AU9531998A - Cyclopeptide derivatives - Google Patents

Description

Cyclopeptide derivatives The invention relates to compounds of the formula I HO 0 NH HN,A-BC 5 in which A is Gly, Ala or NH-NH-CO, where the amino acids mentioned can also be derivatized, 10 B is a radical of the formula II NRI H'N N'R2 | H (CO)m

(CH

2 )n N H 0 C is - (CO) p- (CH2) q- (CO)r- or - (CO) p-CH=CH- (CO)r-, m, p, r in each case independently of one another are 15 0 or 1, n, q in each case independently of one another are 1, 2, 3 or 4, Ri and R2 in each case independently of one another are H or alkyl, R' and R 2 together are also oI R7 R8 R9 or 10 RY, RS, RS, 10R 20 Ll -2 Rio in each case independently of one another are H, alkyl, Ar, OR , Hal, NO 2 , NR6 R , NHCOR, 6 6 6 CN, NHSO 2 R , COOR or COR X is H, Hal, alkyl or Ar, 5 Ar is phenyl which is unsubstituted or mono-, di- or trisubstituted by R 3 , R 4 or R 5 or is unsubstituted naphthyl, R3, R4, R5 in each case independently of one another are R 6, OR 6, Hal, NO 2 , NR R 6', NHCOR 6, CN, NHSO 2 R' 10 COOR or COR6, R , R in each case independently of one another are H, alkyl, phenyl or benzyl, and Hal is F, Cl, Br or I, and if there are radicals of optically active amino 15 acids and amino acid derivatives, both the D and the L forms are included, and their salts. Similar compounds of cyclic peptides are dis closed, for example, in DE 43 10 643 or EP 0 683 173. 20 The invention is based on the object of finding novel compounds having valuable properties, in particu lar those which can be used for the production of medicaments. It has been found that the compounds of the 25 formula I and their salts have very valuable pharmaco logical properties combined with good tolerability. They act especially as integrin inhibitors, in particu lar inhibiting the interactions of the av-, 0 3 - or Ps-integrin receptors with ligands, such as, for 30 example, the binding of fibrinogen to the 0 3 -integrin receptor. The compounds show particular activity in the case of the integrins ai43 1 , aXvP 3 , avp 5 , arIb3 and avP 6 and av38. This action can be demonstrated, for example, 35 by the method which is described by J.W. Smith et al. in J. Biol. Chem. 265, 12267-12271 (1990). The dependence of the origin of angiogenesis on the interaction between vascular integrins and extracellular matrix proteins is described by

UI

-3 P.C. Brooks, R.A. Clark and D.A. Cheresh in Science 264, 569-71 (1994). The possibility of the inhibition of this interaction and thus for the initiation of apoptosis 5 (programmed cell death) of angiogenic vascular cells by a cyclic peptide is described by P.C. Brooks, A.M. Montgomery, M. Rosenfeld, R.A. Reisfeld, T.-Hu, G. Klier and D.A. Cheresh in Cell 79, 1157-64 (1994). Compounds of the formula I which block the 10 interaction of integrin receptors and ligands, such as, for example, of fibrinogen on the fibrinogen receptor (glycoprotein IIb/IIIa), prevent, as GPIIb/IIIa anta gonists, the spread of tumour cells by metastasis. This is confirmed by the following observations: 15 The spread of tumour cells from a local tumour into the vascular system takes place through the forma tion of microaggregates (microthrombi) by interaction of the tumour cells with blood platelets. The tumour cells are screened by protection in the microaggregate 20 and are not recognized by the cells of the immune system. The microaggregates can fix to vascular walls, by means of which a further penetration of tumour cells into the tissue is facilitated. 25 Since the formation of the microthrombi is mediated by fibrinogen binding to the fibrinogen receptors on activated blood platelets, the GPIIa/IIIb antagonists can be regarded as effective metastasis inhibitors. The compounds of the formula I can be employed 30 as pharmaceutical active compounds in human and veterinary medicine, in particular for the prophylaxis and/or therapy of diseases of the circulation, thrombosis, myocardial. infarct, arteriosclerosis, inflammations, apoplexy, angina pectoris, oncoses, 35 osteolytic diseases such as osteoporosis, pathologically angiogenic diseases such as, for example, inflammations, ophthalmological diseases, diabetic retinopathy, macular degeneration, myopia, ocular histoplasmosis, rheumatoid arthritis, osteo

_S'

