their preparation and use as pharmaceutical compositions as integrin antagonists, especially as a4ßl and/or a4ß7 and/orocupl 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 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 autoimmune disorders. Indeed, 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- 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

hence essential to leukocyte migration and attractive therapeutic targets for interven- tion 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 (VCAM-1) and very late antigen 4 (VLA-4, u, 4Pl integrin), as well as the interaction between mucosal addressin cell adhesion molecule 1 (MAdCAM-1) and a4ß7 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 VCAM-1/VLA-4 and/or MAdCAM-1/047 integrin mediated interactions could predominantly medi- ate 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).

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 endothelium activated by such cytokines as interleukin 1 (IL-1) and tumor necrosis factor a (TNF-a), as well as by lipopolysaccharide (LPS), via nuclear factor xB (NF- lcB) 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- flammatory reactions, and the development of autoimmune disorders. Integrins VLA- 4 and a4ß7 both function as leukocyte receptors for VCAM-1.

The integrin 0. 4 ? i 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. VLA-4 binds to different primary sequence determinants, such as a QIDSP motif of VCAM- 1 and an ILDVP sequence of the major cell type-specific adhesion site of the alterna- tively 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 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).

Furthermore, compounds containing a bisarylurea moiety as a substituent were dis- closed as a4pi integrin receptor antagonists : WO 96/22966, WO 97/03094, WO 99/33789, WO 99/37605. However, no aminobenzoic acids or aminocycloalkyl- carboxylic acids or homologues thereof or heterocyclics analogues thereof with a4 (3 integrin receptor antagonists activity have been described.

3- [ [ [ (phenylacetyl) amino] acetyl] amino]-benzoic acid has been described in Bio- chemistry, Vol. 26, No. 12, 1987, 3385 as a substrate for Vlactamases. N- (4-amino- phenylacetylglycyl)-4-aminophenylacetic acid has been described in J. fur prakt.

Chem., 4. Reihe, Band 27, 1965, 63 without giving a pharmaceutical use. Nl- [4- (eth- oxycarbonyl) phenyl]-N2-(phenylacetyl)-a-glutamine and N2-benzoyl-Nl- [4- (ethoxy- carbonyl) phenyl]-a-glutamine and related compounds have been described in Min- erva Medica, 58 (86), 1967, 3651 and NL 6510006 as antisecretory agents. (S)-4- [ [4- carboxy-l-oxo-2- [ (phenylacetyl) amino] butyl] amino]-benzeneacetic acid has been described in Drugs Exp. Clin. Res. Suppl. 1, XIII, 1987, 57 as antitumor agent. N- [2- [ [4-aminosulfonyl) phenyl] amino]-2-oxoethyl]-N-ethylbenzeneacetamide has been described in Eur. J. Med. Chem.-Chim. Ther. 12 (4), 1977, 387 with schistosomicide activity. N- (2-phenylacetylamino-acetylamino)-benzoic acid ethyl ester has been described in Yakugaku Zasshi 79, 1959, 1606 in decomposition studies of penicil-

lins. Japanese publication Hei 11-269135 describes 3-aminosubstituted benzoic acid derivatives as selectin inhibitors.

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 (X4PI integrin antagonistic activity, the compounds of the present in- vention may also be used as a4ß7 or agpl 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) : wherein Rl represents a 4-to 9-membered saturated, unsaturated or aromatic cyclic residue, which can contain 0 to 3 heteroatoms selected independently from the group N, S and O, wherein the cyclic residue Rl can be annulated with a 4-to 8-mem- bered saturated, unsaturated or aromatic cyclic residue, which can contain 0 to 2 heteroatoms selected independently from the group N, S and O,

and wherein the cyclic residue R'and/or a ring annulated to the cyclic residue Rl is substituted by 1 to 2 substituents -R1-1-R1-2R1-3-Z, wherein Rl-l represents a bond, -O-, -S-, NR1-4, C1-C10 alkyl, C2-C10 alkenyl, C2-CIo alkynyl, C6 or Cio aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 het- eroatoms selected from the group oxygen, nitrogen or sulfur, wherein Rl-l can optionally be substituted by 1 to 2 substituents se- lected from the group R1-5, wherein Rl-5 represents hydrogen, Cl-Clo alkyl, C2-CIo alkenyl, C2-Clo alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 het- eroatoms selected from the group oxygen, nitrogen or sulfur, wherein Rl-5 can optionally be substituted by 1 to 3 substituents se- lected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cy- cloalkyl, halogen, nitro, cyano, oxo, Rl-2 represents a bond,-O-,-S-, NRl-4, C1-C10 alkyl, C2-C10 alkenyl, C2-CIO alkynyl, wherein Rl-2 can optionally be substituted by Cl-Clo alkyl, C2-C10 alkenyl, C2-Clo alkynyl or Rus, wherein Rl-6 represents hydrogen, Cl-Clo alkyl, C2-Clo alkenyl, C2-CIo alkynyl, C6 or Cl0 aryl, C3-C7 cycloalkyl or a 4-9-membered

saturated or unsaturated heterocyclic residue containing up to 3 het- eroatoms selected from the group oxygen, nitrogen or sulfur, wherein Rl-6 can optionally be substituted by 1 to 3 substituents se- lected from the group C1-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, oxo, RlX can optionally be hydrogen, Cl-Clo alkyl, C2-C10 alkenyl or C2-C1o alkynyl, Rl-3 represents a bond, C1-C1 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, wherein Rl-3 can optionally be substituted by Cl-Clo alkyl, C2-C10 alkenyl, C2-Clo alkynyl or Rl-7, wherein R1-7 represents hydrogen, Cl-Clo alkyl, C2-Clo alkenyl, C2-C10 alkynyl, C6 or Clo aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 het- eroatoms selected from the group oxygen, nitrogen or sulfur, wherein Rl-7 can optionally be substituted by 1 to 3 substituents se- lected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, oxo, with the proviso that, where Rl-3 is a bond, R1-2 is not a heteroatom, and with the proviso that Rl-l and Rl-2 are not both heteroatom at the same time,

Z represents-C (O) ORZ-1, -C(O)NRZ-2RZ-3, -SO2NRZ-2RZ-3, -SO(ORZ-1), -S02 (ORZ-1),-P(O)RZ-1(ORZ-3), -PO(ORZ-1)(ORZ-3) or 5-tetrazolyl, wherein Rz-2 is hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or Clo aryl,-C (O) RZ4 or-SO2RZ-4, wherein RZ-4 is Cl-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or Cto aryl, wherein RZ-4 can optionally be substituted by 1 to 3 substituents se- lected from the group halogen, nitro, cyano, oxo, RZ-l and RZ-3 are identical or different and represent hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl, wherein RZ-1 and RZ-3 can optionally be substituted by 1 to 3 substitu- ents selected from the group C1-C4 alkyl, Cl-C4 alkyloxy, halogen, ni- tro, cyano, the cyclic residue Rl and/or a ring annulated to the cyclic residue formed by Rl can optionally be substituted by 0 to 2 substituents Rl-8, halogen, nitro, amino, cyano and oxo, wherein Rl-8 can independently be selected from the group of Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, phenoxy, phenylamino, C3-C6 cycloalkyl, and

represents hydrogen, Cl-Clo alkyl, C2-Clo alkenyl, C2-Clo alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsatu- rated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 3 radicals R-', wherein R2-1 represents C1 alkyl, trifluormethyl, trifluormethoxy, -OR2-2, -SR2-2, NR2-3R2-4, -C(O)R2-2, S(O)R2-2, -SO2R2-2, -CO2R2-2, -OC(O)R2-2, -C(O)NR2-3R2-4, -NR2-2C(O)R2-3, -SO2NR2-3R2-4, NR2-2SO2R2-3, -NR2- 2C(O)NR2-3R2-4, -NR2-2C(O)OR2-3, -OC(O)NR2-3R2-4, halogen, cyano, nitro or oxo, wherein R2-2 represents hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl which can optionally be substituted by 1 substituent selected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, and wherein R2-3 and R2-4 are identical or different and represent hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or Clo aryl, or Ra-3 and R2-4 together form a 4-7-membered ring, which includes the nitrogen atom to which R2-3 and R2-4 are bonded and which contains up to 2 ad- ditional heteroatoms selected from the group oxygen, nitrogen or sulfur and which contains up to 2 double bonds, and if R2 is alkyl, R2 together with the cyclic residue R1 and D can form a

