GB2369357A - Aliphatic, cyclic amino carboxylic acids as integrin antagonists - Google Patents

Abstract

The present invention relates to compounds of the general formula (I), <EMI ID=1.1 HE=11 WI=79 LX=718 LY=965 TI=CF> <PC>processes for their preparation, pharmaceutical compositions containing them as well as their use for the production of pharmaceutical compositions for the treatment of inflammatory diseases.

Description

- 1 Aliphatic'cyclic amino carboxvlic acids as intern antagonists The

present invention relates to compounds of formula (I), 5 R6 X-A Cyc y[CR3R41n z (I) their preparation and use as pharmaceutical compositions as integrin antagonists, especially as a4,6 and/or 407 and/or ogre integrin antagonists and in particular for the production of pharmaceutical compositions suitable for the inhibition or the pre 10 vention of cell adhesion and cell-adhesion mediated disorders. Examples are the treatment and the prophylaxis of atherosclerosis, asthma, chronic obstructive pulmo nary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple scle rosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other in flammatory, autoimmune and immune disorders.

Adhesive interactions between the leukocytes and endothelial cells play a critical role in leukocyte trafficking to sites of inflammation. These events are essential for nor mal host defense against pathogens and repair of tissue damage, but can also contrib ute to the pathology of a variety of inflammatory and autoirnmune disorders. Indeed, 20 eosinophil and T cell infiltration into the tissue is known as a cardinal feature of al lergic inflammation such as asthma.

The interaction of circulating leukocytes with adhesion molecules on the luminal surface of blood vessels appears to modulate leukocyte transmigration. These vascu 25 lar cell adhesion molecules arrest circulating leukocytes, thereby serving as the first step in their recruitment to infected or inflamed tissue sites. Subsequently, the leuko cytes reaching the extravascular space interact with connective tissue cells such as fibroblasts as well as extracellular matrix proteins such as fibronectin, laminin, and collagen. Adhesion molecules on the leukocytes and on the vascular endothelium are 30 hence essential to leukocyte migration and attractive therapeutic targets for interven tion in many inflammatory disorders.

J - 2 Leukocyte recruitment to sites of inflammation occurs in a stepwise fashion begin ning with leukocyte tethering to the endothelial cells lining the blood vessels. This is followed by leukocyte rolling, activation, firm adhesion, and transmigration. A num 5 her of cell adhesion molecules involved in those four recruitment steps have been identified and characterized to date. Among them, the interaction between vascular cell adhesion molecule 1 (VCAM-1) and very late antigen 4 (VLA-4, ably integrin), as well as the interaction between mucosal addressin cell adhesion molecule 1 (MAdCAM-1) and a407 integrin, has been shown to mediate the tethering, rolling, 10 and adhesion of lymphocytes and eosinophils, but not neutrophils, to endothelial cells under a physiologic flow condition. This suggests that the VCAM-1 / VLA-4 and/or MAdCAM-1 / a4,B7 integrin mediated interactions could predominantly medi ate a selective recruitment of leukocyte subpopulations in viva. The inhibition of this interaction is a point of departure for therapeutic intervention (A. J. Wardlaw, J. Al 15 lergy Clin. Immunol. 1999, 104, 917- 26).

VCAM-1 is a member of immunoglobulin (Ig) superfamily and is one of the key regulators of leukocyte trafficking to sites of inflammation. VCAM-1, along with intracellular adhesion molecule 1 (ICAM-1) and E-selectin, is expressed on inflamed 20 endothelium activated by such cytokines as interleukin 1 (IL-l) and tumor necrosis factor a (TNF-a), as well as by lipopolysaccharide (LPS), via nuclear factor cB (NF cB) dependent pathway. However, these molecules are not expressed on resting en dothelium. Cell adhesion mediated by VCAM-1 may be involved in numerous physiological and pathological processes including myogenesis, hematopoiesis, in 25 flammatory reactions, and the development of autoimmune disorders. Integrins VLA-4 and OC407 both function as leukocyte receptors for VCAM-1.

The integrin ably is a heterodimeric protein expressed in substantial levels on all circulating leukocytes except mature neutrophils. It regulates cell migration into tis 30 sues during inflammatory responses and normal lymphocyte trafficking. VLA-4 binds to different primary sequence determinants, such as a Q SP motif of VCAM

J - 3 1 and an ILDVP sequence of the major cell type-specific adhesion site of the alterna tively spliced type III connecting segment domain (CS1) of fibronectin.

In viva studies with neutralizing monoclonal antibodies and inhibitor peptides have 5 demonstrated a critical role for a4 integrins interaction in leukocyte-mediated in flammation. Blocking of VLA-4/ligand interactions, thus, holds promise for thera peutic intervention in a variety of inflammatory, autoimmune and immune diseases (Zimmerman, C.; Exp. Opin. Ther. Patents 1999, 9, 129-133).

10 Furthermore, compounds containing a bisarylurea moiety as a substituent were dis closed as a4,B integrin receptor antagonists: WO 96/22966, WO 97/03094, WO 99/33789, WO 99/37605. However, no aminocycloalkyl carboxylic acids or homologues thereof or heterocyclics analogues thereof with a4,B integrin receptor antagonists activity have been described.

None of these compounds have been described in relation to the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders.

Further to their ably integrin antagonistic activity, the compounds of the present in 20 vention may also be used as a4,B7 or ag,B integrin antagonists.

An object of the present invention is to provide new, alternative, aminobenzoic acids or aminocycloalkylcarboxylic acids or homologues thereof or heterocyclic analogues thereof derived integrin antagonists for the treatment of inflmnmatory, autoimmune 25 and immune diseases.

