DE19736772A1 - New guanidino-substituted bi:cyclic peptide compounds - Google Patents

Abstract

Cyclic peptides of formula (I), in which all optically active aminoacid (or derivative) residues may be in D- as well as L-form, and their salts, enantiomers and diastereomers are new. A = Gly, Ala or NHNHCO (where the aminoacids are optionally derivatised); B = guanidino-substituted aminoacid residue of formula (i); C = -(CO)p-(CH2)q-(CO)r- or -(CO)p-CH=CH-(CO)r-; m, p, r = 0 or 1; n, q = 1-4; R1, R2 = H or alkyl; or R1 + R2 = -C(R7)=C(R8)- or ortho-phenylene substituted by R9 and R10; R7-R10 = H, alkyl, Ar, OR6, Hal, NO2, NR6R'6, NHCOR6, CN, NHSO2R6, COOR6 or COR6; X = H, Hal, alkyl or Ar; Ar = phenyl (optionally substituted by 1-3 R3, R4 or R5) or is unsubstituted naphthyl; R3-R5 = R6, OR6, Hal, NO2, NR6R'6, NHCOR6, CN, NHSO2R6, COOR6, or COR6; R6,R'6 = H, alkyl, phenyl or benzyl; Hal = F, Cl, Br or I. An Independent claim is included for the preparation of (I).

Description

The invention relates to compounds of the formula I.

wherein
A Gly, Ala or NH-NH-CO,
the amino acids mentioned can also be derivatized,
B is a radical of formula II

C - (CO) _p - (CH ₂ ) _q - (CO) _r - or - (CO) _p -CH = CH- (CO) _r -,
m, p, r each independently of one another 0 or 1,
n, q each independently of one another 1, 2, 3 or 4,
R ¹ and R ^{2 are} each independently of one another H or alkyl,
R ¹ and R ² together also

R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each independently of one another H, alkyl, Ar, OR ⁶ , Hal, NO ₂ , NR ⁶ R ^{6 '} , NHCOR ⁶ , CN, NHSO ₂ R ⁶ , COOR ⁶ or COR ⁶ ,
XH, Hal, alkyl or Ar,
Ar is unsubstituted or mono-, di- or trisubstituted by R ³ , R ⁴ or R ⁵ phenyl or unsubstituted naphthyl,
R ³ , R ⁴ , R ⁵ each 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 '} each independently of one another are H, alkyl, phenyl or benzyl, and
Hal F, Cl, Br or I
mean,
where, as far as residues of optically active amino acids and amino acid derivatives are concerned, both the D and the L forms are included,
as well as their salts.

Similar compounds of cyclic peptides are e.g. B. from DE 43 10 643 or EP 0 683 173.  

The invention was based on the object, new compounds with valuable len properties to find, especially those that are used to manufacture of drugs can be used.

It has been found that the compounds of the formula I and their salts have very valuable pharmacological properties with good tolerability. Above all, they act as integrin inhibitors, in particular inhibiting the interactions of the α _V -, β ₃ - or β ₅ -integrin receptors with ligands, such as. B. the binding of fibrinogen to the β ₃ integrin receptor. The compounds are particularly effective in the case of the integrins α _V β ₁ , α _V β ₃ , α _V β ₅ , α _IIb β ₃ and α _V β ₆ and α _V β ₈ .

This effect can e.g. B. can be detected by the method that by J.W. Smith et al. in J. Biol. Chem. 265, 12267-12271 (1990) will.

The dependence of the development of angiogenesis on the change effect between vascular integrins and extracellular matrix protein is from P.C. Brooks, R.A. Clark and D.A. Cheresh in Science 264, 569-71 (1994).

The possibility of inhibiting this interaction and thus to Initiation of apoptosis (programmed cell death) angiogenic vascular Cells by a cyclic peptide is from 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 formula I, the interactions of integrin receptors and ligands, such as. B. from fibrinogen to the fibrinogen receptor (glycoprotein IIb / IIIa) prevent GPIIb / IIIa antagonists from spreading tumor cells through metastasis. This is evidenced by the following observations:
The spread of tumor cells from a local tumor into the vascular system occurs through the formation of micro-aggregates (microthrombi) through the interaction of the tumor cells with platelets. The tumor cells are shielded by the protection in the micro-aggregate and are not recognized by cells of the immune system.

The micro-aggregates can stick to the walls of the vessel, causing further penetration of tumor cells into the tissue is facilitated. Since the formation of the microthrombi by fibrinogen binding to the fibri nogenreceptors mediated on activated platelets, can GPIIa / IIIb antagonists who are regarded as effective metastasis inhibitors the.

