WO2002030874A2 - Aliphatic, cyclic amino carboxylic acids as integrin antagonists - Google Patents

Abstract

The present invention relates to compounds of the general formula (I), (I)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

Aliphatic, cyclic amino carboxylic acids as integrin antagonists

The present invention relates to compounds of formula (I),

R— X-A- -Cyc — Y— [CR³R⁴]-

(I)

their preparation and use as pharmaceutical compositions as integrin antagonists, especially as a _\ and/or α₄β₇ and/or agβx integrin antagonists and in particular for the production of pharmaceutical compositions suitable for the inhibition or the pre- vention of cell adhesion and cell-adhesion mediated disorders. Examples are the treatment and the prophylaxis of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, 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 normal host defense against pathogens and repair of tissue damage, but can also contribute to the pathology of a variety of inflammatory and autoimmune disorders. Indeed, eosinophil and T cell infiltration into the tissue is known as a cardinal feature of allergic 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- lar cell adhesion molecules arrest circulating leukocytes, thereby serving as the first step in their recruitment to infected or inflamed tissue sites. Subsequently, the leukocytes reaching the extravascular space interact with connective tissue cells such as fϊbroblasts as well as extracellular matrix proteins such as fibronectin, laminin, and collagen. Adhesion molecules on the leukocytes and on the vascular endothelium are hence essential to leukocyte migration and attractive therapeutic targets for intervention in many inflammatory disorders. 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- ber 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 (NCAM-1) and very late antigen 4 (VLA-4, α₄βj integrin), as well as the interaction between mucosal addressin cell adhesion molecule 1 (MAdCAM-1) and α₄β₇ integrin, has been shown to mediate the tethering, rolling, and adhesion of lymphocytes and eosinophils, but not neutrophils, to endothelial cells under a physiologic flow condition. This suggests that the NCAM-1 / NLA-4 and/or MAdCAM-1 / α₄β₇ integrin mediated interactions could predominantly mediate a selective recruitment of leukocyte subpopulations in vivo. The inhibition of this interaction is a point of departure for therapeutic intervention (A. J. Wardlaw, J Al- lergy Clin. Immunol. 1999, 104, 917-26).

NCAM-1 is a member of immunoglobulin (Ig) superfamily and is one of the key regulators of leukocyte trafficking to sites of inflammation. NCAM-1, along with intracellular adhesion molecule 1 (ICAM-1) and E-selectin, is expressed on inflamed endothelium activated by such cytokines as interleukin 1 (IL-1) and tumor necrosis factor α (TΝF-α), as well as by lipopolysaccharide (LPS), via nuclear factor KB (ΝF- KB) dependent pathway. However, these molecules are not expressed on resting endothelium. Cell adhesion mediated by NCAM-1 may be involved in numerous physiological and pathological processes including myogenesis, hematopoiesis, in- flammatory reactions, and the development of autoimmune disorders. Integrins

NLA-4 and α₄β₇ both function as leukocyte receptors for NCAM-1.

The integrin α₄βι is a heterodimeric protein expressed in substantial levels on all circulating leukocytes except mature neutrophils. It regulates cell migration into tis- sues during inflammatory responses and normal lymphocyte trafficking. NLA-4 binds to different primary sequence determinants, such as a QIDSP motif of VCAM- 1 and an ILDNP sequence of the major cell type-specific adhesion site of the alternatively spliced type III connecting segment domain (CS-1) of fibronectin.

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

Furthermore, compounds containing a bisarylurea moiety as a substituent were disclosed as α₄βj 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 α₄βι 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 a _\ integrin antagonistic activity, the compounds of the present in- vention may also be used as α₄β₇ or α₉βι 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 inflammatory, autoimmune and immune diseases.

