SK262002A3 - Cyclic peptide derivatives as inhibitors of integrin alpha'v'beta'6' - Google Patents

Abstract

Cyclic peptides (I), containing 8 amino acid residues and optionally further alpha ,omega -aminocarboxylic acid residues, are new. Cyclic peptides of formula Cyclo-(Arg-X 1-Asp-X 1-X 2-X 3-X 4-X 5-X 6-R 1) (I) and their salts and solvates are new. X 1Ser, Gly or Thr; X 2Leu, Ile, Nle, Val or Phe; X 3Asp, Glu, Lys or Phe; X 4Gly, Ala or Ser; X 5Leu, Ile, Nle, Val or Phe; X 6Arg, Har or Lys; R 1one or more omega -aminocarboxylic acid residues, having a length of 50-2500 mu m; or is absent; the amino acid residues are optionally derivatized and include D- as well as L-forms (in the case of optically active aminoacids). ACTIVITY : Antithrombotic; Cardiant; Antiarteriosclerotic; Cytostatic; Osteopathic; Antiinflammatory; Antibacterial; Antipsoriatic; Vulnerary; Cerebroprotective; Antianginal; Antidiabetic; Ophthalmological; Antiarthritic; Antirheumatic; Antiulcer; Vasotropic; Nephrotropic; Neuroprotective. MECHANISM OF ACTION : alpha vbeta 6-Integrin receptor inhibitor. Cyclo-(Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu) (Ia), Abu : 4-aminobutyric acid had IC 500.03 times that of the standard compound Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-NH 2(i.e. 75 nM) in an alpha vbeta 6/fibronectin receptor binding assay.

Description

The invention relates to novel peptides of the formula I cyclo (Arg-X ¹ -Asp-X ² -X ³ -X ⁴ -X ^with -X ⁶ -R ¹ ) (I) wherein

X ¹ Ser, Gly or Thr

X ² Leu, clay, no, val or phe,

X ³ Asp, Glu, Lys or Phe,

X ⁴ Gly, Ala or Ser,

X ^with Leu, White, No, Val or Phe,

X ^e Arg, Har or Lys,

R ¹ is absent or represents one or more α-aminocarboxylic acid residues, the α-amino carboxylic acid residue having a length of 500 to 2500 µm, said amino acids also being derivatized, including D and L forms and optically active amino acid residues and their physiologically acceptable salts and solvates.

The invention is based on the discovery of novel compounds which have valuable properties and can be used in the manufacture of medicaments.

It has been found that the compounds of the invention and their salts have very valuable pharmacological properties along with good tolerability.

The peptides of the invention can be used as potent integrin receptor inhibitors and thereby to treat various diseases and pathological findings.

BACKGROUND OF THE INVENTION

Other integrin / 5 inhibitors are described in DE 19858857, and other such types have also been described by S. Kraft et al. (J. Biol. Chem. 274, 1979-1985, 1999). The compounds of the invention may be considered as a selective invention with respect to the above literature.

Integrins belong to the family of heterodimeric class I trans-membrane receptors, which play an important role in many cell-matrix or cell-cell adhesion processes (Tuckwell et al., 1996, Symp. Soc. Ex. Biol. 47). They can roughly be divided into three classes:

- integrins which represent receptors for the extracellular matrix,

- integrins that are activatable on leukocytes and are triggered by inflammatory processes, as well as

integrins that affect the cellular response in wound healing and other pathological processes (Marshall and Hart, Semin. Cancer Biol. 7, 191 (1996)).

Integrins, in natural ligands such as

o / 2>, «y / 3 _t oC Z3 a

For example, the fibronectin or vitronectin are in the 3-vSe. Soluble RGD-containing peptides are able to inhibit the interaction of each of these integrins with the corresponding natural ligands. Integrin is a relatively rare integrin (Busk et al., J. Biol. Chem. 267 (9), p. 5790, 1992), which produces and preferably binds natural matrix molecules of fibronetine and tenascin in epithelial tissue repair processes. Want et al., Am J. Respir. Cell Mol Biol 15 (5), 664 (1996). Physiological and pathological functions are not yet fully understood, but it is believed that this integrin plays an important role in the physiological processes and diseases (e.g., inflammation, wound healing, tumors) in which epithelial cells are involved. So take Invest. Dermatol. 106 (1), p. 42, 1996), it can be concluded that other pathological phenomena of skin such as psoriasis are influenced by agonists and antagonists of this integrin in addition to wound healing and inflammation processes. In addition, Respir has. Cell Mol. Biol. 12 (5), p. 547, 1995), so that the corresponding agonists / antagonists of this integrin could be successfully employed in respiratory diseases such as bronchitis, asthma, pulmonary fibrosis and airway tumors. It is also known that οζζ / β are important for the intestinal epithelium, so that the corresponding integrin agonists / antagonists could be used in the treatment of inflammations, tumors and injuries of the stomach and intestinal tract.

The dependence of angiogenesis on the interaction of vascular integrins and extracellular matrix proteins has been described by P.C. Brooks, R.A. Clark and D.A. Cheresh (Science 264: 569-571, 1994).

It is therefore an object of the invention, in addition to the already known natural high molecular weight ligands and antibodies which are difficult to treat therapeutically and diagnostically, to find potent, specific and selectively low molecular weight ligands prec / β / 3, preferably peptides which could be used for these therapeutic areas. , but also as diagnostic or reagent agents.

SUMMARY OF THE INVENTION

It has been found that the above-described peptide compounds of the invention and their salts as soluble molecules act on cells having said receptor, or when they bind to surfaces, represent artificial ligands for mediated cell adhesion. In particular, they act as inhibitors of ΆΆ integrin, inhibiting in particular the interaction of the receptor with other ligands, such as fibronectin binding. This action can be demonstrated, for example, by the method described by J.W. Smith et al. (J. Biol. Chem. 265: 12267-12271, 1990).

In addition, it has been found that the novel compounds have very valuable pharmacological properties along with good tolerability and can be used as medicaments. This will be described more precisely.