The compounds of formula I can be used as active pharmaceutical ingredients in the Human and veterinary medicine are used, especially for pro phylaxis and / or therapy of circulatory diseases, thrombosis, myocardial infarction, arteriosclerosis, inflammation, apoplexy, angina pectoris, tumor diseases, osteolytic diseases such as osteoporo se, pathologically angiogenic diseases such. B. inflammation, oph thalmological diseases, diabetic retinopathy, macular dege generation, myopia, ocular histoplasmosis, rheumatoid arthritis, osteo arthritis, rubeotic glaucoma, ulcerative colitis, Crohn's disease, athe rosclerosis, psoriasis, angiogenesis and restenosis after angioplasty, vi oral infection, bacterial infection, fungal infection, with acute renal say and in wound healing to support the healing processes.

The compounds of formula I can act as antimicrobial sub punches are used in operations where biomaterials, implanta te, catheter or pacemaker can be used.

They have an antiseptic effect. The effectiveness of antimicrobial activi can be done by P. Valentin-Weigund et al., in Infection and Im munity, 2851-2855 (1988).

Since the compounds of formula I inhibitors of fibrinogen binding and so that ligands of the fibrinogen receptors on platelets represent can be used as diagnostics for the detection and localization of thrombi vascular system can be used in vivo provided, for example be substituted by a radioactive or UV-detectable residue.

The compounds of formula I can also be used as inhibitors of fibrinogen binding as effective tools for studying the metabolism of platelets in different activation stages or of intracellular signaling mechanisms of the fibrinogen receptor. The detectable unit of a "label" to be installed, e.g. B. an isotope label by ³ H, allows, after binding to the receptor, to investigate the mechanisms mentioned.

The abbreviations of amino acid residues listed above and below stand for the residues of the following amino acids:
Ala: Alanine

Arg: arginine

Gly: glycine

H ₂ N-CH-COOH.

Furthermore, below mean:
Ac: acetyl
Boc: tert-butoxycarbonyl
CBZ or Z: benzyloxycarbonyl
DCCl: dicyclohexylcarbodiimide
DMAP: 4-dimethylaminopyridine
DMF: dimethylformamide
EDCl: N-ethyl-N, N '- (dimethylaminopropyl) carbodiimide
Et: ethyl
FCA: fluorescein carboxylic acid
Fmoc: 9-fluorenylmethoxycarbonyl
HOBt: 1-hydroxybenzotriazole
HONSu: N-hydroxysuccinimide
MBHA: 4-methyl-benzhydrylamine
Me: methyl
Mtr: 4-methoxy-2,3,6-trimethylphenyl sulfonyl
NMM: N-methylmorpholine
OBzl: benzyl ester
Oct: octanoyl
OEt: ethyl ester
OMe: methyl ester
OtBu: tert-butyl ester
POA: phenoxyacetyl
Sal: salicyloyl
TFA: trifluoroacetic acid
Trt: trityl (triphenylmethyl).

Provided the above amino acids in several enantiomers Shapes can occur, are above and below, z. B. as Be Part of the compounds of formula I, all of these forms and theirs Mixtures (e.g. the DL forms) included. Furthermore, the Ami nonacids, e.g. B. as part of compounds of formula I with ent speaking protective groups known per se.

So-called prodrug- Including derivatives, i. H. with z. B. alkyl or acyl groups, sugars or oligopeptides modified compounds of formula I, which in Or quickly to the active compounds according to the invention will split.

This also includes biodegradable polymer derivatives of the invention appropriate connections, as z. B. in Int. J. Pharm. 115, 61-67 (1995) is described.

Amino acids, the configuration of which is not specified, 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 their salts, characterized in that

• (a) a compound of formula III
  HZ-OH III
  wherein
  or
  means
  and X, A, B and C have the meanings given in claim 1,
  or treating a reactive derivative of a compound of formula III with a cyclizing agent,
  or
• b) releases a compound of formula I from one of its functional derivatives by treatment with a solvolysing or hydrogenolysing agent,
  and / or that a basic or acidic compound of the formula I is converted into one of its salts by treatment with an acid or base.

Above and below, the radicals X, A, B, C, R ¹ , R ² , m, n, p, q and Z have the meanings given in the formulas I, II and III, unless expressly stated otherwise.

In the above formulas, X preferably represents H, Hal or alkyl, in particular H, Cl or CH ₃ .

In the above formulas, alkyl has 1-6 C atoms and is preferred as for methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, also for pentyl, 1-, 2- or 3-methylbutyl, 1,1-, 1,2- or 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or 4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or 1,2,2-tri methylpropyl. Alkyl is particularly preferably methyl.

R ⁷ , R ⁸ , R ⁹ and R ^{10 are} preferably H.

The amino acids and amino acid residues mentioned can also be derivatized, the N-methyl, N-ethyl, N-propyl, N-benzyl or C _α - methyl derivatives being preferred.