The present invention therefore relates to compounds of the general formula (I): R6 X-A-Cyc Y-[CR R4ln Z (I wherein

Cyc represents a 5- or 6-membered carbocycle, which can optionally be substituted with up to two residues RCYc, 5 wherein the residues RCYc can independently be selected from the group con sisting of halogen, trifluoromethyl, amino, nitro and cyano A represents an amide moiety of the structure 10 -NRA-iC(0)- or-C(O)NRA -, wherein RA-' represents hydrogen or C'-C o alkyl, Z represents-C(O)ORZ,-C(O)NRZ 2Rz 3,-SO2NRz 2R2 3,-SO(ORz), 15 -SO2(ORZ-), -P(o)RZ- (oRZ-3) or-PO(ORZ)(ORz 3), wherein Rz2 is hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 aL cynyl, C3-C6 cycloalkyl, C6 or C 0 atoll, -C(o)RZ-4 or-SO2RZ4, 20 wherein RZ4 is C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C o aryl, RZ' and RZ-3 are independently selected from the group hydrogen, C -C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3 -C6 cycloalkyl, C6 or C 0 aryl 25 or benzyl, wherein RZ- and RZ-3 can optionally be substituted by 1 to 3 substitu-

ents selected from the group C -C4 alkyl, Cl-C4 alkoxy, halogen, nitro, cyano, R3 represents oR3-, NR3-2R3-3'

- 5 wherein R3-' represents hydrogen or C -C4 alkyl, and R3-2 and R3-3 are independently selected from the group hydrogen, Ct-C4 alkyl and acyl, s or R3 represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, Cl- C4 alkyl, C3-C6 cycloalkyl, halogen, trifluoromethyl, nitro or cyano, wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and Cl-C4 alkyl can optionally be substituted with O to 2 substituents indepen dently selected from the group group C -C4 alkyl, C3-C6 cycloalkyl, C'-C4 alkoxy, halogen, nitro, cyano, carboxy, trifluormethoxy, 15 NR3-4R3-s wherein R3- 4 and R35 are independently selected from the group hydrogen, Cl-C4 alkyl and acyl, 20 R4 represents oR4-l, NR4-2R4-3, wherein R4-l represents hydrogen or C -C4 alkyl, and R4-2 and R4-3 are independently selected from the group hydrogen, Cl-C4 alkyl and acyl, or R4 represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, Cl-C4 alkyl, C3-C6 cycloalkyl, halogen, trifluoromethyl, nitro or cyano,

- 6 wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and C1-C4 alkyl can optionally be substituted with O to 2 substituents in-

dependently selected from the group group C -C4 allcyl, C3-C6 cyclo-

alkyl, C -C4 alkoxy, halogen, nitro, cyano, carboxy, trifluormethoxy, 5 NR4-4R4-5

wherein R4 and R4-s are independently selected from the group hydrogen, C -C4 alkyl and acyl, 10 or R3 and R4 together with the carbon atom to which they are attached form a 5- to 7-membered ring, which can contain up to three heteroatoms selected from the group N. O and S. R6 represents phenyl or a 5- to 6-membered aromatic heterocyclic residue 1 S containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen and sulfur, which is substituted by -NR6-2C(o)NR6-3R6-4 and can furthermore optionally be substituted by halogen, wherein R6-2 and R6-3 are independently selected from the group hydrogen or C -C4 alkyl, or together form a group O:H O and wherein R6-4 represents phenyl,

- 7 wherein R6 can optionally be substituted by 1-2 substituents selected from the group C -C4 alkyl, C -C4 alkoxy, halogen, nitro, trifluoromethyl, trifluoro-

methoxy or cyano, S n represents an integer 2, 3 or 4, X represents bond or -CRX- Rx-2-, wherein RX- and RX-2 can be independently selected from the group hydro 10 yen, C1-C4 alkyl, C2- C4 alkenyl, C2-C4 alkynyl, Y represents an amide moiety of the structure -NRY- C(0)- or-C(O)NRY -, wherein RY-i represents hydrogen or C -C4 alkyl, and pharmaceutically acceptable salts thereof.

20 In a preferred embodiment, the present invention relates to compounds of general formula (I), wherein Cyc represents a 5- membered carbocycle.

In another preferred embodiment, the present invention relates to compounds of gen eral formula (I), wherein the moiety A-Cyc-Y represents a y-amino acid.

In another preferred embodiment, the present invention relates to compounds of gen eral formula (I), wherein B' ' represents a bond and Z represents COORS, wherein RZ-i has the meaning indicated above.

- 8 In another preferred embodiment, the present invention relates to compounds of gen-

eral formula (I), wherein R6 represents phenyl, which is substituted by NHC(0)NHR6=, wherein R6-4 is substituted with methyl or trifluoromethoxy.

5 In another preferred embodiment, the present invention relates to compounds of gen eral formula (I), wherein n is 3.

In another preferred embodiment, the present invention relates to compounds of gen eral formula (I), wherein X represents bond.

A process for preparation of compounds of general formula (I) has also been found, which comprises reaction of compounds of general formula (I') O Rib R6 X-Nit Cyc kNH (I) R5 wherein Cyc, X, R5, R6 and Rig have the abovementioned meaning, 20 with compounds of the general formula (I") o AGJ(CR4R5- Z (I)

wherein 25 R4 R5 and Z have the abovementioned meaning and AG represents an activating group,

- 9 - in inert solvents.

In the context of the present invention alkyl stands for a straight-chain or branched alkyl residue, such as methyl, ethyl, n-propyl, iso-propyl, npentyl. If not stated oth 5 erwise, preferred is C -C o alkyl, very preferred is C -C6 alkyl.

Alkenyl and alkinyl stand for straight-chain or branched residues containing one or more double or triple bonds, e.g. vinyl, allyl, isopropinyl, ethinyl. If not stated oth erwise, preferred is C -C o alkenyl or alkinyl, very preferred is C -C6 alkenyl or 1 0 alkinyl.

Cycloalkyl stands for a cyclic alkyl group such as cyclopropyl, cyclobutyl, cyclo-

pentyl, cyclohexyl or cycloheptyl. Preferred is C3-C7 cycloaLkyl.

15 Halogen in the context of the present invention stands for fluorine, chlorine, bromine or iodine. If not specified otherwise, chlorine or fluorine are preferred.

Carbocycle stands for a ring consisting of carbon atoms.

20 Heteroaryl stands for a monocyclic heteroaromatic system containing 4 to 9 ring at oms, which can be attached via a carbon atom or eventually via a nitrogen atom within the ring, for example, furan-2-yl, furan-3-yl, pyrrol-1-yl, pyrrol-2-yl, pyrrol-3 yl, thienyl, thiazolyl, oxazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidyl or pyridazinyl. A saturated or unsaturated heterocyclic residue stands for a heterocyclic system con taining 4 to 9 ring atoms, which can contain one or more double bonds and which can be attached via a ring carbon atom or eventually via a nitrogen atom, e.g. tetra hydrofur-2-yl, pyrrolidine-1- yl, piperidine-1-yl, piperidine-2-yl,, piperidine-3-yl, 30 piperidine-4- yl, piperazine-1-yl, piperazine-2-yl morpholine-1-yl, 1,4-diazepine-1-yl or 1,4-dihydropyridine-1-yl.

