MXPA99005778A - Cyclic peptide derivatives - Google Patents

Abstract

Compounds of the formula (I) Cyclo-(Arg-X-Asp-R1) in which X means Gly, Ala or NH-NH-CO, R1 means a radical of formula (II), and R2, R3, and R4 have the meanings given in claim 1, and their salts, can be used as integrin inhibitors, in particular for the prevention and treatment of circulatory diseases, in cases of thrombosis, cardiac infarction, coronary heart disease, arteriosclerosis, pathological symptoms that are sustained or propagated by angiogenesis, and in tumour therapy.

Description

DERIVATIVES OF CYCLIC PEPTIDES Field of Invention The invention relates to the compounds of formula I: Cyclo- (Arg-X-Asp-R1) (I), where X represents Gli, Ala or NH-NH-CO, wherein said amino acids can also be derivatized and the amino acid residues are linked together by a peptide-type binding through the a-amino and carboxyl groups, RJ is a remainder of formula II Ref .: 30579 Ref: 030579 R, R- R 'independently represent H, A, Ar, R -Ar, Het R ° -Het A represents alkyl of 1 to 6 C atoms, Ar represents phenyl unsubstituted or mono-, di- or trisstituted with R7, R8 or R9, or unsubstituted naphthyl, R- represents alkylene of 1 to 6 C atoms, R6, R6 'independently represent H, A, benzyl or phenyl, R7, R8, R9 independently represent R6, OR6, Hal, N02, NR6R6 ', NHCOR6, CN, NHS02R6, COOR6 or COR6, Hal represents F, Cl, Br or I Het represents a heterocycle of one or two nuclei containing 1 to 4 N, O and / or S atoms and which may be unsubstituted or mono, di or trisubstituted with Hal, A, NR6R6, CN or N02, including, in the case of amino acid residues and optically active amino acid derivatives, both D and L forms, and its salts Background of the Invention German cyclic peptides are described in German Patent No. 4,310,643 or in European Patent No. 0 683 173.

The aim of the invention was to develop new compounds with valuable properties. In particular compounds that can be used in the manufacture of medicines.

Description of the invention It was found that the compounds of formula I and their salts possess valuable pharmacological properties and are well tolerated. First of all, they act as inhibitors of integrin, in particular by inhibiting the interaction of integrin, av, ß3, or ßs receptors with ligands, such as, for example, the binding of fibrinogen to the β3 integrin receptor. These compounds have a particular effect in the case of integrins avß ?, avß3, 0C? I ß3 and also avß6 and ocvßs • This effect can be checked, for example, according to the method described by JW Smith and collaborated in J. Biol Chem. 265, pages. 12267 to 12271 (1990).

P.C. Brooks, R. A. Clark and D. A. Cheresh describe in Science 264, pages. 569-71 (1994), the dependence of the beginning of the angiogenesis of the interaction between the vascular integrins and the extracellular matrix proteins.

The possibility of inhibiting this exchange and thus introducing apotosis (programmed cell death) of angiogenic vascular cells by a cyclic peptide is described in Cell 79, pages 1157-64 (1994), by PC Brooks, AM Montgomery, M. Rosenfeld, RA Reisfeld, T. Hu, G. Klier and DA Cheresh.

The compounds of formula I that block the interaction of integrin receptors with ligands such as, for example, the binding of fibrinogen to the fibrinogen receptor (glycoprotein to Ilb / IIIa), act as GPIIb / IIIA antagonists and they prevent the proliferation of tumor cells by metastasis. This concept is supported by the following observations: The spread of tumor cells from a local tumor to the vascular system occurs through the formation of microaggregates (microtomes) by interaction of tumor cells with platelets. The tumor cells are protected and camouflaged in the microaggregate and are not recognized by the cells of the immune system.

The microaggregates can be fixed to the walls of the vessels, which facilitates the internalization of the tumor cells in the tissue. Since the formation of microthrombi is facilitated by the binding of fibrinogen to fibrinogen receptors on activated platelets, GPIIa / IIIb antagonists can be considered active inhibitors of metastasis.

The compounds of formula I can be used in medicine and veterinary medicine as active substances of drugs, in particular for the prophylaxis and / or therapy of thrombosis, myocardial infarction, arteriosclerosis, inflammations, stroke, angina pectoris, tumor diseases, osteolytic diseases such as osteoporosis, pathological angiogenic diseases such such as inflammations, ophthalmological diseases, diabetic retinopathy, macular degeneration, myopia, ocular histoplasmosis, rheumatic arthritis, osteoarthritis, rubeotic glaucoma, ulcerative colitis, Morbus Crohn, atherosclerosis, psoriasis, restenosis after angioplasty, viral infection, bacterial infection, fungal infection, acute renal insufficiency and in wound healing to help healing processes.

The compounds of formula I can be used as substances with an antimicrobial effect in operations in which biological materials, implants, a catheter or a pacemaker are used. In these cases they act as antiseptic substances. The effect of the antimicrobial activity can be checked according to the procedure described by P. Valentin-Weigund et al in "Infection and Im unity, pages 2851 to 2855 (1988).

Since the compounds of formula I inhibit the binding of fibrinogen and, therefore, constitute ligands of the fibrinogen receptors located on the platelets, these can be used in vivo as diagnostic agents for the arrest and localization of thrombi in the system. vascular, as long as they are replaced, for example, with a radioactive residue or detectable with ÜV radiation.