4 arthritis, rubeotic glaucoma, ulcerative colitis, Crohn's disease, atherosclerosis, psoriasis, angiogenesis and restenosis after angioplasty, viral infection, bacterial infection, fungal infection, in 5 acute kidney failure and in wound healing for supporting the healing processes. The compounds of the formula I can be employed as substances having antimicrobial activity in opera tions where biomaterials, implants, catheters or heart 10 pacemakers are used. They have an antiseptic action here. The efficacy of the antimicrobial activity can be demonstrated by the procedure described by P. Valentin-Weigund et al., in Infection and Immunity, 2851-2855 (1988). 15 Since the compounds of the formula I are inhibitors of fibrinogen binding and thus ligands of the fibrinogen receptors on blood platelets, they can be used in vivo as diagnostics for the detection and localization of thrombi in the vascular system, pro 20 vided they are substituted, for example, by a radio active or UV-detectable residue. The compounds of the formula I can be used as inhibitors of fibrinogen binding and as effective auxiliaries for the study of the metabolism of blood 25 platelets in different activation stages or of intra cellular signal mechanisms of the fibrinogen receptor. The detectable unit of a "label" to be incorporated, e.g. isotopic labelling by 3 H, allows, after binding to the receptor, the mechanisms mentioned to be 30 investigated. The abbreviations of amino acid residues listed above and below stand for the radicals of the following amino acids: Ala alanine NH2 2 35 HC-CH-COOH LL w111, -5 Arg arginine NH2

NH--CHCH

2 CH2-CH-COOH HN=

NH

2 5 Gly glycine

H

2 N-CH-COOH Furthermore, the following have the meanings 10 below: Ac acetyl Boc tert-butoxycarbonyl CBZ or Z benzyloxycarbonyl DCCI dicyclohexylcarbodiimide 15 DMAP 4-dimethylaminopyridine DMF dimethylformamide EDCI N-ethyl-NN'-(dimethylaminopropyl)carbo diimide Et ethyl 20 FCA fluoresceincarboxylic acid Fmoc 9 -fluorenylmethoxycarbonyl HOBt 1-hydroxybenzotriazole HONSu N-hydroxysuccinimide MBHA 4-methylbenzhydrylamine 25 Me methyl Mtr 4-methoxy-2,3,6-trimethylphenylsulfonyl NMM N-methylmorpholine OBzl benzyl ester Oct octanoyl 30 OEt ethyl ester OMe methyl ester OtBu tert-butyl ester POA phenoxyacetyl Sal salicyloyl ALI 5 TFA trifluoroacetic acid Trt trityl (triphenylmethyl).

-6 If the abovementioned amino acids can occur in several enantiomeric forms, then above and below, e.g. as a constituent of the compounds of the formula I, all these forms and also their mixtures (e.g. the DL forms) 5 are included. Furthermore, the amino acids can be pro vided, e.g. as a constituent of compounds of the formula I, with corresponding protective groups which are known per se. So-called prodrug derivatives are also included 10 in the compounds according to the invention, i.e. com pounds of the formula I, modified with, for example, alkyl or acyl groups, sugars or oligopeptides, which are rapidly cleaved in the body to the active compounds according to the invention. 15 These also include biodegradable polymer derivatives of the compounds according to the invention, as is described, for example, in Int. J. Pharm. 115, 61-67 (1995). Amino acids whose configuration is not specifi 20 cally indicated have the (S) or (L) configuration. The invention further relates to a process for the preparation of compounds of the formula I according to Claim 1 and of their salts, characterized in that (a) a compound of the formula III 25 H-Z-OH III in which HO 0 Z is

NH-C-B-A

-NH X 0 OH -B-C-N H

HN-A

W~

-7 X O OH -C-N H

HN-A-B

X

N

0 OH -N H

HN-A-B-C

and X, A, B, and C have the meanings indicated in Claim 1, or a reactive derivative of a compound of the 5 formula III is treated with a cyclizing agent, or b) a compound of the formula I is set free from one of its functional derivatives by treating with a solvolysing or hydrogenolysing agent, 10 and/or in that a basic or acidic compound of the formula I is converted into one of its salts by treating with an acid or base. Above and below, the radicals X, A, B, C, Ri,

R

2 , m, n, p, q and Z have the meanings indicated in the 15 formulae I, II and III, if not expressly stated otherwise. In the above formulae, X is preferably H, Hal or alkyl, in particular H, Cl or CH 3 . In the above formulae, alkyl has 1-6 C atoms 20 and is preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, and further 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 25 3,3-dimethylbutyl, 1- or 2-ethylbutyl, 1-ethyl 1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or 1,2,2-trimethylpropyl. Alkyl is particularly preferably methyl. R', R', R' and R" 0 are preferably H.

-8 The amino acids and amino acid residues men tioned can also be derivatized, the N-methyl, N-ethyl, N-propyl, N-benzyl or Ca-methyl derivatives being preferred. 5 Additionally preferred, in particular, are the methyl, ethyl, propyl, butyl, tert-butyl, neopentyl or benzyl esters of the side chain carboxyl group, and further also derivatives of arginine, which can be substituted on the -NH-C(=NH)-NH 2 group by an acetyl, benzoyl, 10 methoxycarbonyl or ethoxycarbonyl radical. R6 and R6' are preferably, for example, H, methyl or ethyl, and further benzyl or phenyl.

