids and amino acid derivatives, both the D and L forms and their salts are included. An enantiomer or diastereomer of a compound according to claim 1 having the general formula (I). A compound according to claim 1 selected from the group consisting of a) (8S, 14S) -2- (8- (3-guanidinopropyl) -3,6,9,12-tetraoxo-2,7,1013-tetraazabicyclo [13.3.1] nonadeca-16,18,19-triene -14-yl) acetic acid, b) (9S, 15S) -2- (9- (3-guanidinopropyl) -3,7,10,13-tetraoxo-2,8-) 11,14-tetraazabicyclo [14.3.1] eicosan-17,19,20-trien-15-yl) acetic acid, (8S, 14S) - (8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo-2,7,10,13-tetraazabicyclo [13.3.1] nonadeca-1 (18)), 15 (19), 16-trien-14-yl) acetic acid, (d) (6S, 12S) - (6- (3-guanidinopropyl) -4,7,10-trioxo-2,5,8,11-tetraazabicyclo [11.3.1] heptadeca-1 (17), 13,15- trien-12-yl) acetic acid and salts thereof. A process for the preparation of a compound of formula I, wherein the individual symbols are as defined in claim 1 or salts thereof according to claim 1, characterized in that a) the compound of formula III H-Z-OH (III) wherein is Z group of formula NH — C-B-A — NH X OH - B HN-A- X cl 3%; C N? AJ H r hn-a-b— or OH h-a-b-c- and X, A, B and C are as defined above, or a reactive derivative of a compound of formula III is treated with a cyclizing agent, or b) releasing a compound of formula I from its functional derivative by treatment with a solvolysis or hydrogenolysis agent and / or converting a basic compound or acidic compound of formula I by treatment with an acid or base into its salt. A process for the preparation of a pharmaceutical composition, characterized in that the compound according to claim 1 and / or its physiologically acceptable salt is formulated in a dosage form with at least one solid, liquid or semi-solid carrier or excipient. Pharmaceutical composition, characterized in that it contains as active substance at least one compound of the formula I according to claim 1 and / or a physiologically acceptable salt thereof. Compound according to claim 1 and its physiologically acceptable salts as integrin inhibitors for the treatment of diseases such as, in particular, circulatory diseases, thrombosis, heart attack, coronary heart disease, arteriosclerosis, apoplexy, angina pectoris, cancer, osteoporosis, inflammations, infection and restenosis after angioplasty. Use of the compounds according to claim 1 of formula I and / or their physiologically acceptable salts in pathological processes which are ongoing and spread by angiogenesis. Use of the compounds according to claim 1 and / or their physiologically acceptable salts for the manufacture of a medicament. Use of the compounds according to claim 1 of the general formula I and their physiologically acceptable salts for combating diseases.