- 10 If not specified otherwise, in the context of the present invention heteroatom stands preferably for O. S. N or P. 5 Surprisingly, the compounds of the present invention show good integrin antagonis tic activity. They are therefore suitable especially as OC40 and/or a4 7 and/or aged integrin antagonists and in particular for the production of pharmaceutical composi tions for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders. Examples are the treatment and the prophylaxis of atherosclerosis, asthma, 10 chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, trans plant rejection and other inflammatory, autoimmune and immune disorders.

The integrin antagonists of the invention are useful not only for treatment of the 15 physiological conditions discussed above, but are also useful in such activities as purification of integrins and testing for activity.

For the treatment of the above-mentioned diseases, the compounds according to the invention can exhibit non-systemic or systemic activity, wherein the latter is pre 20 ferred. To obtain systemic activity the active compounds can be administered, among other things, orally or parenterally, wherein oral administration is preferred.

For parenteral administration, forms of administration to the mucous membranes (i.e. buccal, lingual, sublingual, rectal, nasal, pulmonary, conjunctival or intravaginal) or 25 into the interior of the body are particularly suitable. Administration can be carried out by avoiding absorption (i.e. intracardiac, intra-arterial, intravenous, intraspinal or intralumbar administration) or by including absorption (i.e. intracutaneous, subcuta neous, percutaneous, intramuscular or intraperitoneal administration).

30 For the above purpose the active compounds can be administered per se or in admini stration forms.

- 11 Suitable administration forms for oral administration are, inter alla, normal and en teric-coated tablets, capsules, coated tablets, pills, granules, pellets, powders, solid and liquid aerosols, syrups, emulsions, suspensions and solutions. Suitable admini 5 stration forms for parenteral administration are injection and infusion solutions.

The active compound can be present in the administration forms in concentrations of from 0.001 - 100 % by weight; preferably the concentration of the active compound should be 0.5 - 90% by weight, i.e. quantities which are sufficient to allow the speci 10 fled range of dosage.

The active compounds can be converted in the known manner into the abovemen tioned administration forms using inert non-toxic pharmaceutically suitable auxili aries, such as for example excipients, solvents, vehicles, emulsifiers and/or disper 1 5 sants.

The following auxiliaries can be mentioned as examples: water, solid excipients such as ground natural or synthetic minerals (e.g. talcum or silicates), sugar (e.g. lactose), non-toxic organic solvents such as paraffins, vegetable oils (e.g. sesame oil), alcohols 20 (e.g. ethanol, glycerol), glycols (e.g. polyethylene glycol), emulsifying agents, dis persants (e.g. polyvinylpyrrolidone) and lubricants (e.g. magnesium sulphate).

In the case of oral administration tablets can of course also contain additives such as sodium citrate as well as additives such as starch, gelatin and the like. Flavour en 25 hancers or colorants can also be added to aqueous preparations for oral adrninistra tion. For the obtainment of effective results in the case of parenteral administration it has generally proven advantageous to administer quantities of about 0. 001 to 100 mg/kg, 30 preferably about 0.01 to 1 mg/kg of body weight. In the case of oral administration

- 12 the quantity is about 0.01 to 100 mg/kg, preferably about 0.1 to 10 mg/kg of body weight. It may nevertheless be necessary to use quantities other than those mentioned above, 5 depending on the body weight concerned, the method of administration, the indivi dual response to the active compound, the type of preparation and the time or interval of administration.

Suitable pharmaceutically acceptable salts of the compounds of the present invention 10 that contain an acidic moiety include addition salts formed with organic or inorganic bases. The salt forming ion derived from such bases can be metal ions, e.g., alumi num, alkali metal ions, such as sodium of potassium, alkaline earth metal ions such as calcium or magnesium, or an amine salt ion, of which a number are known for this purpose. Examples include ammonium salts, arylalkylamines such as dibenzylamine 15 and N,N-dibenzylethylenediamine, lower alkylamines such as methylamine, t butylamine, procaine, lower alkylpiperidines such as Nethylpiperidine, cycloalkyl amines such as cyclohexylamine or dicyclohexylamine, 1-adamantylamine, benza thine, or salts derived from amino acids like arginine, Iysine or the like. The physio logically acceptable salts such as the sodium or potassium salts and the amino acid 20 salts can be used medicinally as described below and are preferred.

Suitable pharmaceutically acceptable salts of the compounds of the present invention that contain a basic moiety include addition salts formed with organic or inorganic acids. The salt forming ion derived from such acids can be halide ions or ions of 25 natural or unnatural carboxylic or sulfonic acids, of which a number are known for this purpose. Examples include chlorides, acetates, trifluoroacetates, tartrates, or salts derived from amino acids like glycine or the like. The physiologically acceptable salts such as the chloride salts, the trifluoroacetic acid salts and the amino acid salts can be used medicinally as described below and are preferred.

- 13 These and other salts which are not necessarily physiologically acceptable are useful in isolating or purifying a product acceptable for the purposes described below.

The salts are produced by reacting the acid form of the invention compound with an 5 equivalent of the base supplying the desired basic ion or the basic form of the inven tion compound with an equivalent of the acid supplying the desired acid ion in a me dium in which the salt precipitates or in aqueous medium and then lyophilizing. The free acid or basic form of the invention compounds can be obtained from the salt by conventional neutralization techniques, e.g., with potassium bisulfate, hydro-chloric 10 acid, sodium hydroxide, sodium bicarbonate, etc. The compounds according to the invention can form non covalent addition compounds such as adducts or inclusion compounds like hydrates or clathrates. This is known to the artisan and such compounds are also object ofthe present invention.

The compounds according to the invention can exist in different stereoisomeric forms, which relate to each other in an enantiomeric way (image and mirror image) or in a diastereomeric way (image different from mirror image). The invention relates to the enantiomers and the diastereomers as well as their mixtures. They can be separated 20 according to customary methods.

The compounds according to the invention can exist in tautomeric forms. This is known to the artisan and such compounds are also object of the present invention.

- 14 General compound synthesis The synthesis of compounds according to the general formula (I) can be illustrated by the following scheme 1: Rs 0 Ra O IR8 Ste B R6 X No AGJLcyc N pG1 Step A R:X_NJLCyc-NO P as, 11 111 R IV

O O R OIR8 AG [CR R n 2 _ Cyc-N Cyc-N H O V OPG 6 IN [CRtR /X IN St iC R R 0 OPGz Step D O Ret 5 V111

Scheme 1 By coupling of the amines (II) with the carboxylic acids or activated derivatives (III), followed by removal of the protecting group PGi the amides (V) can be obtained.