OR

6 is preferably, for example, hydroxyl or methoxy. COR6 is alkanoyl and is preferably formyl, acetyl, 15 propionyl, butyryl, pentanoyl or hexanoyl. Ar is phenyl which is unsubstituted, preferably - as indicated - monosubstituted, specifically preferably phenyl, o-, m- or p-tolyl, o-, m- or p ethylphenyl, o-, m- or p-propylphenyl, o-, m- or 20 p-isopropylphenyl, o-, m- or p-tert-butylphenyl, o-, m or p-trifluoromethylphenyl, 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-(trifluoromethoxy) 25 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-, m- or p-benzyloxycarbonylphenyl, 0-, m- or 30 p-(carboxymethyloxy)phenyl, 0-, m- or p-(methoxycarbonylmethyloxy)phenyl, 0-, m- or p-(methoxycarbonyl-ethyloxy)phenyl, o-, m- or p-(N,N dimethylamino)phenyl, 0-, m- or p-(N-ethylamino) phenyl, o-, m- or p-(N,N-diethylamino)phenyl, o-, m- or 35 p-fluorophenyl, o-, m- or p-bromophenyl, o-, m- or p-chlorophenyl, o-, m- or p-difluoromethoxy)phenyl, o-, m- or p-(fluoromethoxy)phenyl, 0-, m- or p-formylphenyl, o-, m- or p-acetylphenyl, o-, m- or p-propionylphenyl, o-, m- or p-butyrylphenyl, o-, m- or

OU

- 9 p-pentanoylphenyl, o-, m- or p-(methylsulfonamido) phenyl, O-, m- or p-phenoxyphenyl, a-, m- or p-methylthiophenyl, 0-, m- or p-methylsulfinylphenyl, o-, m- or p-methylsulfonylphenyl or naphthyl. 5 Amino protective group preferably denotes acetyl, propionyl, butyryl, phenylacetyl, benzoyl, toluyl, POA, methoxycarbonyl, ethoxycarbonyl, 2, 2

,

2 -trichloroethoxycarbonyl, Boc, 2-iodoethoxy carbonyl, CBZ ("carbobenzoxy"), 4-methoxybenzyloxy 10 carbonyl, Fmoc, Mtr or benzyl. The compounds of the formula I have at least two chiral centres and can therefore occur in various stereoisomeric forms. The formula I includes all these forms. 15 Accordingly, the invention relates in particular to those compounds of the formula I in which at least one of the radicals mentioned has one of the preferred meanings indicated above. Some preferred groups of compounds can be expressed by the following 20 subformulae Ia, Ib and Ic, which correspond to the formula I and in which in Ia X is H, alkyl or Hal, Rf , R2 are H, m is O, 25 n is 3, p, r are 1, and q is 2 or 3, and in Ib X is H, alkyl or Hal, 30 Ri, R2 are H, m is 0, n is 3, p is 1, r is 0, and 35 q is 1, and in Ic X is H, alkyl or Hal, -10

R

1 and R 2 together are or m is 1 n is 2 5 p, r are 1, and q is 2. The compounds of the formula I and also the starting substances for their preparation are otherwise prepared by methods known per se, such as are described 10 in the literature (e.g. in the standard works such as Houben-Weyl, Methoden der Organischen Chemie [Methods of Organic Chemistry), Georg-Thieme-Verlag, Stuttgart), namely under reaction conditions which are known and suitable for the reactions mentioned. In this case, use 15 can also be made of variants which are known per se but not mentioned here in greater detail. If desired, the starting substances can also be formed in situ such that they are not isolated from the reaction mixture, but are immediately reacted further 20 to give the compounds of the formula I. The compounds of the formula I can be prepared, for example, according to the following schemes 1 and 2: 77 Lu -- 11 Scheme 1: 0 0 NO2 NO 2

O.IL'*NH

4

NH

2 V 1. BocGy-OH MoO 0 2. HCIldioxane a

NO

2 3. Boc-Arg(Mtr)-OH NH NO 2 NN Gly-Arg(Mr)-Boc VI Vu MViO 1. HCIldioxane rN02 o2. Cyclization o o o orNH 3. Hydrolysis HO 0 4. Mtr removal iy H 0 0 N CH2)q q=2 or 3 NH NH NN

ALL

- 12 Scheme 2: Mo O HMoO Ra moo 0C H 3 - BO20 0 *T a 0

NO

2

NO

2

NH

2 NH-SoC Ix x MoO O CH, o o MOO OCH 3 NH2 NH NH-Soc NH-Soc Xi HOOC XII mo %O CH3 K.. ~Removal of the su-O 0 NH Bocand the H-Arg(Mtr)-GIy-Ot Bu NH-OC 0 Bu-O group NH H N 0 0 NH N N H H MoO 0 CHii HO 0 CH HO O 1Cclization YH IN Hdrolysi O IN 0.L 3. Mtr removal IN ~~ _TN H NN 0 O NHNH 0 H H N NH2 H XIV Is