In addition, the peptide compounds of the invention may be used as diagnostics for detecting pathological conditions in the epithelial system and in addition can localize in vivo and in vitro if they are provided with suitable markers (e.g., a biotinyl residue) according to the prior art.

The invention also encompasses combinations with at least one other active compound or conjugates with other active agents, such as cytotoxic active agents, as well as radiolabelled conjugates for X-ray therapy or PET diagnosis, or such fusion proteins with protein markers such as GFP or antibodies, or therapeutic proteins such as IL-2.

Particularly effective are compounds of formula I wherein the octapeptide sequence is cyclo (Arg-X ¹ -Asp-X ³ -X ³ -X ⁴ -X ⁵ -X ⁶ ) and wherein X ² , X ³ , X ⁴ , X ⁵ , and X ⁶ are as defined above, extended by R ^x . The effect of ring expansion is shown in FIG. 1 for example cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg) [EMD 271588]. As a comparative. compound 'sa' mentioned ·

Ac-Arg-Thr-Asp-Leu-Asp-Ser-Leu-Arg-NH4 [EMD cyclo (Arg-Gly-Asp-Leu-D-Asp-Ala-Leu-Arg-R1 ⁾ ]

2711293]. Some peptides have a distance from R ¹ (calculated) and a Q value (IC [compound] / IC _s [EMD 271293] and - log Q given in the table.

R ^x 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 I see 870 0,036 1,444 AEE 1078 0,038 1,420 Gly-Gly-Gly 1110 0.03 1,523 Abu-abu 1235th 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 graphical representation is shown in FIG. 1. Particularly active compounds are obtained when the spacer length R ¹ reaches approximately 500 µm.

Particularly preferred compounds are those compounds of formula (I) wherein R ¹ is one or more N-aminocarboxylic acid residues, wherein the α-aminocarboxylic acid residue has a length of 600 to 2000 µm.

-Aminocarboxylic acid radicals are understood to mean residues of any -aminocarboxylic acid, with particularly preferred amino acids selected from Ala, Asn, Asp,

I

Arg, Cys, Gln, Glu, Hcy, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val, and H N- (CH CHO) - (CH) -COOH, where

2 2 m 2 n m, n each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, provided that m + n is greater than 0.

The invention therefore relates in particular to derivatives of peptides of the general formula I cyclo (Arg-X ^and --Asp-X ² -X ³ -X ⁴ -X ^with -X ⁶ -R ¹ ) (I) wherein

X ¹ Ser, Gly or Thr

X ² Leu, clay, no, val or phe,

X ³ Asp, Glu, Lys or Phe,

X ⁴ Gly, Ala or Ser,

X ^with Leu, White, No, Val or Phe,

X ⁶ Arg, Har or Lys,

R ¹ is absent or is 1 to 10 amino acids selected from the group consisting of Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hcy, His,

Hse, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr,

Val and H N- (CH CHO) - (CH) -COOH, where is

2 2 m 2 n m, n independently of each other 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,

10, 11, or 12, provided that m + n is greater than 0.

wherein said amino acids may also be derivatized, including D and L forms and optically active amino acid residues, and physiologically acceptable salts and solvates thereof.

‘I

Some preferred groups of compounds of formula (I) are the following compounds of formulas Ia to Ih, wherein in particular the not shown symbols have the meanings given in formula (I) in the formula

and ) X ¹ Gly or Thr, b ) X ¹ Gly or Thr, X ² Leu, C ) X ¹ Gly or Thr, X ² leu, x ³ Asp or D-Asp, D ) X ¹ Gly or Thr, X ² Leu, x ³ Asp or D-Asp, x ⁴ Gly or Ala. e ) X ¹ Gly or Thr, X ² Leu, x ³ Asp or D-Asp, x ⁴ Gly or Ala. x ^s Leu, F ) x ¹ Gly or Thr, x ² Leu, x ³ Asp or D-Asp, x ⁴ Gly, Ala or Ser, Χ ^Ξ Leu, x ⁶ Arg,

(g) X ¹ Gly or Thr;

X ² Leu,

X ³ Asp or D-Asp,

X ⁴ Gly, Ala or Ser,

X ⁵ Leu,

X ⁶ Arg,

R ¹¹ 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 N- (CH CHO) - (CH) -COOH, where z '2 2' xn '2' n is ⁹ m, n each independently 0, 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10, 11, or 12, provided that m + n is greater than 0.

(h) X ¹ Gly or Thr

X ² Leu,

X ³ Asp or D-Asp

X ⁴ Gly, Ala or Ser,

X ⁵ Leu,

X ^e Arg,

R ^{11 is from} 1 to 6 amino acids selected from the group consisting of Gly, β-Ala, Abu or Aha, and salts thereof.

In particular, the invention relates 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-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-# -Ala), and physiologically acceptable salts thereof.

Amino acid residue abbreviations are the following amino acid residues:

Abu 4-Aminobutyric acid AEE H N- (CH CHO) -CH -COOH 2 ^x 2 2 ⁹ 2 2 I see 6-aminohexanoic acid, 6-aminocaproic acid Aib alpha-aminoisobutyric acid Ala alanine same time asparagine Asp Arg BGL aspartic acid arginine C-alpha-tert-butylglycine

Cys cysteine

Dab 2,4-diaminoisobutyric acid Dap gin 2,3-diaminopropionic acid glutamine GLP Glu pyroglutamic acid glutamic acid Gly Har glycine homoarginine Hcy homocysteine

His histidine homo-Phe homo-phenylalanine Hse homo-serine ile. izoleycín Leu leucine Lys lysine Met methionine Nal naphth-2-yl alanine Not norleucine Orn ornithine Pen penicillamine Phe phenylalanine PHG phenylglycine 4-Hal-Phe 4-halogenfenylalanin for proline Ser serine Thr threonine TIS triisopropylsilane [sic] Trp tryptophan Tyr tyrosine wall valine