10 Coupling with the carboxylic acids or activated derivatives (VI), and, if necessary, followed by removal of the protecting group PG2, affords carboxylic acids of type (VIII).

In the above scheme AG stands for hydroxyl or a suitable activating group forming 15 an activated carboxylic acid derivative. Activated carboxylic acids derivatives of this type are known to the person skilled in the art and are described in detail in standard textbooks such as, for example in (i) Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg Thieme Verlag, Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

20 The carboxylic acid is preferably activated symmetrical anhydride or as mixed anhydride, such as, for example, AG = iso-butyl-carbonate or by a coupling agents such as, for example dicyclohexylcarbodiimid (DCC), 1ethyl-3-(3'-dimethylamino

- 15 propyl)carbodiimidexHCI (EDCI), 2-(7-aza-3-oxido- 1H- 1,2,3benzotriazol-1 -yl) 1,1,3,3-tetramethyluronium hexafluorophosphate. Other activated carboxylic acid derivatives such as, for example symmetric anhydrides, halides, or activated esters e.g. succinyl, pentafluorophenyl or N-hydroxybenzotriazole esters may also be 5 employed.

In the above scheme PIG stands for a suitable protecting group of the amino group that is stable under the respective reaction conditions. Protecting groups of this type are known to the person skilled in the art and are described in detail in T. W. Greene, 10 P. G. Wuts, Protective Groups in Organic Synthesis, 3r ea., John Wiley, New York, 1999. The amino group is preferably protected by carbamates, PEG being for example tert-butyloxycarbonyl (Boo), 9-fluorenylmethyloxycarbonyl (FMOC) or benzyloxy carbonyl (Cbz- / Z-) or other oxycarbonyl derivatives.

15 In the above scheme PG2 stands for a suitable protecting group of the carboxyl group or COOPG2 stands for the carboxylic group attached to a polymeric resin suitable for solid phase synthesis. Protecting groups of this type are known to the person skilled in the art and are described in detail in T. W. Greene, P. G. Wuts, Protective Groups in Organic Synthesis, 3r ea., John Wiley, New York, 1999. The carboxyl group is 20 preferably esterified, pG2 being C -alkyl such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, a C3 7 cycloalkyl such as, for example, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclo pentyl, cyclohexyl, an aryl such as, for example, phenyl, benzyl, tolyl or a substituted derivative thereof.

Step A Formation of the amides (IV) can take place by reacting an activated form of the re spective carboxylic acid (III), such as an isobutylcarbonate or N-hydroxybenzo triazole ester - with the desired amine (II) or an acceptable salt thereof.

- 16 iso-Butylcarbonates can be prepared in situ by reaction of the Nprotected amino acid (III) with iso-butylchloroformate as described below. Activated derivatives of the acids (III) such as other anhydrides, halides, esters e.g. succinyl, N-hydroxybenzo triazole or pentafluorophenyl esters or activated carboxylic acids obtained by the 5 reaction with coupling agents such as, for example dicyclohexylcarbodiimid (DCC), 1-ethyl-3-(3'-dimethylaminopropyl) carbodiimidexHCI (EDCI), 2-(7-aza-3-oxido 1 H-1,2,3-benzotriazol-1 -yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate may also be employed.

10 1-Hydroxy-lH-benzotriazol ester of (III) can be prepared, for example, by the re action of the 1-hydroxy-lH-benzotriazol with the carboxylic acids (III) in presence of an coupling agents such as, for example, dicyclohexylcarbodiimid (DCC), 1-ethyl-3 (3'-dimethylaminopropyl) carbodiimidexHCl (EDCI), 2-(7-aza-3-oxido-lH-1,2,3 benzotriazol-1-yl)-1,1, 3,3-tetramethyluronium hexafluorophosphate. Further activat 15 ed derivatives of the acids (III) such as other anhydrides, halides, esters e.g. succinyl or pentafluorophenyl esters or activated carboxylic acids obtained by the reaction with coupling agents such as, for example dicyclohexylcarbodiimid (DCC), 1-ethyl 3 -(3 '-dimethylaminopropyl) carbodiimidexHCl (EDCI), 2-(7-aza-3 -oxide- 1 H- 1,2,3 benzotriazol-1-yl)1,1,3,3-tetramethyluronium hexafluorophosphate may also be 20 employed.

For example, amides oftype (IV) can be prepared as follows: 1) Mixed anhydride procedure A solution of the carboxylic acid derivative (III) and of N-methylmorpholine in an inert solvent was cooled to -15 C and isobutyl chloroformate was added and stirred at 0 C. The amine (II) in an inert solvent was added at -15 C. The solution was stir red at 0 C, and at r.t. and was evaporated. The residue was redissolved in ethyl 30 acetate, washed with aqueous acid and base, dried and evaporated. If necessary the

- 17 product was purified by trituration or by flash-chromatography or used without fur ther purification.

2) 1-Hydroxy-lH-benzotriazol ester procedure A solution of carboxylic acid, 1-hydroxy-lH-benzotriazol (HOBt) and 1-ethyl-3-(3' dimethylaminopropyl)carbodiimidexHC1 (EDCI) in an inert solvent is stirred at r.t ARer addition of the amine and a non-nucleophilic base such as ethyldiisopropyl amine or potassium carbonate stirring is continued at r.t. or elevated temperature.

10 After evaporation, the residue was redissolved in ethyl acetate, washed with aqueous acid and base, dried and evaporated. If necessary the product was purified by trituration or by flash-chromatography or used without further purification.

The above reactions and their implementation are well known to the person skilled in 15 the art and are described in detail in standard textbooks such as, for example, in (i) Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg Thieme Verlag, Stuttgart or Stuttgart or (ii) Comprehensive Organic Synthe sis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

20 Compounds of general formula (II) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives.

When more than one choice of reaction methods exist, the person skilled in the art is able to choose the appropriate pathway according to selectivity and possible use of 25 protecting groups such as described in T. W. Greene, P. G. Wuts, Protective Groups in Organic Synthesis, 3r ea., John Wiley, New York, 1999.

Step B The removal of protecting group PA can be performed, depending on the nature of 30 PG, either by an acid such as trifluoroacetic acid for example in the case PGi is tert butyloxycarbonyl (Boo), a base such as piperidine for example in the case PA is 9

- 18 fluorenylmethyloxycarbonyl (FMOC) or by catalytic hydrogenation for example in the case PA is benzyloxycarbonyl (Cbz- / Z-).