The present invention therefore relates to compounds of the general formula (I):

R— X— A- -Cyc — Y— [CR³R⁴]-

(I)

wherein Cyc represents a 5- or 6-membered carbocycle, which can optionally be substituted with up to two residues R^cyc,

wherein the residues R^cyc can independently be selected from the group consisting of halogen, trifluoromethyl, amino, nitro and cyano

A represents an amide moiety of the structure

-NR^C^- or-C^ ^¹-

wherein R ,A-^"1 represents hydrogen or C o alkyl,

Z represents -C(O)OR^z-¹, -C(O)NR^z"2R^z_3, -SO₂NR²-²R^2"3, -SOiOR^2"1), -SOaCOR^2"1), -P(O)R^z"1(OR^z"3) or -POCOR^XOR^2"3),

wherein R^z"2 is hydrogen, Ci -C4 alkyl, C -C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆ or C₁₀ aryl, -C(O)R^2"4 or -SO₂R^2"4,

wherein R^z"4 is C1-C4 alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆ or C₁₀ aryl,

R^2"1 and R^2"3 are independently selected from the group hydrogen, Cι-C alkyl, C₂ -C₆ alkenyl, C₂ -C₆ alkynyl, C₃ -C₆ cycloalkyl, C₆ or Cio aryl or benzyl,

wherein R^2"1 and R^2'3 can optionally be substituted by 1 to 3 substitu- ents selected from the group Cι-C₄ alkyl, Cι-C alkoxy, halogen, nitro, cyano,

R³ represents OR^3"1, NR^3"2R 3-3 wherein R ^" represents hydrogen or C;,-C alkyl, and

R^3"2 and R^3"3 are independently selected from the group hydrogen, d-C₄ alkyl and acyl,

or

R³ represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, Cι-C₄ alkyl, C₃-C₆ cycloalkyl, halogen, trifluoromethyl, nitro or cyano,

wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and - alkyl can optionally be substituted with 0 to 2 substituents independently selected from the group group Cι-C₄ alkyl, C -C₆ cycloalkyl, C₁-C₄ alkoxy, halogen, nitro, cyano, carboxy, trifluormethoxy,

-NR^3'4R^3"5,

wherein R^3"4 and R^3"5 are independently selected from the group hydrogen, - alkyl and acyl,

R⁴ represents OR^4"1 , NR^4"2R^4"3,

wherein R^4"1 represents hydrogen or Cι-C₄ alkyl, and

R^4"2 and R^4"3 are independently selected from the group hydrogen, Cι-C₄ alkyl and acyl,

or

R⁴ represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, Cι-C alkyl, C -C₆ cycloalkyl, halogen, trifluoromethyl, nitro or cyano, wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and C1-C₄ alkyl can optionally be substituted with 0 to 2 substituents independently selected from the group group -C₄ alkyl, C₃-C₆ cycloalkyl, -C₄ alkoxy, halogen, nitro, cyano, carboxy, trifluormethoxy, -NR⁴-⁴R^4"5,

wherein R^4"4 and R^4"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 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 containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen and sulfur,

which is substituted by -NR^6"2C(O)NR^6"3R^6"4 and can furthermore optionally be substituted by halogen,

wherein R^6"2 and R^6"3 are independently selected from the group hydrogen or Cι-C₄ alkyl, or together form a group

and wherein R , 6-4 represents phenyl, wherein R^6"4 can optionally be substituted by 1-2 substituents selected from the group Cι-C₄ alkyl, C]-C₄ alkoxy, halogen, nitro, trifluoromethyl, trifluoro- methoxy or cyano,

n represents an integer 2, 3 or 4,

X represents bond or -CR^X_1R^X"2-

wherein R^x_1 and R^x"2 can be independently selected from the group hydro- gen, C1-C4 alkyl, C₂ - C₄ alkenyl, C2-C4 alkynyl,

Y represents an amide moiety of the structure

-NR ■ Y^Y-^"1^lC-V(O-Y).- o »-r. - nC/(nOu)NmRY"-l

wherein R^γ_1 represents hydrogen or C C₄ alkyl,

and pharmaceutically acceptable salts thereof.

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 general formula (I), wherein the moiety A-Cyc-Y represents a γ-amino acid.

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

1 1 7 1 eral formula (I), wherein R ^" represents a bond and Z represents COOR ^" , wherein R ^" has the meaning indicated above. In another preferred embodiment, the present invention relates to compounds of general formula (I), wherein R⁶ represents phenyl, which is substituted by -NHC(O)NHR^6"4, wherein R^6"4 is substituted with methyl or trifluoromethoxy.