The compounds of formula I, as inhibitors of fibrinogen binding, can also be used as effective auxiliary agents for the study of the metabolism of platelets in their different activation states or of the receptor's intracellular signaling mechanisms. of fibrinogen. The detectable unit of a marker (or tracer) introduced, for example, isotopic labeling with 3H, allows to study the mentioned mechanisms after producing the binding to the receptor.

The abbreviations of amino acid residues that are indicated in this text correspond to the remains of the following amino acids: Wing Alanine AMP - Rest of aminomet ilphenyl Asn Asparagine Asp Aspartic acid Arg Arginine Cis Cysteine Gln Glutamine Glu Glutamic acid Glycyl Glycine His Histidine Homo- Phe Homo- phenylalanine Ile Isoleucine Leu Leucine Lis Lysine Met Methionine Nle Norleucine Orn Ornithine Phe Phenylalanine Phg Phenylglycine 4 -Hal -Phe 4 -halogen-phenylalanine Pro Proline Ser - Serine Thr Threonine Trp Triptophan Tir Tyrosine Val Val ina.

The rest 3-AMP has the following structure In addition, the remains indicated below mean the following: Ac Acetyl BOC Terbutoxycarbonyl CBZ or Z Benzyloxycarbonyl DCC1 Dicyclohexylcarbodiimide DMF Dimethylformamide EDC1 N-ethyl-N, N'- (dimethylaminopropyl) carbodiimide Et Ethyl FCA Acid fluorscarboxylic Fmoc 9-fluorenylmethoxycarbonyl HOBt 1-hydroxybenzotriazole Me Methyl MBHA-methyl-benzhydrylamine Mtr 4 -methoxy -2, 3, 6-trimethylphenylsulfonyl HONSu N-hydroxysuccinimide Obzl Benzyl ester OtBu Terbutyl ester Oct Octanoyl OMe Methyl ester Oet Ethyl ester POA Phenoxyacetyl Salicyloyl salt TFA Trifluoroacetic acid Trt Trityl (triphenylmethyl) If the amino acids indicated above have several enantiomeric forms, then the present text also covers - for example, as a component of the compounds of formula I - all these forms and their mixtures (for example, the DL forms). In addition, the amino acids can be protected - for example as components of the compounds of formula I with known protecting groups.

Compounds of the invention also include so-called "prodrugs", that is, compounds of formula I that have been derivatized with, for example, alkyl or acyl groups, sugars or oligopeptides, which are rapidly cleaved in the body thus releasing the active compounds of the invention.

Also included in this group are the biodegradable polymers of the compounds of the invention, as described, for example, in Int. J. Pharm. 115, pages 61 to 67 (1995).

Amino acids whose configuration is not explicitly indicated have a (S) or (L) configuration.

Another object of the invention is a process for preparing the compounds of formula I, according to claim 1, and their salts, characterized in that a) a compound of formula III is treated H-Z-OH (III) where represents -Arg-X-Asp-R1- -X-Asp-Rx-Arg- -Asp-R1-Arg-X or -R1-Arg-X-Asp-, and X and R1 have the meanings indicated in claim 1, or a reactive derivative of a compound of formula II with a cyclization agent, b) a compound of formula I is released from one of its functional derivatives by treatment with a solvotic or hydrogenolysis agent, and / or a base or acid of formula I is transformed into one of its salts by treatment with an acid or a base.

The X, R1, R2, R3 and R4 residues mentioned in this text have the meanings indicated for formulas I, II and III, unless otherwise indicated.

In the formulas indicated above, alkyl preferably represents methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tertbutyl, then also pentyl, 1-, 2- or 3-methylbutyl, 1,1-, 1,2- or 2, 2-dimethylpropyl, 1-etylpropyl, hexyl, 1-, 2-, 3- or -methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2, 3-, or 3, 3-dimethyl butyl, 1- or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or 1, 2, 2 -trimethylpropyl.

R2 and R3 represent, independently of one another, preferably, for example, H or A, then also Ar or R5-Ar.

R4 preferably represents, for example, H, A, Ar or R5-Ar, then also Het or R5-Het.

When R represents alkyl, then a methyl group of said alkyl chain may also be replaced by N, O or S.

Alkylene preferably represents methylene, ethylene, propylene, butylene, pentylene or hexylene.

R5-Ar preferably represents benzyl or phenethyl.

The amino acids and amino acid residues indicated may also be derivatized, with the N-methyl, N-ethyl, N-propyl, N-benzyl or Ca-methyl derivatives being preferred. Also preferred are Asp and Glu derivatives, in particular the methyl, ethyl, propyl, butyl, tertbutyl, neopentyl or benzyl esters of the carboxyl groups of the side chains, then also the derivatives of Arg, whose NH- group C (= NH) -NH2- may be substituted with an acetyl, benzoyl, methoxycarbonyl moiety.

R preferably represents, for example, H, methyl or ethyl, then benzyl or phenyl.

OR * preferably represents, for example, hydroxy or methoxy.

COR6 represents alkanoyl, with formyl, acetyl, propionyl, butyryl, pentanoyl or hexanoyl being preferred.