R3 represents hydrogen, Cl-Coo alkyl, C2-Clo alkenyl, C2-Clo alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsatu- rated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, wherein R3 can optionally be substituted by 1 to 3 radicals R3-1, and wherein R3 can furthermore be single-foldedly substituted by C3-C7 cycloalkyl, C6 or Clo. aryl, C4-C9 heteroaryl or a heterocyclic residue contain- ing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can be annulated with a phenyl ring, and which can optionally be substituted by 1 to 3 radicals R3-1, wherein R represents Cl-C4 alkyl, trifluormethyl, trifluonnethoxy, -SR NR3-3R3-4, -C(O)R3-2, S(O)R3-2, -SO2R3-2, -OC(O)R3-2, _C (O)NR3-3R3-4, -NR3-2C(O)R3-3, -SO2NR3-3R3-4, -NR3-2C(O)NR3-3R3-4, -NR3-2C(O)OR3-3, -OC(O)NR3-3R3-4, -CO2R3-5, halogen, cyano, nitro or oxo, wherein R3-2 represents hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl which can optionally be substituted by 1 substituent selected from the group C1-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, and wherein R3-3 and R3-4 are identical or different and represent hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or CIO aryl, benzyl or 9-fluorenylmethyl, or

R3-3 and R34 together form a 4-7-membered ring, which includes the nitrogen atom to which R3-3 and R3-4 are bonded and which contains up to 2 additional heteroatoms selected from the group oxygen, nitrogen or sulfur and which contains up to 2 double bonds, and wherein R3-5 represents Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or Clo aryl R4 represents hydrogen, Cl-Clo alkyl, C2-Cso alkenyl, C2-Clo alkynyl, C6 or Cl0 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsatu- rated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 3 radicals R4-1 and which can furthermore be single-foldedly substituted by C3-C7 cyclo- alkyl, C6 or Clo aryl, C4-Cg heteroaryl or a heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 3 radicals R4-1, wherein R4-l represents Cl-C4 alkyl, trifluormethyl, trifluormethoxy,-oR4-2, -SR4-2, NR4-3R4-4, -C(O)R4-2, S(O)R4-2, -SO2R4-2, -OC(O)R4-2, -C(O)NR4-3R4-4, -NR4-2C(O)R4-3, -SO2NR4-3R4-4, NR4-2SO2R4-3, -NR4-2C(O)NR4-3R4-4, -NR4-2C(O)OR4-3, -OC(O)NR4-3R4-4, -CO2R4-5, halogen, cyano, nitro or oxo, wherein R4-2 represents hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl

which can optionally be substituted by 1 substituent selected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, and wherein R4-3 and R44 are identical or different and represent hydrogen, CI-4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl, or R4-3 and R44 together form a 4-7-membered ring, which includes the nitrogen atom to which R4-3 and R4-4 are bonded and which contains up to 2 additional heteroatoms selected from the group oxygen, nitrogen or sulfur and which contains up to 2 double bonds, and wherein R4-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl Rs represents hydrogen, Cl-Clo alkyl, C2-Clo alkenyl, C2-CIO alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsatu- rated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 3 radicals R5-1. and which can furthermore be single-foldedly substituted by C3-C7 cycloalkyl, C6 or C10 aryl, C4-C9 heteroaryl or a heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 3 radicals R5-1. wherein R5-l represents Cl-C4 alkyl, trifluormethyl, trifluormethoxy, -OR5-2, -SR5-2 NR5-3R5-4, -C(O)R5-2, S(O)R5-2, -SO2R5-2, -CO2R5-2, -OC(O)R5-2, -C(O)NR5-3R5-4, -NR5-2C(O)R5-3, -SO2NR5-3R5-4, NR5-2SO2R5-3,

-NR5-2C(O)NR5-3R5-4, -NR5-2C(O)OR5-3, -OC(O)NR5-3R5-4, halogen, cyano, nitro or oxo, wherein R5-2 represents hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl which can optionally be substituted by 1 substituent selected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, and wherein R5-3 and R54 are identical or different and represent hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl, or R5-3 and R5-4 together form a 4-7-membered ring, which includes the nitrogen atom to which R5-3 and Rs4 are bonded and which contains up to 2 additional heteroatoms selected from the group oxygen, nitrogen or sulfur and which contains up to 2 double bonds, R6 represents phenyl or a 5-to 6-membered aromatic heterocyclic residue containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen or sulfur, which can optionally be annulated with a 5-to 8-membered saturated or un- saturated cyclic residue containing up to 2 heteroatoms independently se- lected from the group oxygen, nitrogen or sulfur, and which can optionally be independently substituted by 1 to 3 radicals R-1 and which can furthermore be single-foldedly substituted by C3-C7 cyclo- alkyl, C6 or C10 aryl, C4-Cg heteroaryl or a heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

wherein the latter cyclic substituents can themselves optionally be substituted by 1 to 3 radicals R6-1, wherein R represents Cl-C4 alkyl, trifluormethyl, trifluormethoxy,-OR6-4, -SR6-2, NR6-3R6 4,-C (O) R6-2, S (O) R6-2, -SO2R6-2, -CO2R6-2, -OC(O)R6-2, -C(O)NR6-3R6-4, -NR6-2C(O)R6-2, -SO2NR6-3R6-4, -NR6-2C(O)NR6-3R6-4, -NR6-2C(O)OR6-4, -OC(O)NR6-3R6-4, halogen, cyano, nitro or oxo, wherein R6-2 represents hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or Clo aryl which can optionally be substituted by 1 to 3 substituents selected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, C3-C6 cycloalkyl, halogen, nitro, cyano, and wherein R6-3 and R64 are identical or different and represent hydrogen, Cl-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group Cl-C4 alkyl, phenyl, C3-C7 cycloalkyl, Cl-C4 alkyloxy, halogen, nitro, cyano, or R6-3 and R64 together form a 4-7-membered ring, which includes the nitrogen atom to which R6-3 and R64 are bonded and which contains up to 2 additional heteroatoms selected from the group oxygen, nitrogen or

sulfur and which contains up to 2 double bonds, which can optionally be substituted by 1 to 2 substituents selected from the group Cl-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, Cl-C4 alkyloxy, halogen, ni- tro, cyano, oxo, and in case that Ri represents a 3-amino benzoic acid derivative and R 6-1 represents-OR6-4,-C (O) NR6-3R6 Or-NR6-2C (O) R6-4, then R6-4 represents C6 or C10 aryl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, wherein the ring formed by R6-3 and R6-4 can optionally be substituted by 1 to 2 substituents selected from the group Cl-C4 alkyl, phenyl, C3-C7 cycloalkyl, Cl-C4 alkyloxy, halogen, nitro, cyano, or R3 and R4 or R4 and R together form a 4-7-membered saturated or unsatu- rated ring containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group Cl-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, Cl-C4 alkyloxy, halogen, nitro, cyano, oxo and which can be fused with a 3-7 membered homocyclic or heterocyclic, saturated, unsaturated or aromatic ring, A represents-C (O)-,-C (O)-C (O)-,-SO-,-SO2-,-PO-,-PO2-, 2-pyri- midyl, 4-pyrimidyl, 2-pyridyl, 2-imidazolyl, 4-imidazolyl, 2-benz- imidazolyl or a ring selected from the following group :

wherein the abovementioned ring systems can optionally be substituted by Cl-C4 alkyl, Cl-C4 alkoxy, halogen, nitro, amino, cyano, X represents-CRX-1RX-2-, wherein RX-l and RX-2 can be independently selected from the group hydro- gen, C 1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, or together with R6 form a 4-7-membered ring, which can contain up to 2 heteroatoms independently selected from the group oxygen, nitrogen or sulfur and containing up to 2 double bonds, which can optionally be substituted by 1 to 2 substituents selected from the group Cl-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, Cl-C4 alkyloxy, halogen, nitro, cyano, oxo,