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

In another preferred embodiment, the present invention relates to compounds of general 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 (P)

wherein

Cyc, X, R⁵, R⁶ and R⁸ have the abovementioned meaning,

with compounds of the general formula (T ')

wherein

R⁴' R⁵ and Z have the abovementioned meaning and AG represents an activating group, 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, n-pentyl. If not stated oth- erwise, preferred is Cι-C₁₀ alkyl, very preferred is -Cβ 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 otherwise, preferred is C₁-C₁₀ alkenyl or alkinyl, very preferred is C]-C₆ alkenyl or alkinyl.

Cycloalkyl stands for a cyclic alkyl group such as cyclopropyl, cyclobutyl, cyclo- pentyl, cyclohexyl or cycloheptyl. Preferred is C -C cycloalkyl.

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.

Heteroaryl stands for a monocyclic heteroaromatic system containing 4 to 9 ring atoms, 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 containing 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, piperidine-4-yl, piperazine-1-yl, piperazine-2-yl morpholine-1-yl, 1,4-diazepine-l-yl or 1,4-dihydropyridine-l-yl. If not specified otherwise, in the context of the present invention heteroatom stands preferably for O, S, N or P.

Surprisingly, the compounds of the present invention show good integrin antagonistic activity. They are therefore suitable especially as α₄βi and/or ₄β and/or α₉β_! integrin antagonists and in particular for the production of pharmaceutical compositions for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders. Examples are the treatment and the prophylaxis of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, autoimmune and immune disorders.

The integrin antagonists of the invention are useful not only for treatment of the 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- 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 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, subcutaneous, percutaneous, intramuscular or intraperitoneal administration).

For the above purpose the active compounds can be administered per se or in administration forms. Suitable administration forms for oral administration are, inter alia, normal and en- teric-coated tablets, capsules, coated tablets, pills, granules, pellets, powders, solid and liquid aerosols, syrups, emulsions, suspensions and solutions. Suitable admini- 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- fied range of dosage.

The active compounds can be converted in the known manner into the abovementioned administration forms using inert non-toxic pharmaceutically suitable auxiliaries, such as for example excipients, solvents, vehicles, emulsifiers and/or disper- 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 (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- hancers or colorants can also be added to aqueous preparations for oral administration.

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, preferably about 0.01 to 1 mg/kg of body weight. In the case of oral administration 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, depending on the body weight concerned, the method of administration, the individual 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 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., aluminum, 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 and N,N-dibenzylethylenediamine, lower alkylamines such as methylamine, t- butylamine, procaine, lower alkylpiperidines such as N-ethylpiperidine, cycloalkyl- amines such as cyclohexylamine or dicyclohexylamine, 1-adamantylamine, benza- thine, or salts derived from amino acids like arginine, lysine or the like. The physiologically acceptable salts such as the sodium or potassium salts and the amino acid salts can be used medicinally as described below and are prefened.

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 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. 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 equivalent of the base supplying the desired basic ion or the basic form of the invention compound with an equivalent of the acid supplying the desired acid ion in a medium 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

_ 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 of the 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 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.

General compound synthesis

The synthesis of compounds according to the general formula (I) can be illustrated by the following scheme 1 :

Scheme 1

By coupling of the amines (II) with the carboxylic acids or activated derivatives (III), followed by removal of the protecting group PG¹ the amides (V) can be obtained. 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 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 Nerlag, Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991. The carboxylic acid is preferably activated symmetrical anhydride or as mixed anhydride, such as, for example, AG = wo-butyl-carbonate or by a coupling agents such as, for example dicyclohexylcarbodiimid (DCC), l-ethyl-3-(3'-dimethylamino- propyl)carbodiimidexHCl (EDCI), 2-(7-aza-3-oxido-lH-l,2,3-benzotriazol-l-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 employed.

In the above scheme PG 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, P. G. Wuts, Protective Groups in Organic Synthesis, 3^r ed., John Wiley, New York,

1999. The amino group is preferably protected by carbamates, PG¹ being for example tert-butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbonyl (FMOC) or benzyloxy- carbonyl (Cbz- / Z-) or other oxycarbonyl derivatives.