Ar represents unsubstituted phenyl, preferably - as indicated above - monosubstituted, in particular preferably phenyl, o-, m- or p-tolyl, o-, m- or p-ethylphenyl, or-, m- or m- or p-propylphenyl, o-, m- or p-isopropyl-phenyl, o-, m- or p-tert-butylphenyl, o-, m- or p-trifluoromethyl-phenyl, o-, m- or p-hydroxy phenyl, or -, m- or p-nor trofenil, o-, m- or p-aminophenyl, o-, m- or p- (N-methylamino) -phenyl, o-, m- or p-acetamidophenyl, or-, m- or p- (trifluoromethoxy) -phenyl, o- - or p-methoxyphenyl, o- m- or p-et oxy phenyl, o-, m- or p-carboxyphenyl, o-, m- or p-methoxycarbonylphenyl, o-, - or p-ethoxycarbonyl phenyl, o-, m- or p-benzyloxycarbonylphenyl, o-, m- or p- (carboxymethyloxy) -phenyl, o-, m- or p- (methoxycarbonyl-ethyloxy) -phenyl, or -, m- or p- (methoxycarbonyl-etyloxy) -phenyl, o-, m- or p- (N, N-dimethylamino) -phenol, o-, m- or p- (N-et i lamino ) -phenyl, o-, m- or p- (N, N-di-ethylamino) -phenyl, o-, m- or p-flurophenyl, or-, m- or p-bromophenyl, o-, m- or p-chlorophenyl, o-, m- or p- (di fluoromethoxy) -phenyl, o-, m- or p- (fluoromethoxy) -phenyl, or-, - or p-formylphenyl, o-, m- or p-acetylphenyl, o-, m- or p-propionyl phenyl, o-, m- or p-butyryl phenyl, o-, m- or p-pentanoyl phenyl, or-, m- or p- (phenyl-sulfonamidocarbonyl) -phenyl, o-, m- or p-phenoxyphenyl, o-, - or p-methyl thiophenyl, o-, m- or p-methylsulfinylphenyl, o-, m- or p -meti lsul fonilphenyl or naphthyl.

Het preferably represents 2- or 3-furyl, 2- or 3-thienyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, 4- or 5-imidazolyl, 1-, 3-, 4- or 5 -pyrazoloi, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5-isothiazolyl, 2-, 3- or 4 -pyridyl, 2-, 4-, 5- or 6-pyrimidinyl, then preferably 1, 2, 3-triazol-l-, -4- or -5-yl, 1, 2, -t-riazol-1 -, -3- or -5-yl, 1- or 5-tetrazolyl, 1, 2, 3 -oxa zol-4 - or -5-yl, 1,2,4-oxadiazol-3- or 5-yl, 1, 3, A -thiadiazol-2 - or -5-yl, 1, 2,4-diazodiazol-3- or -5-yl, 1,2,3-thiadiazol-4-y-5-yl, 2-, 3-, 4-, 5- or 6-2H-thiopyranyl, 2- 3- or -4H-thiopyranyl, 3- or 4-pyridazinyl, pyrazinyl, 2-, 3 -, A-, 5-, 6- or 7-benzofuryl, 2-, 3-, 4-, 5-, 6 - or 7-benz otienyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 1-, 2-, 4- or 5-benzimidazolyl, 1-, 3-, 4- , 5-, 6- or 7-benzopyrazolyl, 2-, 4-, 6- or 7-benzoxazolyl, 3-, 4-, 5-, 6- or 7-benzothiazolyl, 2-, 4-, 5- , 6- or 7-benzisothiazolyl, 4-, 5-, 6- or 7-benzo-2, 1,3-oxodiazolyl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl , 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 3-, 4-, 5-, 6-, 7- or 8-ci cl olini lo, 2-, 4- , 5-, 6-, 7- or 8 -quinazolinyl. The heterocyclic moieties may also be partially or totally hydrogenated. Thus Het can also represent, for example, 2, 3-dihydro-2 -, 3-, -4- or 5-furyl, 2,5-dihydro-2 -3-, -4- or -5-furyl, tetrahydro-2 - or -3-furyl, 1,3-dioxolanyl-yl, tetrahydro-2- or -3-thienyl, 2,3-dihydro-l-, -2-, -3-, -4- or -5-pyrrolyl, 2, 5-dihydro-1 -, -2-, -3-, -4- or -5-pyrrolyl, 1-, 2- or 3-pyrrolidinyl, tertrahydro-1-, - 2- or -4-imidazolyl, 2,3-dihydro-1-, -2- -3-, -4- or -5 -pira zolyl, t -hydro-1 -, -3- or -4-pyrazolyl, 1 , 4-dihydro-1-, -2-, -3- or -4-pyridyl, 1, 2, 3, 4 -1etrahydro-1-, -2-, -3-, -4-, -5- or -6-pyridyl, 1-, 2-, 3- or -piperidinyl, 2-, 3- or 4 -mor folinyl or, tert-rahydro-2 -, -3- or -4-pyranyl, 1,4-dioxanyl , 1,3-dioxan-2 -, 4- or -5-yl, hexahydro-1--3- or -4-pyridazinyl, hexahydro-1-, -2-, -4- or -5-pi rimidinyl, 1-, 2- or 3-piperazinyl, 1, 2, 3, 4-tet rahydro-1, -2-, -3-, -4-, -5-, -6-, -7- or 8 -quinolyl , 1, 2, 3,, tetrahydro-1, -2-, -3-, -4-, -5-, -6-, -7-u -8 -i soquinoleil.