In addition, the abbreviations they have

next meaning: Ac acetyl Boe tert-butoxycarbonyl BSA bovine serum albumin CBZ or Z benzyloxycarbonyl DCC1 dicyclohexylcarbodiimide DCM. dichloromethane DIEA diethylamine DMF dimethylformamide EDC1 N-ethyl-N, N '- (dimethylaminopropyl) carbodiimide et ethyl FCA fluorescein carboxylic acid

FITC fluorosceinizotiokyanát Fmoc 9-fluorenylmethoxycarbonyl FTH fluoresceintiomočovina HOBt 1-Hydroxybenzotriazole. ' HONSu N-hydroxysuccinimide Me methyl MBHA 4-methylbenzhydrylamine Mtr 4-methoxy-2,3,6-trimethylphenylsulfonyl OBu tert-butyl Oct octanoyl OMe methyl ester OEt ethyl PBF 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl pmc 2,2,5,7,8-pentamethyl-6-sulfonyl POA phenoxyacetyl Sal saliciloyl TBS ++ tris buffered brine with a divalent cation TBSA TBS + BSA TBTU 2- (lH-benzotriazole-1-yl) -1,1,3-tetramethyluronium tetrafluoroborate TFA trifluoroacetic acid Trt trityl (triphenylmethyl).

While the above amino acids may be in several enantiomeric forms, all of these forms as well as mixtures thereof (e.g., DL forms) include the invention. In addition, amino acids can be protected by well-known protecting groups.

The invention also relates to hydrates and solvates, for example alcoholates of the compounds of formula I.

The compounds of the invention also include so-called prodrug derivatives, acyl groups, i.e., alkyl or sugars or oligopeptides-altered compounds of the formula I which are rapidly cleaved in the body into the active compounds of the invention. This group also includes biodegradable polymer derivatives of the compounds of the invention described in the literature (for example, J. Pharm. 115, pp. 61-67 (1995)).

Said amino acids or amino acid residues, for example NH groups or terminal amide groups, may also be derivatized, with N-methyl, N-ethyl, N-propyl, N-benzyl or C 6 -methyl derivatives being preferred. Asp and Glu derivatives are especially methyl-, ethyl-, propyl-, butyl-, tert-butyl-, neopentyl- or benzyl esters of side-carboxyl groups and are additionally preferred derivatives of Arg which may be substituted on the -NH-C (= NH) group -NH ₂ - is acetyl, benzoyl, methoxycarbonyl or ethoxycarbonyl.

The invention also encompasses compounds of the invention which are coupled to one another via the peptide via alpha-amino and alpha-carboxyl groups (head-tail bond), and are cyclic compounds having a functional side chain, e.g. SH, and linked in the following way - side e.g. SS (disulfide) head - minor minor - tail

The invention also includes derivatives containing known markers which facilitate the detection of peptides. Examples of such derivatives include radiolabeled, biotinylovary or fluorescently labeled peptides.

Fluorescent dye groups are preferably 7-acetoxycoumarin-3-yl, fluorescein-5- (or 6-) yl, 2 ', 7'-dichlorofluorescein-5- (and 6-) yl, dihydrotetramethylrodamine-

4-yl, tetramethylrodamine-5- (and 6-) yl, 4,4-difluoro-5,7-dimethyl-

4-boron-3a, 4a-diaza-s-indacen-3-ethyl or 4,4-difluoro-5,7diphenyl-4-boron-3a, 4a-diaza-s-indacen-3-ethyl.

Suitable groups of functionalized fluorescent dyes that can be used as reagents in the preparation of the compounds of Formula I of the invention are described, for example, in Handbook of Fluorescent Probes and Research Chemicals, 5th Edition, 1992-1994, RP Haughland Molecular Probes Inc .

The invention encompasses not only the peptides described above but also compositions and compositions which, in addition to the compounds of the invention, contain such other pharmacologically active compounds or adjuvants which may favorably influence the primary pharmacological action of the peptides of the invention.

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

Preferably, the peptides of the invention are prepared by a solid phase method followed by separation and purification as described, for example, by Jonczyk and Meienhofer (Peptides, Proc. 8. Am. Pept. Symp. Eds. V. Hruby and DHRich, Pierce Comp. III, pp. 73-77 (1983) or Angew. Chem. 104: 375 (1992) or Merrifield (J. Am. Chem. Soc. 94: 3102 (1972)).

The peptides of the invention can be prepared by solid phase (manually or using automated synthesizers) Fmoc strategy using acid labile side protecting groups and purification by chromatography

RP-HPLC. Peak homogeneity can be measured by chromatography

RP-HPLC and compound identity using FAb-MS.

Peptides may also be prepared by methods known per se for the preparation of amino acids and peptides known from the literature (e.g., Novabiochem - 1999 Catalog and Peptide Synthesis Handbook of Calbiochem-Novabiochiem GmbH, D-65796 Bad Soden) and from many standard literature and published patent publications. .

Successive coupling and fragment condensation can be used. Various N-terminal and C-terminal and side protecting groups can be used, which are preferably chosen to be orotogonally cleavable. The coupling steps may be carried out using various condensing agents such as carbodiimides, carbodiimidazoles, uronium type reagents such as TBTU, using mixed anhydride and acid halide methods or active esters. Activated esters are preferably formed in situ, for example by the addition of HOBt or N-hydroxysuccinimide.

Similarly, cyclization of a linear precursor molecule having side protecting groups can be carried out using the coupling reactions described in the literature (for example, DE 43 10 643; or Houben-Weyl, 1c, Volume 15/11, p. 1 to 806 (1974).

Various resins and anchoring functions can be used in solid phase preparation. The resins may, for example, be based on polystyrene or polyacrylamide; suitable anchoring functions are Wang, o-chlorotrityl for the preparation of peptide acids, aminoxanthenoxy anchors, for example for the preparation of peptide amines.

Biotinylated or fluorescently labeled peptides / proteins can likewise be prepared by methods known per se (e.g.

E.A. Bayer and M. Wilchek, Methods of Biochemical Analysis, Volume 26, The Use of the Avidin-Biotin Complex as a Tool in

Molecular Biology; and Handbook of Fluorescent Probes and Research Chemicals, 6th Edition, 1996, R.P. Haugland Milecular Probes Inc .; or WO 97/14716).