In the above scheme PG 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, 3^rd ed., John Wiley, New York, 1999. The carboxyl group is preferably esterified, PG² being -_δ-alkyl such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, a C .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 respective carboxylic acid (III), such as an /so-butylcarbonate or N-hydroxybenzotriazole ester - with the desired amine (II) or an acceptable salt thereof. ..yo-Butylcarbonates can be prepared in situ by reaction of the N-protected amino acid (III) with t^'jrø-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 reaction with coupling agents such as, for example dicyclohexylcarbodiimid (DCC), l-ethyl-3-(3'-dimethylaminopropyl)carbodiimidexHCl (EDCI), 2-(7-aza-3-oxido- 1H- 1 ,2,3-benzotriazol-l -yl)- 1 , 1 ,3,3-tetramethyluronium hexafluorophosphate may also be employed.

1-Hydroxy-lH-benzotriazol ester of (III) can be prepared, for example, by the reaction of the 1-hydroxy-lH-benzotriazol with the carboxylic acids (III) in presence of an coupling agents such as, for example, dicyclohexylcarbodiimid (DCC), l-ethyl-3- (3'-dimethylaminopropyl)carbodiimidexHCl (EDCI), 2-(7-aza-3-oxido-lH-l,2,3- benzotriazol-l-yl)-l,l,3,3-tetramethyluronium hexafluorophosphate. Further activat- 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-oxido-lH-l,2,3- benzotriazol-l-yl)-l,l,3,3-tetramethyluronium hexafluorophosphate may also be employed.

For example, amides of type (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 iso-butyl chloroformate was added and stirred at 0°C. The amine (II) in an inert solvent was added at -15°C. The solution was stirred at 0°C, and at r.t. and was evaporated. 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.

2) 1-Hydroxy-lH-benzotriazol ester procedure

A solution of carboxylic acid, 1-hydroxy-lH-benzotriazol (HOBt) and l-ethyl-3-(3'- dimethylaminopropyl)carbodiimidexHCl (EDCI) in an inert solvent is stirred at r.t. After 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. 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 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 Nerlag, Stuttgart or Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

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 protecting groups such as described in T. W. Greene, P. G. Wuts, Protective Groups in Organic Synthesis, 3^rd ed., John Wiley, New York, 1999.

Step B

The removal of protecting group PG¹ can be performed, depending on the nature of PG¹, either by an acid such as trifluoroacetic acid for example in the case PG¹ is tert- butyloxycarbonyl (Boc), a base such as piperidine for example in the case PG¹ is 9- fluorenylmethyloxycarbonyl (FMOC) or by catalytic hydrogenation for example in the case PG¹ is benzvloxycarbonyl (Cbz- / Z-).

Step Cl

Formation of the amides (Nil) can take place by reacting the respective carboxylic acids (NI) - activated by a coupling agent such as DCC and HOBt; EDCI and HOBt or HATU - 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 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.

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

Step D 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 the above solvents may be used. An alternative synthesis of compounds according to the general formula (I), wherein n = 3,5,7,... represents substituted dicarboxyhc acids, can be illustrated by the following scheme 2.

Scheme 2

The coupling of amides (V) with the carboxylic acid anhydrides (IX) affords carboxylic acids of type (VIII).

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 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 Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

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

Examples

Abbreviations

AcOH acetic acid

Boc tert-butyloxycarbonyl

DCC dicyclohexylcarbodiimid

DCM dichloromethane

DIPEA diisopropylethylamine

EDCI l-ethyl-3-(3'-dimethylaminopropyl)carbodiimidexHCl eq. equivalents

EtOAc ethyl acetate

FC flash chromatography

GC gas chromatography

HATU 2-(7-aza-3-oxido-lH-l,2,3-benzotriazol-l-yl)-l,l,3,3-tetramethyluro- nium hexafluorophosphate