Further preferred are in particular the methyl, ethyl, propyl, butyl, tert-butyl, neopentyl or benzyl ester of the side chain carboxy group, furthermore also derivatives of arginine which is based on the -NH-C (= NH) -NH ₂ group may be substituted with an acetyl, benzoyl, methoxycarbonyl or ethoxycarbonyl radical.

R ⁶ and R ⁶ , preferably mean z. B. H, methyl or ethyl, also benzyl or phenyl.

OR ⁶ preferably means z. B. hydroxy or methoxy.

COR ⁶ is alkanoyl and preferably means formyl, acetyl, propionyl, butyryl, pentanoyl or hexanoyl.

Ar is unsubstituted, preferably - as indicated - monosub substituted phenyl, in particular preferably phenyl, o-, m- or p-tolyl, o-, m- or p-ethylphenyl, o-, m- or p-propylphenyl, o-, m- or p- Isopropylphenyl, o-, m- or p-tert-butylphenyl, o-, m- or p-trifluoro methylphenyl, 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) 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, o-, m- or p- (carboxymethyloxy) phenyl, o-, m- or p- (methoxycarbonyl methyloxy) phenyl, o-, m- or p- (methoxycarbonyl-ethyloxy) phenyl, o-, m- 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-fluorine phenyl, o-, m- or p-bromophenyl, o-, m- or p-chlorophenyl, o-, m- or p- (difluoromethoxy) phenyl, o-, m- or p- (fluoromethoxy) phenyl, o-, m- or p-formylphenyl, o-, m- or p-acetylphenyl, o-, m- or p-propionyl phenyl, o-, m- or p-butyrylphenyl, o-, m- 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-, m- or p- Methylsulfonylphenyl or naphthyl.

Amino protecting group preferably means acetyl, propionyl, butyryl, Phenylacetyl, benzoyl, toluyl, POA, methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, Boc, 2-iodoethoxycarbonyl, CBZ ("Carbo benzoxy "), 4-methoxybenzyloxycarbonyl, Fmoc, Mtr or benzyl.  

The compounds of formula I have at least two chiral centers and can therefore exist in various stereoisomeric forms. Formula I encompasses all of these forms.

Accordingly, the invention relates in particular to those compounds of the formula I in which at least one of the said radicals has one of the preferred meanings indicated above. Some preferred groups of compounds can be expressed by the following sub-formulas Ia, Ib and Ic, which correspond to the formula I and in which
in Ia
XH, alkyl or Hal,
R ¹ , R ² H,
m 0,
n 3,
p, r 1, and
q 2 or 3, and
in Ib
XH, alkyl or Hal,
R ¹ , R ² H,
m 0,
n 3,
p 1,
r 0, and
q 1, and
in Ic
XH, alkyl or Hal,
R ¹ and R ² together

m 1
n 2
p, r 1, and
q 2
mean.

The compounds of formula I and also the starting materials for their manufacture position are otherwise produced by methods known per se, as described in literature (e.g. in standard works such as Houben-Weyl, Methods of Organic Chemistry, Georg-Thieme-Verlag, Stuttgart) be are written, namely under reaction conditions that are genan Implementations are known and suitable. You can also from known variants not mentioned here in detail use ma chen.

If desired, the starting materials can also be formed in situ, so that they are not isolated from the reaction mixture, but immediately further reacted to the compounds of formula I.

The compounds of formula I can, for example, according to the following Schemes 1 and 2 are produced:  

Scheme 1

Scheme 2

The important 3-amino-3- (3-nitro-phenyl) propionic acid building block from Sche ma 1 is produced according to J. Org. Chem. 25, 1758 (1960) from 3 nitro benzaldehyde, malonic acid and ammonium acetate. In the synthesis of analogue compounds, the corresponding nitrobenzal is used dehyde derivatives.

Compounds of formula I can preferably by cyclization of Compounds of formula III under the conditions of peptide synthesis be preserved. It is convenient to work with the usual metho the peptide synthesis, as z. B. in Houben-Weyl, 1.c., volume 15 / II, Pages 1 to 806 (1974).

The reaction is preferably carried out in the presence of a dehydration by means of e.g. B. a carbodiimide such as DCCl or EDCl, further z. B. Propane phosphonic anhydride (cf. Angew. Chem. 92, 129 (1980)), diphenyl phosphorylazide or 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, in an inert solvent, e.g. B. a halogenated hydrocarbon such as dichloromethane, an ether such as tetrahydrofuran or dioxane, one Amide such as DMF or dimethylacetamide, a nitrile such as acetonitrile, in Di methyl sulfoxide or in the presence of these solvents, at temperatures between about -10 and 40, preferably between 0 and 30 °. To the in tramolecular cyclization prior to intermolecular peptide binding it is advisable to work in dilute solutions.