Y represents bond,-C (O)-,-S (O)-,-S02-,-O-,-S-,-CRY-1RY-2-, or -NRY-3 wherein RY-1, RY-2, RY-3 can be independently selected from the group bond, hydrogen, Cl-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, and can optionally be substituted by 1 to 2 substituents independently selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, Cl-C4 alk- yloxy, halogen, nitro, cyano, oxo, D represents N or CRD-l, wherein RD-l can be independently selected from the group bond, hydrogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, and RD-1 can optionally be substituted by 1 to 2 substituents independently selected from the group Cl-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo, with the proviso that, where D represents-N-, Y does not represent-O-or -S-, and the compound is not one of the following : 3-[[[(phenyl- acetyl) amino] acetyl] amino]-benzoic acid ; N- (4-aminophenylacetylglycyl)-4- aminophenylacetic acid ; Nl- [4- (ethoxycarbonyl) phenyl]-N2- (phenylacetyl)-a- glutamine ; N2-benzoyl-N'- [4- (ethoxycarbonyl) phenyl]-a-glutamine ; (S)-4- [ [4-carboxy-l-oxo-2- [ (phenylacetyl) amino] butyl] amino]-benzeneacetic acid ; N-[2-[[4-aminosulfonyl) phenyl] amino]-2-oxoethyl]-N-ethylbenzeneacet- amide ; N- (2-phenylacetylamino-acetylamino)-benzoic acid ethyl ester, and pharmaceutically acceptable salts thereof.

In a preferred embodiment, the present invention relates to compounds of general formula (I), wherein Rl represents a 4-to 6-membered saturated, unsaturated or aromatic cyclic residue, which can contain 0 to 3 heteroatoms selected independently from the group N, S and O, wherein the cyclic residue Rl can be annulated with a 5-to 6-mem- bered saturated, unsaturated or aromatic cyclic residue, which can contain 0 to 2 heteroatoms selected independently from the group N, S and O, and wherein the cyclic residue Rl and/or a ring annulated to the cyclic residue Rl is substituted by 1 to 2 substituents-R1-1-R1-2-R1-3-Z, wherein RI-1 represents a bond, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl or C6 aryl, wherein Rl-l can optionally be substituted by 1 substituent selected from the group Rl-5, wherein Rl-5 represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl or C6 aryl, Rl-2 represents a bond, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl Rl-3 represents a bond, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl

Z represents-C (O) ORZ-,-C (o) NRZ-2RZ-3 or 5-tetrazolyl, wherein RZ-I, Rz-2 and RZ-3 are identical or different and represent hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl or benzyl, the cyclic residue Rl and/or a ring annulated to the cyclic residue formed by RI can optionally be substituted by 0 to 2 substituents Rl-8, halogen, nitro, amino, cyano and oxo, wherein Rl-8 can independently be selected from the group of Cl-C4 alkyl, Cl-C4 alkyloxy, phenyl, phenoxy, phenylamino, R2 represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6 aryl, C5-C6 cycloalkyl, and if R2 is alkyl, R2 together with the cyclic residue Rl and D can form a 5- to 6-membered ring, R3 represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6 aryl, Cs-C6 cycloalkyl or a 5-6-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 radical R3-1, and wherein R3 can furthermore be single-foldedly substituted by C3-C7 cycloalkyl, C6 aryl, C4-C9 heteroaryl or a heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can be annulated with a phenyl ring,

wherein R3-1 represents trifluormethyl, trifluormethoxy,-OR3-2,-SR3-2, NR3-IRI-4, -NR3-2C(O)OR3-3, -CO2R3-5, halogen, cyano, nitro or oxo, wherein R3-2 represents hydrogen or Cl-C4 alkyl, and wherein R3-3 and R3-4 are identical or different and represent hydrogen, Cl-C4 alkyl or benzyl or 9-fluorenyhnethyl, and wherein R3-5 represents Cl-C4 alkyl, R4 represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 or C6 aryl, R5 represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl or C6 aryl, which can optionally be substituted by 1 radical R5-l, wherein R5-1 represents trifluormethyl, trifluormethoxy, -OR5-2, -SR5-2, NR5-3R5-4, halogen, cyano, nitro or oxo, wherein R5-2, R5-3 and R5-4 are identical or different and represent hydrogen or Cl-C4 alkyl, R6 represents phenyl or a 5-to 6-membered aromatic heterocyclic residue containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen or sulfur, and which can optionally be independently substituted by 1 to 3 radicals R6- wherein R6-1 represents-NR6-2C(O)NR6-3R6-4,

wherein R6-2 and R6-3 are identical or different and represent hydrogen or C1-C4 alkyl, and wherein R64 represents C6 aryl, which can optionally be substituted by 1-2 substituents selected from the group Cl-C4 alkyl, Cl-C4 alkyloxy, halogen, nitro, cyano, or R3 and Ri or R4 and RS together form a 5-6-membered saturated or unsatu- rated ring containing up to 2 nitrogen atoms, A represents-C (O)-, -SO-, -SO2-, X represents-CRX-1Rx-2, wherein R-1 and R-2 can be independently selected from the group hydro- gen, C1-C4 alkyl, Y represents-C (O)-, D represents-N-, and pharmaceutically acceptable salts thereof.

In another preferred embodiment, the present invention relates to compounds of gen- eral formula (I), wherein Rl represents a 5-to 6-membered saturated, unsaturated or aromatic cyclic residue,

which can contain 0 to 3 heteroatoms selected independently from the group N and S, wherein the cyclic residue Rl can be annulated with a 5-membered un- saturated or aromatic cyclic residue, which contains 1 nitrogen atom, and wherein the cyclic residue Rl and/or a ring annulated to the cyclic residue Rl is substituted by 1 to 2 substituents -R1-1-R1-2-R1-3-Z, wherein Rl-l represents a bond or Cl alkyl, wherein Rl-l can optionally be substituted by cyclopentyl, Rl-2 represents a bond, Rl-3 represents a bond, Z represents-C (O) ORZ-1 or 5-tetrazolyl, RZ'1 represents hydrogen, Cl-C2 alkyl or benzyl, the cyclic residue R1 can optionally be substituted by 0 to 2 substituents Rl-8, halogen and nitro, wherein R 1-8 can independently be selected from the group of Cl-C4 alkyloxy, phenoxy and phenylamino,

R2 represents hydrogen or Cl-C3 alkyl, or and if R2 is alkyl, R2 together with the cyclic residue R1 and D can form a piperidine ring, R3 represents hydrogen or Cl-C4 alkyl, which can optionally be substituted by 1 radicalR3-1, wherein R3-l represents NR3-3R34 or-NR3-2C(O)OR3-3, wherein R3-2 and R3-4 represent hydrogen, R3-3 represents hydrogen, benzyl or 9-fluorenylmethyl, R4 represents hydrogen, W represents hydrogen or C3 alkyl, which can optionally be substituted by 1 radical R5-1, wherein R5-l respresents -OR5-2, wherein R5-2 represents Cl alkyl, R6 represents phenyl, and which is substituted by 1 radical R6-

wherein R6-1 represents-NR6-2C (o) NR6-3R6-4, wherein R6-2 represents hydrogen, and wherein R6-3 represents hydrogen and R6-4 represents C6 aryl, which is substituted by 1 substituent Cl alkyl, A represents-C (O)-, X represents -CRX-1RX-2-, wherein el and RX-2 represent hydrogen, Y represents-C (O)-, D represents N, and pharmaceutically acceptable salts thereof.