The term "amino protecting group" preferably represents acetyl, propionyl, butyryl, phenylacetyl, benzoyl, toluyl, POA, methoxycarbonyl, ethoxycarbonyl, 2,2,2-tri chloroethoxycarbonyl, BOC, 2-iodoethoxycarbonyl, CBZ (carbobenzoxy), -methoxobenzyloxycarbonyl, FMOC, Mtr or benzyl.

The compounds of formula I can have a plurality of chiral centers, which is why they can have different terrestrial forms. Formula I covers all these forms.

Therefore, a particular object of the invention are those compounds of formula I in which at least one of the mentioned moieties has one of the preferred meanings indicated above. Some preferred groups of compounds can be represented by the partial formulas a to Ih, which correspond to formula I and in which the radicals which are not explicitly listed have the meanings indicated for formula I, namely: in R 'R- independently represent H or A, R- represents H, A, Ar, Het or R5-Het, and R * R6' represent H or A; in b) R 'R; independently represent H or A, R * represents H, A, Ar, R5-Ar, Het or R5-Het, R < R 'represent H or A, and Ar represents phenyl unsubstituted or monosubstituted with R7; in c) R2, R3 independently represent H or A, R "represents H, A, Ar, R5-Ar, Het or R5-Het, R6, R6 'represent H or A, Ar represents phenyl unsubstituted or monosubstituted with R7 , and Het represents a single-core aromatic or saturated heterocycle, containing 1 or 2 N or O atoms and which may be unsubstituted or mono- or di-substituted with Hal, A, NR6R6 ', CN or N02; in d) R 'R' independently represent H or A, R * represents H, A, Ar or R5-Ar, represent H or A, and Ar represents phenyl unsubstituted or monosubstituted with R7; in e X represents Gli or Ala, R 'independently represents H or A, represents H, A, Ar or R5-Ar, R < R 'represent H or A, and Ar represents phenyl unsubstituted or monosubstituted with R7.

In general, both the compounds of formula I and the starting materials for their preparation are prepared according to known methods, as described in the literature (for example, in certain works such as Houben Weyl's, "Methoden der organischen Chemie" (Method of Organic Chemistry), Georg-Thieme-Verlag, Stuttgart) and under reaction conditions that are known and suitable for the reactions mentioned. You can also make use of known variants of these methods that are not detailed in this text.

If desired, the starting materials can be prepared in situ, but in such a way that instead of isolating them from the reaction mixture they are directly reacted to form the compounds of formula I.

The compounds of formula I can be obtained preferably by cyclization of compounds of formula III under the conditions of a peptide synthesis. For this it is convenient to work according to the usual methods of peptide synthesis, such as, for example, those described in Houben-Weyl, l.c, Vol. 15/11, pages 1 to 806 (1974).

The reaction is conveniently carried out in the presence of a dehydrating agent, for example, of a carbodiimide such as DCC1 or EDC1, also, for example, of propanophosphonic anhydride (see Angew.Chem.92, page 129 (1980)). , dif enylphosphorylazide or 2-ethoxy-N-ethoxycarbonyl-1-1,2-dihydroquinoline, in an inert solvent such as, for example, a halogenated hydrocarbon such as dichloromethane, an ether such as tetrahydrofuran or dioxane, an amide such as DMF or dimethylacetamide, a nitrile such as acetonitrile, in di-ethyl-sulfoxide or in the presence of mixtures of these solvents, at temperatures between about -10 and 40, preferably between 0 ° and 30 ° C. To favor intramolecular cyclization against intermolecular peptide binding it is convenient work with diluted solutions.

The reaction times vary, depending on the conditions used, between a few minutes and 14 days.

Instead of the compounds of formula III, it is also possible to use the derivatives of the compounds of formula III, preferably a carboxylic acid which has been previously activated, or an acid halide, a symmetrical or mixed anhydride or an activated ether. This type of residues, which are used to activate the carboxyl group in typical acylation reactions, are described in the literature (for example, in works such as the one by Houben-Weyl, "Methoden der organischen Chemie" (Methods of Organic Chemistry), Georg-Thieme -Verlag, Stuttgart).

Activated esters are preferably prepared in situ, for example by adding HOBt or N-hydroxysuccinimide.

Generally, the reaction is carried out in an inert solvent, and when a carboxylic acid halide is employed, it is worked in the presence of an acid trapping agent, preferably an organic base such as triethylamine, dimethylaniline, pyridine or quinoline The addition of an alkali metal or alkaline earth metal hydroxide, carbonate or bicarbonate or another alkaline or alkaline earth metal salt of a weak acid, preferably a salt of potassium, sodium, calcium or cesium can also be favorable.

The starting compounds of formula III are usually novel and can be prepared according to known methods of the synthesis of peptides.

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

As starting substances for solvolysis or hydrogenolysis, those substances are preferred which instead of one or more free amino groups and / or free hydroxyl groups contain correspondingly protected amino and / or hydroxyl groups, preferably those which in Instead of an H atom attached to an N atom, they carry an amino protecting group, for example those that respond to formula I, but instead of a group of NH2 carry a group of NHR '(where R' is a group amino protector, such as BOC or CBZ).

