AU772782B2 - Cyclic peptide derivatives as inhibitors of integrin alphavbeta6 - Google Patents

Description

WO 01/05810 1 PCT/EP00/06188 Cyclic peptide derivatives as inhibitors of the integrin OcvP The invention relates to novel peptide derivatives of the formula I cyclo(Arg-X-Asp-X 2

-X

3

-X

4

-X

5

-X

6

-R

l

I

in which

X

1 is Ser, Gly or Thr,

X

2 is Leu, Ile, Nle, Val or Phe,

X

3 is Asp, Glu, Lys or Phe,

X

4 is Gly, Ala or Ser,

X

5 is Leu, Ile, Nle, Val or Phe,

X

6 is Arg, Har or Lys,

R

1 is absent or is one or more o-aminocarboxylic acid residue(s), the (-aminocarboxylic acid residue(s) having a length of 500 to 2500 pm, where the amino acids mentioned can also be derivatized, the D and the L forms of the optically active amino acid residues are included, and their physiologically acceptable salts and i. solvates.

The invention seeks to provide finding novel compounds S having valuable properties, in particular those which can be used for the production of medicaments.

It has been found that the compounds according to the invention and their salts have very valuable .0 pharmacological properties together with good tolerability.

The peptides according to the invention can be employed as efficacious inhibitors of the avp integrin receptor 2 and thus for the treatment of various diseases and pathological findings.

Other inhibitors of the integrin o46P are described in DE 19858857 and by S. Kraft et al. in J. Biol. Chem.

274, 1979-85 (1999). The compounds according to the invention are to be considered as a selection invention with respect to the application mentioned.

Integrins belong to the family of heterodimers of Class I transmembrane receptors which play an important role in numerous cell-matrix or cell-cell adhesion processes (Tuckwell et al., 1996, Symp. Soc. Exp. Biol.

47). They can be roughly divided into three classes: the P1 integrins, which are receptors for the extracellular matrix, the P2 integrins, which are activatable on leucocytes and are "triggered" during inflammatory processes, and the o, integrins, which influence the cell response during wound-healing and other pathological processes (Marshall and Hart, 1996, Semin. Cancer Biol. 7, 191).

The integrins c 5 x 1 XIIb 3 8sP1 v 1 43, 5vP5, CvP8 and avP 6 all bind to the Arg-Gly-Asp (RGD) peptide sequence in natural ligands, such as, for example, fibronectin or vitronectin. Soluble RGD-containing peptides are able to inhibit the interaction of each of these integrins with the corresponding natural ligands. o06P is a relatively rare integrin (Busk et al., 1992 J.

Biol. Chem. 267(9), 5790), which is formed to an increased extent in repair processes in epithelial tissue and preferably binds the natural matrix molecules fibronectin and tenascin (Wang et al., 1996, Am. J. Respir. Cell Mol. Biol. 15(5), 664). The physiological and pathological functions of av 6 are still not precisely known; it is suspected, however, that this integrin plays an important role in physiological processes and disorders (e.g.

inflammation, wound healing, tumours) in which 3 epithelial cells are involved. Thus 0CP 6 is expressed on keratinocytes in wounds (Haapasalmi et al., 1996, J.

Invest. Dermatol. 106(1), 42), from which it is to be assumed that in addition to wound-healing processes and inflammation, other pathological skin events, such as, for example, psoriasis, can also be influenced by agonists or antagonists of the said integrin. uvP 6 furthermore plays a role in the respiratory tract epithelium (Weinacker et al., 1995, Am. J. Respir. Cell Mol. Biol. 12(5), 547), so that appropriate agonists/antagonists of this integrin could be successfully employed in respiratory tract disorders, such as bronchitis, asthma, pulmonary fibrosis and respiratory tract tumours. Finally, it is known that 0oPs also plays a role in the intestinal epithelium, so that appropriate integrin agonists/antagonists could be used in the treatment of inflammation, tumours and wounds of the stomach/intestinal tract.

The dependence of the formation of angiogenesis on the interaction between vascular integrins and extracellular matrix proteins is described by P.C. Brooks, R.A. Clark and D.A. Cheresh in Science 264, 569-71 (1994).

9 The present seeks to provide therefore, (in addition to the previously known natural high molecular weight ligands and antibodies, which are therapeutically and diagnostically difficult to handle), potent, specific and "00 selective low molecular weight ligands for oCVP 6 S preferably peptides, which can be used for the therapeutic o** areas mentioned, but also diagnostic or reagent.

It has been found that the peptide compounds according to the invention and their salts, as soluble molecules, exert an effect on cells which carry the said receptor, or, if they are bound to surfaces, are artificial ligands for av 6 -mediated cell adhesion. Above all, they 4 act as cvB 6 integrin inhibitors, where they particularly inhibit the interactions of the receptor with other ligands, such as, for example, the binding of fibronectin. This action can be detected, for example, by the method which is described by J.W. Smith et al. in J. Biol. Chem. 265, 12267-12271 (1990).

It has furthermore been found that the novel substances have very valuable pharmacological properties together with good tolerability and can be employed as medicaments. This is described more precisely further below.

The peptide compounds according to the invention can furthermore be used in vivo and in vitro as diagnostics for the detection and localization of pathological conditions in the epithelial system, if they are equipped with appropriate markers the biotinyl radical) according to the prior art.

The invention also comprises combinations with at least one other active compound and/or conjugates with other active compounds, such as cytotoxic active compounds and conjugates with radiolabels for X-ray therapy or PET diagnosis, but also fusion proteins with marker proteins such as GFP or antibodies, or therapeutic proteins such as IL-2.

Particularly active compounds are those of the formula I in which an octapeptide sequence cyclo(Arg-X 1 -Asp-X 2 X -X4-X-X6) in which the radicals X 1

X

2

X

3

X

4

X

5 and

X

6 have the meanings indicated, is expanded by R 1 The effect of the ring expansion is shown in Fig. 1 by the example of cyclo(Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg) [EMD 271588]. Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-NH2 [EMD 271293] serves as a comparison compound.

Some cyclo(Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-R) peptides, the distance from R 1 (calculated) and the value Q (IC50[substance]/IC 0 [EMD 271293]) and -log Q are given in the following table.