In another preferred embodiment, the present invention relates to compounds of gen- eral formula (I), wherein Rl represents phenyl, and wherein the phenyl is substituted by 1 to 2 substituents-Rl-l-Rl-2 -R1-3-Z,

wherein Rl-l represents a bond or Cl alkyl, Rl-2 represents a bond, Rl-3 represents a bond, In another preferred embodiment, the present invention relates to compounds of gen- eral formula (I), wherein Z represents-C (O) ORZ~I RZ-1 represents hydrogen, Cl-C2 alkyl or benzyl, R represents hydrogen, represents hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C6 aryl, CS-C6 cycloalkyl or a 5-6-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 radical R 3-1 and wherein R3 can furthermore be single-foldedly substituted by C3-C7 cycloalkyl, C6 aryl, C4-C9 heteroaryl or a heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can be annulated with a phenyl ring,

wherein R3-l represents trifluormethyl, trifluormethoxy,-oR3-2,-SR3-2, -NR3-3R3-4, -NR3-2C(O)OR3-3, -CO2R3-5, halogen, cyano, nitro or oxo, wherein R3-2 represents hydrogen or Cl-C4 alkyl, and wherein R3-3 and R3-4 are identical or different and represent hydrogen, Cl-C4 aIkyl or benzyl or 9-fluorenylmethyl, and wherein R3-5 represents C1-C4 alkyl, R4 represents hydrogen, R5 represents hydrogen, R6 represents phenyl, and which is substituted by 1 radical Run wherein R6-1 represents -NR6-2C(O)NR6-3R6-4, wherein R6-2 represents hydrogen, and wherein R6-3 represents hydrogen and R6-4 represents C6 aryl, which is substituted by 1 substituent Cl alkyl, or R3 and R4 or R4 and R5 together form a 5-6-membered saturated or a un- saturated ring containing up to 2 nitrogen atoms,

A represents-C (O)-, X represents -CRX-1RX-2-, wherein RX-1 and RX-2 represent hydrogen, Y represents-C (O)-, D represents N, and pharmaceutically acceptable salts thereof.

In a more preferred embodiment, the present invention relates to compounds of gen- eral formula (I), wherein Rl represents phenyl, which is 1, 4-substituted by a substituent -R1-1-R1-2-R1-3-Z, wherein R1-1, Rl-2 and Rl-3 represent bonds.

In another more preferred embodiment, the present invention relates to compounds of general formula (I), wherein Rl represents phenyl,

which is 1, 3-substituted by a substituent -R1-1-R1-2-R1-3-Z, wherein Rl-l represents-CH2-, Rl-2 and Rl-3 represent bonds.

In another more preferred embodiment, the present invention relates to compounds of general formula (I), wherein Rl represents a 5-membered heterocycle.

In another more preferred embodiment, the present invention relates to compounds of general formula (I), wherein Rl represents a cyclohexyl ring.

In a very preferred embodiment, the present invention relates to compounds of general formula (I), wherein R6 represents A preferred process for preparation of compounds of general formula (VII)

has also been found, which comprises reaction of carboxylic acids of general formula (V) or activated derivatives thereof, with compounds of the general formula (VI) in the presence of a coupling agent and a base in inert solvents, which will be de- scribed in more detail in the descriptive part of the specification.

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 Cl-Coo alkyl, very preferred is Cl-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 Cl-Coo alkenyl or alkinyl, very preferred is Cl-C6 alkenyl or alkinyl.

Cycloalkyl stands for a cyclic alkyl group such as cyclopropyl, cyclobutyl, cyclo- pentyl, cyclohexyl or cycloheptyl. Preferred is C3-C7 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.

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. C4-Cg heteroaryl also stands for a 4 to 9-membered ring, wherein one or more of the carbon atoms are replaced by heteroatoms.

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, piperidine-4-yl, piperazine-1-yl, piperazine-2-yl morpholine-1-yl, 1, 4-diazepine-1-yl or 1, 4-dihydropyridine-1-yl.

If not specified otherwise, in the context of the present invention heteroatom stands preferably for O, S, N or P.

Annulated describes 1, 1- or 1, 2-fused ring systems, e. g. spiro systems or systems with a [0]-bridge. If not stated otherwise, substituents described for the"parent"ring system (the ring to which the annulated ring is attached) can be also present on the annulated ring.

Derivative stands for a compound that is derived from the parent compound by ex- change of one or more hydrogen atoms by other functional groups.

Surprisingly, the compounds of the present invention show good integrin antagonis- tic activity. They are therefore suitable especially as a4ßl and/or a4ß7 and/or agßl 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, 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 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, subcuta- neous, percutaneous, intramuscular or intraperitoneal administration).

For the above purpose the active compounds can be administered per se or in admini- stration 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- station 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 abovemen- tioned administration forms using inert non-toxic pharmaceutically suitable auxili- aries, 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 administra- 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, 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 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 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 and NN-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 physio- logically acceptable salts such as the sodium or potassium salts and the amino acid salts can be used medicinally as described above 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 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 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, hydrochloric 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 :

0 o 1-2-1-3 Of R R 0 R 0 PGN AG + H R_ Step A PGIiN p Rd/\R3 wD R4 Ra Rz IN Ruz (III) O R (IV) Six OH VU () Step B H'IN D R au 3 R R Step C Nez 0 -OPGz O -OH Rs Ri-2 R-3 5 Ryz Ri-3 R O RR'---StepC (V) y 4 3 1 O R (Vil) (Vill) Scheme 1 By coupling of the carboxylic acids or activated derivatives (II) with the amines (III) (D = nitrogen), followed by removal of the protecting group PGI the amides (V) can be obtained. Coupling with the carboxylic acids (VI) followed by removal of the protecting group PG affords carboxylic acids of type (VIII). Further examples with different A, Y and D groups as defined in formula (I) are described below.

In the above scheme the depicted ring in formulas (III)- (V), (VII) and (VIII) as well as in scheme 3 represents a cyclic moiety formed by Rl. 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 Verlag, Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991. The carboxylic acid is preferably activated as mixed anhydride, such as, for example, AG = iso-butyl-carbonate ; as N-carboxyanhydride (Rs and AG =-CO-) ; or by a coupling agents such as, for example dicyclohexyl- carbodiimid (DCC), l-ethyl-3- (3'-dimethylaminopropyl) carbodiimidexHCl (EDCI), 2- (7-aza-3-oxido-1H-1, 2, 3-benzotriazol-1-yl)-l, 1, 3, 3-tetramethyluronium hexa- fluorophosphate. Other activated carboxylic acid derivatives such as, for example symmetric anhydrides, halides, or activated esters e. g. succinyl or pentafluorophenyl esters may also be employed.

In the above scheme PGl 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, 3d ed., John Wiley, New York, 1999. The amino group is preferably protected by carbamates, PGl being for example tert-butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbonyl (FMOC) or benzyloxy- carbonyl (Cbz-/Z-) or other oxycarbonyl derivatives.

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, 3d ed., John Wiley, New York, 1999. The carboxyl group is preferably esterified, PG2 being Cl 6-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 (I can take place by reacting an activated form of the re- spective carboxylic acid (II), such as a N-carboxyanhydride or an iso-butylcarbonate with the desired amine (III) or an acceptable salt thereof.

N-carboxyanhydrides of (II) are commercially available or can be prepared for ex- ample by the reaction of the Bis- (N-tert-butyloxycarbonyl) protected derivative of (II) with thionylchloride and pyridine in dimethylformamide or by the reaction of the free amino acid of (II) with phosgene or with phosgene equivalents such as diphos- gene, triphosgene or methylchloroformate. Iso-butylcarbonates can be prepared in situ by reaction of the N-protected amino acid (II) with iso-butylchloroformate as described below. Activated derivatives of the acids (II) 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 dicyclohexyl- carbodiimid (DCC), l-ethyl-3- (3'-dimethylaminopropyl) carbodiimidexHCl (EDCI), 2- (7-aza-3-oxido-1H-1, 2, 3-benzotriazol-1-yl)-1, 1, 3, 3-tetramethyluronium hexafluo- rophosphate may also be employed.

For example, amides of type (IV) can be prepared as follows : 1) N-carboxyanhydride procedure A solution/suspension of the amine (III), the N-carboxyanhydride of (II) and catalytic amounts of 4- (N, N'-dimethylamino) pyridine in an inert solvent was refluxed for 0. 5-14 days with exclusion of moisture. The product was either isolated by filtration or by aqueous workup employing standard procedures. If necessary the product was purified by trituration or by flash-chromatography or used without further purifica- tion.