. F

The compounds of formula (I) may further be prepared by liberation from their functional derivatives by solvolysis, in particular by hydrolysis or hydrogenolysis. For solvolysis or for hydrogenolysis, suitable starting materials are compounds having, instead of one or more free amino or hydroxyl groups, corresponding protected amino or hydroxyl groups, preferably compounds having, instead of one H-atom which is linked to an N-atom, an amino protecting group, or which have a hydroxyl protecting group instead of a hydrogen atom in a hydroxyl group. The same is true for carboxylic acids which may be protected by replacing their -CO-OH hydroxyl group with a protecting group, for example an ester group.

The term amino protecting group refers to groups which are suitable for protecting the amino group from chemical reactions, which are removable when the desired chemical reaction is generally known and (to block) are readily carried out elsewhere in the molecule. The term hydroxyl-protecting group is likewise generally known and refers to groups which are suitable for protecting (blocking) a hydroxyl group from chemical reactions, but which are readily removable when the desired chemical reaction at another site of the molecule is carried out. Depending on the protecting group used, the release of the compounds of the formula I from their functional derivatives is successful, for example, with strong acids such as, for example, trifluoroacetic acid and perchloric acid, but also with other strong inorganic acids such as hydrochloric or sulfuric acid. trichloroacetic acid or benzesulfonic acid with sulfonic acids such as or p-toluenesulfonic acid.

Hydrogenolytically removable protecting groups (e.g. CBZ or benzyl groups) can be cleaved, for example, by treatment with hydrogen in the presence of a catalyst (e.g. a light metal catalyst such as palladium, preferably on a support such as carbon).

Typical protecting groups for the N-terminal groups and for the amino side groups are Z, BOC, Fmoc, for the terminal

C-groups or, for Asp or Glu, O-prim-alkyl groups (e.g. ethoxy), O-tert-alkyl side chains are methoxy or (e.g. OBut) or

O-benzyl. For example, Z, BOC, NO ₂ , Mtr, Pmc or Pbf are suitable for protecting the guanidine moiety of Arg. Alcohol groups may be protected with tert-alkyl or trityl groups.

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

The trityl group of histidine, asparagine, is used to protect the amino acids glutamine and cysteine. The protecting group is cleaved off with the product trifluoroacetic acid according to the desired final acid / 10% thiophenol, whereby the trityl group is cleaved from said amino acids.

Using the trifluoroacetic acid / anisole or trifluoroacetic acid / thianisole system, only the trityl group is cleaved from histidine, asparagine and glutamine, but remains on the Cys side chain.

Hydrogenolytically removable protecting groups (e.g. CBZ or benzyl groups) can be cleaved, for example, by treatment with hydrogen in the presence of a catalyst (e.g. noble metal catalyst such as palladium, preferably on a support such as carbon). Suitable solvents are the abovementioned solvents, in particular, for example, alcohols such as methanol or ethanol or amides such as dimethylformamide.

The hydrogenolysis is generally carried out at a temperature of about 0 ° to 100 ° C, at a pressure of about 0.1 to 20 MPa, preferably at a temperature of about 20 ° to 30 ° C, at a pressure of about 0.1 to 1 MPa. Hydrogenolysis of the CBZ group, for example, performs well on 5-10% palladium on carbon in methanol or using ammonium formate (instead of hydrogen) on palladium on carbon. in methanol / dimethylformamide at 20 ° to 30 ° C.

As mentioned above, they also include their physiologically acceptable peptides according to the salt, which the invention can be prepared by methods known per se. The base of formula (I) can be converted into an appropriate acid addition salt by acid, for example by reaction of an equivalent amount of base and acid in an inert solvent such as ethanol and subsequent evaporation of the solvent. Suitable acids for this reaction are, in particular, those which give physiologically acceptable salts.

Inorganic acids such as sulfuric, nitric, hydrohalic acids such as hydrochloric or hydrobromic acids, phosphoric acids such as orthophosphoric acid, sulfamic acid and organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic monounsaturated or sulfonic or sulfuric acids may be used. , acetic, propionic, pivalic, polyunsaturated carboxylic acids such as formic, diethylacetic, malonic, succinic, pimelic, fumaric, maleic, lactic, tartaric, apple, lemon, gluconic, ascorbic, nicotinic, isonicotonic, ethanesulfonic, methanesulfonic, methanesulfonic, methanesulfonic,

2-hydroxyethanesulfonic, benzenesulfonic, p-toluenesulfonic, naphthalenesulfonic and naphthalenedisulfonic and laurylsulfonic acids. Salts with physiologically unacceptable acids, for example picrates, can be used to isolate or purify the compounds of formula I.

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

The peptide compounds of the invention can be used, as mentioned above, as pharmaceutically active compounds in human and veterinary medicine, in particular for the prophylaxis or therapy of disorders in which epithelial cells are involved. Especially for the treatment of inflammation, for wound healing, for the treatment of organs of the respiratory tract and for the treatment of stomach and intestines, for example for the treatment of apoplexy, angina, tumors, osteolytic diseases such as osteoporosis, pathological angiogenic diseases such as fibrosis, ophthalmological degenerative , pulmonary diseases, diabetic retinopathy, myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthritis, rubeotic glaucoma, ulcerative colitis, psoriasis, renal restenosis, nephritis, sclerosis.

The invention relates to

Crohn's disease, atherosclerosis, after angioplasty, acute failure of microbial infection, and scattered peptide compounds of the above general formula I and their physiologically acceptable salts as drugs, diagnostic and reagents.

In particular, the invention relates to corresponding medicaments as inhibitors for the treatment of diseases which are directly or not directly caused by the expression of integrin receptors, in particular pathogenic angiogenic diseases, thromboses, heart attack, coronary heart disease, arteriosclerosis, tumors, osteoporosis, inflammation, infections as well as infections as wound healing processes.