Preference is also given to starting substances which have a hydroxyl-protecting group instead of the H atom of a hydroxyl group, for example those which correspond to formula I, but instead of a hydroxyphenyl group carry a group of hydroxyl groups. R "O-phenyl (where R" is a hydroxyl protecting group).

There can also be several groups, the same or different, of protected amino and / or hydroxyl in the starting material molecule. If the protective groups present are different from each other, then they can be excised in many cases selectively.

The term "amino protecting group" is known and refers to the groups that are adapted to protect (block) an amino group from chemical reactions, but which can easily be cleaved after the desired chemical reaction has been carried out in another place in the molecule. Typical examples of these groups are, in particular, the unsubstituted or substituted groups of acyl, aryl, aralkoxymethyl or aralkyl. Since the amino protecting groups are cleaved after the desired reaction (or the sequence of reactions) has elapsed, the type and size thereof is not a critical point; however, those groups from 1 to 20, in particular from 1 to 8, C atoms are preferred. In the context of the process of the present invention, the expression "acyl group" has a very broad interpretation. It encompasses acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids such as, in particular, the alkoxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonyl groups. Examples of acyl groups of this type are alkanoyl groups such as acetyl, propionyl and butyryl; aralkanoyl such as phenylacetyl; aroyl such as benzoyl or toluyl; aryl oxal canoil such as POA; alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOC, 2-iodoethoxycarbonyl; to which loxycarboni such as CBZ ("carbobenzoxy"), 4-methoxybenzyloxycarbonyl, FMOC; arylsulfonyl such as Mtr. Preferred amino protecting groups are BOC and Mtr, then CBZ, Fmoc, benzyl and acetyl.

The term "hydroxyl protecting group" is also known and refers to groups that are adapted to protect a hydroxyl group from chemical reactions, but which are easily cleaved once the desired chemical reaction that has been carried out has been completed. in a different place on the molecule. Typical examples of these groups are the substituted or unsubstituted groups of aryl, aralkyl or acyl mentioned above, and also the alkyl groups. The nature and size of the hydroxyl protecting groups do not constitute a critical point, as these are re-cleaved once the desired chemical reaction (or sequence of chemical reactions) has been completed; however, groups of 1 to 20, in particular of 1 to 10, atoms are preferred. Examples of hydroxyl protecting groups are, inter alia, benzyl, p-nor t-robenzoyl, p-toluenesulfonyl, tertbutyl and acetyl, benzyl and tertbutyl being particularly preferred groups. The COOH groups of the aspartic and glutamic acids are preferably protected as terbutyl esters (for example, Asp (OBut)).

The release of the compounds of formula I from their functional derivatives is carried out - depending on the protective group used - for example, with strong acids, conveniently with TFA or perchloric acid, but also with other strong inorganic acids such as the acid hydrochloric or sulfuric, with strong organic carboxylic acids such as trichloroacetic acid, or with sulfonic acids such as benzenesulonic or p-toluenesulfonic. It is possible to carry out the reaction in an additional inert solvent, but the presence thereof is not indispensable in all cases.

As inert solvents, organic solvents such as, for example, carboxylic acids such as acetic acid, ethers such as tetrahydrofuran or dioxane, amides such as DMF, halogenated hydrocarbons such as methanol, can be used. Ethanol or isopropanol, and water.

Mixtures of these solvents can also be used. The TFA is preferably used in excess and without the additional addition of other solvents; the perchloric acid is used in the form of a mixture composed of acetic acid and 70% perchloric acid in a ratio of 9: 1. The reaction temperatures for the cleavage are conveniently between about 0 and 50 ° C, preferably between 15 and 30 ° C (room temperature).

The groups BOC, OBut and Mtr can be cleaved preferably with, for example, TFA in dichloromethane or with about 3N HCl to 5N in dioxane, at a temperature comprised between 15 and 30 ° C; The FMOC group can be cleaved using a solution of 5.50% dimethylamine, diethylamine or piperidine in DMF, at a temperature between 15 and 30 ° C.

The trityl group is used to protect the amino acids histidine, asparagine, glutamine and cysteine. The cleavage of said group is carried out, according to the desired final product, with 10% TFA / thiophenol, thus cleaving the trityl group of all the mentioned amino acids, while with TFA / anisole or FA / thioanisole only the trityl group of His, Asn and Gln, the one of the lateral chain of the Cis remaining intact.

Protective groups which are separated by hydrogenolysis (for example, CBZ or benzyl) can be cleaved, for example, by treatment with hydrogen in the presence of a catalyst (for example, a noble metal catalyst such as palladium, conveniently on a support like coal). Suitable solvents are the same as mentioned above, in particular alcohols such as methanol or ethanol, or amides such as DMF. In general, the hydrogenolysis is carried out at temperatures between 0 and 100 ° C and pressures between 1 and 200 bar, preferably at temperatures between 20 and 30 ° C and pressures between 1 and 10 bar. The hydrogenolysis of the CBZ group proceeds favorably with 5-10% of pd / C in methanol or with ammonium formate (instead of hydrogen) on pd / C in methanol / DMF, temperatures between 20 and 30 ° C.