The response time is between depending on the conditions used a few minutes and 14 days.

Instead of compounds of the formula III, derivatives of Ver compounds of the formula III, preferably a preactivated carboxylic acid, or a carboxylic acid halide, a symmetrical or mixed type hydride or an active ester can be used. Such residues for activation tion of the carboxy group in typical acylation reactions are in the Literature (e.g. in standard works such as Houben-Weyl, methods of Organic Chemistry, Georg-Thieme-Verlag, Stuttgart). Activated esters are conveniently formed in situ, e.g. B. by addition HOBt or N-hydroxysuccinimide.  

The reaction is usually carried out in an inert solvent Use of a carboxylic acid halide in the presence of an acid bin preferably an organic base such as triethylamine, Dimethylaniline, pyridine or quinoline.

Also the addition of an alkali or alkaline earth metal hydroxide, carbonate or bicarbonate or other salt of a weak acid Alkali or alkaline earth metals, preferably of potassium, sodium, calci um or cesium can be cheap.

The starting materials of formula III are usually new. You can go after known methods of peptide synthesis.

The compounds of the formula I can also be obtained by to get them from their functional derivatives by solvolysis, in particular Hydrolysis, or released by hydrogenolysis.

Preferred starting materials for solvolysis or hydrogenolysis are those which, instead of one or more free amino and / or hydroxyl groups, contain corresponding protected amino and / or hydroxyl groups, preferably those which, instead of an H atom, have an N atom is connected to carry an amino protecting group, e.g. B. those which correspond to the formula I, but instead of an NH ₂ group contain an NHR 'group (in which R' is an amino protecting group, for example Boc or CBZ).

Preference is furthermore given to starting materials which instead of the H atom Hydroxy group carry a hydroxy protecting group, e.g. B. those that the Correspond to formula I, but instead of a hydroxyphenyl group an R''O-phenyl contain group (wherein R '' means a hydroxy protecting group).

Several - identical or different - protected amino and / or Hydroxy groups present in the molecule of the starting material his. If the existing protecting groups are different from each other they can be split off selectively in many cases.  

The term "amino protecting group" is well known and refers to on groups that are suitable, an amino group before chemical order to protect (block) settlements that are easily removable, after the desired chemical reaction elsewhere in the Molecule has been carried out. Typical of such groups are ins special unsubstituted or substituted acyl, aryl, aralkoxymethyl or Aralkyl groups. Since the amino protecting groups according to the desired Reaction (or sequence of reactions) to be removed is their type and size the rest not critical; however, preference is given to those with 1-20, in particular special 1-8 carbon atoms. The term "acyl group" is related with the present procedure in the broadest sense. He around includes aliphatic, araliphatic, aromatic or hetero cyclic carboxylic acids or sulfonic acids derived acyl groups so such as especially alkoxycarbonyl, aryloxycarbonyl and especially aral koxycarbonyl groups. Examples of such acyl groups are alkanoyl such as acetyl, propionyl, butyryl; Aralkanoyl such as phenylacetyl; Aroyl like Ben zoyl or toluyl; Aryloxyalkanoyl such as POA; Alkoxycarbonyl such as methoxy carbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, Boc, 2- Iodoethoxycarbonyl; Aralkyloxycarbonyl such as CBZ ("carbobenzoxy"), 4- Methoxybenzyloxycarbonyl, Fmoc; Arylsulfonyl such as Mtr. Preferred ami Protective groups are Boc and Mtr, also CBZ, Fmoc, Benzyl and Acetyl.

The term "hydroxy protecting group" is also well known and refers to groups that are suitable for a hydroxy group to protect chemical reactions, which are easy to remove after the desired chemical reaction elsewhere in the Molecule has been carried out. They are typical of such groups unsubstituted or substituted aryl, aralkyl or Acyl groups, also alkyl groups. The nature and size of the hydroxy Protecting groups are not critical as they are chemical-based Reaction or sequence of reactions are removed again; are preferred Groups with 1-20, especially 1-10 carbon atoms. Examples of hydroxy protecting groups are a. Benzyl, p-nitrobenzoyl, p-toluenesulfonyl, tert.- Butyl and acetyl, with benzyl and tert-butyl being particularly preferred. The COOH group is preferably protected in the form of its tert-butyl ester.  