HOBt N-hydroxybenzotriazole monohydrate

HPLC high performance liquid chromatography

ICAM-1 intracellular adhesion molecule 1

IL-1 interleukin 1

LPS lipopolysaccharide

MAdCAM-1 mucosal addressin cell adhesion molecule 1

MeOH methanol

MeCN acetonitril min. minutes

M.p. melting point

NF-κB nuclear factor KB

NMR nuclear magnetic resonance n.d. not determined

PE light petroleum (b.p. 40-60 °C) r.t. room temperature

Rf TLC: R_f value = distance spot traveled / distance solvent front traveled

TFA trifluoroacetic acid THF tetrahydrofurane

TLC thin layer chromatography

TNF-α tumor necrosis factor α t_R retention time determined by HPLC NCAM-1 vascular cell adhesion molecule 1

NLA-4 very late antigen 4 (α₄βj integrin)

General remarks

In the examples below, all quantitative data, if not stated otherwise, relate to percent- ages by weight.

Flash chromatography was carried out on silica gel 60, 40-63 μm (E. Merck, Darmstadt, Germany).

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

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

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

Precursor synthesis

Example I: N-(4-Ammophenyl)-N'-(2-methylphenyl)urea

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) R_f 0.32; 1H-NMR (400 MHz, D₆-DMSO) δ 2.10 (s, 3H); 4.76 (s, 2H); 6.59 (mc, 2H); 6.89 (mc, 1H); 7.07-7.15 (m, 4H); 7.73 (s, 1H); 7.85 (mc, 2H); 8.50 (s, 1H).

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

2-Methylphenylisocyanate (7.57 g, 59.83 mmol) was added dropwise at 0 °C to a solution of (4-amino-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. J Med. Chem. 1999, 42, 2504 - 2526) in 120 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 (19.20 g, 54.00 mmol). M.p. 200-202 °C; TLC (PE/EtOAc 1/1) R_f 0.65; 1H NMR (400 MHz, D₆-DMSO) δ 1.39 (s, 9H); 2.24 (s, 3H); 4.06 (d, J=6 Hz, 2H); 6.93 (mc, 1H); 7.12- 7.17 (m, 4); 7.32 (mc, 1H); 7.40 (mc, 2H); 7.85 (mc, 1H); 7.90 (s, 1H); 8.98 (s, 1H). Example III: N-[4-(Aminomethyl)phenyl]-N'-(2-methylphenyl)urea

To a solution of tert-butyl 4-({[(2-methylphenyl)amino]carbonyl}amino)benzylcarb- amate (2.00 g, 5.63 mmol) in CH₂C1₂ (120 mL) TFA (36 mL) was added at 0 °C and stirred 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) R_f 0.14; 1H NMR (400 MHz, D₆-DMSO) δ 2.24 (s, 3H); 3.97 (q, J=5 Hz, 2H); 6.96 (mc, 1H); 7.13-7.19 (m, 2); 7.36 (mc, 2H); 7.51 (mc, 2H); 7.81 (mc, 2H); 8.06 (s, 1H); 8.08 (s, 3H); 9.23 (s, 1H).

Compound synthesis

Examoles 1-11 were prepared by the following general procedure.

Scheme 3 Step A:

Example IN: tert-Butyl-(lS^*,3.R^*)-3-({[4-({[(2-methylρhenyl)amino]carbonyl}- amino)phenyl]amino}carbonyl)cyclopentylcarbamate

(IS ,3R )-3-[(tert-Butoxycarbonyl)amino]cyclopentanecarboxylic acid (Lit.: Marco- Contelles, Jose; Bernabe, Manuel; Tetrahedron Lett. 1994, 35, 6361-6364) (2.00 g, 8.72 mmol) was dissolved in DMF (15 mL), HOBT (1.47 g, 9.60 mmol), EDCI (1.84 g, 9.60 mmol) and DIPEA (3.38 g, 26.17 mmol) were added at r.t. and stirred for 2 h. Ν-(4-Aminophenyl)-Ν'-(2-methylphenyl)urea (2.18 g, 9.60 mmol dissolved in 25 mL DMSO) was added and stirring was confined for 12 h. The reaction mixture was hydrolysed with ice, tert-butyl-(lS^*,3i?^*)-3-({[4-({[(2-methylphenyl)amino]carb- onyl}amino)phenyl]amino}carbonyl)cyclopentylcarbamate (3.32 g, 7.34 mmol) was collected by filtration, washed with water and isolated. M.p. 188-190 °C. ESI-MS: 453 [M+H]⁺