A base of formula I can be converted into its salt by the addition of an acid, for example, by reaction of equivalent amounts of the base and the acid in an inert solvent such as ethanol and then by evaporation of the latter. For this reaction, acids that form physiologically acceptable salts are particularly suitable. Therefore, inorganic acids such as, for example, sulfuric acid, nitric acid, hydrocides such as hydrochloric or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid, then also organic acids can be used, in particular the aliphatic, alicyclic, araliphatic, aromatic or heterocyclic mono- or polybasic carboxylic, sulfonic- or sulfuric acids such as, for example, formic, acetic, propionic, pivalic, diethylactic, malonic, succinic, pimelic, fumaric, maleic, lactic, tartaric, malic, citric, gluconic, ascorbic, nicotinic, isonic, methanesulfonic or ethanesulfonic, ethanedi sulonic, 2-hydroxyethanesulonic, benzenesulonic, p-toluensosulonic, naphthalenemonosulfonic, naphthalenedisulfonic and lauryl sulfuric. Salts of physiologically unacceptable acids, for example picrates, can be used to isolate and / or purify the compounds of formula I.

An acid of formula I can also be converted into one of its physiologically acceptable metal or ammonium salts by treatment with a base. Among the salts, sodium, potassium, magnesium, calcium and ammonium salts are particularly preferred, and also substituted ammonium salts, for example dimethylammonium, diethylammonium or diisopropylammonium salts, monoethanolamine or cyclohexylammonium salts. , dicyclohexyl ammonium, dibenzylethylenediamonium, and also, for example, salts formed with arginine or lysine.

The invention also relates to the use of the compounds of formula I and / or their physiologically acceptable salts for preparing pharmaceutical preparations, in particular by a non-chemical route. For these purposes, the compounds can be brought into a suitable dosage form, together with at least one solid, liquid and / or semi-liquid auxiliary excipient or auxiliary product and, optionally, in combination with one or more additional active substances.

The invention also relates to pharmaceutical preparations containing at least one compound of formula I and / or one of its physiologically acceptable salts.

These preparations can be used in medicine and veterinary medicine as a medicine. Among the excipients there may be mentioned organic or inorganic substances which are suitable for enteral (for example, oral), parenteral or topical application or for application in the form of inhalant solution in aero s_p1, and which do not react with the new compounds. Examples of these excipients are water, vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glycerin triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc and petrolatum. For oral administration, in particular, tablets, pills, dragees, capsules, powders, granules, syrups, juices or drops are used for the rectal application of suppositories, parenteral administration of solutions, preferably oily solutions or aqueous, and also suspensions, emulsions or implants, and for topical application ointments, creams or powders. The new compounds can also be lyophilized and the resulting lyophilized products can be used, for example, for the preparation of injectable preparations. The preparations mentioned can be sterilized and / or contain auxiliary substances such as lubricants, preservatives, stabilizers and / or wetting agents, emulsifying agents, salts for influencing the osmotic pressure, pH regulating substances, dyes, taste-correcting substances and / or various additional active substances such as, for example, one or several vitamins.

For the application in the form of aerosol inhaling solution, aerosols containing the active substance either dissolved or suspended in a carrier gas or a mixture of gases (for example, C02 or chlorofluorocarbons) can be used. The particles of active substances usually have a micron size, and one or more additional solvents which are physiologically acceptable, such as ethanol, can also be added. Inhalant solutions can be administered using conventional inhalers.

The compounds of formula I and their physiologically acceptable salts can be used as integrin inhibitors to combat diseases, in particular thromboses, myocardial infarction, coronary diseases, arteries, islets, tumors, osoporosis, inflammations and infections.

The compounds of formula I, according to claim 1, and / or their physiologically acceptable salts can also be used in the pathological processes that are propagated or maintained by angiogenesis, in particular, in the case of tumors or rheumatoid arthritis For these treatments, the substances of formula I of the invention are generally administered analogously to the other known and commercial peptides, in particular analogously to the compounds described in US Pat. 4,472,305, preferably in doses between approximately 0.05 and 500 mg, in particular between 0.5 and 100 mg per dosage unit. The daily dose is preferably comprised between about 0.01 and 2 mg / kg of body weight. However, the particular dose for each patient depends on a wide variety of factors, for example, the effectiveness of the particular compound used, age, body weight, general state of health, sex, diet , of the time and method of administration, of the rate of excretion, of the drug combination of the severity of the particular disease to which the therapy is applied. Parenteral administration is preferred.

The new compounds of formula I can also be used as integrin ligands in the preparation of columns for affinity chromatography in order to obtain integrins at the pure state. To do this, the ligand - that is, a compound of formula I - is covalently bound to a polymeric support via a binding function, such as, for example, the carboxyl group of Asp.

As polymer support materials, the fixed phases based on polymers which have preferably hydrophilic properties and which are known and customary in the chemistry of peptides are suitable. Examples of these fixed phases are crosslinked polyazugars such as cellulose, sepharose or Sephadex®, acrylamides, polymers based on polyethylene glycol or Tentakel® polymers.

The preparation of the materials for affinity chromatography, intended for the purification of integrins, is carried out under the known and usual conditions of amino acid condensation.

The compounds of formula I have one or several chiral centers, whereby they can be obtained as racemates or in their optically active forms. The obtained racemates can be separated into their enantiomers by known chemical or mechanical methods. The diastereomers are formed from the racemic mixture, preferably by reaction with an optically active separation agent. Examples of agents such as the D and L forms of tartaric acid, diacetyl tartaric acid, dibenzoyl-1 tartaric acid, mandelic acid, malic acid, lactic acid, or the various optically active fluorosulphonic acids such as ß-canphosulonic acid. It is also convenient to separate the enantiomers through a column filled with an optically active separation agent (for example dini trobenz oil-phenylglycine); As the mobile phase, for example, a mixture of hexane / i sopropanol / to ketone can be used in a volume ratio of, for example, 82: 15: 3.