2) Mixed anhydride procedure A solution of the carboxylic acid derivative (II) 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 (III) 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 fur- ther purification.

Compounds of general formula (II) are commercially available, known or can be prepared by customary methods starting from known a-amino acids or precursors for customary a-amino acid synthesis. For the preparation process according to the in- vention, the amino group is in this case blocked by a conventional protective group PGI.

In the a-position to the carboxyl group, these carboxylic acid derivatives can have substituents such as described under R3 and R4, for example, hydrogen, a Cl-Cl0- alkyl, a C3-C7-cycloalkyl, an aryl, an alkenyl residue, or an alkinyl residue. The alkyl, alkenyl and cycloalkyl residues and the benzyl residue can be introduced by reaction of the ester of the starting compounds with the appropriate alkyl, alkenyl, cycloalkyl or benzyl halides in basic medium, if the corresponding derivatives are not commercially available. The alkinyl residue can be introduced, for example, by reac- tion of the bromo ester of the present starting compound with an appropriate acet- ylide anion. In the case of the phenyl residue the starting materials used are prefer- ably the corresponding a-phenyl-a-aminocarboxylic acid derivatives and, if neces- sary, the other substituents at the a-C atom to the terminal carboxyl group are intro- duced via the appropriate alkyl halide.

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 Verlag, Stuttgart or Stuttgart or (ii) Comprehensive Organic Synthe- sis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

If the substituents themselves should be substituted, e. g. by R', appropriate reactive groups should be present in the substituent to allow further functionalization. These reactive groups should be inert to the reaction conditions of the previous step. For this purpose, the substituent can also be unsaturated to allow further functionalization such as palladium catalyzed C-C-coupling reactions (e. g. Heck-reaction or Sonoga- shira-reaction), eventually followed by hydrogenation (scheme 2) : Scheme 2 In the abovementioned scheme PG4 stands for a protecting group of the carboxyl group as described under PG2, hal stands for a leaving group such as a halogen, tosyl, mesyl or triflate, [Pd] stands for a Palladium (0) or Palladium (II) moiety. PG3 stands for a protecting group of the amino group such as described under PG1.

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, 3rd ed., John Wiley, New York, 1999.

If the substituent R3 or R4 in the a-position to the carboxylic group carry an ap- propriate substituted aryl or heteroaryl unit, another method for insertion of an addi- tional substituent are the C-C-coupling reactions as described under the synthesis of precursors (VI).

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

In case Rl-l, Rl-2 andlor Rl-3 are methylen groups, the carbon chain can be elongated by Amdt-Eistert-reaction and optionally be derivatized by common methods for a- derivatization of carboxylic acids such as nucleophilic substitution.

In case, Y is different from carbonyl and/or D is different from nitrogen-as defined in formula (1)-the respective compounds (IV) can be prepared as follows : For example, in case Y and D form an sulfinamide, or sulfonamide, they may be pre- pared by reacting the respective sulfinylchlorides or sulfonylchlorides with the de- sired amine (III) or an acceptable salt thereof.

For example, in case Y and D form an ether or thioether, the O-C or S-C-bonds are formed via alkylation of the corresponding alcohols or thiols with alkylating agents such as alkyl halides, alkyl tosylates and the like. The thioether can be converted into the corresponding sulfoxides or sulfones by oxidation with reagents like mCPBA or hydrogen peroxide.

In case Y and D form a carbon-nitrogen-bond or a nitrogen-carbon-bond, the bond is established by reductive amination via the corresponding aldehyde or ketone and the corresponding amine in the presence of a reducing agent such as sodium cyanoboro- hydride. In the case Y and D form a carbon-nitrogen bond in which the nitrogen atom is attached to an aromatic ring, the amine group-Y-NR2H can be coupled to the aro- matic ring by an Buchwald reaction employing an halogen or triflate substituted aromatic residue and a suitable catalyst such as, for example Pd (0) or Pd (II) with phospine ligands such as triphenylphosphine, 2, 2'-bis- (diphenylphosphino)-1, 1'-bi- naphthyle (BINAP) or 1, 1'-bis-(diphenylphosphino) ferrocene (dppf) together with an appropriate base such as, for example cesium carbonate or cesium fluoride.

In case Y and D form a carbon-carbon-bond, the bond may be established by Wittig reaction of the corresponding ketone or aldehyde and the corresponding phospho- nium ylide followed by reduction of the double bond, e. g. by catalytic hydrogenation.

In case Y is carbonyl and D is a carbon moiety, the bond may be formed by a Grignard type reaction of the corresponding aldehyde of Y and the corresponding Grignard-reagent of D, followed by the oxidation of the resulting alcohol to the ke- tone, e. g. by Swern-oxidation or Jones-oxidation.

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 Verlag, Stuttgart Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991.

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 d ed., John Wiley, New York, 1999.

Step B The removal of protecting group PGl can be performed, depending on the nature of Pagi, either by an acid such as trifluoroacetic acid (for example in the case PGl is tert- butyloxycarbonyl (Boc)), a base such as piperidine (for example in the case PGl is 9- fluorenylmethyloxycarbonyl (FMOC)) or by catalytic hydrogenation (for example in the case Pu1 ils benzyloxycarbonyl (Cbz-/Z-)).

Step C 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 HATU-with the desired amines (V) or an acceptable salt thereof. Activated de- rivatives of the acids (VI) such as anhydrides, 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 ethylisopropylamine 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.

For example, biphenyl substituted acetic acid derivatives can be prepared by means of an aryl-aryl coupling of the respective phenyl acetic acid derivatives and a suitable phenyl system.

Possible coupling reactions are, for example, the reaction of two unsubstituted phenyl groups in the presence of Aids and an acid (Scholl reaction), the coupling of the two phenyl iodides in the presence of copper (Ullmann reaction), the reaction of the unsubstituted carboxylic acid derivative with a phenyldiazonium compound un- der basic conditions (Gomberg-Bachmann reaction) or coupling with participation of organometallic reagents such as coupling of a phenyl halide with an organometallic phenyl compound in the presence of a palladium compound, for example a Pd (0), a Pd (II) or a Pd (IV) compound, and of a phosphane such as triphenylphosphane (e. g.

Suzuki reaction).

Bisarylureas can be prepared by coupling of an amino phenyl acetic acid derivative and a phenylisocyanate. Bisarylamides can be prepared by coupling of an amino phenyl acetic acid and an activated benzoic acid derivative such as described under Step A. Bisarylcarbamates can be prepared by coupling of an isocyanato phenyl ace- tic acid ester and a phenol derivative followed by saponification as described in Step D.

In case, A-as defined in formula (I)-is different from carbonyl, the respective com- pounds (IV) can be prepared as follows : For example, in case A forms a sulfinamide, sulfonamide, they may be prepared as described under Step A. Oxalic amides can be prepared by the same means as the amides described above. Phosphinic acid amides and phosphonic acid amides can be prepared by coupling of activated phosphinic/phosphonic acids with amines (V). In case A is a heteroaromatic or aromatic system, the respective compounds (IV) can be prepared by nucleophilic substitution of the respective fluorosubstituted systems with a suitable amine (V).

Step D The removal of the protecting group PG 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.

In case PG2 stands for polymeric resin, the removal can take place using strong acid such as trifluoroacetic acid in dichloromethane.

Examples Abbreviations AcOH acetic acid Boc tert-butyloxycarbonyl DCC dicyclohexylcarbodiimid GC gas chromatography DIPEA diisopropylethylamine EDCI l-ethyl-3-(3 «-dimethylaminopropyl) carbodiimidexHCl eq. equivalents FC flash chromatography HATU 2- (7-aza-3-oxido-1H-1, 2, 3-benzotriazol-1-yl)-1, 1, 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 min. minutes M. p. melting point NF-KB nuclear factor KB NMR nuclear magnetic resonance n. d. not determined r. t. room temperature Rf TLC : Rf value = distance spot traveled/distance solvent front traveled TFA trifluoroacetic acid THF tetrahydrofurane TLC thin layer chromatography

TNF-a tumor necrosis factor a tR retention time determined by HPLC VCAM-1 vascular cell adhesion molecule 1 VLA-4 very late antigen 4 (aryl 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, 40-63pm (E. Merck, Darm- stadt, Germany).