The invention also relates to suitable pharmaceutical compositions comprising at least one medicament of formula I and optionally a carrier or excipients.

The invention furthermore relates to the use of the peptide compounds or their physiologically acceptable salts according to the invention and to the instructions for the manufacture of medicaments for the treatment of disorders which are indirectly or directly based on expression of the [beta] 3 integrin receptor, particularly for pathological angiogenic disorders, thrombosis. heart attack, coronary heart disorders, arteriosclerosis, tumors, osteoporosis, inflammation, infections, and to affect wound healing.

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

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

As a rule, the compounds of the formula I according to the invention are used in doses similar to the commercially known peptides, in particular in analogy to the compounds of U.S. Pat. No. 4,472,305, preferably at a dose of about 0.05 to 500 mg, in particular 0.5 to 100 mg per dosage. unit. The daily dose is preferably about 0.01 to 20 mg / kg body weight. The dose for each individual depends on a variety of factors, such as the efficacy of the particular compound used, age, body weight, general health, sex, diet, time and route of administration, elimination rate, drug combination and the severity of the disease. . Parenteral administration is preferred.

The invention also relates to compounds of formula I which can be used in analytical biology and molecular biology.

Novel compounds of formula I wherein X is a fluorescent dye group bonded via -CONH, -C00, -NH-C (= S) -NH, -NH-C (= O) -NH, -SO _with NH or -NHCO can be used as diagnostic markers in the FACS (Fluorescence Activad Cell Sorter) technique and in fluorescence microscopy.

The use of labeled compounds in fluorescence microscopy is described in the literature (eg, Y.-L.Wang and D.L. Taylor Fluorescence Microscopy of Living Cells in Culture, Part A + B, Academic Press, Inc. 1989).

The novel compounds of the invention can also be used as integrin ligands for the production of affinity chromatography columns for the production of purified integrins. A complex of an Avidin-deivatized carrier such as Sepharose and the novel compounds of formula I is formed in a manner known per se (as described, for example, by EA Bayer and M. Wilchek, Methods of Biochemical Analysis, Volume 26, The Use of the Avidin-Biotin Complex as a Tool in Molecular Biology). Suitable polymeric carrier materials for this purpose are the polymeric solid phases known from peptide chemistry, preferably with hydrophilic properties, for example crosslinked polycucars such as cellulose, Sepharose or Sephadex ^R , acrylamides, polyethylene glycol-based polymer or Tentacelpolymer ¹ *.

The invention also encompasses recombinant DNA sequences which comprise regions which encode regions of peptides having the peptide structural motifs of the invention.

Such DNA can be transferred by particles to cells as described by Ch. Andree et al. (Proc. Natl. Acad. Sci.

12188-12192, 1994) or may enhance cell transfer by other co-agents such as liposomes (A.I. Aronsohn and

J.A. Hughes, J. Drug Targeting 5, p. 163-169 (1997).

A transporter of such DNA could therefore be used in yeast via Bacculoviruses or in mammalian cells to produce peptides.

When infected _{(by an} animal or human organism by said recombinant DNA), the infected peptides can finally bind the infected peptides directly to the integrin receptor, for example tumor cells, and block them.

However, the corresponding recombinant DNA, which can be prepared by known and conventional methods, can also be, for example, in the form of viral DNA which contains regions which encode the viral envelope protein. By infecting the host organism with such recombinant, especially non-pathogenic viruses, host cells that express integrin can advantageously be attacked (targeting).

Suitable viruses are, for example, adenoviruses, which are often used as vectors for foreign genes in mammalian cells. Because of their many properties, they are suitable for gene therapy, as stated

S.J. Watkins et al. (Gene Therapy 4, pp. 1004-1012, 1997) and also J. Engelhard et al. (1993, Hum. Gene Ther. 4: 759-769). As reported by A. Fasbender et al. (J. Clin. Invest. 102: 184-193, 1998), the common problem of gene therapy with viral and non-viral vectors is the limited efficiency of gene transfer. By the above-described additional integrin ligand sequence, in the adenovirus coat protein, an improvement in the transfer of, for example, the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) cDNA can be achieved.

Similar to T. Tanaka et al. (Cancer Research 58, 3362-3369, 1998), DNA for the sequences of the invention for transfecting cells using retroviral or adenoviral vectors can be used instead of angiostatin DNA.

The peptides of the invention may also be used in a liposome complex from a lipid / peptide / DNA system made to transfect cell cultures from a liposome complex consisting of a lipid / DNA system (without peptide), for use in human gene therapy. The preparation of the liposome complex from the lipid / DNA / peptide system has been described, for example, by S.L. Hart et al. (Lipid-Mediated Enhancement of Transfection by Non-Viral Integrin-Targeting Vector, Human Gene Therapy 9, pp. 575-585, 1998).

A lipid / peptide / DNA liposome complex can be prepared, for example, from the following stock solutions: 1 µg / µl lipofectin (equimolar mixture of DOTMA (= N- [1- (2,3-dioleyloxy) propyl] -N, N, N-trimethylammonium chloride) and DOPE (dioleylphosphatidyl ethanolamine), 10 µg / µl plasmid DNA, and 100 µg / ml peptide, for this purpose, both DNA and peptide are dissolved in the cell culture medium.

The liposome complex is prepared by mixing the three components at a particular weight ratio (lipid: DNA: peptide, for example, 0.75: 1: 4). The liposome-DNA complex has already been described for human gene therapy (Caplen N.J. et al., Liposome-Mediated CFTR gene transfer to the nasal epithelium of cystic fibrosis patients, Nature Medicine 1, pp. 39-46, 1995).

The invention therefore also relates to the use of appropriately modified recombinant DNA from gene-releasing systems, in particular from the virus-DNA system, for the treatment of diseases which are directly or not directly caused by expression of [beta] / [beta] 3 integrin receptors, in particular pathogenic angiogenic diseases, thromboses. cardiac infarction, coronary heart disease, arteriosclerosis, tumors, osteoporosis, inflammation, infections as well as to influence wound healing processes.

The invention is illustrated, but not limited, by the following examples. Temperatures are always given in degrees Celsius.