5 R Distance [pm] Q -log Q 0 0 47 -1.672 Gly 370 2.1 -0.322 Abu 617 0.028 1.553 Gly-Gly 740 0.05 1.301 Aha 870 0.036 1.444 Aee 1078 0.038 1.420 Gly-Gly-Gly 1110 0.03 1.523 Abu-Abu 1235 0.03 1.523 Gly-Gly-Gly-Gly 1480 0.033 1.481 Aha-Aha 1740 0.036 1.444 Gly-Gly-Gly-Gly-Gly 1850 0.046 1.337 Gly-(Gly) 4 -Gly 2220 0.05 1.301 The graphic representation can be seen in Fig. 1.

Particularly active compounds are obtained even if the spacer length R 1 has reached approximately 500 pm.

Particularly preferred compounds of the formula I are those in which R 1 is one or more o-aminocarboxylic acid residue(s), the n-aminocarboxylic acid residue(s) having a length of 600 to 2500 pm, very particularly preferably those having a spacer length of 600 to 2000 pm.

o-Aminocarboxylic acid residue(s) is/are understood as meaning any o-aminocarboxylic acid, among which the amino acids below selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hey, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val and H 2

N-(CH

2

CH

2 0)m- (CH 2 )n-COOH, where m, n in each case independently of one another are 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, with the proviso that m n is 0, are particularly preferred.

6 The invention thus preferably relates to peptide derivatives according to Claim 1, of the formula I Cyclo (Arg-X-Asp-X -X -X4-X5-X6-R in which

X

1

X

2

X

3

X

4

X

5

X

6

R

1 m,n is Ser, Gly or Thr, is Leu, Ile, Nle, Val or Phe, is Asp, Glu, Lys or Phe, is Gly, Ala or Ser, is Leu, Ile, Nle, Val or Phe, is Arg, Har or Lys, is absent or is 1-10 amino acids selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hcy, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val and

H

2 N- (CH 2

CH

2 0)m- (CH 2 )n-COOH, in each case independently of one another are 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, with the proviso that m n is 0, where the amino acids mentioned can also be derivatized, the D and the L forms of the optically active amino acid residues are included, and their physiologically acceptable salts.

Some preferred groups of compounds can be expressed by the following subformulae Ia to Ih, which correspond to the formula I and in which the radicals not designated in greater detail have the meaning [sic] indicated in the formula I, but in which in a) X 1 is Gly or Thr; in b) X 1

X

2 is Gly or Thr, is Leu; 7 in c) X 1 is Gly or Thr,

X

2 is Leu,

X

3 is Asp or D-Asp; in d) X 1 is Gly or Thr,

X

2 is Leu,

X

3 is Asp or D-Asp,

X

4 is Gly or Ala; in e) X 1 is Gly or Thr,

X

2 is Leu,

X

3 is Asp or D-Asp,

X

4 is Gly or Ala,

X

5 is Leu; in f) X 1 is Gly or Thr,

X

2 is Leu,

X

3 is Asp or D-Asp,

X

4 is Gly, Ala or Ser,

X

5 is Leu,

X

6 is Arg; in g) X 1 is Gly or Thr,

X

2 is Leu,

X

3 is Asp or D-Asp,

X

4 is Gly, Ala or Ser,

X

5 is Leu,

X

6 is Arg;

R

1 is 1-10 amino acids selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hey, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val and H 2 N- (CH 2

CH

2 0)m- (CH 2 )n-COOH, m,n in each case independently of one another are 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, with the proviso that m n is 0; 8in h) xi is Gly or Thr, *2 is Leu, *3 is Asp or D-Asp, *4 is Gly, Ala or Ser, x 5 is Leu, *6 is Arg; Ri is 1-6 amino acids selected from the group consisting of Gly, 13-Ala, Abu or Aha; and their salts.

The invention relates in particular to peptide compounds selected from the group consisting of cyclo (Arg-Gly-Asp-Leu-Asp-Ala-Leu-Arg-Gly-Gly-Gly), cyclo (Arg-Gly-Asp-Leu-Asp-Gly-Leu-Arg-Gly-Gly-Gly), cyclo (Arg-Gly-Asp-Leu-D-Ala-Ala-Leu-Arg-Gly-Gly-Gly), cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Gly-Gly-Gly), cyclo(Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-A-bu-Abu), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aha-Aha), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aha), cyclo(Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aee), cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu), cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-3-Ala), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-3-Ala), and their physiologically acceptable salts.

The abbreviations of amino acid residues mentioned above and below stand for the radicals of the following amino acids: Abu 4-aminobutyric acid Aee H 2 N- (CH 2

CH

2 O) 2

-CH

2

COOH

Aha 6-aminohexanoic acid, 6-aminocaproic acid Aib ax-aminoisobutyric acid Ala alanine Asn asparagine 9- Asp Arg Bgl Cys Dab Dap Gin Gip Glu Gly Har Hcy His hono- Phe Hse Ile Leu Lys Met Nal Nie Orn Pen Phe Phg 4-Hal-Phe Pro Ser Thr

TIS

Trp Tyr Val aspartic acid arginine C-alpha- tert-butyiglycine cysteine 2, 4-diaminobutyric acid 2, 3-diaminopropionic acid glut amine pyroglutamic acid glutamic acid glycine homoarginine homocyst eime histidine homophenylalanine homoserine isoleucine leucine lysine methionine naphth-2 -ylalanine norleucine orni thine penicillamine phenylalanine phenyiglycime 4-halophenylalanine proline serine threonine triisopropylsilane (sic] tryptophan tyrosine valine.

In addition, the following have the meanings below: Ac acetyl BOC tert-butoxycarbonyl* BSA bovine serum albumin 10 CBZ or Z DCCl

DCM

DIEA

DMF

EDC1 Et

FCA

FITC

Fmoc

FTH

HOBL

Me

MBHA

Mtr HONSu OBut Oct OMe OEt Pbf Pmc

POA

Sal

TBS++

TBSA

TBTU

benzyloxycarbonyl dicyclohexylcarbodiimide dichioromethane diethylamine dimethylformamide N-ethyl-N,N'-(dimethylaminopropyl)carbodiimide ethyl fluoresceincarboxylic acid fluorescein isothiocyaate 9-fluorenylmethoxycarbonyl fluoresceinthiourea l-hydroxybenzotriazole methyl 4-methylbenzhydrylamine 4-methoxy-2,3,6-trimethylphenylsulfonyl N-hydroxysuccinimide tert-butyl ester octanoyl methyl ester ethyl ester 2,2,4,6,7-pentamethyldihydrobenzofuran-5sulfonyl 2,2,5,7,8-pentamethylchroman-6-sulfonyl phenoxyacetyl salicyloyl tris buffered saline with divalent cations TBS BSA 2-(1H-benzotriazol-1-yl)-1,1,3tetramethyluronium tetrafluoroborate trifluoroacetic acid trityl (triphenylmethyl).