LL/J

-13 The important 3-amino-3-(3-nitrophenyl) propionic acid unit from Scheme 1 is prepared according to J. Org. Chem. 25, 1758 (1960) from 3 nitrobenzaldehyde, malonic acid and ammonium acetate. 5 In the synthesis of analogous compounds the corresponding nitrobenzaldehyde derivatives are used. Compounds of the formula I can preferably be obtained by cyclization of compounds of the formula III under the conditions of a peptide synthesis. In this 10 case, the reaction is expediently carried out by customary methods of peptide synthesis, such as are described, for example, in Houben-Weyl, 1.c., Volume 15/II, pages 1 to 806 (1974). The reaction preferably takes place in the 15 presence of a dehydrating agent, e.g. of a carbodiimide such as DCCI or EDCI, and further, for example, propanephosphonic anhydride (cf. Angew. Chem. 92, 129 (1980)), diphenylphosphoryl azide or 2-ethoxy-N-ethoxy carbonyl-1,2-dihydroquinoline, in an inert solvent, 20 e.g. 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 dimethyl sulfoxide or in the presence of these solvents, at temperatures between 25 approximately -10 and 40, preferably between 0 and 30*. In order to promote intramolecular cyclization ahead of intermolecular peptide bonding, it is expedient to work in dilute solutions. Depending on the conditions used, the reaction time is 30 between a few minutes and 14 days. Instead of compounds of the formula III, it is also possible to employ derivatives of compounds of the formula III, preferably a preactivated carboxylic acid, or a carboxylic acid halide, a symmetrical or mixed 35 anhydride or an active ester. Radicals of this type for the activation of the carboxyl group in typical acylation reactions are described in the literature AL% (e.g. in the standard works such as Houben-Weyl, Methoden der Organischen Chemie [Methods of Organic - 14 Chemistry], Georg-Thieme-Verlag, Stuttgart). Activated esters are expediently formed in situ, e.g. by addition of HOBt or N-hydroxysuccinimide. As a rule, the reaction is carried out in an 5 inert solvent, when using a carboxylic acid halide in the presence of an acid-binding agent, preferably of an organic base such as triethylamine, dimethylaniline, pyridine or quinoline. The addition of an alkali metal or alkaline earth metal 10 hydroxide, carbonate or bicarbonate or of another salt of a weak acid of the alkali metals or alkaline earth metals, preferably of potassium, sodium, calcium or caesium, may also be favourable. As a rule, the starting substances of the 15 formula III are novel. They can be prepared by known methods of peptide synthesis. The compounds of the formula I can further be obtained by setting them free from their functional derivatives by solvolysis, in particular hydrolysis, or 20 by hydrogenolysis. Preferred starting substances for the solvolysis or hydrogenolysis are those which, instead of one or more free amino and/or hydroxyl groups, contain corresponding protected amino and/or hydroxyl 25 groups, preferably those which, instead of an H atom which is bonded to an N atom, carry an amino protective group, e.g. those which correspond to the formula I but instead of an NH 2 group contain an NHR' group (in which R' is an amino protective group, e.g. Boc or CBZ). 30 Starting substances are furthermore preferred which, instead of the H atom of a hydroxyl group, carry a hydroxyl protective group, e.g. those which corres pond to the formula I but instead of a hydroxyphenyl group contain an R"O-phenyl group (in which R" is a 35 hydroxyl protective group). A plurality of - identical or different - pro tected amino and/or hydroxyl groups can also be present AL in the molecule of the starting substance. If the - -15 protective groups present are different from one another, in many cases they can be selectively removed. The expression "amino protective group" is generally known and relates to groups which are 5 suitable for protecting (for blocking) an amino group from chemical reactions, but which are easily removable after the desired chemical reaction has been carried out at other positions in the molecule. Typical of such groups are, in particular, unsubstituted or substituted 10 acyl, aryl, aralkoxymethyl or aralkyl groups. Since the amino protective groups are removed after the desired reaction (or reaction sequence), their nature and size is otherwise not critical; however, those having 1-20, in particular 1-8, C atoms are preferred. The 15 expression "acyl group" is to be interpreted in the widest sense in connection with the present process. It includes acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic acids or sulfonic acids and also, in particular, 20 alkoxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonyl groups. Examples of acyl groups of this type are alkanoyl such as acetyl, propionyl, butyryl; aralkanoyl such as phenylacetyl; aroyl such as benzoyl or toluyl; aryloxyalkanoyl such as POA; alkoxy 25 carbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, Boc, 2-iodoethoxy carbonyl; aralkyloxycarbonyl such as CBZ ("carbo benzoxy"), 4-methoxybenzyloxycarbonyl, Fmoc; aryl sulfonyl such as Mtr. Preferred amino protective groups 30 are Boc and Mtr, and further CBZ, Fmoc, benzyl and acetyl. The expression "hydroxyl protective group" is likewise generally known and relates to groups which are suitable for protecting a hydroxyl group from 35 chemical reactions, but which are easily removable after the desired chemical reaction has been carried out at other positions in the molecule. Typical of such groups are the abovementioned unsubstituted or substituted aryl, aralkyl or acyl groups, and fur