Table 1: The following example was prepared according to the general procedure

No Structure Name M.p:(°C)

amino } carbonyl)cyclopentyl carbamate Step B:

Example VI: (1R^*,3S )-3-Amino-N-[4-({[(2-methylphenyl)amino]carbonyl}amino)- phenyljcyclopentanecarboxamid

tert-Butyl-(lS^*,3i?^*)-3-( {[4-( {[(2-methylphenyl)amino]carbonyl} amino)phenyl]amin- 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 (li-^*,3S^*)-3-amino-N-[4-({[(2-methyl- 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

Step Cl:

Example VIII: Benzyl N²-[(9H-fluoren-9-ylmethoxy)carbonyl]- ^-[(lR^*^^^*)^-

({[4-({[(2-methylphenyl)amino]carbonyl}amino)phenyl]amino}carbonyl)cyclo- pentyl]-Z-glutaminate

FMOC-L-Glutamic acid benzyl ester (359 mg, 0.78 mmol) was dissolved in DMF (4 mL), HOBT (144 mg, 0.94 mmol), EDCI (180 mg, 0.94 mmol) and DIPEA (-J4U mg, l.b8 mmoij were added. After stirring for 2 h at r.t., (li?^*,3S )-3-amino-N- [4-({[(2-methylphenyl)amino]carbonyl}amino)phenyl]cyclopentanecarboxamide (291 mg, TFA salt dissolved in 2 mL DMF) was added and stirring was continued for 12 h. The reaction mixture was hydrolysed with ice, N¹-[(lS^*,3i?^*)-3-({[4-({[(2- methylphenyl)amino]carbonyl}amino)phenyl]amino}carbonyl)cyclopentyl]-α-gluta- mine (435 mg, 0.55 mmol) was collected by filtration, washed with water and isolated. ESI-MS: 838 [M+HCOO]⁺ , -.

Step D:

Example 1: N¹-[(lS^*,3 -^*)-3-({[4-({[(2-Methylphenyl)amino]carbonyl}amino)- phenyl]amino}carbonyl)cyclopentyl]-α-glutamine

Benzyl N²-[(9H-fluoren-9-ylmethoxy)carbonyl]- N¹-[(li-^*,3S^*)-3-({[4-({[(2-methyl- phenyl)amino]carbonyl}amino)phenyl]amino}carbonyl)cyclopentyl]-i.-glutaminate (210 mg, 0.27 mmol) was dissolved in THF / water (1/1) and KOH (148 mg, 2.65 mmol) was added at r.t. The reaction mixture was stirred at 55 °C for 2 h. MTBE (5 mL) was added, the phases were separated, the aqueous phase was acidified (pH < 2) and extracted with EE (4*5 mL). The combined organic layers were dried, evaporated and N¹-[(lS^*,3i?^*)-3-({[4-({[(2-Methylphenyl)amino]carb- onyl}amino)phenyl]amino}carbonyl)cyclopentyl]-α-glutamine (31 mg, 0.06 mmol) was isolated as crystalline material. ESI-MS: 482 [M+H]⁺ Table 3: The following examples were prepared according to the general procedure

No Structure Procedure Name ESI-MS M.p. (°C)

°C)

°C)

No Structure Procedure Name ESI-MS M.p. (°C)

3-Methyl-5-{[(lS ,3R )-3-({[4-({[(2-methylphenyl)-

17 C2 amino]carbonyl}amino)phenyl]amino}carbonyl)- 557.4 221-228 cyclopentyl] amino } -5 -oxo-3 -phenylpentanoic acid [M+H]⁺