Step C1 5 Formation of the amides (VII) can take place by reacting the respective carboxylic acids (VI) - activated by a coupling agent such as DCC and HOBt; EDCI and HOBt or KATU - or its symmetrical anhydride with the desired amines (V) or an acceptable salt thereof. Activated derivatives of the acids (VI) such as halides, and esters e.g. succinyl or pentafluorophenyl esters may also be employed.

For example, amides (VII) can be prepared as follows: A solution of carboxylic acid, HOBt and EDCI in an inert solvent is stirred at r.t After addition of the amine and a non-nucleophilic base such as ethyldiisopropyl amine stirring is continued at r.t. or elevated temperature. The reaction mixture is 15 poured into water and worked up by standard procedures.

Compounds of general formula (VI) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives.

20 Bisarylureas can be prepared by coupling of an amino phenyl acetic acidderivative and a phenylisocyanate. Cyanuric acid derivatives can be prepared by treatment of ureas with a-chlorocarbonyl isocyantes.

Step D 25 The removal of the protecting group pG2 can be performed either by an acid such as trifluoroacetic acid or an base such as potassium hydroxide or lithium hydroxide, depending on the nature of PG2. Reactions are carried out in aqueous, inert organic solvents such as alcohols e.g. methanol or ethanol, ethers e.g. tetrahydrofurane or dioxane or polar aprotic solvents e.g. dimethylformamide. If necessary, mixtures of 30 the above solvents may be used.

- 19 An alternative synthesis of compounds according to the general formula (I), wherein n = 3,5,7,... represents substituted dicarboxylic acids, can be illustrated by the following scheme 2.

o R8 Cyc-N [C 3 X-I [CR3R41;; /x l H Step C2 K R6 Rs OR V 5 Scheme 2 The coupling of amides (V) with the carboxylic acid anhydrides (IX) affords carboxylic acids of type (VIII).

10 Step C2 Formation of the amides (VIII) can take place by reacting the respective carboxylic acid anhydrides (IX) with the desired amines (V) or an acceptable salt thereof.

The above reaction and their implementation are well known to the person skilled in 15 the art and are described in detail in standard textbooks such as, for example, in (i) Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg Thieme Verlag, Stuttgart or Stuttgart or (ii) Comprehensive Organic Synthe sis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

20 Compounds of general formula (IX) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives.

- 20 Examples

Abbreviations AcOH acetic acid Boc tert-butyloxycarbonyl 5 DCC dicyclohexylcarbodiimid DCM dichloromethane DIPEA diisopropylethylamine EDCI 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimidexHCI eq. equivalents 10 EtOAc ethyl acetate FC flash chromatography GC gas chromatography HATU 2-(7-aza-3 -oxido- 1 H- 1,2,3 -benzotriazol- 1 -yl)- 1,1,3,3 tetramethyluro nium hexafluorophosphate 15 HOBt N-hydroxybenzotriazole monohydrate HPLC high performance liquid chromatography ICAM-1 intracellular adhesion molecule 1 IL-1 interleukin 1 LPS lipopolysaccharide 20 MAdCAM-1 mucosal addressin cell adhesion molecule 1 MeOH methanol MeCN acetonitril min. minutes M.p. melting point 25 NF-,cB nuclear factor cB NMR nuclear magnetic resonance n.d. not determined PE light petroleum (b.p. 40-60 C) r.t. room temperature 30 Rf TLC: Rf value = distance spot traveled / distance solvent front traveled TFA trifluoroacetic acid

t - 21 THF tetrahydrofurane TLC thin layer chromatography TNF-a tumor necrosis factor a tR retention time determined by HPLC 5 VCAM-1 vascular cell adhesion molecule 1 VLA-4 very late antigen 4 ( 40 integrin) General remarks In the examples below, all quantitative data, if not stated otherwise, relate to percent 10 ayes by weighs.

Flash chromatography was carried out on silica gel 60, 403 (E. Merck, Darm-

stadt, Germany).

15 Thin layer chromatography was earned out, employing silica gel 60 F254 coated alu minum sheets (E. Merck, Darmstadt, Germany) with the mobile phase indicated.

Melting points were determined in open capillaries and are not corrected.

20 The mass determinations were earned out using the electron spray ionization (ESI) method employing loop injection or split injection via a HPLC system.

- 22 Precursor synthesis Example I: N-(4-Aminophenyl)-N'-(2-methylphenyl) urea N N: H H 5 2-Methylphenylisocyanate (24.6 g, 184.9 mmol) was added dropwise at 0 C to a solution of 1,4-diamino benzene (20.00 g, 184.9 mmol) in 1000 mL EtOAc. After stirring for 2 h at r.t. the product was collected by filtration (42.7 g, 177.0 mmol).

M.p. >300 C; TLC (PE/EtOAc 1/4) Rf 0.32; H-NMR (400 MHz, D6-DMSO) 2.10 (s, 3H); 4.76 (s, 2H); 6.59 (me, 2H); 6.89 (me, 1H); 7.07-7.15 (m, 4H); 7. 73 (s, 1H); 10 7.85 (me, 2H); 8.50 (s, 1H).

Example II: tert-Butyl 4-( {[(2-methylphenyl)amino]carbonyl} amino)benzyl carbamate J - H J

2-Methylphenylisocyanate (7.57 g, 59.83 mmol) was added dropwise at 0 C to a solution of (4-arnino-benzyl)-carbamic acid tert-butyl ester (13.30 g, 59.83 mmol; prepared analoguous to: Moloney, Gerard P.; Martin, Graeme R.; Mathews, Neil; Milne, Aynsley; Hobbs, Heather; et al. Med. Chem. 1999, 42, 2504 - 2526) in 120 20 mL DCM. The reaction was heated under reflux for 16 h, cooled to r.t. and the precipitated product was collected by filtration and dried in vacuum (19.20g, 54.00 mmol). M.p. 200-202 C; TLC (PE/EtOAc 1/1) Rf 0.65; H NMR (400 MHz, D6-DMSO) 1.39 (s, 9H); 2.24 (s, 3H); 4.06 (d, J=6 Hz, 2H); 6.93 (me, 1H); 7.12 7.17 (m, 4); 7.32 (me, 1H); 7.40 (me, 2H); 7.85 (me, 1H); 7.90 (s, 1H); 8.98 (s, 1H).