TFA

Trt If the abovementioned amino acids can occur in two or more enantiomeric forms, all these forms and their mixtures (for example the DL forms) are included above and below. In addition, the amino acids can be provided with appropriate protective groups which are known per se.

I 11 The invention also relates to the hydrates and solvates, e.g. alcoholates, of these compounds.

So-called prodrug derivatives are included in the compounds according to the invention, that is compounds modified with, for example, alkyl or acyl groups, sugars or oligopeptides, which are cleaved rapidly in the body to give the active compounds according to the invention. 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).

The amino acids and amino acid residues mentioned, such as, for example, the NH functions or terminal amide functions, can also be derivatized, the N-methyl, Nethyl, N-propyl, N-benzyl or Ca-methyl derivatives being preferred. Derivatives which are additionally preferred are those of Asp and Glu, in particular the methyl, ethyl, propyl, butyl, tert-butyl, neopentyl or benzyl esters of the side chain carboxyl groups, and in addition also derivatives of Arg, which can be substituted on the -NH-C(=NH)-NH 2 group by an acetyl, benzoyl, methoxycarbonyl or ethoxycarbonyl radical.

In the compounds according to the invention, which are linked to one another in peptide fashion via the a-amino and a-carboxy groups (head-tail linkage), those cyclic compounds are also contained which, e.g. in the presence of a functional side chain, such as, for example, an SH group, are linked in the following way side-side e.g. S-S (disulfide), head-side side-tail.

Furthermore, derivatives additionally included in the compounds according to the invention are those which 12 consist of the actual peptides according to the invention and known marker compounds which make it possible to detect the peptides easily. Examples of such derivatives are radiolabelled, biotinylated or fluorescence-labelled peptides.

A fluorescent dye radical is preferably 7-acetoxycoumarin-3-yl, fluorescein-5-(and/or 6-)yl, 6-)yl, dihydrotetramethylrosamin-4-yl, tetramethylrhodamin-5-(and 6-)yl, 4,4difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3ethyl or 4 4 -difluoro-5,7-diphenyl-4-bora-3a,4a-diazas-indacene-3-ethyl.

Suitable functionalized fluorescent dye radicals which can serve as reagents for the preparation of the compounds of the formula I according to the invention are described, for example, in "Handbook of Fluorescent Probes and Research Chemicals, 5 th Edition, 1992-1994, by R.P. Haughland, Molecular Probes, Inc.".

The invention comprises not only the peptides mentioned but also mixtures and preparations which, in addition to these compounds according to the invention, also contain other pharmacological active compounds or adjuvants which can influence the primary pharmacological action of the peptides according to the invention in a desired manner.

The compounds according to the invention and also the starting substances for their preparation are otherwise prepared by methods which are known per se and frequently employed, such as are described in the literature 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. Use can also be made in this case of variants which are known per se.

13 Preferably, the peptides according to the invention can be prepared by means of solid-phase synthesis and subsequent removal and purification, as was described, for example, by Jonczyk und Meienhofer (Peptides, Proc.

8 th Am. Pept. Symp., Eds. V. Hruby and D.H. Rich, Pierce Comp. III, pp. 73-77, 1983, or Angew. Chem. 104, 1992, 375) or according to Merrifield Am. Chem. Soc. 94, 1972, 3102).

The peptides according to the invention can be prepared on a solid phase (manually or in an automated synthesizer) in an Fmoc strategy using acid-labile side protective groups and purified by means of RP-HPLC. The peak homogeneity can be measured by RP-HPLC and the substance identity by means of FAB-MS.

Otherwise, the peptides can be prepared by customary methods of amino acid and peptide synthesis, such as is known, for example, from Novabiochem 1999 Catalog Peptide Synthesis Handbook of Calbiochem-Novabiochem GmbH, D-65796 Bad Soden, from numerous standard works and published patent applications.

Stepwise couplings and fragment condensations can be utilized. Different N-terminal, C-terminal and side protective groups can be used, which are preferably selected to be orthogonally cleavable. Coupling steps can be carried out using different condensing reagents such as carbodiimides, carbodiimidazole, those of the uronium type such as TBTU, mixed anhydride methods, and acid halide or active ester methods. Activated esters are expediently formed in situ, for example by addition of HOBt or N-hydroxysuccinimide.

The cyclization of a linear precursor molecule having side protective groups can likewise be carried out using such condensation reactions, as is described, for example, in DE 43 10 643 or in Houben-Weyl, l.c., Volume 15/II, pages 1 to 806 (1974).

14 Different resins and anchor functions can be utilized in the solid-phase peptide synthesis. Resins can be based, for example, on polystyrene or polyacrylamide, anchor functions such as Wang, o-chlorotrityl are utilizable for the preparation of peptide acids, aminoxanthenoxy anchors, for example for the preparation of peptide amides.

Biotinylated or fluorescence-labelled peptides/proteins can likewise be prepared by standard methods (for example E.A. Bayer and M. Wilchek in Methods of Biochemical Analysis Vol. 26 The Use of the Avidin- Biotin Complex as a Tool in Molecular Biology; and Handbook of Fluorescent Probes and Research Chemicals, 6 th Edition, 1996, by R.P. Haugland, Molecular Probes, Inc.; or alternatively WO 97/14716).

Of course, the peptides according to the invention can also be liberated by solvolysis, in particular hydrolysis, or by hydrogenolysis of their functional derivatives. 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 groups, preferably those which, instead of an H atom which is connected to an N atom, carry an amino protective group or which, instead of the H atom of a hydroxyl group, carry a hydroxyl protective group. The same applies to carboxylic acids which can be protected by substitution of their -CO-OH hydroxyl function by means of a protective group, for example as an ester.

The expression "amino protective group" is generally known and relates to groups which are suitable for protecting (or 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. The expression "hydroxyl protective group" is likewise generally known and 15 relates to groups which are suitable for protecting a hydroxyl group from chemical reactions, but which are easily removable after the desired chemical reaction has been carried out at other positions in the molecule. The liberation of the compounds from their functional derivatives takes place depending on the protective group utilized for example using strong acids, expediently using TFA or perchloric acid, but also using other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids such as trichloroacetic acid or sulfonic acids such as benzene- or p-toluenesulfonic acid. Hydrogenolytically removable protective groups (for example CBZ or benzyl) can be removed, for example, by treatment with hydrogen in the presence of a catalyst (for example of a noble metal catalyst such as palladium, expediently on a support such as carbon).