Chromatographic HPLC analysis (retention time R.) was performed using the following systems:

5 µm LichroSpher 60 RP-Select B (250-4) column with a 50 minute gradient from 0 to 80% 2-propanol in water / 0.3% trifluoroacetic acid at a flow rate of 1 mL / min and detection at 215 nm .

Mass spectrometry (MS): EI (electron impact ionization) M * ·;

FAB (fast atom bombardment) (M + H) Examples

Example 1

Preparation and purification of peptides of the invention t

The peptides of the invention are essentially prepared and purified using the Fmoc strategy to protect acid labile side chains on acid labile resins using commercially available continuous flow peptide synthesizers as described by Haubner et al. (J. Am. Chem. Soc. 118, 17703 (1996)).

Cycling (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu) [EMD 272914]

[Sic] 2.7 g of Fmoc-Abu-OH (Bachem B-1910) are suspended in [sic] 150 ml of dichloromethane and dissolved to clarity using ml of dimethylformamide. 5.6 ml of diisopropylethylamine are added.

The solution is poured onto 12.0 g of o-chlorotryl chloride polystyrene resin (1.14 mmol / g, Bachem D-196512) and the batch is shaken at room temperature.

After five hours, the resin was filtered off with suction and washed with 300 ml of DMC / MeOH / DIEA (17/2/1) each, DCM, DMF, DCM and MeOH. After removal of the solvents, 14.55 g of the Fmoc-amino acid resin is obtained. Three times the Fmoc assay demonstrates a mean load of 541 Aimol / g Fmoc-Abu-O-oCITrt-resin.

Subsequently, it is subjected to 0.7 g

Fmoc-Abu-O-oCITrt-polystyrene resin to the coupling step by double coupling technique 2 x each with 0.30 g TBTU, 0.315 ml ethyldiisopropylamine and Fmoc-amino acids in 4.1 ml dimethylformamide using a commercially available synthesis apparatus and in a manner known per se (apparatus) and Milligen 9050 PpeSynthetizer ™, 1987) · in each case for 30 minutes. Washing is performed in dimethylformamide for 10 minutes, cleavage in piperidine / dimethylformamide (1: 4 v / v) for 5 minutes and N-terminal acetylation (capping) is performed using acetic anhydride / pyridine / dimethylformamide (2: 3 v / v): 15) 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) are used. , then Fmoc-Arg (Pmc) finally Fmoc-ABu. After removal of the Fmoc protecting group from the Fmoc-Abu-Arg- (Pmc) -Thr (But) -Asp (OBut) -Leu-D-Asp (OBut) -AlaLeu-Arg (Pmc) -Abu-0-oCITrt-polystyrene resin After washing with dimethylformamide and isopropanol followed by drying in vacuo at room temperature 2: 0.9 g

Abu-Arg (Pmc) -Thr (But) -Asp (OBut) -Leu-D-Asp (OBut) -Ala-Leu-Arg (Pmc) -Abu-O-oClTrt-polystyrene resin.

Treatment of this peptidyl resin with 20 ml of trifluoroethanol / dichloromethane / acetic acid (2: 6: 2, v / v) for two hours at room temperature, filtration, concentration in vacuo and trituration with diethyl ether affords

0.19 g on the side chain protected

Abu-Arg (Pmc) -Thr (But) -Asp (OBut) -Leu-D-Asp (OBut) -Ala-Leu-Arg (Pmc) -Abu-OH.

¹ and added in dropwise a solution of this product in DMF (100 mg peptide / DMF 15 ml) to a stirred solution of TBTU / HOBt / DIPEA (10:10:11 eq) in 50 ml of dimethylformamide / 100 mg of peptide for 30 minutes and further stirring cyclization is achieved for one hour. After concentration and precipitation with water, 0.15 g of crude cyclo (Arg (Pmc) -Thr (But) -Asp (OBut) -Leu-D-Asp (OBut) -Ala-Leu-Arg (Pmc) -Abu-) is obtained. Abu). Treatment with trifluoroacetic acid / water / TIS (94: 3: 3, v / v) for two hours at room temperature, concentration in vacuo and trituration with diethyl ether gave a precipitate of 85 mg of cyclo (Arg-Thr-Asp-Leu-D-Asp-). Ala-Leu-Arg-Abu-Abu).

The product was purified by RP-HPLC on Lichrosorbe RP 18 (250-25.7 µm, Merck KGaA) in 0.3% trifluoroacetic acid using a gradient of 4% to 24% 2-propanol over 1 hour at a flow rate of 10 mL / mih and the eluate is evaluated by a UV flow photometer at 215 and 254 nm. 51 mg of product are obtained; FAB 1067; Rt [sic] 19.0.

Similarly, the following compounds were prepared:

code (EMD) sequence ΜΟ1. by weight. (G / mol) FAB rt [Sic] 271312 cycling (RTDLDSLR) 957.1 958 15,10 271578 cycling (RTDLdSLR) 957.1 957 17.51 271579 cycling (RTDLČLALR) 941.1 941 16.64 271586 cycling (RGDLDSLR) 913.0 913 17.95 271587 cycling (RGDLdSLR) 913.0 913 17.24