UJ

- 16 thermore also alkyl groups. The nature and size of the hydroxyl protective groups is not critical, since they are removed again after the desired chemical reaction or reaction sequence; groups having 1-20, in particular 5 1-10, C atoms are preferred. Examples of hydroxyl pro tective groups are, inter alia, benzyl, p-nitrobenzoyl, p-toluenesulfonyl, tert-butyl and acetyl, benzyl and tert-butyl being particularly preferred. The COOH group is preferably protected in the form of its tert-butyl 10 ester. The compounds of the formula I are set free from their functional derivatives - depending on the protective group used - e.g. using strong acids, expediently using TFA or perchloric acid, but also 15 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- or p-toluenesulfonic acid. The presence of an additional inert solvent is possible, but not always 20 necessary. Suitable inert solvents are preferably 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, and further also alcohols such as 25 methanol, ethanol or isopropanol, and also water. Mix tures of the abovementioned solvents are furthermore suitable. TFA is preferably used in an excess without addition of a further solvent, perchloric acid in the form of a mixture of acetic acid and 70% perchloric 30 acid in the ratio 9:1. The reaction temperatures for the cleavage are expediently between approximately 0 and approximately 500, preferably the cleavage is carried out between 15 and 30* (room temperature). The groups Boc, OtBu and Mtr can, for example, 35 preferably be removed using TFA in dichloromethane or using approximately 3 to 5 N HCl in dioxane at 15-30*, the Fmoc group using an approximately 5 to 50% solution of dimethylamine, diethylamine or piperidine in DMF at 15-30*. w - 17 The trityl group is employed for the protection of the amino acids histidine, asparagine, glutamine and cysteine. Removal is carried out, depending on the desired final product, using TFA/10% thiophenol, the 5 trityl group of all amino acids mentioned being removed, when using TFA/anisole or TFA/thioanisole only the trityl group of histidine, asparagine and glutamine being removed, while it remains on the cysteine side chain. 10 Hydrogenolytically removable protective groups (e.g. CBZ or benzyl) can be removed, for example, by treating with hydrogen in the presence of a catalyst (e.g. of a noble metal catalyst such as palladium, expediently on a support such as carbon). Suitable 15 solvents here are those indicated above, in particular, for example, alcohols such as methanol or ethanol or amides such as DMF. As a rule, the hydrogenolysis is carried out at temperatures between approximately 0 and 100* and pressures between approximately 1 and 200 bar, 20 preferably at 20-30" and 1-10 bar. Hydrogenolysis of the CBZ group is readily carried out, for example, on 5 to 10% Pd/C in methanol or using ammonium formate (instead of hydrogen) on Pd/C in methanol/DMF at 20-30*. 25 A base of the formula I can be converted into the associated acid addition salt using an acid, for example by reaction of equivalent amounts of the base and of the acid in an inert solvent such as ethanol and subsequent evaporation. In particular, acids which 30 yield physiologically acceptable salts are suitable for this reaction. Thus inorganic acids can be used, e.g. sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid, and 35 furthermore organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic mono or polybasic carboxylic, sulfonic or sulfuric acids, LALI e.g. formic acid, acetic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, - 18 pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methane- or ethanesulfonic acid, ethanedisulfonic acid, 5 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenemono- and disulfonic acids, laurylsulfuric acid. Salts with physiologically unacceptable acids, e.g. picrates, can be used for the isolation and/or purification of the compounds of the 10 formula I. On the other hand, an acid of the formula I can be converted into one of its physiologically acceptable metal or ammonium salts by reaction with a base. Possible salts in this case are, in particular, the 15 sodium, potassium, magnesium, calcium and ammonium salts, and furthermore substituted ammonium salts, e.g. the dimethyl-, diethyl- or diisopropylammonium salts, monoethanol-, diethanol- or diisopropylammonium salts, cyclohexyl- or dicyclohexylammonium salts, dibenzyl 20 ethylenediammonium salts, and furthermore, for example, salts with arginine or lysine. The invention further relates to the use of the compounds of the formula I and/or their physiologically acceptable salts for the production of pharmaceutical 25 preparations, in particular in a non-chemical way. In this context, they can be brought into a suitable dose form together with at least one solid, liquid and/or semi-liquid excipient or auxiliary and, if appropriate, in combination with one or more other active compounds. 30 The invention furthermore relates to pharmaceu tical preparations comprising at least one compound of the formula I and/or one of its physiologically accept able salts. These preparations can be used as medicaments 35 in human or veterinary medicine. Suitable excipients are organic or inorganic substances which are suitable for enteral (e.g. oral) or parenteral administration, topical application or administration in the form of an inhalation spray and do not react with the novel Lu