Typical protective groups for N termini and for sideposition amino groups are Z, BOC, Fmoc, those for Ctermini or the Asp or Glu side chains are O-prim-alkyl (for example OMe or OEt), O-tert-alkyl (for example OBut) or OBenzyl. Z, BOC, NO 2 Mtr, Pmc or Pbf, for example, is suitable for the guanidino function of the Arg. Alcoholic functions can be protected by benzyl radicals, tert-alkyl radicals or trityl groups.

The groups BOC, OBut and Mtr can preferably be removed, for example, using TFA in dichloromethane or using approximately 3 to 5 N HC1 in dioxane at 15-300, the FMOC [sic] group using an approximately 5 to solution of dimethylamine, diethylamine or piperidine in DMF at 15-300.

The trityl group is employed, for example, for the protection of the amino acids histidine, asparagine, glutamine and cysteine. Removal is carried out, depending on the desired final product, using thiophenol, the trityl group being removed from all 16 amino acids mentioned, when using TFA/anisole or TFA/thioanisole only the trityl group of His, Asn and Gin is removed, whereas that on the Cys side chain remains.

Hydrogenolytically removable protective groups (for example CBZ or benzyl) can be removed, for example, by treatment with hydrogen in the presence of a catalyst (for example of a noble metal catalyst such as palladium, expediently on a support such as carbon).

Suitable solvents in this case 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 1000 and pressures between approximately 1 and 200 bar, preferably at 20-300 and 1-10 bar. Hydrogenolysis of the CBZ group takes place readily, for example, on 5 to 10% Pd/C in methanol or using ammonium formate (instead of hydrogen) on Pd/C in methanol/DMF at 20-300.

As already mentioned, the peptides according to the invention include their physiologically acceptable salts, which can likewise be prepared by standard methods. Thus, a base of a compound according to the invention 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. Acids which are suitable for this reaction are in particular those which yield physiologically acceptable salts. Thus, inorganic acids can be used, for example sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acids such as orthophosphoric acid, sulfamic acid, furthermore organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic mono- or polybasic carboxylic, sulfonic or sulfuric acids, e.g. formic acid, acetic acid, 1 1111 17 propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, 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, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenemono- and -disulfonic acids and laurylsulfuric acid. Salts with physiologically unacceptable acids, e.g. picrates, can be used for the isolation and/or purification of the compounds according to the invention. On the other hand, an acid of the compounds according to the invention 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 sodium, potassium, magnesium, calcium and ammonium salts, furthermore substituted ammonium salts, for example the dimethyl-, diethyl- or diisopropylammonium salts, monoethanol-, diethanol- or diisopropylammonium salts, cyclohexyl- or dicyclohexylammonium salts, dibenzylethylenediammonium salts, furthermore, for example, salts with arginine or lysine.

The peptide compounds according to the invention can be employed, as already mentioned, as pharmaceutical active compounds in human and veterinary medicine, in particular for the prophylaxis and/or therapy of disorders in which epithelial cells are involved.

Particularly to be emphasized in this context are disorders or inflammations or wound-healing processes of the skin, the respiratory tract organs and the stomach and intestinal region, thus, for example, apoplexy, angina pectoris, oncoses, osteolytic diseases such as osteoporosis, pathological angiogenic diseases such as, for example, inflammation, fibrosis, in particular pulmonary fibrosis, opthalmological diseases, diabetic retinopathy, macular degeneration, II 18 myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, rubeotic glaucoma, ulcerative colitis, Crohn's disease, atherosclerosis, psoriasis, restenosis after angioplasty, in acute kidney failure, nephritis, microbial infections and multiple sclerosis.

The invention accordingly relates to peptide compounds of the formulae defined above and below and in the claims including their physiologically acceptable salts as medicaments, diagnostics or reagents.

The invention in particular relates to appropriate medicaments as inhibitors for the control of disorders which are indirectly or directly based on expression of the aIP6 integrin receptor, thus in particular in pathological angiogenic disorders, thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, inflammation, infections and for influencing wound-healing processes.

[lacuna] also relates to appropriate pharmaceutical preparations, which contain at least one medicament of the formula I and, if appropriate, vehicles and/or excipients.

The invention furthermore relates to the use of the peptide compounds and/or their physiologically acceptable salts according to the claims and the description for producing a medicament for the control of disorders which are based indirectly or directly on expression of the aIv6 integrin receptor, thus in particular in pathological angiogenic disorders, thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, inflammation, infections and for influencing wound-healing processes.

The medicaments according to the invention or pharmaceutical preparations comprising them can be used in human or veterinary medicine. Possible vehicles are organic or inorganic substances which are suitable for enteral oral) or parenteral administration, or 19 topical application or for administration in the form of an inhalation spray and do not react with the novel compounds, for example water, vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum 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, furthermore suspensions, emulsions or implants, are used for parenteral administration, and ointments, creams or powders are used for topical application. The novel compounds can also be lyophilized and the lyophilizates obtained can be used, for example, for the production of injection preparations. The preparations indicated can be sterilized and/or can contain excipients such as lubricants, preservatives, stabilizers and/or wetting agents, emulsifiers, salts for influencing the osmotic pressure, buffer substances, colorants, flavourings and/or [lacuna] more further active compounds, e.g. one or more vitamins.

For administration as an inhalation spray, sprays can be used which contain the active compound either in dissolved form or suspended in a propellant or propellant mixture (for example CO 2 or fluorochlorohydrocarbons). The active compound is expediently used in this case in micronized form, where one or more additional physiologically tolerable solvents can be present, e.g. ethanol. Inhalation solutions can be administered with the aid of customary inhalers.

As a rule, the substances according to the invention can be administered in analogy to other known, commercially available peptides (for example described in US-A-4 472 305), preferably in doses between approximately 0.05 and 500 mg, in particular between and 100 mg, per dose unit. The daily dose is 20 preferably between approximately 0.01 and 20 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 weight, general state of health, sex, on the diet, on the time and route of administration, and on the excretion rate, pharmaceutical combination and severity of the particular disorder to which the therapy relates. Parenteral administration is preferred.

Furthermore, the novel compounds of the formula I can be used in analytical biology and molecular biology.