271588 cycling (RGDLdALR) 897.0 897 19.60 271589 cycling (RGDLdGLR) 882,9 883 16.68 271590 cycling (RGDLd- → Ala-LR) 897.0, 897 15.47 271591 cycling (RGDLdALRG) 954.1 954 16.79 271592 cycling (RGDLdALRGG) 1011,1 1011 17.83 271593 cycling (RGDLdALRGGG) 1,068.2 1068 17.79 271594 cycling (RGDLdALRGGGGGG) 1,239.3 1239 17,30 272914 cycling (RGDLdALR-Abu-Abu) 1,067.2 1067 19,00 272966 cycling (RTDLdALRGGG) 1,112.2 1113 17.14 272967 cycling (RTDLdGLRGGG) 1,098.2 1099 19,00 272968 cycling (RTDLDALRGGG) 1,112.2 1113 17.39 272969 cycling (RTDLDGLRGGG) 1,098.2 1099 18.88 272970 cycling (RGDLDALRG) 954.1 955 17.32 272971 cycling (RaDLdALRGGGj 1,082.2 1083 18.26 272972 cycling (RGDLaALRGGG) 1,082.2 1083 18.23 272958 cycling (RGDLdALR-Aha-Aha) 1,123.3 1123 19.38 272959 cycling (RGDLdALR-Aha) 1010,2 1010 20.63 272960 cycling (RGDLdAĽR-Aee) 1042.2 1042 18.92 272961 cycling (RGDLdALRGGGG) 1,125.2 1125 17.56 272962 cycling (RGDLdALRGGGGG) 1,182.3 1182 17.52 272963 cycling (RGDLdGLRGGG) 1,054.1 1054 15.59 272964 cycling (RGDLDALRGGG) 1,068.2 1069 16.84 272965 cycling (RGDLDGLRGGG) 1,054.1 1055 16.03 273028 cycling (RTDLdALR-Aha) 1,072.2 n.d. 19,30 273033 cycling (RTDLČLALR-Abu) 1,044.2 n.d. 16.72 273035 cycling (RGDLdALR-Abu) 1000,1 n.d. 16.96 273038 cycling (RTDLdALR-Aha-Aha) 1,185.4 n.d. 20.46 273040 cycling (RTĎLdALR-Abu-Abu) 1,111.3 n.d. 18.75

X 2 TFA

304219 cycling (RTDLdALR- → Ala) 1012,1 1012 19.4 304218 cycling (RGDLdALR- / 3ala) 968.1 968 18.7 329400 cycling (RGDLdALR) 811.9 812 21.15 329412 cycling (RTDLdALR-Abu-Abu) 1,111.3 1112 20.31 329402 cycling (RGDLdALAGGG) 983.1 984 20.48

329399 cyclo (NMeArg- GDLaALRGGG) 1,038.2 1039 19.66 329398 cyclo (R-NMeGly- DLaALRGGG) 1,038.2 1039 19.76 329397 cyclo (RGD-NMeLeuaALRGGG) 1,038.2 1039 20.22 329396 cyclo (RGDL- [D-NMeAla] - ALRGGG) 1,038.2 1039 20.55 329395 cyclo (RGDLa-NMeAla LRGGG) 1,038.2 1039 21.35 329394 cyclo (RGDLaA-NMeLeu- RGGG) 1,038.2 1039 18.87 329393 cyclo (RGDLaLA-NMeArg- 1,038.2 1039 20.64

GGG) cyclo (RGDLdAAR) cyclo (RGDLdAARGGG)

The amino acid nomenclature is governed by the Eur Directive. J. Biochem. 138, Str. 9-37, 1984.

index = D-amino acid

n.d. = not determined

Example 2

The test receptor binding systémucé ^ / ^ / f ibronektin _(Made peptides of the invention bind in solution together with competitively acting fibronectin to immobilized receptor determined, the Q value as a measure of the selectivity of binding of the test peptide to the delta value Q is calculated from the quotient of the IC _sq test peptide and standards. standards were used as the linear _{Ac-RTDLDSLR-NH2} (code EMD 271293) (lit./patent's Pytela et al ·, Science 231, pp. 1559, 1986).

The binding test shall be carried out as follows:

l

Immobilization of the soluble receptor on the microtiter plates is performed after dilution of the protein solution in TBS ++ and subsequent incubation overnight at 4 ° C (100 µl / well). Non-specific binding sites are blocked by incubation (two hours at 37 ° C) with 3% (w / v) BSA in TBS ++ (200> 11 / well). Excess BSA is removed three times by washing with TBSA ++. The peptides are sterile (1:10) in TBSA ++ diluted and incubated with immobilized integrase (50 µl peptide + 50 µl per well; two hours at 37 ° C) along with biotinylated fibronectin 2 µg / ml. Unbound fibronectin and peptides are removed three times by washing with TBSA ++. Detection of bound fibronectin is performed by incubation (one hour at 37 ° C) with alkaline phosphatase coupled with antibiotic antibody (Biorad) (1: 20,000 in TBSA ++; 100 µl / well). After washing three times with TBSA ++, a colorimetric determination is made by incubating (10-15 minutes at 25 ° C in the dark) with substrate solution (5 mg nitrophenyl phosphate, 1 ml ethanolamine, 4 ml water; 100 µl / well). The enzyme reaction is terminated by the addition of 0.4 M sodium hydroxide solution (100> 11 / well). The color intensity is determined at 405 nm in an ELISA reader and compared to zero. Wells not coated with the receptor are used as zero. Ac-RTDLDSLR-NH ₂ is used as standard. The IC _of the test peptides are subtracted from the graph and from which, together with the IC _having the value of the standard peptide, the Q value of the detected peptides of the invention both.

Q = IC _of test peptide / IC _of standard. The Q value is calculated as the center of the repeat tests.

The results of the described test are shown in the following table.

Table II

Results of the ibronectin receptor binding assay

code (EMD) sequence Q value = IC 50 test peptide EMD 271293 50 271293 Ac-NH-RTDLDSLR 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) 0.03 272,963th cyclo (RGDLdGLRGGG) 0.084 271590 cyclo (RGDLd- / 3Äla-LR) 233 271591 cyclo (RGDLdALRG) 2.1. 271592 cyclo (RGDLČLALRGG) 0.05 271594 cyclo (RGDLdALRGGGGGG) 0, O5 272914 cycling (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 cyclo (RGDLdALRGGGG) 0,033 272962 cyclo (RGDLdALRGGGGG) 0,046 273035 cycling (RGDLdALR-Abu) 0,028 271312 cyclo (RTDLDSLR) 8 271578 cyclo (RTDLdSLR) 104 271579 cyclo (RTDLdALR) 12 272966 cyclo (RTDLdALR-GGG) 0.05

272967 cycling (RTDLdGLR-GGG) 0.15 272968 cycling (RTDLDALR-GGG) 0.14 272969 cycling (RTDLDGLR-GGG) 0.18 273028 cycling (RTDLdALR-Aha) 0,015 273033 cycling (RTDLdALR-Abu) 0,043 273038 cycling (RTDLdALR-Aha-Aha) 0,015 273040 cycling (RTDLdALR-Abu-Abu) 0,014 272971 cycling (RaDLdALR-GGG) 8.2

The following examples illustrate pharmaceutical compositions:

Example A.