U-

- 19 compounds, for example water, vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glyceryl triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum 5 jelly. In particular, tablets, pills, coated tablets, capsules, powders, granules, syrups, juices or drops are used for oral administration, suppositories are used for rectal administration, solutions, preferably oily or aqueous solutions, and furthermore suspensions, 10 emulsions or implants are used for parenteral adminis tration, and ointments, creams or powders are used for topical application. The novel compounds can also be lyophilized and the lyophilizates obtained used, for example, for the production of injection preparations. 15 The preparations indicated can be sterilized and/or can contain auxiliaries such as lubricants, preservatives, stabilizers and/or wetting agents, emulsifiers, salts for affecting the osmotic pressure, buffer substances, colorants, flavourings and/or one or more other active 20 compounds, e.g. one or more vitamins. For administration as an inhalation spray, sprays can be used which contain the active compound either dissolved or suspended in a propellant or propellant mixture (e.g. CO 2 or chlorofluorohydrocarbons). 25 Expediently, the active compound is in this case used in micronized form, it being possible for one or more additional physiologically tolerable solvents, e.g. ethanol, to be present. Inhalation solutions can be administered with the aid of customary inhalers. 30 The compounds of the formula I and their physiologically acceptable salts can be used as integrin inhibitors in the control of illnesses, in particular of diseases of the circulation, thromboses, cardiac infarct, coronary heart diseases, 35 arteriosclerosis, apoplexy, angina pectoris, tumours, osteoporosis, inflammations, infections and restenosis after angioplasty. Lt The compounds of the formula I according to Claim 1 and/or their physiologically acceptable salts

LUJ

- 20 are also used in pathological processes which are supported or propagated by angiogenesis, in particular in tumours or rheumatoid arthritis. In this case, the substances according to the 5 invention can as a rule be administered in analogy to other known, commercially available peptides, but in particular in analogy to the compounds described in US-A-4 472 305, preferably in doses between approxi mately 0.05 and 500 mg, in particular between 0.5 and 10 100 mg per dose unit. The daily dose is preferably between approximately 0.01 and 2 mg/kg of body weight. The specific dose for each patient depends, however, on all sorts of factors, for example on the efficacy of the specific compound employed, on the age, body 15 weight, general state of health, sex, on the diet, on the time and route of administration, on the excretion rate, pharmaceutical combination and severity of the particular disorder to which the therapy applies. Parenteral administration is preferred. 20 The compounds of the formula I can furthermore be used as integrin ligands for the preparation of columns for affinity chromatography for the preparation of integrins in pure form. The ligand, i.e. a compound of the formula I, is in 25 this case covalently coupled to a polymeric support via an anchor function, e.g. the free carboxyl group. Suitable polymeric supports are the polymeric solid phases, preferably having hydrophilic properties, known per se in peptide chemistry, for example 30 crosslinked polysugars such as cellulose, Sepharose or Sephadex*, acrylamides, polymer based on polyethylene glycol or Tentakel polymers The materials for affinity chromatography for integrin purification are prepared under conditions 35 such as are customary and known per se for the conden sation of amino acids. The compounds of the formula I contain at least two chiral centres and can therefore be present in racemic or in optically active form. Racemates obtained -21 can be separated into the enantiomers mechanically or chemically by methods known per se. Preferably, diastereomers are formed from the racemic mixture by reaction with an optically active resolving agent. 5 Suitable resolving agents are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids such as 10 -camphorsulfonic acid. Also advantageous is an enantiomer resolution with the aid of a column packed with an optically active resolving agent (e.g. dinitrobenzoylphenylglycine); a suitable eluent is, for example, a mixture of hexane/isopropanol/acetonitrile, 15 e.g. in the volume ratio 82:15:3. Of course, it is also possible to obtain opti cally active compounds of the formula I by the methods described above by using starting substances which are already optically active. 20 All temperatures above and below are indicated in *C. Room temperature is 22*C. In the following examples, "customary working up" means: if necessary, water is added, the mixture is adjusted, if necessary, to a pH of between 2 and 10, depending on the 25 constitution of the final product, and extracted with ether or dichloromethane, the organic phase is separated off, dried over sodium sulfate, filtered and evaporated, and the residue is purified by chromatography on silica gel and/or by crystallization. 30 RT = retention time (minutes) on HPLC in the following systems: Column: Lichrosorb@ RP 18 (250 x 4; 5 pm) Eluent A: 0.1% TFA in water Eluent B: 0.1% TFA in 90% acetonitrile, 10% water 35 Flow rate: 1 ml/min Gradient: 20 - 95% B/50 min Detection at 215 nm. The separation of the diastereomers is prefer ably carried out under the conditions indicated.

-22 Mass spectrometry (MS) FAB (Fast Atom Bombard ment) (M+H)* Example 1 HO O NH 0 NH t-N H N 0