The novel compounds of the formula I, where X is a fluorescent dye radical linked via a -CONH-, -COO-, -S0 2 NH- or -NHCO- bond, can be used as diagnostic markers in the FACS (Fluorescence Activated Cell Sorter) technique and fluorescence microscopy.

The use of labelled compounds in fluorescence microscopy is described, for example, by Wang and D.L. Taylor in "Fluorescence Microscopy of Living Cells in Culture, parts A B, Academic Press, Inc. 1989".

The novel compounds according to the invention can also be used as integrin ligands for the preparation of columns for affinity chromatography for the preparation of integrins in pure form. The complex of an avidinderivatized support material, e.g. Sepharose, and the novel compounds is formed by methods known per se (for example E.A. Bayer and M. Wilchek in Methods of Biochemical Analysis Vol 26 The Use of the Avidin- Biotin Complex as a Tool in Molecular Biology).

Suitable polymeric support materials here are the polymeric solid phases having preferably hydrophilic properties and known per se in peptide chemistry, for example crosslinked polysugars such as cellulose, ~-II 21 Sepharose or Sephadex®, acrylamides, polymers based on polyethylene glycol or Tentakel polymers®.

The invention finally also includes recombinant DNA sequences which contain sections which code for peptide regions which have the peptide structural motifs according to the invention.

DNA of this type can be transferred to cells by particles, as is described in Ch. Andree et al. Proc.

Natl. Acad. Sci. 91, 12188-12192 (1994), or the transfer to cells can be increased by other excipients, such as liposomes Aronsohn and J.A. Hughes J.

Drug Targeting, 5, 163-169 (1997)).

The transfer of a DNA of this type could accordingly be utilized in yeasts, by means of baculoviruses or in mammalian cells for the production of the peptide substances of this invention.

If an animal or human body is infected with a recombinant DNA of this type, the peptides according to the invention finally themselves formed by the infected cells can immediately bind to the 0vP 6 integrin receptor, for example of tumour cells, and block it.

Corresponding recombinant DNA, which can be prepared by known and customary techniques, can, for example, however also be present in the form of virus DNA which contains sections which code for the virus coat protein. By infection of a host organism with recombinant, preferably non-pathogenic viruses of this type, host cells which express the integrin aI6 can preferably be attacked (targeting).

Suitable viruses are, for example, adenoviral types which have been used a number of times already as vectors for foreign genes in mammalian cells. A number of properties make them good candidates for gene therapy, as can be seen from S.J. Watkins et al. Gene 22 Therapy 4, 1004-1012 (1997) (see also J. Engelhardt et al. Hum. Gene Ther. 4, 759-769 (1993)).

As can be found in A. Fasbender et al. J. Clin. Invest.

102, 184-193 (1998), a common problem in gene therapy by means of viral and non-viral vectors is the limited efficiency of the gene transfer. Using the additional ligand sequence for ovP 6 integrin described above in the coat protein of the adenoviruses, an improvement in the transfer, for example, of cystic fibrosis transmembrane conductance regulator (CFTR) cDNA can be achieved.

In a manner similar to that in the work of T. Tanaka et al. Cancer Research 58, 3362-3369 (1998), instead of the DNA for angiostatin the DNA for the sequences of this invention can also be utilized for cell transfections by means of retroviral or adenoviral vectors.

The peptides according to the invention can also be employed within a liposome complex of lipid/peptide/DNA for a transfection of cell cultures together with a liposome complex consisting of lipid/DNA (without peptide) for use in gene therapy in man. The preparation of a liposome complex from lipid/DNA/peptide is described, for example, in Hart S.L. et al. 1998: Lipid-Mediated Enhancement of Transfection by a Non-Viral Integrin-Targeting Vector.

Human Gene Therapy 9, 575-585.

A liposome complex of lipid/peptide/DNA can be prepared, for example, from the following stock solutions: 1 pg/gl of lipofectin (equimolar mixture of DOTMA 2 ,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride) and DOPE (dioleyl phosphatidylethanolamine)), 10 gg/ml of plasmid DNA and 100 pg/ml of peptide. For this, both DNA and peptide are dissolved in cell culture medium.

The liposome complex is prepared by mixing the three components in a specific weight ratio (lipid:DNA:peptide, for example 0.75:1:4). Liposomal II_ 1_~ 23 DNA complexes for gene therapy in man have already been described (Caplen N.J. et al. 1995: Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis Nature Medicine 1, 39-46).

The invention thus also relates to the use of accordingly modified recombinant DNA of gene-releasing systems, in particular virus DNA, for the control of diseases which are based indirectly or directly on expression of av 6 Ps integrin receptors, thus in particular in pathological angiogenic disorders, thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, inflammation, infections and for influencing wound-healing processes.

Above and below, all temperatures are indicated in oC.

The HPLC analyses (retention time Rt) were carried out in the following systems: Column 5 pm LichroSpher 60 RP-Select B (250-4), with a S. 50-minute gradient from 0 to 80% 2-propanol in water/0.3% trifluoroacetic acid, at 1 ml/min flow and detection at 215 nm.

Mass spectrometry El (electron impact ionization) M FAB (fast atom bombardment)

(M+H)

The invention will now be described with reference to the following, non-limiting Examples.

0..0 Example 1 Preparation and purification of peptides according to the invention: In principle, the preparation and purification were carried out by means of Fmoc strategy with protection of acid-labile side chains on acid-labile resins using a commercially obtainable "continuous flow" peptide synthesizer according to Haubner et al. Am. Chem.

Soc. 118, 1996, 17703).

24 Cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu) [EMD 272914] 2.7 g of Fmoc-Abu-OH (Bachem B-1910) was [sic] suspended in 150 ml of methylene chloride and dissolved until clear using 10 ml of DMF. 5.6 ml of diisopropylethylamine were added, the solution was poured onto 12.0 g of o-chlorotrityl chloride polystyrene resin (1.14 mmol/g, Bachem D-196512) and the batch was shaken at room temperature.

After 5 hours, the resin was filtered off with suction and washed with 300 ml each of DCM/MeOH/DIEA 17/2/1, DCM, DMF, DCM and MeOH. After removal of the solvents, 14.55 g of Fmoc-amino acid resin were obtained. A triplicate Fmoc determination showed, on average, a loading of 541 mol/g of Fmoc-Abu-O-oClTrt-resin.