Of course it is also possible to obtain optically active formula I compounds by applying the methods described above and employing optically active starting substances.

All temperatures of the present text are given in ° C. In the examples that follow, the expression "is worked (or treated) in a usual manner" means the following: if necessary, water is added, if necessary the pH is adjusted between 2 and 10 according to the constitution of the final product, it is extracted with ethyl acetate or dichloromethane, the phases are separated, the organic phase is dried over sodium sulphate, concentrated by evaporation and purified by chromatography on silica gel and / or by recrystallization. The Rf values are given on silica gel; mobile phase: ethyl acetate / methanol 9: 1 TR = _ retention time (minutes) on HPLC and under the following conditions: [A] Column: Lichrosorb® RP 18 (250 x 5 um); Eluent A: 0.1% TFA in water Eluent B: 0.1% TFA in 90% acetonitrile, 10-water Flow: 1 ml / min Gradient: 20-95% B / 50 min Detection at 215 nm.

The separation of the diastereomers is preferably carried out under the indicated conditions. Mass spectrometry (MS): FAB (fast atom bombardment) (M + H) + Use 1 Reacting equimolar amounts of methyl (R, S) -2-bromo-2-phenyl-acetate and 3-hydroxymethyl-aniline yields N- (3-hydroxymethyl phenyl) -amino-phenyl-methyl acetate. By reaction with thionyl chloride, methyl N- (3-chloromethylpheneyl) -amino-phenyl-acetate is obtained and by subsequent reaction with sodium azide the N- (3-azidomethyl-phenyl) -amino-phenyl-methyl acetate is obtained. ("TO") .

A solution of 9.2 g of "A" in 350 ml of ethyl acetate in the presence of 1 g of Pd / C (5%) is hydrogenated for 35 minutes. After removing the catalyst and the solvent, the N- (3-aminometylphenyl) -amino-phenyl-methyl acetate ("B) is obtained in the oil state, TR 19.5, FAB 271.

By reaction of "B" with dibenzyl anhydride, methyl N- (3-benzyloxycarbonyl-aminomethylphenyl) -amino-phenyl-acetic acid is obtained, which is then hydrolyzed with KOH / methanol to give the N- (3) acid. -benzyloxycarbonyl-aminomethylphenyl) -amino-phenyl-acetic acid (= N- (Z -3-AMO) -amino-phenyl-acetic acid). By reaction with 1 equivalent of H-Arg (Mtr) -gli-OtBu, of DCC1 and of HOBt in dichloromethane, Z-3-AMP-Phg-Arg (Mtr) -gli-OtBu is obtained. The removal of protective group Z is carried out by catalytic hydrogenation as indicated above, by subsequent petidic coupling with BOC-Asp (OBzl) -OH the BOC-Asp (OBzl) -3-AMP-Phg- is obtained. Arg (Mtr) -Gli-OtBu.

After cleaving the BOC protecting group and the terbutyl ester in HCl / dioxane, the H-Asp (OBzl) -3-AMP-Phg-Arg (Mtr) -Gli-OH is obtained, and then the cyclo- compound (Asp ( Obzl) -3-AMP-Phg-Arg (Mtr) -Gli) After saponification of the ester, cleaving the protecting group Mtr in 98% trichloroacetic acid, purifying and separating by HPLC, the compounds are obtained.

Cyclo- (Asp-3-AMP-L-phg-Arg-Gli) and cyclo- (Asp-3-AMP-D-Phg-Arg-Gli).

Both compounds are characterized as follows: TR 15.5; FAB 567 and TR 12.5; FAB 567, values that can be freely assigned to both diastereomers.

Analogously, the compounds are obtained from 2-bromo-3-methyl-butyric acid, Cyclo- (Asp-3-AMP-L-Va] -Arg-Gli) and Cyclo- (Asp-3-AMP-D-Val-Arg-Gli), from 2-bromoacetic acid, the compound cyclo- (Asp-3-AMP-Gli-Arg-Gli) and from 2-bromo-3-phenyl-propionic acid, the compounds Cyclo- (Asp-3-AMP-L-Phe-Arg-Gli) and cyclo- (Asp-3-AMP-D-Phe-Arg-Gil).

The following examples relate to pharmaceutical preparations.

Example A: injection bottles The pH of a solution of 100 g of an active substance of formula I and 5 g of disodium hydrogen phosphate in 3 1 of bidistilled water is adjusted to 6.5 with 2N hydrochloric acid, then filtered under sterile conditions, this is introduced solution inside the bottles, it is lyophilized and finally the bottles are closed under sterile conditions. Each bottle for injection contains 5 mg of the active substance.

Example B: suppositories A mixture composed of 20 g of an active substance of formula I, 100 g of soy lecithin and 1400 g of cocoa butter is melted, the melt is poured into the molds and allowed to cool. Each suppository contains 20 mg of active substance.