Injection ampoules

A solution of 100 g of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Ab) and 5 g of disodium hydrogen phosphate in 3 l of double-distilled water and adjusted to pH 6.5 with 2N hydrochloric acid is filtered and filled into vials, lyophilized under sterile conditions and the sterile vials are sealed. Each vial contains 5 mg of active ingredient.

Example B.

lace,

A mixture of 20 g of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-LeuArg-Abu-Abu) is melted with 100 g of soya lecithin and 1400 g of cocoa butter, poured into molds and allowed to cool. Each suppository contains 20 mg of active ingredient.

- 32 Example C.

solution

Prepare a solution of 1 g of cyclo (Arg-Thr-Asp-Leu-D-Asp-AlaLeu-Arg-Abu-Abu), 9.38 g of sodium dihydrogen phosphate dihydrate,

28.48 g of disodium hydrogen phosphate with 12 molecules of water and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. The pH of the solution is adjusted to 6.8, made up to 1 liter and sterilized by irradiation. This solution can be used as eye drops.

Example D.

ointment

Mix 500 mg of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-ArgAbu-Abu) with 99.5 g of petroleum jelly under aseptic conditions.

Example E.

tablets

From a mixture of 1 kg of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-ArgAbu-Abu), 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate Tablets are conventionally compressed so that each tablet contains 10 mg of active ingredient.

Examples F.

I '' ¹ * ¹

Film-coated tablets

Similar to Example E, tablets are compressed and then coated in a conventional manner with a coating of sucrose, potato starch, talc, tragacanth and dye.

Example G.

Capsules 2 kg of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-Arg-Abu-Abu) are filled into ¹ hard gelatin capsule in a manner known per se such that each capsule contains 20 mg of the active ingredient .

Example H.

ampoule

A solution of 1 kg of cyclo (Arg-Thr-Asp-leu-D-Asp-Ala-Leu-Arg-AbuAbu) in 60 L of double distilled water is sterile filtered and filled into ampoules, lyophilized under sterile conditions and the ampoules are sealed . Each ampoule contains 10 mg of active ingredient.

Example I.

Inhalation spray

Dissolve 14 g of cyclo (Arg-Thr-Asp-Leu-D-Asp-Ala-Leu-ArgAbu-Abu) in 10 L of isotonic sodium chloride solution and fill into conventional commercial spray canisters with a pump mechanism. The solution may be sprayed into the mouth or nose. Each spray (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg.

Industrial Applicability>

Peptide compounds which, as integrin ligand, are useful in the manufacture of pharmaceutical compositions for the treatment of diseases based on the expression and pathological function of integrin receptors.

- 34 TF ^> - 2jdo2_

Claims (9)

I * ' Cyclic peptide derivatives of the general formula I cyclo (Arg-X ¹ -Asp-X ² -X ³ -X ¹ -X ⁵ -X ^e -R ^{: 1} -) (I) wherein means X ¹ Ser, Gly or Thr X ² Leu, clay, no, val or phe, X ³ Asp, Glu, Lys or Phe, X ⁴ Gly, Ala or Ser, X ⁵ Leu, clay, no, val or phe, X ^e Arg, Har or Lys, R ¹ is absent or one or more δ-aminocarboxylic acid residues, wherein the aminocarboxylic acid residue is 500 to 2500 µm in length, said amino acids may also be derivatized, including D and L forms and optically active amino acid residues and their physiologically acceptable salts and solvates. 2. A cyclic peptide of the formula I cyclo ^{(Arg-X: L-Asp-X 2} -X ³ -X ¹ -X ⁵ -X ⁶ -R ^L) (I) wherein X ¹ Ser, Gly or Thr, x ² Leu, clay, no, val or phe, x ³ Asp, Glu, Lys, or Phe, x * Gly, Ala or Ser, x ⁵ Leu, clay, no, val or phe, X® Arg, Har or Lys, R ¹ missing or 1 to 10 amino acids selected from the group including Ala, Asn, Asp, Arg, Cys, Gln, Glu, Hcy, His, Hse, Ile, Leu, Lys, Met, Pen, Phe, Pro, Ser, Thr, Trp, Tyr, Val, and H N- (CH) CHO) - (CH) -COOH, wherein m, n are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, provided that: m + n is greater than 0. wherein said amino acids may also be derivatized, including D and L forms and optically active amino acid residues, and physiologically acceptable salts and solvates thereof. The cyclic peptide derivative according to claim 1 or 2, 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-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-Ae), 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-.-Ala), and their physiologically acceptable salts. The cyclic peptide derivatives of the general formula I according to claim 1 and 2a, the compounds according to claim 3 and their physiologically acceptable salts and solvates as medicaments. Medicaments according to claim 4 as inhibitors for the treatment of diseases based on expression and pathological function Medicaments according to claim 5 for the treatment of thromboses, heart attack, coronary heart disease, arteriosclerosis, tumors, osteoporosis, fibrosis, inflammations, infections, psoriasis, as well as to affect wound healing. A pharmaceutical composition comprising at least one medicament according to claims 5 and 6 and optionally a carrier or excipient and optionally other active substances. • Use of a cyclic peptide derivative according to claims 1 to 3, or a physiologically acceptable salt thereof, for the manufacture of a medicament for the treatment of diseases based on the expression and pathological function of integrin receptors. V é Use of a cyclic peptide derivative according to claim 8 for the manufacture of a medicament for the treatment of coronary heart disease, osteoporosis, fibrosis, inflammation, to affect wound healing. thromboses, heart attack, arteriosclerosis, tumors, infections, psoriasis and also