- 23 Example III: N-[4-(Aminomethyl)phenyl]-N'-(2-methylphenyl)urea ó:N N -NH2* x TFA To a solution of tert-butyl 4-({[(2-methylphenyl)amino] carbonyl}amino)benzylcarb 5 amate (2.00 g, 5.63 mmol) in CH2Cl: (120 mL) TFA (36 mL) was added at 0 C and stinted for 2 h at r.t.. The reaction mixture was evaporated and the product was collected (2.72 g, TFA salt). M.p. 142-143 C; TLC (PE/EtOAc 3/2) Rf 0.14; H NMR (400 MHz, D6-DMSO) 2. 24 (s, 3H), 3.97 (q, J=5 Hz, 2H); 6.96 (me, 1H); 7.13-7.19 (m, 2); 7.36 (me, 2H); 7.51 (me, 2H); 7.81 (me, 2H); 8.06 (s, 1H); 8.08 (s, 10 3H); 9. 23 (s, 1H).

- 24 Compound synthesis Examples 1-11 were prepared by the following general procedure.

[art Jet AG NtO : J:N:NJ(; Step B 9\N;:NJ

1 \ = F

AG 3 4 Step C2 IX [CR R In: \ O Vl [ O O JL[CR3R4]n Step C1 NJ:N; H: O O NJ:N': JL[CR3R4]ffOPG Step D O ó NtN:; 3f\ N:NgL[CR R4 5 Scheme 3

- 25 Step A: Example IV: tert-Butyl-(lS,3R)-3-({[4-({[(2-methylphenyl) amino]carbonyl}-

amino)phenyl]amino} carb onyl)cycl op entylc arb arn ate o 9; N o s (1S, 3R)-3-[(tert-Butoxycarbonyl)amino]cyclopentanecarboxylic acid (Lit.: Marco-

Contelles, Jose; Bernabe, Manuel; Tetrahedron Lett. 1994, 35, 6361-6364) (2.00 g, 8.72rnmol) was dissolved in DMF (lSmL), HOBT (1.47g, 9.60 mmol), EDCI (1.84 g, 9.60 rnmol) and DIPEA (3.38 g, 26.17 mmol) were added at r. t. and stirred 10 for 2 h. N-(4-Aminophenyl)-N'-(2-methylphenyl)urea (2. 18 g, 9.60 rnmol dissolved in 25 rnL DMSO) was added and stirnng was confined for 12 h. The reaction mixture was hydrolysed with ice, tertbutyl-(lS,3R)-3-({[4-({[(2-methylphenyl)amino]carb onyl}arnino)phenyl] amino}carbonyl)cyclopentylcarbamate (3.32 g, 7.34 rnmol) was collected by filtration, washed with water and isolated. M.p. 188-190 C. ESI-MS: 15 453 [M+H]

Table 1: The following example was prepared according to the general procedure NoStructure Name M.p. ( C) H R tert-Butyl (1S,3R)-3-({[4 N)N: ({[(2-methylphenyl)amino]- 180-182 VH H carbonyl} amino)benzyl] amino} carbonyl)cyclopentyl carbamate

- 26 Step B: Example VI: ( 1 R,3S)-3-Amino-N-[4-( {[(2-methylphenyl)amino] carbonyl} amino) phenyl]cyclopentanecarboxamid H H H tert-Butyl-(lS,3R)-3( {[4-( {[(2-methylphenyl)amino]carbonyl} amino)phenyl]arnin-

o}carbonyl)cyclopentylcarbamate was added to TFA (616 mL) at -5 C and stirred for 0.75 h at r.t.. TFA was removed under vacuum, the residue was triturated with MTBE and DCM and dried, was collected (1R,3S)-3-amino-N[4-({[(2-methyl 10 phenyl)amino]carbonyl}amino)phenyl] cyclopentanecarboxamid (23.87 g, TFA salt).

ESI-MS: 353 [M+H]'

Table 2: The following example was prepared according to the general procedure No Structure Name Àx TFA (1R,3S)-3-Amino-N-[4-({[(2-methyl NiN 2 phenyl)amino]carbonyl}amino)benzyl] H H cyclopentanecarboxamide Step C1: Example VIII: Benzyl N2-[(9H-fluoren-9-ylmethoxy)carbonyl]- N -[(1R, 3S)-3 ( {[4-( {[(2-methylphenyl)amino]carbonyl} amino)phenyl]amino} carbonyl)cyclo pentyl] -L- glut aminate HN HN 4,

- 27 FMOC-L-Glutamic acid benzyl ester (359 ma, 0.78 mmol) was dissolved in DMF (4 mL), HOBT (144 ma, 0.94 mmol), EDCI (180 ma, 0.94 mmol) and DIPEA (240 ma, 1.88 mmol) were added. Alter stirring for 2 h at r.t., (1R, 3S)-3-amino-N [4-( {[(2-methylphenyl)amino]carbonyl} amino)phenyl] cyclopentanecarboxamide - 5 (291 ma, TEA salt dissolved in 2 mL DMF) was added and stirring was continued for 12 h. The reaction mixture was hydrolysed with ice, Ni-[(1S,3R)-3-( [4-({[(2 methylphenyl)amino]carbonyl} amino)phenyl]amino} carbonyl)cyclopentyl]-a-gluta mine (435 ma, 0.55 mmol) was collected by filtration, washed with water and isolated. ESI- MS: 838 [M+HCO0]+ Step D: Example 1: N -[(1S,3R)-3-( {[4-( {[(2Methylphenyl)amino]carbonyl} amino) phenyl]amino} carbonyl)cyclopentyl]-aglutarnine 0 0 H OH Benzyl N2-[(9H-fluoren-9-ylmethoxy)carbonyl]- Nl [(1R,3S)-3-({[4-({[(2-methyl phenyl)amino]carbonyl} amino)phenyl]amino} carbonyl)cyclopentyl]-L-glutaminate (210mg, 0.27 mmol) was dissolved in THE / water (1/1) and KOH (148mg, 2.65 mmol) was added at r.t. The reaction mixture was stirred at 55 C for 2 h. 20 MTBE (5 mL) was added, the phases were separated, the aqueous phase was acidified (pH c 2) and extracted with BE (4*5 mL). The combined organic layers were dried, evaporated and N -[(1S,3R)-3-({[4-({[(2-Methylphenyl)amino]carb onyl} amino)phenyl] amino} carbonyl)cyclopentyl] -a-glutamine (31 ma, 0.06 mmol) was isolated as crystalline material. ESI-MS: 482 [M+H]+

- 28 t =+ =.

tb1 ' t1 ' 1 À. 5, c,., c b.