Example C: solution A solution is prepared with 1 g of active substance of formula I, 9.38 g of NaH2P0 x 2 H20, 28.48 g of NaH2P04 x 12 H20, 0.1 g of benzalkonium chloride and 940 ml of bidistilled water. The pH is adjusted to 6.8, brought to a volume of 1 1 and sterilized by irradiation. The solution can be used in the form of eye drops.

Example D: ointment Under aseptic conditions, 500 mg of an active substance of formula I are mixed with 99.5 g of vanillin.

Example E: tablets A mixture composed of 1 kg of an active substance of 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 into tablets, such that each tablet contains 10 mg of the active substance.

Example F: dragees The tablets are formed analogously to that described in Example E and then they are coated in the usual manner with a bath of sucrose, potato starch, talc, tragacanth and dye.

Example G: capsules With 2 kg of an active substance of formula I, hard gelatin capsules are filled in such a way that each capsule contains 20 mg of the active substance.

Example H: ampoules A solution of 1 kg of an active substance of formula I in 60 1 of bidistilled water is filtered under sterile conditions. The ampules are filled with this solution and then lyophilized and closed under sterile conditions. Each vial contains 10 mg of the active substance.

Example I: aerosol inhalant solution 14 g of an active substance of formula I are dissolved in 10 1 of isotonic NaCl solution. With this solution, commercial containers that have a vaporizing mechanism are filled. The solution can be inhaled through the mouth or nose. Each spray (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (10)

Claims

1. Compounds of formula I Cyclo- (Arg-X-Asp-R1) (I) characterized because X represents Gli, Ala or NH-NH-CO, wherein said amino acids can also be derivatized and the amino acid residues are linked together by a peptide-type binding through the a-amino and a-amino groups. carboxy lo, RJ is a remainder of formula II R2, R3, R4 independently represent H, A, Ar, R5-Ar, Het or R5-Het, A represents alkyl of 1 to 6 carbon atoms, Ar represents phenyl unsubstituted or mono-, di- or trisstituted with R ~, R8 or R9, or unsubstituted naphthyl, R- represents alkylene of 1 to 6 C atoms, R6, R (independently represent H, A, benzyl or phenyl, R- independently represent R6, OR6, Hal, N02, NR6R6 ', NHCOR6, CN, NHS02Rb, COOR6 or COR' Hal represents F, Cl, Br or I and Het represents a heterocycle of one or two nuclei containing 1 to 4 N, O and / or S atoms and which may be unsubstituted or mono, di or trisubstituted with hal, A, NR6R6, CN or N02, including, in the case of the remains of amino acids and derivatives of optically active amino acids, both the D-forms and the L-forms, and their salts.

2. An enantiomer or diastereomer of a compound of formula I, according to claim 1.

3. Compounds of formula I, according to claim 1, characterized in that they are: a) Cyclo- (Arg-Gl? -Asp-3-AMP-L-Phg); b) Cyclo- (Arg-Glí-Asp-3-AMP-D-Phg); c) cyclo- (Arg-Gli-Asp-3-AMP-L-Val); d) cyclo- (Arg-Gli-Asp-3-AMP-D-Val); e) Cyclo- (Arg-Gli-Asp-3-AMP-Phe); f) cyclo- (Arg-Gli-Asp-3-AMP-D-Phe); and g) cyclo- (Arg-Gli-Asp-3-AMP-Gli); and its salts.

4. A process for preparing the compounds of formula I, according to claim 1, and their salts, characterized in that (a) a compound of formula III is treated H-Z-OH (III where represents -Arg-X-Asp-R1- -X-Asp-R1-Arg- -Asp-Rx-Arg-X or -R1-Arg-X-Asp-, and X and R1 have the meanings indicated in claim 1, or a reactive derivative of a compound of formula II with a cyclization agent, (b) a compound of formula I is released from one of its functional derivatives by treatment with a solvolysis or hydrogenolysis agent, and / or a base or acid of formula I is transformed into one of its salts by treatment with an acid or a base.

5. A process for obtaining pharmaceutical preparations, characterized in that a compound of formula I according to claim 1 is carried, and / or one of its salts physiologically acceptable to a suitable dosage form, together with at least one excipient or solid, liquid or semi-liquid auxiliary product.

6. A pharmaceutical preparation, characterized in that it contains at least one compound of formula I, according to claim 1, and / or one of its salts acceptable from the physiological point of view.

7. Compounds of formula I, according to claim 1, and their physiologically acceptable salts characterized as they act as integrin inhibitors and fight thrombosis, myocardial infarction, coronary diseases, arteriesclerosis, tumors, osteoporosis , inflammations and infections.

8. The use of the compounds of formula I, according to claim 1, and / or their physiologically acceptable salts in the pathological processes that are propagated or maintained by angiogenesis.

9. The use of the compounds of formula I, according to claim 1, and / or of their physiologically acceptable salts for preparing a medicament.

10. The use of the compounds of formula I, according to claim 1, and / or their physiologically acceptable salts to combat diseases. Summary of the Invention The compounds of formula I are described Cyclo- (Arg-X-Asp-R1) (I) where X represents Gli, Ala or NH-NH-CO, RJ is a remainder of formula II and R2, R3, R4 have the meanings indicated in claim 1 and its salts can be used as integrin inhibitors, in particular for the prophylaxis and treatment of diseases of the circulatory system, thrombosis, myocardial infarction, coronary heart disease, arteriosclerosis, pathological processes that are spread or sustained by angiogenesis, in the therapy of tumors.