0.7 g of Fmoc-Abu-O-oClTrt-polystyrene resin was successively subjected to a coupling step in a doublecoupling technique 2 x with 0.30 g each of TBTU, 0.315 ml of ethyldiisopropylamine and Fmoc-amino acid in 4.1 ml of DMF in a commercial synthesis apparatus and a typical procedure (Milligen 9050 Pep Synthesizer

TM

apparatus and handbook, 1987), for 30 minutes in each case. Washing steps were carried out in DMF for minutes, cleavage steps in piperidine/DMF (1:4 vol) for minutes, and N-terminal acetylations (capping) were carried out with acetic anhydride/pyridine/DMF (2:3:15 vol) for 15 minutes.

The amino acids Fmoc-Arg(Pmc), then Fmoc-Leu, then Fmoc-Ala, then Fmoc-D-Asp(OBut), then Fmoc-Leu, then Fmoc-Asp(OBut), then Fmoc-Thr(But), then Fmoc-Arg(Pmc) and finally Fmoc-Abu were used. After cleavage of the Fmoc protective group from the Fmoc-Abu-Arg(Pmc)- Thr(But)-Asp(OBut)-Leu-D-Asp(OBut)-Ala-Leu-Arg(Pmc)- Abu-O-oClTrt-polystyrene resin, it was washed with DMF and isopropanol and, after drying in vacuo at room temperature, 0.9 g of Abu-Arg(Pmc)-Thr(But)-Asp(OBut)- 25 Leu-D-Asp(OBut)-Ala-Leu-Arg(Pmc)-Abu-O-oClTrtpolystyrene resin was obtained.

By treatment of this peptidyl resin with 20 ml of trifluoroethanol/dichloromethane/acetic acid (2:6:2 vol) for 2 hours at room temperature, filtration, concentration in vacuo and trituration with diethyl ether, 0.19 g of the side chain-protected peptide Abu-Arg(Pmc)-Thr(But)-Asp(OBut)-Leu-D-Asp(OBut)-Ala- Leu-Arg(Pmc)-Abu-OH was obtained.

By dropwise addition of a solution of this product in DMF (100 mg of peptide/15 ml of DMF) to a stirred solution of TBTU/HOBt/DIPEA (10:10:11 equivalents) in ml of DMF/100 mg of peptide in the course of 30 minutes and further stirring for 1 hour, cyclization was achieved. After concentration and precipitation with water, 0.15 g of crude cyclo(Arg(Pmc)-Thr(But)- Asp(OBut)-a-L-D-Asp(OBut)-Ala-Leu-Arg(Pmc)-Abu-Abu) was obtained. After treatment with trifluoroacetic acid/water/TIS (94:3:3 vol) for 2 hours at room temperature, concentration in vacuo and trituration with diethyl ether, a precipitate of 85 mg of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu) was obtained.

Purification of the product was carried out by RP-HPLC on Lichrosorb RP 18 (250 25, 7 pm, Merck KGaA) in 0.3% TFA using a gradient of 4% to 24% 2-propanol over one hour at 10 ml/min and assessment of the eluate by means of a UV flow-through photometer at 215 and 254 nm. 51 mg of product were obtained, FAB 1067; Rt [sic] 19.0.

The following products were prepared analogously: Code Sequence MW FAB Rt (EMD) (g/mol) [sic] 271312 cyclo(RTDLDSLR) 957.1 958 15.10 271578 cyclo(RTDLdSLR) 957.1 957 17.51 26 Code Sequence MW FAB Rt (EMD) (g/mol) [sic] 271579 cyclo(RTDLdALR) 941.1 941 16.64 271586 cyclo(RGDLDSLR) 913.0 913 17.95 271587 cyclo(RGDLdSLR) 913.0 913 17.24 271588 cyclo(RGDLdALR) 897.0 897 19.60 271589 cyclo(RGDLdGLR) 882.9 883 16.68 271590 cyclo(RGDLd-p-Ala-LR) 897.0 897 15.47 271591 cyclo(RGDLdALRG) 954.1 954 16.79 271592 cyclo(RGDLdALRGG) 1011.1 1011 17.83 271593 cyclo(RGDLdALRGGG) 1068.2 1068 17.79 271594 cyclo(RGDLdALRGGGGGG) 1239.3 1239 17.30 272914 cyclo(RGDLdALR-Abu-Abu) 1067.2 1067 19.00 272966 cyclo(RTDLdALRGGG) 1112.2 1113 17.14 272967 cyclo(RTDLdGLRGGG) 1098.2 1099 19.00 272968 cyclo(RTDLDALRGGG) 1112.2 1113 17.39 272969 cyclo(RTDLDGLRGGG) 1098.2 1099 16.88 272970 cyclo(RGDLDALRG) 954.1 955 17.32 272971 cyclo(RaDLdALRGGG) 1082.2 1083 18.26 272972 cyclo(RGDLaALRGGG) 1082.2 1083 18.23 272958 cyclo(RGDLdALR-Aha-Aha) 1123.3 1123 19.38 272959 cyclo(RGDLdALR-Aha) 1010.2 1010 20.63 272960 cyclo(RGDLdALR-Aee) 1042.2 1042 18.92 272961 cyclo(RGDLdALRGGGG) 1125.2 1125 17.56 272962 cyclo(RGDLdALRGGGGG) 1182.3 1182 17.52 272963 cyclo(RGDLdGLRGGG) 1054.1 1054 15.59 272964 cyclo(RGDLDALRGGG) 1068.2 1069 16.84 272965 cyclo(RGDLDGLRGGG) 1054.1 1055 16.03 273028 cyclo(RTDLdALR-Aha) 1072.2 n.d. 19.30 273033 cyclo(RTDLdALR-Abu) 1044.2 n.d. 16.72 273035 cyclo(RGDLdALR-Abu) 1000.1 n.d. 16.96 273038 cyclo(RTDLdALR-Aha-Aha) 1185.4 n.d. 20.46 273040 cyclo(RTDLdALR-Abu-Abu) 1111.3 n.d. 18.75 x 2 TFA 304219 cyclo(RTDLdALR-PAla) 1012.1 1012 19.4 304218 cyclo(RGDLdALR-PAla) 968.1 968 18.7 1329400 cyclo(RGDLdAR) 811.9 812 121.15 27 Code Sequence MW FAB Rt (END) (g/Mol) [sic] 329412 cyclo(RTDLdALR-Abu-Abu) 1111.3 1112 20.31 329402 icyclo(RGDLdALAGGG) 983.1 984 20.48 329399 cyclo(NI~eArg- 1038.2 1039 19.66 GDLaALRGGG) 329398 cyclo(R-NMeGly- 1038.2 1039 19.76 ______DLaALRGGG) 326397 cyclo(RGD-N~eLeu- 1038.2 1039 20.22 ______aALRGGG) 329396 cyclo(RGDL-[D-N~eAla]- 1038.2 1039 20.55

______ALRGGG)

329395 cyclo(RGDLa-NMeAla- 1038.2 1039 21.35 329394 cyclo(RGDLaA-Nl~eLeu- 1038.2 1039 18.87

RGGG)____

329393 cyclo(RGDLaAL-N~eArg- 1038.2 1039 20.64 GGG)__ cyclo (RGDLdAAR)____ (RGDLdAARGGG)____ Nomenclature for amino acids according to Eur. J.