The liberation of the compounds of formula I from their radio tional derivatives succeed - depending on the protective group used - e.g. B. with strong acids, suitably with TFA or perchloric acid, but also with other strong inorganic acids such as hydrochloric acid or sulfur acid, strong organic carboxylic acids such as trichloroacetic acid or Sulfonic acids such as benzene or p-toluenesulfonic acid. The presence of an egg Additional inert solvent is possible, but not always conducive. Suitable inert solvents are preferably organic, for example carboxylic acids such as acetic acid, ethers such as tetrahydrofuran or dioxane, amides such as DMF, halogenated hydrocarbons such as Dichloromethane, also alcohols such as methanol, ethanol or isopropanol, as well as water. Mixtures of the aforementioned Lö also come means in question. TFA is preferably in excess without addition another solvent used, perchloric acid in the form of a Mixture of acetic acid and 70% perchloric acid in a ratio of 9: 1. The reaction temperatures for the cleavage are advantageously between about 0 and about 50 °, preferably between 15 and 30 ° (Room temperature).

The groups Boc, OtBu and Mtr can e.g. B. preferably with TFA in Di chloromethane or with about 3 to 5 N HCl in dioxane at 15-30 ° the Fmoc group with an approximately 5 to 50% solution of Di methylamine, diethylamine or piperidine in DMF at 15-30 °.

The trityl group is used to protect the amino acids histidine, asparagine, Glutamine and cysteine are used. The separation takes place, depending on ge desired end product, with TFA / 10% thiophenol, the trityl group is split off from all the amino acids mentioned, when using TFA / anisole or TFA / thioanisole only the trityl group of histidine, Aspa split off ragin and glutamine, whereas on the cysteine side chain remains.

Hydrogenolytically removable protective groups (e.g. CBZ or benzyl) can e.g. B. by treatment with hydrogen in the presence of a kata lysators (z. B. a noble metal catalyst such as palladium, appropriate on a carrier such as coal). As a solvent  NEN the above, especially z. B. alcohols such as Methanol or ethanol or amides such as DMF. The hydrogenolysis is in usually at temperatures between about 0 and 100 ° and pressure between rule about 1 and 200 bar, preferably at 20-30 ° and 1-10 bar guided. Hydrogenolysis of the CBZ group succeeds e.g. B. good at 5 to 10% Pd / C in methanol or with ammonium formate (instead of what serstoff) on Pd / C in methanol / DMF at 20-30 °.

A base of formula I can with an acid in the associated acid addition salt can be transferred, for example by reaction equi valent amounts of the base and the acid in an inert solvent such as ethanol and subsequent evaporation. For this implementation men in particular acids, the physiologically acceptable Sal ze deliver. So inorganic acids can be used, e.g. B. Sulfuric acid, nitric acid, hydrohalic acids such as chlorine hydrochloric acid or hydrobromic acid, phosphoric acids such as ortho phosphoric acid, sulfamic acid, also organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyc misch mono- or polybasic carboxylic, sulfonic or sulfuric acids, e.g. B. Formic acid, acetic acid, propionic acid, pivalic acid, diethyl acetic acid acid, malonic acid, succinic acid, pimelic acid, fumaric acid, malein acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, Ascorbic acid, nicotinic acid, isonicotinic acid, methane or ethanesulfone acid, ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfone acid, p-toluenesulfonic acid, naphthalene mono- and disulfonic acids, lauryl sulfuric acid. Salts with physiologically unacceptable acids, e.g. B. Picrates, can be used to isolate and / or purify the compounds of formula I can be used.

On the other hand, an acid of the formula I can be reacted with a Ba se in one of their physiologically harmless metal or ammonium salts are transferred. The Na come in particular as salts trium, potassium, magnesium, calcium and ammonium salts into consideration, further substituted ammonium salts, e.g. B. the dimethyl, diethyl or di  isopropyl ammonium salts, monoethanol, diethanol or diisopropyl ammonium salts, cyclohexyl, dicyclohexylammonium salts, dibenzyl ethylenediammonium salts, further z. B. salts with arginine or lysine.

The invention further relates to the use of the compounds of formula I and / or their physiologically acceptable salts provision of pharmaceutical preparations, especially on non- chemical way. You can do this together with at least one solid, liquid and / or semi-liquid carrier or auxiliary and counter possibly in combination with one or more other active ingredients a suitable dosage form.

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

These preparations can be used as medicinal products in human or veteri used in medicine. Organic or inorganic substances in question that are suitable for enteral (e.g. oral), parenteral, topical application or for an application in the form of a Inhalation sprays are suitable and do not react with the new compounds ren, for example water, vegetable oils, benzyl alcohols, alkylene glycol le, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as Lac tose or starch, magnesium stearate, talc, petroleum jelly. For oral application tablets, pills, coated tablets, capsules, pul ver, granules, syrups, juices or drops, for rectal use Sup positions, for parenteral use solutions, preferably oily or aqueous solutions, also suspensions, emulsions or implanta for topical application of ointments, creams or powder. The new Compounds can also be lyophilized and the lyophilizates obtained e.g. B. can be used for the preparation of injectables. The on given preparations can be sterilized and / or auxiliary substances such as Lubricants, preservatives, stabilizers and / or wetting agents, emulsifiers Ren, salts to influence the osmotic pressure, buffer substances, Contain color, taste and / or several other active ingredients, for. B. one or more vitamins.  