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- 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- lowing sets of primers: (U-VCAMdl-3) 5'-CCA TAT GGT 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 sec, 55 °C for 45 sec, 72 °C for 2 min, repeating 15 cycles. After the purification of the PCR product, the fragment was digested with Kpnl-Avrll. The digested fragment was ligated into pBluescript IISK(-) (Strategene), which was linearized by digesting with Kpnl-Xhol. The ligation was followed by transformation to a Dam Dcm methylase-free E. coli strain SCSI 10 (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 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-1 (pHIO) was constructed by ligation of 0.9 kb fragment from Avrll/Klenow/Bcll digests of pH7 into Sall/Klenow/BamHI digests of pMelBacB (Invitrogen). Recombinant baculovirus was generated by using Bac-N-Blue™ Trans- fection kit (Invitrogen) according to the manufacture's instruction. The recombinant virus was amplified by infection to High-Five™ 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 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 hours at room temperature. The cells were pelleted again and washed once in fresh Express Five™ serum free medium. The cells were pelleted again and finally, resus- penαeu in zυυ mi oi rresh 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 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.

Preparation of VCAM-1 coated micro titer plates

Recombinant human VCAM-1 (extracellular domains 1-3) was dissolved at 1.0 μg/ml in PBS. Each well of the microtiter plates (Nalge Nunc International, Fluoro- nunc Cert, 437958) was coated with 100 μl of substrate or for background control with buffer alone for 15 hours at 4 C. After discarding the substrate solution, the wells were blocked using 150 μl per well of block solution (Kirkegaard Perry Laboratories, 50-61-01) for 90 minutes. The plate was washed with wash buffer containing 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 mM KC1 and 2 raM MnCl₂ just before addition of the assay.

In Vitro Assay using Ramos cells

Preparation of fluorescence labeled Ramos cells:

Ramos cells (American Type Culture Collection, Clone CRL-1596) were cultured in 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 μg/ml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37 °C with 5% CO₂.

Ramos cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25 μM of 5(-and -6)-carboxyfluorescein diacetate, succinimidyle ester (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 pellet 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-HCl (pH 7.4), 137 mM NaCl, 27 mM KCl, 4 mM glucose, 0.1 % bovine serum albumin (BSA, Sigma,

A9647) and 2 mM MnCl₂.

Assay procedure (Ramos cells)

The assay solution containing each test compounds or 5 μg/ml anti-CD49d monoclonal antibody (Immunotech, 0764) was transferred to the VCAM-1 coated plates. The final concentration of each test compounds was 5 μM, 10 μM or various concentrations ranging from 0.0001 μM to 10 μM using a standard 5 -point serial dilution. The assay solution containing the labeled Ramos cells was transferred to the 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-1 was analyzed by percent binding calculated by the formula:

100 x ( FTS - FBG ) / ( FTB - FBG ) = % binding, where FTB is the total fluores- cent intensity from VCAM-1 coated wells without test compound; FBG is the fluorescent intensity from wells with anti-CD49d monoclonal antibody and FTS is the fluorescent intensity from wells containing the test compound of this invention. In vitro activity:

In the Ramos-VCAM-1 assay the observed IC₅₀ value ranges are indicated in

Table 4.

Table 4: A > 10 μM > B > 1 μM > C

Claims

Claims

1. Compounds of the general formula (I),

R— X— A— Cyc -Y— [CR³R⁴]-

(I)

wherein

Cyc represents a 5- or 6-membered carbocycle, which can optionally be substituted with up to two residues R^cyc,

wherein the residues R^cyc can independently be selected from the group consisting of halogen, trifluoromethyl, amino, nitro and cyano

A represents an amide moiety of the structure

-NR^A_1C(O)- or -C(O)NR^A_1-

wherein R^A represents hydrogen or - o alkyl,

Z represents -C(O)OR^z_1, -C(O)NR^z"2R^z"3, -SO₂NR^z"2R^z"3, -SO(OR^z_1),

-SO₂(OR^z"1), -P(O)R^z"1(OR^z"3) or -PO OR^XOR^2"3),

wherein R^z"2 is hydrogen, C1-C4 alkyl, ^₂-^₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆ or C₁₀ aryl, -C(O)R^z"4 or -SO₂R^z"4,

wherein R^z"4 is C1-C4 alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆ or C₁₀ aryl, R^2"1 and R^z"3 are independently selected from the group hydrogen, C₁-C₄ alkyl, C -C₆ alkenyl, C -C₆ alkynyl, C₃ -C₆ cycloalkyl, C₆ or C₁₀ aryl or benzyl,

wherein R^z_1 and R^z"3 can optionally be substituted by 1 to 3 substitu- ents selected from the group -C₄ alkyl, CrC₄ alkoxy, halogen, nitro, cyano,