4, - 8 a >, _ = E Z = O K C = 0 O K

_ _ I A _ '- _ '_ I A

a o E E TO I A.- O O A,

a, = a, a a _,, K a Mu 4 _ 4' - E q O O O O -O a=) 0 _ L I( i =74 rmo Z

- 29 (ú.. = = ==

1 _ Y y Y. _, y 5 3 Y g y Y I Y _ 5

', ' I,. ,,, '. /

O0::D: 6:

1 I4zI I4zI I I4zI ZI

- 30 o Z if:i;,:'ll' ZI ZI ZI ZI ZI

| 0 ZI ZI

Z _1 In

- 31 V = hi'

- 32 In vitro assay: adhesion of Ramos cells to immobilized VCAM-1 (domains 1-3) Preparation of VCAM-1 (extracellular domains 1-3) Complementary DNA (cDNA) encoding 7-domain form of VCAM-1 (GenBank ac 5 cession #M60335) was obtained using Rapid-ScreenTM cDNA library panels (OriGene Technologies, Inc) at Takara Gene Analysis Center (Shiga, Japan). The primers used were 5'-CCA AGG CAG AGT ACG CAA AC-3' (sense) and 5'- TGG CAG GTA TTA TTA AGG AG-3' (antisense). PCR amplification of the 3- domain VCAM-1 cDNA was perform using Pfu DNA polymerase (Stratagene) with the fol 10 lowing sets of primers: (IJ-VCAMdl-3) 5'-CCA TAT GOT ACC TGA TCA ATT TAA AAT CGA GAC CAC CCC AGA A-3'; (L-VCAMdl-3) 5'-CCA TAT AGC AAT CCT AGG TCC AGG GGA GAT CTC AAC AGT AAA-3'. PCR cycle was 94 C for 45 see, 55 C for 45 see, 72 C for 2 min. repeating 15 cycles. After the purifi cation of the PCR product, the fragment was digested with KpnI- AvrII. The digested 15 fragment was ligated into pBluescript IISK(-) (Strategene), which was linearized by digesting with KpnI-XhoI. The ligation was followed by transformation to a Dam/Dcm methylase-free E. cold strain SCS110 (Strategene) to create the donor plasmid pHH7. To direct VCAM-1 molecule into the insect cell secretory pathway, the VCAM-1 coding sequence was fused to signal peptide sequence of honeybee 20 melittin. The resulting melittin-VCAM fusion was placed in correct orientation to the baculovirus polyhedrin promoter. Baculovirus transfer vector containing first 3-do main form VCAM-l (pH10) was constructed by ligation of 0.9 kb fragment from AvrII/KlenowtBclI digests of pH7 into SalI/Klenow/BamHI digests of pMelBacB (Invitrogen). Recombinant baculovirus was generated by using Bac-N-BlueTM Trans 25 fection kit (Invitrogen) according to the manufacture's instruction. The recombinant virus was amplified by infection to High-FiveiM insect cells for 5 - 6 days, and virus titer was determined by plaque assay.

High-Five_ insect cells were pelleted in a 225 ml conical tube by centrifugation at 30 1000 rpm for 5 min. After discarding the supernatant, the pellet was resuspended in 1.5 x 109 pfu (MOI = 5) of high-titer virus solution, followed by incubation for 1.5

- 33 hours at room temperature. The cells were pelleted again and washed once in fresh Express FiveTM serum free medium. The cells were pelleted again and finally, resus pended in 200 ml of fresh Express Five TM medium, transferred to a 1,000 ml shaker flask, and incubated in a shaker at 27 C, 130 rpm, for 48 hours before the culture 5 supernatant was collected. The purification of 3-domain form of VCAM-1 from the culture supernatant was performed by one-step anion exchange chromatography.

Protein concentration was determined by using Coomassie protein assay reagent (Pierce) according to the manufacture's instruction.

10 Preparation of VCAM-1 coated microtiter plates Recombinant human VCAM1 (extracellular domains 1-3) was dissolved at 1.0 g/ml in PBS. Each well of the microtiter plates (Nalge Nunc International, Fluoro nunc Cert, 437958) was coated with 100 pi of substrate or for background control

15 with buffer alone for 15 hours at 4 C. After discarding the substrate solution, the wells were blocked using 150 Ill per well of block solution (Kirkegaard Perry Labo ratories, 50-61-01) for 90 minutes. The plate was washed with wash buffer contain ing 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 rnM KCl and 2 mM MnCl2 just be fore addition of the assay.

In Vitro Assay using Hamos cells Preparation of fluorescence labeled Ramos cells: Ramos cells (American Type Culture Collection, Clone CRL- 1596) were cultured in 25 RPMI 1640 medium (Nikken Bio Medical Laboratory, CM1101) supplemented with 10% fetal bovine serum (Hyclone, A-1119-L), 100 U/ml penicilin (Gibco BRL, 15140-122) and 100 glml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37 C with 5% CO2.

30 Ramos cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25 M of 5(-and-6)-carboxyfluorescein diacetate, succinimidyle ester

- 34 (CFSE, Dojindo Laboratories, 345-06441) for 20 min at room temperature while gently swirling every 5 min. After centrifugation at 1000 rpm for 5 min. the cell pel let was resuspended with adhesion assay buffer at a cell density of 4 x 106 cells/ml.

The adhesion assay buffer was composed of 24 mM Tris-HC1 (pH 7.4), 137 mM 5 NaCl, 27 mM KCl, 4 mM glucose, 0.1 % bovine serum albumin (BSA, Sigma, A9647) and 2 mM MnCI2.

Assay procedure (Ramos cells) 10 The assay solution containing each test compounds or 5 g/ml anti-CD49d mono clonal antibody (Immunotech, 0764) was transferred to the VCAM-1 coated plates.

The final concentration of each test compounds was 5 EM, 10 EM or various con centrations ranging from 0.0001 EM to 10 EM using a standard 5-point serial dilu tion. The assay solution containing the labeled Ramos cells was transferred to the 15 VCAM-1 coated plates at a cell density of 2 x 105 cells per well and incubated for 1 hour at 37 C. The non-adherent cells were removed by washing the plates 3 times with wash buffer. The adherent cells were broken by addition of 1 % Triton X-100 (Nacalai Tesque, 355-01). Released CFSC was quantified fluorescence measurement in a fluorometer (Wallac, ARVO 1420 multilabel counter).

The adhesion of Ramos cells to VCAM-l was analyzed by percent binding calcu lated by the formula: 100 x ( FTS - FBG) / ( FTB - FBG) = % binding, where FTB is the total fluores 25 cent intensity from VCAM-1 coated wells without test compound; FBG is the fluo rescent intensity from wells with anti-CD49d monoclonal antibody and FTS is the fluorescent intensity from wells containing the test compound of this invention.