Thin layer chromatography was carried 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.

All retention times are indicated in minutes and, if not stated otherwise, were deter- mined by high-performance liquid chromatography (HPLC) by means of UV detec- tion at 210/250 nm and a flow rate of 1 ml/min. An acetonitrile/water mixture with 0. 1% trifluoroacetic acid (vol./vol.) was used as eluent with a linear gradient of : 0 min. = 0% acetonitrile, 25 min. = 100% acetonitrile, 31 min = 100 % acetonitrile, 32 min 0% acetonitrile, 38 min 0% acetonitrile. Two methods were used : for method A a LiChrospher 100 RP-18, 5 um, 250x4mm (E. Merck, Darmstadt, Germany) col- umn and for method B a Purospher RP-18e, 5pm, 250x4mm (E. Merck, Darmstadt, Germany) column was used.

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 1 : 2-{4-[(2-Toluidinocarbonyl) amino] phenyl} acetic acid

To a solution of 2- (4-aminophenyl) acetic acid (108. 8 g, 0. 72 mol) in CH2C12 (1. 0 1) and triethylamine (120 ml) was added a solution of 2-methylphenyl isocyanate (90. 5 ml, 0. 72 mol) in CH2C12 (500 ml) dropwise at r. t.. After stirring for 18 h at r. t., water (2. 5 1) and CH2C12 (2. 0 1) were added and the layers were separated. The or- ganic layer was extracted with water (3 x 400 ml). The combined aqueous layers were concentrated to 3. 0 1 and acidified to pH 2 by the addition of concentrated aqueous HC1. The precipitate was collected by filtration, washed with cold water and dried in an exsiccator over concentrated H2SO4 affording 166. 5 g (82%) white solid.

M. p. 205-206°C ; TLC (CH2Cl2/MeOH 9 : 1) : Rf 0. 14. 1H-NMR (400 MHz, D6- DMSO) : 12. 21 (br s, 1H), 9. 11 (s, 1H), 8. 00 (s, 1H), 7. 83 (d, 7. 6 Hz, 1H), 7. 40 (d, 8. 5 Hz, 2H), 7. 17-7. 12 (m, 4H), 6. 96-6. 92 (m, 1H), 3. 48 (s, 2H), 2. 24 (s, 3H).

Example II : 2-{4-[(2-Toluidinocarbonyl) amino] phenyl} acetyl-L-leucine methyl ester A solution of 2-{4-[(2-toluidinocarbonyl) amino] phenyl} acetic acid (1. 96 g, 6. 89 mmol), HOBt (1. 16 g, 7. 58 mmol) and EDCI in 70 ml dimethylformamide was stirred 90 min at r. t.. After addition of L-leucine methyl ester hydrochloride (1. 25 g, 6. 89 mmol) in dimethylformamide (20 ml) and ethyldiisopropylamine (5. 75 ml, 34. 5 mmol) stirring at r. t. was continued for 18 h. The reaction mixture was poured into water (350 ml) and extracted with ethyl acetate (4x150 ml). The combined organic layers were washed with 0. 1 N aqueous HCl, saturated aqueous Na2C03, brine, dried (MgSO4) and evaporated. Yield : 2. 49 g (88%) white solid. M. p. 166-168°C ; TLC

(CH2Cl2/MeOH 9 : 1) : Rf 0. 56 ; 1H-NMR (400 MHz, D6-DMSO) : 8. 96 (s, 1H), 8. 42 (d, 7. 7 Hz, 1H), 7. 89 (s, 1H), 7. 84 (d, 7. 44 Hz, 1H), 7. 38 (d, 8. 5 Hz, 2H), 7. 18-7. 11 (m, 4H), 6. 96 (m, 1H), 4. 30-4. 23 (m, 1H), 3. 61 (m, 3H), 3. 43-3. 36 (m, 2H), 2. 24 (s, 3H), 1. 67-1. 45 (m, 3H), 0. 89 (d, 6. 4 Hz, 3H), 0. 82 (d, 6. 4 Hz, 3H).

Example III : 2-{4-[(2-toluidinocarbonyl) amino] phenyl} acetyl-L-leucine

A solution of 2-{4-[(2-toluidinocarbonyl) amino] phenyl} acetyl-L-leucine methyl es- ter (2. 42 g, 5. 88 mmol) and KOH (3. 30 g, 58. 75 mmol) in methanol/water 1 : 1 (180ml) was stirred at 50°C for 5 h. After washing with methyl-tert-butylether (80 ml) the volume of the reaction mixture was reduced until a slight turbidity was observed. The solution was acidified to pH 2 by the addition of 1 N aqueous HC1.

The precipitate was collected by filtration, washed with cold water and dried in vac- uum. Yield : 1. 75 g (72 %) white solid. M. p. : 178-179°C, TLC (CH2Cl2/MeOH/AcOH 9 : 1 : 0. 1) : Rf 0. 16 ; IH-NMR (400 MHz, D6-DMSO) : 12. 51 (br s, 1H), 9. 00 (s, 1H), 8. 25 (d, 8. 0 Hz, 1H), 7. 93 (s, 1H), 7. 83 (d, 7. 5 Hz, 1H), 7. 36 (d, 8. 5 Hz, 2H), 7. 17-7. 12 (m, 4H), 6. 95-6. 91 (m, 1H), 4. 23-4. 17 (m, 1H), 3. 43-3. 32 (m, 2H), 2. 24 (s, 3H), 1. 68-1. 46 (m, 3H), 0. 89 (d, 6. 5 Hz, 3H), 0. 82 (d, 6. 5 Hz, 3H).

Example IV : Methyl 4- ( ( [ (3-methoxypropyl) amino] acetyl} amino) benzoate To a solution of methyl 4-aminobenzoate (10. 0 g, 66. 2 mmol) and triethylamine (10. 1 ml, 72. 8 mmol) in dichloromethane (100 ml) was added a solution of bromo- acetylbromide (6. 34 ml, 72. 8 mmol) in dichloromethane (30 ml) at 0°C. After stir- ring for 18 h at room temperature and 18 h under reflux the reaction mixture was concentrated under vacuum. The residue was taken up in ethyl acetate, washed with 1 N aqueous HC1 and water, dried over MgSO4 and evaporated. Yield 15. 8 g (88%)

of methyl 4- [ (bromoacetyl) amino] benzoate as a pale brown solid. M. p. : 144-146°C, TLC (hexane/ethyl acetate 1 : 1) : Rf 0. 46.

To a solution of methyl 4- [ (bromoacetyl) amino] benzoate (2. 72 g, 10. 0 mmol) in di- methylformamide (20 ml) was added 3-methoxypropylamine (1. 78 g, 20. 0 mmol) and triethylamine (22. 3 ml, 160 mmol). After stirring at room temperature for 18 h, the reaction mixture was concentrated under vacuum and purified by flash chroma- tography (CH2C12/MeOH 9 : 0. 4) affording 1. 81 g (65%) of methyl 4- ( { [ (3-methoxy- propyl) amino] acetyl} amino) benzoate as a pale red solid.

Example V : 4- (lH-tetraazol-5-yl) aniline

To a solution of 4-aminobenzonitrile (11. 8 g, 100 mmol) and triethylamine hydro- chloride (17. 9 g, 130 mmol) in toluene (550 ml) was added sodium azide (8. 45 g, 130 mmol). After stirring for 24 h at 95°C, the reaction mixture was cooled to room temperature and was extraced with water (3x60 ml). The combined aqueous phases were acidified with concentrated aqueous HCl to pH 2-3. The product was collected by filtration, washed with water and dried in vacuum. Yield 9. 59 g (60%) pale brown solid. M. p. : 280-281°C, TLC (CH2CIa/MeOH/AcOH 9 : 1 : 0. 1) : Rf 0. 30 Example VI : Ethyl 1, 2, 3, 4-tetrahydro-6-quinolinecarboxylate 0 0 vOH 1) EtOH, H2SO4 XO N 2) Pd-Mohr, H2 HN 1

A solution of 6-quinolinecarboxylic acid (9. 50 g, 54. 9 mmol) and 2 ml of concen- trated sulfuric acid in ethanol (250 ml) was refluxed for 8 h. The solvent was evapo- rated and the residue was taken up in water. After adjustment of the pH to 8 by the addition of potassium hydroxide the product was collected by filtration and dried in

vacuum. Yield 9. 85 g (89%) of ethyl 6-quinolinecarboxylate as a pale brown solid.