For the application as an inhalation spray, sprays can be used who contain the active ingredient either dissolved or suspended in a propellant gas or propellant mixture (e.g. CO ₂ or chlorofluorocarbons). Appropriately, the active ingredient is used in micronized form, and one or more additional physiologically compatible solvents may be present, for. B. ethanol. Inhalation solutions can be administered using standard inhalers.

The compounds of formula I and their physiologically acceptable Salts can act as integrin inhibitors in fighting diseases, in particular of diseases of the circulatory system, thrombosis, heart attack, coronary heart disease, arteriosclerosis, apoplexy, angina pectoris, Tumors, osteoporosis, inflammation, infection and restenosis after Angioplasty can be used.

The compounds of formula I according to claim 1 and / or their physiolo gisch harmless salts are also used in pathological Processes that are maintained or propagated through angiogenesis, especially for tumors or rheumatoid arthritis.

The substances according to the invention can generally be found in Ana logie zu other known, commercially available peptides, esp special but in analogy to that described in US-A-4,472,305 Compounds are administered, preferably in doses between about 0.05 and 500 mg, especially between 0.5 and 100 mg per dose unit administered. The daily dosage is preferably between around 0.01 and 2 mg / kg body weight. The special dose for everyone However, patients depend on various factors, for example as from the effectiveness of the special compound used, from Age, body weight, general health, gender, from the diet, the time and route of administration, the excretion application rate, drug combination and severity of each Disease to which the therapy applies. The parenteral application is be prefers.  

Furthermore, the compounds of formula I as integrin ligands for Manufacture of columns for affinity chromatography for pure Darst integrins can be used.

The ligand, i.e. H. a compound of formula I, is over a Anchor function, e.g. B. the free carboxy group on a polymeric support covalently coupled.

The polymeric carrier materials which are in themselves in the peptide are suitable Chemically known polymeric solid phases with preferably hydro phile properties, for example cross-linked poly sugars such as Cellu loose, Sepharose or Sephadex®, acrylamides, polymer on polyethylene glycol-based or tentacle polymer®.

The manufacture of materials for affinity chromatography for inte Grin cleaning takes place under conditions such as those for the condensation of Amino acids are common and known per se.

The compounds of formula I contain at least two chiral centers and can therefore be in racemic or optically active form. Racemates obtained can be mecha by methods known per se nisch or chemically separated into the enantiomers. Preferably are from the racemic mixture by reaction with an op table active release agent diastereomers formed. Suitable as a release agent z. B. optically active acids, such as the D and L forms of tartaric acid, Diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, milk acid or the various optically active camphorsulfonic acids such as β-camphorsulfonic acid. Enantiomer separation is also advantageous with the help of an optically active release agent (e.g. dinitrobenzoyl phenylglycine) filled column; as a solvent is z. B. a mixture Hexane / isopropanol / acetonitrile, e.g. B. in a volume ratio of 82: 15: 3.

Of course, it is also possible to use optically active compounds of the formula I. according to the methods described above, by choosing Aus used materials that are already optically active.  

All temperatures above and below are given in ° C. Room temperature means 22 ° C. In the following examples, "customary work-up" means: if necessary, water is added, and if necessary, depending on the constitution of the end product, the pH is between 2 and 10, extracted with ether or dichloromethane, the organic is separated Phase, the organic phase dries over sodium sulfate, filtered, evaporated and purified by chromatography on silica gel and / or by crystallization.
RT = retention time (minutes) with HPLC in the following systems:
Column: Lichrosorb® RP 18 (250 × 4; 5 µm);
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 conditions specified.
Mass spectrometry (MS): FAB (Fast Atom Bombardment) (M + H) ⁺

example 1

Synthesis of the compound to be cyclized

Analogous to Scheme 1, (3R, S) -3-amino-3- (4-methyl-3-nitro-phenyl) propion Acid methyl ester (3R, S-IX) synthesized. This ester is after known method split into the enantiomers and (3S) -3-amino-3- (4-me methyl-3-nitro-phenyl) propionic acid methyl ester (3S-IX) then analogous to Scheme 2 for the Mtr-protected compound 3S-amino-3- (3- {3- [1- (carb oxy-methyl-carbamoyl) -4-guanidino-1S-butylcarbamoyl] -propionyl amino} -4-methyl-phenyl) propionic acid methyl ester (S, S-XIV) implemented.