Biochem. 138, 9-37 (1984) lower case letter D-amino acid n.d. not determined Exampl1e 2: af8 6 fibronec tin receptor binding test: The prepared peptides according to the invention were bonded to the immobilized 00~ 6 receptor together with competitively acting fibronectin in solution and the Q value was determined as a measure of the selectivity of the binding of the peptide to be tested to C06~. The Q value is calculated here from the quotient of the IC 5 0 values of test peptide and a standard. The standard used was the linear Ac-RTDLDSLR-NH 2 (Code END 271293) (ref./patent cf. Pytela et al. Science 231, 1559, 28 (1986)). The binding test was carried out in detail as follows: The immobilization of soluble 4ov 6 receptor on microtitre plates was carried out by dilution of the protein solution in TBS++ and subsequent incubation overnight at 4 0 C (100 l/hollow). Non-specific binding sites were blocked by incubation (2 h, 37 0 C) with 3% BSA in TBS++ (200 il/hollow). Excess BSA was removed by washing three times with TBSA++. Peptides were diluted serially (1:10) in TBSA++ and incubated with the immobilized integrin (50 gl of peptide 50 p1 of ligand per hollow; 2 h; 370C) together with biotinylated fibronectin (2 gg/ml). Unbound fibronectin and peptides were removed by washing three times with TBSA++. The bound fibronectin was detected by incubation (1 h; 37 0 C) with an alkaline phosphatasecoupled anti-biotin antibody (Biorad) (1:20,000 in TBSA++; 100 il/hollow). After washing three times with TBSA++, colorimetric detection was carried out by incubation (10-15 min; 25 0 C, in the dark) with substrate solution (5 mg of nitrophenyl phosphate, 1 ml of ethanolamine, 4 ml of H 2 0; 100 l/hollow). The enzyme reaction was stopped by addition of 0.4 M NaOH (100 gl/hollow). The colour intensity was determined at 405 nm in an ELISA measuring apparatus and made equal to the zero value. Hollows which were not coated with receptor served as a zero value. Ac-RTDLDSLR-NH 2 was employed as a standard. The IC 50 values for the peptides tested were read off from a graph and from this, together with the IC 50 value of the standard peptide, the Q value of the peptide according to the invention was determined.

Q value IC 50 test peptide IC 50 standard Q values were calculated as means from repeat experiments.

The results of the test described are summarized in the Table 1 which follows: 29 Table 1 Results of the aiJs fibronectin receptor binding test Code (END) Sequence Q value test peptide IC5o EMD 271293 271293 Ac-RTDLDSLR-NH 2 1.00 (=75 nM,) 271586 Cyclo(RGDLDSLR) 26 271587 cyclo(RGDLdSLR) 88 271588 cyclo(RGDLdALR) 47 271589 cyclo(RGDLdGLR) 19 272970 cyclo(RGDLDALRG) 6.7 272964 cyclo(RGDLDALRGGG) 0.037 272965 cyclo(RGDLDGLRGGG) 0.16 272972 cyclo(RGDLaALRGGG) 0.05 271593 cyclo(RGDLdALRGGG) 0.03 272963 Cyclo(RGDLdGLRGGG) 0.084 271590 cyclo(RGDLd-P3Ala-LR) 233 271591 cyclo(RGDLdALRG) 2.1 271592 Cyclo(RGDLdALRGG) 0.05 271594 cyclo(RGDLdALRGGGGGG) 0.05 272914 cyclo(RGDLdALR-Abu-Abu) 0.03 272958 cyclo(RGDLdALR-Aha-Aha) 0.036 272959 cyclo(RGDLdALR-Aha) 0.036 272960 cyclo(RGDLdALR-Aee) 0.038 272961 cyc1o(RGDLdALRGGGG) 0.033 272962 cyclo(RGDLdALRGGGGG) 0.046 273035 cyclo(RGDLdALR-Abu) 0.028 271312 cyclo(RTDLDSLR) 8 271578 cyclo(RTDLdSLR) 104 271579 cyclo(RTDLdALR) 12 272966 cyclo(RTDLdALR-GGG) 0.05 272967 cyclo(RTDLdGLR-GGG) 0.15 272968 cyclo(RTDLDALR-GGG) 0.14 272969 cyrI-o(RTDLDGLR-GGG) 0.18 30 Code (EMD) Sequence Q value IC 50 test peptide

IC

50 EMD 271293 273028 cyclo(RTDLdALR-Aha) 0.015 273033 cyclo(RTDLdALR-Abu) 0.043 273038 cyclo(RTDLdALR-Aha-Aha) 0.015 273040 cyclo(RTDLdALR-Abu-Abu) 0.014 272971 cyclo(RaDLdALR-GGG) 8.2 The following preparations: examples relate to pharmaceutical Example A: injection vials A solution of 100 g of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala- Leu-Arg-Abu-Abu) and 5 g of disodium hydrogenphosphate is adjusted to pH 6.5 in 3 1 of double-distilled water using 2 N hydrochloric acid, sterile-filtered, dispensed into injection vials, lyophilized under sterile conditions and aseptically sealed. Each injection vial contains 5 mg of active compound.

Example B: suppositories A mixture of 20 g of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala- Leu-Arg-Abu-Abu) 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.

Example C: solution A solution is prepared from 1 g of cyclo(Arg-Thr-Asp- Leu-D-Asp-Ala-Leu-Arg-Abu-Abu), 9.38 g of NaH 2

PO

4 2 H 2 0, 28.48 g of Na 2 HP0 4 12 H 2 0 and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. The mixture -is adjusted to pH 6.8, made up to 31 1 1 and sterilized by irradiation. This solution can be used in the form of eye drops.