M. p. : 66-67°C, TLC (CH2Cl2/MeOH/AcOH 9 : 0. 5 : 0. 1) : Rf 0. 52 A solution of ethyl 6-quinolinecarboxylate (9. 80 g, 48. 7 mmol) was acidified to pH 2 by the addition of 1N aqueous HC1. After addition of 20% Pd-Mohr catalyst (1. 96 g) the solution was hydrogenated at 60°C under 3 bar of hydrogen pressure for 17 h. The reaction mixture was filtered through celite. The filtrate was evaporated and the residue was taken up in ethyl acetate and water. The pH was adjusted to 10 by the additon of 1 N aqueous potassium hydroxide. The phases were separated and the organic phase was washed with brine, dried over Na2S04 and evaporated. Yield 8. 72 g (87%) of ethyl 1, 2, 3, 4-tetrahydro-6-quinolinecarboxylate as a pale brown solid.

M. p. : 68-70°C, GC-MS : [M+] = 205.

Compound synthesis 0 0 -OPG' IRs 1=2 R1. 3 H if II 0 N R U f-L- H H AG Hw R> > a s I Ra Ra N O R R H H O --OPGZ/I p I OH s =zf2-a /O II (l H H Step B HNNx TR Step C 0 -OPG' f 3/-\ R 1-1 Step D 3 °-OH N NJ 0 H H 0 -OH "yz Ria 0 O R4 R3 H H H

Scheme 3 Step A : General procedure Al (GP Al) : Coupling of amines with Boc-L-leucin-N-carboxy- anhydride : A solution/suspension of 1. 0 eq. of the amine, 1. 0 eq. of Boc-L-leucin-N-carboxyan- hydride and 0. 3 eq. of 4- (N, N'-dimethylamino) pyridine was refluxed for 0. 5-14 days with exclusion of moisture. If a precipitate was formed, the precipitate (product) was collected by filtration. The reaction mixture/filtrate was evaporated to dryness, redissolved in ethyl acetate and washed with 1 N aqueous HC1, saturated aqueous NaHC03 and brine, dried over MgS04 and evaporated. Both solids were combined.

If necessary the product was purified by trituration or by flash-chromatography or used without further purification.

Example 1 : Methyl 4-({Boc-L-leucine} amino) benzoate Methyl 4-aminobenzoate (0. 75 g, 4. 97 mmol) was dissolved in CH2C12 (7 ml). After the addition of Boc-L-leucin-N-carboxyanhydride (1. 28 g, 4. 79 mmol) and 4- (N, N'- dimethylamino) pyridine (180 mg, 1. 49 mmol) the solution was stirred under reflux for 4 days. The precipitate (product) was collected by filtration. The filtrate was evaporated to dryness, redissolved in ethyl acetate and washed with 1 N aqueous HC1, saturated aqueous NaHCO3 and brine, dried over MgS04 and evaporated. Com- bined Yield : 1. 35 (75%) white solid.

General procedure A2 (GP A2) : Coupling of amines with carboxylic acids activated by iso-butyl chloroformate.

A solution of 1. 0 eq. of the carboxylic acid derivative and 1. 0 eq. of N-methyl- morpholine in tetrahydrofurane was cooled to-15°C and 1. 0 eq. of iso-butyl chloro- formate was added dropwise. After 5 min at 0°C, 1. 0 eq. of the amine in tetrahydro- furane was added at-15°C. The solution was stirred for 1 h at 0°C, 1-4 d at r. t. and was evaporated. The residue was redissolved in ethyl acetate, washed with 1 N aque- ous HCl (2x), saturated aqueous NaHCO3 and brine, dried over MgS04 and evapo- rated.

Table 1: Characterization of reaction products according to Step A<BR> Example Structure Procedure Yield [%] Product Rf M.p.[°C] ESI-MS HPLC<BR> No. tR[min] GP A1 75 white solid 0. 52 72-73 309. 1 n. d. > (0If N (CH2CI2/MeOH 9 : 0. 1) [M+H] + 'I Y 2 GP Al, 31 white solid 0. 68 (petrol ether/ethyl n. d. 413. 4 27. 5 i 0 NJ FC : CH2Cl2/MeOH 9 : 0. 3-8 : 2 acetate 8 : 2) [M+H] + MethodB zizi 3 GP Al, 42 pale red 0. 60 (petrol ether/ethyl n. d. 409. 1 25. 4 > O) crude product solid acetate 6 : 4) [M+H] + MethodB 4 GP Al, 39 pale 0. 30 (petrol ether ethyl n. d. 399. 0 25. 4 ^ ., i crude product brown oil acetate 8 : 2) [M+H] + Method B oh 5 GP Al, 52 whitesolid 0. 46 (petrol ether/ethyl 207-209 456. 1 27. 8 o) <Y-o H crude product acetate 8 : 2) [M+H] Method B ''6 Example structure procedure Yield[%] Product Rf M.p. [°C] ESI-MS HPLC<BR> No. tR[min] 6 GP Al, 52 white solid 0. 70 (petrol ether/ethyl n. d. 457. 1 26. 0 crude product acetate 8 : 2) [M+H] Method B _ 7 GP Al, 37 pale 0. 84 (petrol ether/ethyl n. d. 431. 0 [M-27. 1 cri crude product brown acetate 6 : 4) H]-Method B 0 solin SOI1C 8 u-Nm GPA1 (aq. NaHCO3 wash 39 pale 0. 62 n. d. 373. 1 [M-21. 3 j JN H brown (CH2Cl2/MeOH/AcOH H]-Method A N crude product solid 9 : 1 : 0. 1) 9 GP Al, 46 pale 0. 80 (petrol ether/ethyl n. d. 424. 1 25. 9 , 0 HNUs ° crudeproduct yellow acetate 1 : 1) [M+H] + Method B N g solid 10 GP Al, 3 yellow oil 0. 52 (petrol ether/ethyl n. d. 419 [M+H] + n. d. FC : petrol ether/ethyl acetate acetate 1 : 1) 9 : 0. 3-9 : 1 Example structure procedure Yield[%] Product Rf M.p. [°C] ESI-MS HPLC<BR> No. tR[min] 11 0 GPA2, 17 pale 0. 90 (petrol ether/ethyl n. d. 461. 5 n. d. JQ FC : petrol ether/ethyl acetate yellow acetate 6 : 4) [M+H] + 0""Y'N 9 : 1 solid 12 0 GP A2, 89 pale 0. 83 n. d. 414. 3 25. 3 crude product yellow oil (CH2CI2/MeOH/AcOH [M+M + Method B type s 13 GP A2, 35 white solid 0. 36 (petrol ether/ethyl 50-52 590. 4 n. d. t FC : petrol ether/ethyl acetate acetate 6 : 4) [M+H] + 1'H II _I 10 : 1-6 : 4 l \ I OuNH 0 14 as described in the precursor 65% pale red 0. 36 49-50 281. 0 n. d. synthesis solid (CH2CI2/MeOH 9 : 1) [M+H] + H

Step B : General procedure B (GP B) : Cleavage of the Boc-protecting group with trifluoro- acetic acid To a solution of the Boc-protected amine was added 20 vol% trifluoroacetic acid in dichloromethane at 0°C. Stirring was continued at room temperature for 0. 5-24 h.

The solvent was removed at room temperature under reduced pressure. The residue was coevaporated twice with dichloromethane, dried under high vacuum and sub- jected to the reaction step C without further purification.