R³ represents OR^3"1, NR^3_2R^3"3,

wherein R^3"1 represents hydrogen or C₁-C₄ alkyl, and

R^3"2 and R^3"3 are independently selected from the group hydrogen, Cι-C₄ alkyl and acyl,

or

R³ represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, -C₄ alkyl, C₃-C₆ cycloalkyl, halogen, trifluoromethyl, nitro or cyano,

wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and -C₄ alkyl can optionally be substituted with 0 to 2 substituents independently selected from the group group Cι-C₄ alkyl, C -C₆ cycloalkyl, Cr alkoxy, halogen, nitro, cyano, carboxy, trifluor- methoxy, -NR^3"4R^3"5,

wherein R^3"4 and R ^" are independently selected from the group hydrogen, Cι-C₄ alkyl and acyl,

R⁴ represents OR^4-1, NR^R^4-3,

wherein R^4"1 represents hydrogen or C C₄ alkyl, and R^4"2 and R^4"3 are independently selected from the group hydrogen, Ci-Gj alkyl and acvl,

or

R⁴ represents phenyl, benzyl, benzyloxy or phenoxy, thiophenyl, C₁-C₄ alkyl, C₃-C₆ cycloalkyl, halogen, trifluoromethyl, nitro or cyano,

wherein phenyl, benzyl, benzyloxy or phenoxy, thiophenyl and -C₄ alkyl can optionally be substituted with 0 to 2 substituents independently selected from the group group C₁-C₄ alkyl, C₃-C₆ cycloalkyl, Cι-C₄ alkoxy, halogen, nitro, cyano, carboxy, trifluor- methoxy, -NR^4"4R^4"5,

wherein R^4"4 and R^4"5 are independently selected from the group hydrogen, C₁-C₄ alkyl and acyl,

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 containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen and sulfur,

which is substituted by -NR^6"2C(O)NR^6"3R^6"4 and can furthermore optionally be substituted by halogen,

wherein R^6"2 and R^6"3 are independently selected from the group hydrogen or

C₁-C4 alkyl, or together form a group

and wherein R^6"4 represents phenyl,

wherein R^6"4 can optionally be substituted by 1-2 substituents selected from the group -C₄ alkyl, C₁-C₄ alkoxy, halogen, nitro, trifluoromethyl, trifluoro- methoxy or cyano,

n represents an integer 2, 3 or 4,

X represents bond or -CR X^X-^_l¹πRX-2

y 1 y wherein R ^" and R ^" can be independently selected from the group hydro- gen, C1-C4 alkyl, C - C₄ alkenyl, C2-C4 alkynyl,

Y represents an amide moiety of the structure

-NR^Y"1C(O)- or-C^NR ^"1-,

wherein R^γ_1 represents hydrogen or Cι-C₄ alkyl,

and pharmaceutically acceptable salts thereof.

2. Compounds of general formula (I) according to claim 1,

wherein Cyc represents a 5- membered carbocycle.

3. Compounds of general formula (I) according to claim 1 or 2,

wherein me moiety A-Cyc-Y represents a γ-amino acid.

4. Compounds of general formula (I) according to any one of claims 1 to 3,

wherein R^1"1 represents a bond and Z represents COOR^2"1,

7 1 wherein R ^" has the meaning indicated above.

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)NHR^6"4, wherein R^6"4 is substituted with methyl or trifluoromethoxy.

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 formula (?)

wherein Cyc, X, R⁵, R⁶ and R⁸ have the abovementioned meaning,

with compounαs of the general formula (I")

wherein

R⁴' R⁵ 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 manufacture of a medicament.

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

11. The use of a compound according to any one of claims 1 to 7 in the manufacture 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- immune and immune disorders.

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