Example D: ointment 500 mg of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu- Abu) are mixed with 99.5 g of petroleum jelly under aseptic conditions.

Example E: tablets A mixture of 1 kg of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala- Leu-Arg-Abu-Abu), 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is compressed in a customary manner to give tablets in such a way that each tablet contains 10 mg of active compound.

Example F: coated tablets Analogously to Example E, tablets are pressed and are then coated in a customary manner with a coating of sucrose, potato starch, talc, tragacanth and colorant.

Example G: capsules 2 kg of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu- Abu) are dispensed into hard gelatin capsules in a customary manner such that each capsule contains 20 mg of the active compound.

Example H: ampoules A solution of 1 kg of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala- Leu-Arg-Abu-Abu) in 60 1 of double-distilled water is sterile-filtered, dispensed into ampoules, lyophilized under sterile conditions and aseptically sealed. Each ampoule contains 10 mg of active compound.

32 Example I: inhalation spray 14 g of cyclo(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu- Abu) are dissolved in 10 1 of isotonic NaCI 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 puff of spray (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or group of integers or steps but not the exclusion of any other integer or group of integers or steps.

Persons skilled in the art will appreciate that numerous variations and modifications will become apparent. All such variations and modifications which become apparent to oooo persons skilled in the art, should be considered to fall within the spirit and scope that the invention broadly appearing before described.

i The reference to any prior art in this specification is not, and should not be taken as, an acknowledgement or any form of suggestion that the prior art forms part of the common general knowledge in Australia.

ooeo **ooo oooo* ooo o ooo* 1

Claims (11)

1. Peptide derivatives of the formula I cyclo (Arg-X-Asp-X 2 -X 3 -X 4 -X 5 -X 6 -R I in which X 1 is Ser, Gly or Thr, X 2 is Leu, Ile, Nle, Val or Phe, X 3 is Asp, Glu, Lys or Phe, X 4 is Gly, Ala or Ser, X 5 is Leu, Ile, Nle, Val or Phe, X 6 is Arg, Har or Lys, R 1 is one or more amino acid residue(s), selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hcy, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val and H 2 N- (CH 2 CH20) m- (CH 2 n-COOH, wherein m,n in each case independently of one another are 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, with the proviso that m n is 0, and with the further proviso that the w-aminocarboxylic acid residue(s) are having a length of 500 to 2500 pm, where the amino acids mentioned can also be 25 derivatized, Sthe D and the L forms of the optically active amino acid residues are included, and their physiologically acceptable salts and solvates.

2. Peptide derivatives according to Claim 1, of the formula I P:\WPDOCS\CRSETSpc\76692.nw cdai-doc-l6O2O4 -34- cyclo(Arg-X 1 -AspX 2 -X 3 _X 4 -X 5 X 6 -R1) I in which xi is Ser, Gly or Thr, x2 is Leu, Ile, Nie, Val or Phe, x3 is Asp, Glu, Lys or Phe, x4 is Gly, Ala or Ser, is Leu, Ile, Nie, Val or Phe, x6 is Arg, Har or Lys, R, is i-ia amino acid residue(s), selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gin, Glu, Hcy, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val and H 2 N- (CH 2 CH 2 O)m- (CH 2 )m,,COOH, n in each case independently of one another are 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, with the proviso that m n is 0, where the amino acids mentioned can also be derivatized, the D and the L forms of the optically active amino acid residues are included, and their physiologically acceptable salts and solvates.

3. Peptide compounds according to Claim 1 or 2 selected from the group consisting of cyclo (Arg-Giy-Asp-Leu-Asp--Ala-Leu-Arg-Gly-Gly-Gly), cyclo (Arg-Gly-Asp-Leu-Asp-Gly-Leu-Arg-Gly-Gly-Gly), cyclo (Arg-Gly-Asp-Leu-D-Ala-Ala-Leu-Arg-Gly-Gly-Giy), PAWPDOCS\CRSESp=\766 1920.nw climsd-IO2/04 cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-ArgGly-Gly-Gly-Gly), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aha-Aha), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aha), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-Aee), cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu), cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg--Ala), cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-0-Ala), and their physiologically acceptable salts and solvates.

4. A method of treating and/or preventing disorders which are based on an expression and pathological function of xUv9 6 integrin receptors in a subject, comprising administering to the subject an effective amount of a peptide derivative of the formula I, as defined in claim 1 or claim 2, or a peptide compound as defined in claim 3. 20 5. The method according to Claim 4, wherein the disorders are selected from thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, fibroses, inflammation, infections, psoriasis and wound-healing processes.

6. Medicaments comprising one or more peptide compounds Sof the formula I according to Claims 1 or 2 and/or the compounds according to Claim 3, and/or their pysiologically acceptable salts and solvates. 6. Medicament according to Claim 6 as an inhibitor for the control of disorders which are based on an P:WPDOCS\CRN\SESpo\7661920on, Whdo-I6O2/4 -36- expression and pathological function of oYV6 integrin receptors.

8. Medicament according to Claim 7 for the control of thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteo- porosis, fibroses, inflammation, infections, psoriasis and for influencing wound-healing processes.

9. Pharmaceutical preparation, comprising at least one medicament according to one of Claims 7 and 8 and, if appropriate, vehicles and/or excipients and, if appropriate, other active compounds.

10. Use of peptide compounds according to Claims 1 to 3 and/or their physiologically acceptable salts for producing a medicament for the control of disorders which are based on expression and pathological function of CvP6 integrin receptors. 020 *0 *t

11. Use according to Claim 10 for producing a medicament for the control of thromboses, cardiac infarct, coronary heart disorders, arteriosclerosis, tumours, osteoporosis, fibroses, inflammation, infections, 25 psoriasis and for influencing wound-healing processes.

12. Peptide derivatives of the formula I, as defined in Claim 1 or Claim 2, uses thereof, and methods of treating and/or preventing based on an expression and pathological function of uvP0 integrin receptors, substantially as hereinbefore described, with reference to the accompanying Examples. PAWDOCS\CRN\SE~pcw\766192.nmw caiams.dow-161O2/04 -37-

13. Peptide compounds as defined in Claim 3, uses thereof, and methods of treating and/or preventing disorders based on an expression and pathological function of U'03 6 integrin receptors, substantially as hereinbefore described, with reference to the accompanying Examples. DATED this 16th day of February, 2004 MERCK PATENT GMBH By its Patent Attorneys DAVIES COLLISON CAVE