NH

2 .N NH H 5 8S, 14S-IB Synthesis of the compound to be cyclized 10 Methyl (3R,S)-3-amino-3- (4-methyl-3-nitro phenyl)propionate (3R, S-IX) is synthesized analogously to Scheme 1. This ester is cleaved into the enantiomers according to the known method and methyl (3S)-3-amino 3-(4-methyl-3-nitrophenyl)propionate (3S-IX) is then 15 reacted analogously to Scheme 2 to give the Mtr protected compound methyl 3S-amino-3-(3-{3-[l (carboxymethylcarbamoyl) -4-guanidino-1s-butyl carbamoyl]propionylamino)-4-methylphenyl)propionate (S, S-XIV). 20 Cyclization 616 mg of the Mtr-protected compound methyl 3S-amino-3- (3-{3- [1- (carboxymethylcarbamoyl) -4-guan 25 idino-1S-butylcarbamoyl]propionylamino)-4-methyl phenyl)-propionate (S,S-XIV) are dissolved in 80 ml of DMF and diluted with 800 ml of dichloromethane. The mixture is then cooled to -20 0 C and 300 mg of EDCI, 98 mg of DMAP and 0.176 ml of NMM are added successively. It is warmed to room temperature and

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- 23 stirred overnight. The solution is concentrated and the residue is stirred into 200 ml of half-saturated NaHCO 3 solution. The precipitate is filtered off with suction and washed. 400 mg of substance are obtained, which are 5 purified by preparative HPLC. After chromatography, 44 mg of the Mtr-protected cyclic compound methyl (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]acetate, RT 26.1; FAB 716. 10 Hydrolysis and removal of the Mtr protective group The Mtr-protected cyclic compound (8S,14S)-[8 (3- (guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo 15 2,7,10,13-tetraazabicyclo[13.3.1]nonadeca 1(18),15(19),16-trien-14-yl] acetic acid is obtained by hydrolysis in KOH/methanol. 25 mg of this compound are then dissolved in 4.3 ml of 98% strength trifluoroacetic acid and stirred overnight at room 20 temperature. The solution is concentrated in a rotary evaporator and the residue is 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.llnonadeca-1(18),15(19),16-trien-14-yl] acetic 25 acid (8S,14S-IB) are obtained, RT 20.2; FAB 490. Examnles 2-3: HO O NH 0 NH HN H (CH 2 )q 0

NH

2 N NH H

LU

- 24 Example 2 (q = 2): Starting from 85 mg of the Mtr-protected compound ( 8 S,1 4 S)-2-(8-(3-guanidinopropyl)-3 ,6,9,12 5 tetraoxo-2,7,10,13-tetraazabicyclo[13.3.1]nonadeca 16,18,19-trien-14-yl) acetic acid, there are obtained, in analogy to Example 1 by reaction with 14.7 ml of 98% trifluoroacetic acid, 13 mg of (8S,14S)-2-(8-(3 guanidinopropyl)-3,6,9,12-tetraoxo-2,7,10,13 10 tetraazabicyclo[13.3.1]nonadeca-16,18,19-trien-14-yl) acetic acid; RT 17.2; FAB 476. Example 3 (q = 3): 15 Starting from 300 mg of the Mtr-protected compound ( 9 S,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, there are obtained in analogy to Example 1, 74 mg of (9S,15S)-2-(9-(3 20 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: HO O NH 0 NH I CH2 HN O N

NH

2 N NH 25 H Starting from the Mtr-protected compound (6S,1 2 )-[6-(3-guanidinopropyl-4,7,10-trioxo-2,5,8,11 AL/1 tetraazabicyclo[11.3.1]heptadeca-1(17),13,15-trien 12-yl] acetic acid, there are obtained in analogy to

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- 25 Example 1, (6S,12S)-[6-(3-guanidinopropyl-4,7,10 trioxo-2, 5, 8, ll-tetraazabicyclo- [11.3 .1]heptadeca 1(1 7 ),1 3 ,15-trien-12-yl] acetic acid. 5 Examples 5-8: HO N 0 NH C 0 H

(CH

2 )n (CO)m H.N YN RI E'2 The following compounds are synthesized in analogy to Example 1: 10 Ex. No. X C m n R and R 2 5 H -CO-CH=CH-CO- 0 3 H 6 Cl -CO-CH2-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 1l CH The following examples relate to pharmaceutical preparations: Example A: injection vials A solution of 100 g of an active compound of the formula I and 5 g of disodium hydrogenphosphate is -26 adjusted to pH 6.5 using 2 N hydrochloric acid in 3 1 of double-distilled water, sterile-filtered, dispensed into injection vials, lyophilized under sterile condi tions and aseptically sealed. Each injection vial 5 contains 5 mg of active compound. Example B: suppositories A mixture of 20 g of an active compound of the 10 formula I is fused with 100 g of soya lecithin and 1400 g of cocoa butter, poured into moulds and allowed to cool. Each suppository contains 20 mg of active compound. 15 Example C: solution A solution is prepared from 1 g of an active compound of the formula I, 9.38 g of NaH 2 PO4-2H 2 0, 28.48 g of Na 2 HPO4-12H 2 0 and 0.1 g of benzalkonium 20 chloride in 940 ml of double-distilled water. It is adjusted to pH 6.8, made up to 1 1 and sterilized by irradiation. This solution can be used in the form of eye drops. 25 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. 30 Example E: tablets A mixture of 1 kg of active compound of the formula I, 4 kg of lactose, 1.2 kg of potato starch, 35 0.2 kg of talc and 0.1 kg of magnesium stearate is compressed in a customary manner to give tablets such that each tablet contains 10 mg of active compound.