Cyclization

616 mg of the Mtr-protected compound 3S-amino-3- (3- {3- [1- (carboxy methyl-carbamoyl) -4-guanidino-1S-butylcarbamoyl] -propionylamino} -4-methyl-phenyl) -propionic acid methyl ester (S, S-XIV) are dissolved in 80 ml DMF and diluted with 800 ml dichloromethane. It is then cooled to -20 ° C. and 300 mg of EDCl, 98 mg of DMAP and 0.176 ml of NMM are added in succession. It is warmed to room temperature and stirred overnight. The solution is concentrated and the residue is stirred into 200 ml of semi-saturated NaHCO ₃ solution. The precipitate is suctioned off and washed. 400 mg of substance are obtained, which are purified by preparative HPLC. After chromatography, 44 mg of the Mtr-protected cyclic compound (8S, 14S) - [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetra oxo-2,7,10,13 are obtained -tertraazabicyclo [13.3.1] nonadeca-1 (18), 15 (19), 16-trien-14-yl] -acetic acid methyl ester, RT 26.1; FAB 716.

Saponification and cleavage of the Mtr protective group

Saponification in KOH / methanol gives the Mtr-protected cy clical compound (8S, 14S) - [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetra oxo-2,7,10,13-tertraazabicyclo [13.3.1] nonadeca-1 (18), 15 (19), 16-trien-14-yl] vinegar acid. 25 mg of this compound are then in 4.3 ml of 98% trifluoroacetic acid dissolved and overnight at room temperature touched. The solution is spun in and purified by preparative HPLC. 9.5 mg (8S, 14S) - [8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetra oxo-2,7,10,13-tertraazabicyclo [13.3.1] nonadeca-1 (18), 15 (19), 16-trien-14-yl] vinegar Acid (8S, 14S-IB) obtained, RT 20.2; FAB 490.  

Examples 2-3

Example 2 (q = 2)

Starting from 85 mg of the Mtr-protected compound (8S, 14S) -2- (8- (3- Guanidinopropyl) -3,6,9,12-tetraoxo-2,7,10,13-tertraazabicyclo [13.3.1] nona deca-16,18,19-trien-14-yl) acetic acid is obtained analogously to Bei game 1 by reaction with 14.7 ml of 98% trifluoroacetic acid 13 mg (8S, 14S) -2- (8- (3-guanidinopropyl) -3,6,9,12-tetraoxo-2,7,10,13-tertraaza bicyclo [13.3.1] nonadeca-16,18,19-trien-14-yl) acetic acid; RT 17.2; FAB 476.

Example 3 (q = 3)

Starting from 300 mg of the Mtr-protected compound (9S, 15S) -2- (9- (3- Guanidinopropyl) -3,7,10,13-tetraoxo-2,8,11,14-tertraazabicyclo [14.3.1] -eico san-17,19,20-trien-15-yl) acetic acid is obtained analogously to Bei game 1 74 mg (9S, 15S) -2- (9- (3-guanidinopropyl) -3,7,10,13-tetraoxo-2,8,11,14-tertra azabicyclo [14.3.1] eicosan-17,19,20-trien-15-yl) vinegar acid; RT 18.3; FAB 490.  

Example 4

Starting from the Mtr-protected compound (6S, 12S) - [6- (3-guani dinopropyl-4,7,10-trioxo-2,5,8,11-tertaazabicyclo [11.3.1] heptadeca-1 (17), 13,15-trien-12-yl] vinegar Acid is obtained analogously to Example 1 (6S, 12S) - [6- (3-guanidinopropyl-4,7,10-trioxo-2,5,8,11-tertaazabicyclo- [11.3.1] heptadeca-1 (17), 13,15-trien-12-yl] acetic acid.

Examples 5-8

The following compounds are synthesized in analogy to Example 1:

The following examples relate to pharmaceutical preparations.

Example A: Injection glasses

A solution of 100 g of an active ingredient of the formula I and 5 g of disodium Hydrogen phosphate is dissolved in 3 liters of double distilled water with 2N salt acid adjusted to pH 6.5, sterile filtered, filled into injection glasses, lyophilized under sterile conditions and sealed sterile. Every In jection glass contains 5 mg of active ingredient.

Example B: Suppositories

A mixture of 20 g of an active ingredient of the formula I is melted with 100 g soy lecithin and 1400 g cocoa butter, pour into molds and leave cold Each suppository contains 20 mg of active ingredient.

Example C: solution

A solution is prepared from 1 g of an active ingredient of the formula I, 9.38 g of NaH ₂ PO ₄ .2 H ₂ O, 28.48 g of Na ₂ HPO ₄ .12 H ₂ O and 0.1 g of benzalkonium chloride in 940 ml double distilled water. It is adjusted to pH 6.8, made up to 1 l and sterilized by irradiation. This solution can be used in the form of eye drops.

Example D: ointment

500 mg of an active ingredient 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 formula I, 4 kg lactose, 1.2 kg kar Potato starch, 0.2 kg talc and 0.1 kg magnesium stearate are common Formed into tablets in such a way that each tablet contains 10 mg of active ingredient contains.