- 35 In vitro activity: In the Ramos-VCAM-l assay the observed Icso value ranges are indicated in Table 4.

Table4:A>lOpM>B>l M2C No ICso l I A A B 10 A 12 B 15 C 16 C 17 C

Claims (1)

1. - 36 Claims

   1. Compounds of the general formula (I), R6 X-A-Cyc-Y-[CR R41n (I) wherein Cyc represents a 5- or 6-membered carbocycle, 10 which can optionally be substituted with up to two residues RCYc, wherein the residues RCYc can independently be selected from the group consisting of halogen, trifluoromethyl, amino, nitro and cyano 15 A represents an amide moiety of the structure -NRA-IC(0)- or-C(O)NRA I-, wherein RAY represents hydrogen or Cl-Clo alkyl, Z represents-C(O)ORZ I,-C(O)NRz 2Rz-3,-SO2NRz 2Rz 3,-SO(ORz l), -SO2(0RZ-), -P(o)RZ-'(oRZ-3) or-PO(ORZ-I)(ORz-3), wherein Rz-2 is hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 25 C3-C6 cycloalkyl, C6 or Co aryl, '(o)RZ4 or-SO2RZ, wherein Red is C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloaLkyl, C6 or CIO aryl,

   - 37 RZ-I and RZ-3 are independently selected Dom the group hydrogen, ClC4 aLkyl, C2-C6 alkenyl, C2-C6 aLkynyl, C3 -C6 cycloalkyl, C6 or C1O aryl or benzyl, 5 wherein RZ and RZ-3 can optionally be substituted by 1 to 3 substitu ents selected from the group C -C4 alkyl, C,-C4 alkoxy, halogen, nitro, cyano, R3 represents oR3-', NR3 2R3 3, wherein R3-l represents hydrogen or Cl-C4 alkyl, and R3 2 and R3 3 are independently selected from the group hydrogen, Cl-C4 alkyl and acyl, 15 or R3 represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, Cl-C4 alkyl, C3-C6 cycloalkyl, halogen, trifluoromethyl, nitro or cyano, 20 wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and Cl-C4 alkyl can optionally be substituted with O to 2 substituents independently selected Dom the group group C -C4 alkyl, C3-C6 cycloaLkyl, Cl-C4 alkoxy, halogen, nitro, cyano, carboxy, trifluor methoxy, -NR34R3 5, wherein R34 and R3 s are independently selected from the group hydrogen, Cl-C4 alkyl and acyl, R4 represents oR4-i, NR4 2R4-3, wherein R4 l represents hydrogen or Cl-C4 aL cyl, and

   - 38 R4-2 and R4-3 are independently selected from the group hydrogen, C C4 alkyl and acyl, or s R4 represents phenyl, benzyl, ber yloxy or phenoxy, thiophenyl, Cl-C4 alkyl, C3-C6 cycloaLkyl, halogen, trifluoromethyl, nitro or cyano, wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and Cl-C4 10 alkyl can optionally be substituted with 0 to 2 substituents independently selected from the group group C'-C4 alkyl, C3-C6 cycloalkyl, C -C4 alkoxy, halogen, nitro, cyano, carboxy, trifluor methoxy, -NR44R4-5, 15 wherein R44 and R4-5 are independently selected from the group hydrogen, C -C4 alkyl and acyl, or R3 and R4 together with the carbon atom to which they are attached form a 5- to 7membered ring, which can contain up to three heteroatoms selected 20 from the group N. O and S. R6 represents phenyl or a S- to 6-membered aromatic heterocyclic residue containing up to 3 heteroatoms independently selected from the group 25 oxygen, nitrogen and sulfur, which is substituted by -NR6 2C(o)NR6-3R6-4 and can furthermore optionally be substituted by halogen, 30 wherein R6 2 and R6-3 are independently selected from the group hydrogen or C -C4 alkyl, or together form a group

   - 39 oiN4O and wherein R6-4 represents phenyl, s wherein R6 4 can optionally be substituted by 1-2 substituents selected from the group C,C4 alkyl, C,-C4 alkoxy, halogen, nitro, trifluoromethyl, trifluoro methoxy or cyano, 10 n represents an integer 2, 3 or 4, X represents bond or -CRX Rx-2-, wherein RX ' and RX-2 can be independently selected from the group hydro 15 yen, C1-C4 aLkyl, C2- C4 alkenyl, C2-C4 allynyl, Y represents an amide moiety of the structure -NRY- C(0)- or-C(O)NR -, wherein RY-1 represents hydrogen or C,-C4 aLkyl, and pharmaceutically acceptable salts thereof.

   2. Compounds of general formula (I) according to claim 1, wherein Cyc represents a 5- membered carbocycle.

   - 40 3. Compounds of general formula (I) according to claim 1 or 2, wherein the moiety A-Cyc-Y represents a y-amino acid.

   5 4. Compounds of general formula (I) according to any one of claims 1 to 3, wherein R'- represents a bond and Z represents COORS-, wherein RZ- has the meaning indicated above.

   10 5. Compounds of general formula (I) according to any one of claims 1 to 4, wherein R6 represents phenyl, which is substituted by -NHC(o)NHR64, wherein R64 is substituted with methyl or trifluoromethoxy.

   15 6. Compounds of general formula (I) according to any one of claims 1 to 5, wherein n is 3.

   7. Compounds of general formula (I) according to any one of claims 1 to 6, wherein X represents bond.

   8. A process for preparation of compounds of general formula (I) according to any one of claims 1 to 7, which comprises reaction of compounds of general 25 formula (I') R6 X-I Cyc (I) R5 wherein

   l l - 41 Cyc, X, Rs, R6 and Rig have the abovementioned meaning, with compounds of the general formula (I") 5 AGJ:CR4R5_Z (I)

   wherein R4 Rs and Z have the abovementioned meaning and AG represents an activating group, in inert solvents.

   9. The use of a compound according to any one of claims 1 to 7 in the manu facture of a medicament.

   10. The use of a compound according to any one of claims 1 to 7 in the manu facture of a medicarnent for the treatment or the prevention of a condition mediated by integrins.

   20 11. The use of a compound according to any one of claims 1 to 7 in the manu facture of a medicament for the treatment or the prevention of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, auto 25 immune and immune disorders.

   12. Pharmaceutical composition, comprising compounds according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.