UW

- 27 Example F: coated tablets Analogously to Example E, tablets are pressed which are then coated in a customary manner with a 5 coating of sucrose, potato starch, talc, tragacanth and colorant. Example G: capsules 10 2 kg of active compound of the formula I are dispensed into hard gelatin capsules in a customary manner such that each capsule contains 20 mg of the active compound. 15 Example H: ampoule 4 A solution of 1 kg of active compound of the formula I in 60 1 of double-distilled water is sterile filtered, dispensed into ampoules, lyophilized under 20 sterile conditions and aseptically sealed. Each ampoule contains 10 mg of active compound. Example I: inhalation spray 25 14 g of active compound of the formula I are dissolved in 10 1 of isotonic NaCl solution and the solution is dispensed into commercially available spray containers having a pump mechanism. The solution can be sprayed into the mouth or nose. One burst of spray 30 (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg. C-) 00

Claims (10)

1. Compounds of the formula I HO O NH M' lA-B'C 5 in which A is Gly, Ala or NH-NH-CO, where the amino acids mentioned can also be derivatized, 10 B is a radical of the formula II WR1 H,N N'R2 1 H (CO)m (CH 2 )n "N H 0 C is - (CO) p- (CH2) q- (CO) r- or - (CO) p-CH=CH- (CO)r, m, p, r in each case independently of one another are 15 0 or 1, n, q in each case independently of one another are 1, 2, 3 or 4, Ri and R2 in each case independently of one another are H or alkyl, R 1 and R 2 together are also II R , R8, R, or 0 - 29 Rio in each case independently of one another are H, alkyl, Ar, OR 6 , Hal, NO 2 , NR 6 R 6 ', NHCOR , CN, NHSO 2 R 6, COOR or COR, X is H, Hal, alkyl or Ar, 5 Ar is phenyl which is unsubstituted or mono-, di- or trisubstituted by R 3 , R 4 or R 5 or is unsubstituted naphthyl, R3 R4 R5 in each case independently of one another are R , OR , Hal, NO 2 , NR 6 R , NHCOR , CN, NHSO 2 R , 10 COOR 6 or COR 6 , R, R in each case independently of one another are H, alkyl, phenyl or benzyl, and Hal is F, Cl, Br or I, and if there are radicals of optically active amino 15 acids and amino acid derivatives, both the D and the L forms are included, and their salts.

2. An enantiomer or a diastereomer of a compound of the formula according to Claim 1. 20

3. Compounds of the formula I according to Claim 1: a) (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; 25 b) ( 9 S,15S)-2-(9-(3-guanidinopropyl)-3,7,10,13 tetraoxo-2,8,11,14-tetraazabicyclo[14.3.1]eicosan 1 7 ,1 9 ,20-trien-15-yl)acetic acid; c) ( 8 S,1

4 S)-(8-(3-guanidinopropyl)-18-methyl 3 ,6, 9 ,1 2 -tetraoxo-2,7,10,13-tetraazabicyclo[13.3.1] 30 nonadeca-1(18),15(19),16-trien-14-yl)acetic acid; d) (6S,1 2 S)-(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 their salts. 35 4. Process for the preparation of compounds of the formula I according to Claim 1 and of their salts, characterized in that (a) a compound of the formula III LU- - 30 H-Z-OH III in which HO O Z is NH-C-B-A -NH x O 0 OH -B-C-N H HN-A x HOH -C-N Zo H HN-A-B x 0 OH -N H HN-A-B-C 5 and X, A, B and C have the meanings indicated in Claim 1, or a reactive derivative of a compound of the formula III is treated with a cyclizing agent, 10 or b) a compound of the formula I is set free from one of its functional derivatives by treating with a solvolysing or hydrogenolysing agent, and/or in that a basic or acidic compound of the 15 formula I is converted into one of its salts by treat ing with an acid or base.

5. Process for the production of pharmaceutical preparations, characterized in that a compound of the formula I according to Claim 1 and/or one of its 2 i physiologically acceptable salts is brought into a - 31 suitable dose form together with at least one solid, liquid or semi-liquid excipient or auxiliary.

6. Pharmaceutical preparation, characterized in that it contains at least one compound of the formula I 5 according to Claim 1 and/or one of its physiologically acceptable salts.

7. Compounds of the formula I according to Claim 1 and their physiologically acceptable salts as integrin inhibitors for the control of diseases of the 10 circulation, thromboses, cardiac infarct, coronary heart diseases, arteriosclerosis, apoplexy, angina pectoris, tumours, osteoporosis, inflammations, infections and restenosis after angioplasty.

8. Use of compounds of the formula I according to 15 Claim 1 and/or their physiologically acceptable salts in pathological processes which are supported or propagated by angiogenesis.

9. Use of compounds of the formula I according to Claim 1 and/or their physiologically acceptable salts 20 for the production of a medicament.

10. Use of compounds of the formula I according to Claim 1 and/or their physiologically acceptable salts in the control of illnesses.