Example F: coated tablets

Analogously to Example E, tablets are pressed, which are then made in the usual manner Wise with a coating of sucrose, potato starch, talc, tragacanth and dye are coated.

Example G: capsules

2 kg of active ingredient of formula I are in the usual way in hard gelatin capsules filled so that each capsule contains 20 mg of the active ingredient.

Example H: ampoules

A solution of 1 kg of active ingredient of formula I in 60 l of double distilled Water is sterile filtered, filled into ampoules, under sterile conditions lyophilized and sealed sterile. Each ampoule contains 10 mg Active ingredient.  

Example I: Inhalation spray

14 g of active ingredient of the formula I are dissolved in 10 l of isotonic NaCl solution and fills the solution into commercially available spray vessels with a pump mechanism. The solution can be sprayed into the mouth or nose. A spray (about 0.1 ml) corresponds to a dose of about 0.14 mg.

Claims (10)

1. Compounds of formula I.
wherein
A Gly, Ala or NH-NH-CO,
the amino acids mentioned can also be derivatized,
B is a radical of formula II
C- (CO) _p - (CH ₂ ) _q - (CO) _r - or - (CO) _p -CH = CH- (CO) _r -,
m, p, r each independently of one another 0 or 1,
n, q each independently of one another 1, 2, 3 or 4,
R ¹ and R ^{2 are} each independently of one another H or alkyl,
R ¹ and R ² together also
R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each independently of one another H, alkyl, Ar, OR ⁶ , Hal, NO ₂ , NR ⁶ R ^{6 '} , NHCOR ⁶ , CN, NHSO ₂ R ⁶ , COOR ⁶ or COR ⁶ ,
XH, Hal, alkyl or Ar,
Ar is unsubstituted or mono-, di- or trisubstituted by R ³ , R ⁴ or R ⁵ phenyl or unsubstituted naphthyl,
R ³ , R ⁴ , R ⁵ each 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 '} each independently of one another H, alkyl, phenyl or benzyl, and
Hal F, Cl, Br or I
mean,
where, in the case of residues of optically active amino acids and amino acid derivatives, both the D and the L forms are included,
as well as their salts. 2. An enantiomer or a diastereomer of a compound of the formula according to claim 1. 3. Compounds of formula I according to claim 1:

• a) (8S, 14S) -2- (8- (3-guanidinopropyl) -3,6,9,12-tetraoxo-2,7,10,13-tetra azabicyclo [13.3.1] nonadeca-16,18, 19-trien-14-yl) acetic acid;
• b) (9S, 15S) -2- (9- (3-guanidinopropyl) -3,7,10,13-tetraoxo-2,8,11,14-tetra azabicyclo [14.3.1] eicosan-17,19, 20-trien-15-yl) acetic acid;
• c) (8S, 14S) - (8- (3-guanidinopropyl) -18-methyl-3,6,9,12-tetraoxo-2,7,10,13-tetra azabicyclo [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-tetra azabicyclo [11.3.1] heptadeca-1 (17), 13.15 -trien-12-yl) -acetic acid;
  as well as their salts.

4. A process for the preparation of compounds of formula I according to claim 1 and their salts, characterized in that

• (a) a compound of formula III
  HZ-OH III
  wherein
  or
  means
  and X, A, B and C have the meanings given in claim 1,
  or treating a reactive derivative of a compound of formula III with a cyclizing agent,
  or
• b) liberates a compound of the formula I from one of its functional derivatives by treatment with a solvolysing or hydro genolysing agent,

and / or that a basic or acidic compound of the formula I is converted into one of its salts by treatment with an acid or base. 5. Process for the preparation of pharmaceutical preparations, thereby characterized in that a compound of formula I according to An saying 1 and / or one of their physiologically acceptable salts together with at least one solid, liquid or semi-liquid Bring carrier or excipient into a suitable dosage form. 6. Pharmaceutical preparation, characterized by a content at least one compound of formula I according to claim 1 and / or one of their physiologically acceptable salts. 7. Compounds of formula I according to claim 1 and their physiological harmless salts as integrin inhibitors for combating Circulatory disorders, thrombosis, heart attack, coronary artery Heart disease, arteriosclerosis, apoplexy, angina pectoris, Tu moren, osteoporosis, inflammation, infection and restenosis after angioplasty. 8. Use of compounds of formula I according to claim 1 and / or their physiologically acceptable salts in pathologi processes that entertain through angiogenesis or propa be greeded. 9. Use of compounds of formula I according to claim 1 and / or their physiologically acceptable salts for the preparation of a drug. 10. Use of compounds of formula I according to claim 1 and / or their physiologically acceptable salts in the fight disease control.