Step C : <BR> <BR> <BR> <BR> <BR> General procedure (GP C1) : Coupling of amines with 2-f4-[(2-toluidinocarbonyl)-<BR> <BR> <BR> <BR> aminoJphenylacetic acid : A solution of 1. 0 eq. 2- {4- [ (2-toluidinocarbonyl) amino] phenyl} acetic acid, 1. 1 eq.

HOBt and 1. 1 eq. EDCI in DMF was stirred for 2 h at r. t.. After addition of 1. 0 eq. amine e. g. as TFA salt and 3-9 eq. ethylisopropylamine stirring was continued for 18 h at r. t.. The reaction mixture was poured into the 4-fold amount of water. The precipitate was collected by filtration, washed with cold water and dried in vacuum.

If necessary the product was purified by trituration or by flash-chromatography.

Methyl 4-([({4-[(2-toluidinocarbonyl)amino]phenyl}acetyl)L-leucin]a mino)benzoate Methyl 4-[(L-leucin) amino] benzoate trifluoroacetate (3. 81 g, 10. 1 mmol) was reacted according to GP Cl in a total volume of 60 ml of dimethylacetamide. Trituration

with CH2C12 yielded 4. 78 g (90%) pale brown solid. M. p. 250-252°C, TLC (AcOH : MeOH : CH2Cl2 0. 1 : 0. 5 : 9) : Rf 0. 46 ;'H-NMR (400 MHz, D6-DMSO) : 10. 47 (s, 1H), 8. 96 (s, 1H), 8. 39 (d, 7. 7 Hz, 1H), 7. 93-7. 89 (m, 3H), 7. 83 (d, 7. 8 Hz, 1H), 7. 75 (d, 8. 8 Hz, 2H), 7. 37 (d, 8. 4 Hz, 2H), 7. 18-7. 12 (m, 4 H), 6. 95-6. 92 (m, 1H), 4. 49-4. 43 (m, 1H), 3. 82 (s, 3H), 3. 47-3. 38 (m, 2H), 2. 24 (s, 3H), 1. 66-1. 50 (m, 3H), 0. 92 (d, 6. 4 Hz, 3H), 0. 86 (d, 6. 4 Hz, 3H) ; ESI-MS : 531. 3 [M+H] +.

General procedure (GP C2) : Coupling of amines with 2-{4-[(2-toluidinocarbon- yl) aminoZphenylyacetyl-L-leucine In several cases it is advisable to couple the amine (III) directly with with 2-{4-[(2- toluidinocarbonyl) amino] phenyl} acetyl-L-leucine followed by the cleavage of the protecting group PG2, thus omitting steps A and B : A solution of 1. 0 eq. 2-{4-[(2-toluidinocarbonyl) amino] phenyl} acetyl-L-leucine, 1. 1 eq. HOBt and 1. 1 eq. EDCI in DMF was stirred for 2 h at r. t.. After addition of 1. 0 eq. amine (as free amino or as a salt) and 3-9 eq. ethylisopropylamine stirring was continued for 18 h at r. t.. The reaction mixture was poured into the 4-fold amount of water. The precipitate was collected by filtration, washed with cold water and dried in vacuum. If necessary the product was purified by trituration or by flash-chroma- tography.

Table 2 The following eamples were prepared by subsequently applying the general procedures B & C1/C2 as indicated.<BR> <P>Example structure procedure Yield Product Rf M.p. ESI-MS HPLC<BR> No. [%] [°C] tR[min] 15 H R GP C2---170-n. d. 'N q o H H 16 GP C2 220 n. d. 'oleo Fui 17 1) GP B 90 pale 0. 46 250-5313 26. 6 i (1 2) GP Cl, 9 eq. brown (CH2C12/MeOIi/AcOH 252 [M+H] + Method A DIPEA solid 9 : 0. 5 : 0. 1) 18 GP C2 218 n. d. H HAN b 19 GP C2---200-n. d. fslo gNiNm- I i o Nr^ Example structure procedure Yield Product Rf M.p. ESI-MS HPLC<BR> No. [%] [°C] tR[min] 20 H 8, GPC2---222-n. d. n, (A", Or 0 N''N O 1 H H 21 1) GP B 81 pale 0. 74 163-579. 3 25. 5 N GP C1, brown (CH2Cl2/MeOH/AcOH 165 [M+H] + Method B 'H H I --H 9 eq. DIPEA solid 9 : 1 : 0. 1) 22 1) GP B 81 pale 0. 34 139-575. 0 24. 5 H 2) GPC1, brown (CH2Cl2/MeOH/AcOH 141 [M+H] + Method A "IrH H 3) 9 eq. DIPEA solid 9 : 1 : 0. 1) 23 1) GP B 66 white 0. 74 (CH2CI2/MeOH9 : 1) 181-564. 5 24. 0 i o' 2 2) GP Cl : 1. 1 eq. solid 184 [M+H] + MethodA I I H H HATU (no EDCI, HOBt) & 5 eq. DIPEA 24 1) GP B 90 pale 0. 34 205-622. 2 26. 1 N>gNH 2) GP C1, 9 eq. brown (CHzClz/MeOH/AcOH 206 [M+H] + Method B H H DIPEA solid 9 : 0. 5 : 0. 1) Example structure procedure Yield Product Rf M.p. ESI-MS HPLC<BR> No. [%] [°C] tR[min] 25 GP B 30 yellow 0. 66 (CH2ClJMeOH 9 : 1) 191-622. 9 24. 3 1, 2) GP C1, 9 eq. solid 195 [M+H] + Method A H NEO 26 1) GP B 21 white 0. 34 (petrol ether ethyl 204-598. 9 25. 5 cul "r, 2) GP C1, 9 eq. solid acetate 1 : 1) 205 [M+H] + Method B o"DIPEA MHZ 27 1) GP B, 3 eq. 2 white 0. 16 255-541. 2 21. 4 N$NjyHN'Ihiohpenole solid (CH2Cl21MeOH/AcOH 257 [M+H] + Method A added 9. 5 : 0. 5 : 0. 1) 2) GP Cl, 9 eq. DIPEA 28 0. 1) GP B 99 pale 0. 80 90-95 590. 0 25. 0 NA way 2) GP C1, 9 eq. brown (CH2Cl2JMeOHlAcOH [M+H] + Method B N DIPEA solid 9 : 1 : 0. 1) 29 o 1) GP B 1. 6 pale 0. 68 (CH2ClJMeOH n. d. 585. 2 n. d. (2) GP C1, 9 eq. yellow 9 : 0. 5) [M+H] + DIPEA oil H H Example structure procedure Yield Product Rf M.p. ESI-MS HPLC<BR> No. [%] [°C] tR[min] 30 °Y° 1) GP B 73 pale 0. 66 (CH2CIJMeOH 9 : 1) 186-627. 4 26. 2 H 2) GP Cl, 9 eq. brown 189 [M+H] t Method A fft 9 YYr gHtm Y DIPEA solid H H y 31 0 GP C2 n. d. n. d. - ( Su H H 32 GP C2-n. d.-n. d. 33 q° 1) GP B 16 yellow 0. 90 (CH2Cl2/MeOH 9 : 1) 125-580. 2 24. 2 gNt GP Cl, 9 eq. solid 130 [M+H] + Method A Y°)-'DIPEA i 34 H 0 GP C2 n. d. n. d. N'IC /N''\ I'H \ _l0 i H H ! Example structure procedure Yield Product Rf M.p. ESI-MS HPLC<BR> No. [%] [°C] tR[min] 35 1) GP B 97 pale 0. 62 188-756. 4 n. d. 9 GP Cl, 9 eq. brown (CH2Cl2/MeOH/AcOH 189 [M+H] + o H L H H'DIPEA solid 9 : 1 : 0. 1) (D"O'r 36 1) GP B64 white 0. 50 (CH2Cl2/MeOH 9 : 1) 197-547. 0 21. 8 . O nT ? 2) GP Cl, 3 eq. solid 198 [M+Hl+ Method A I i , j j_ Ij g H [M H H DIPEA 37 GP C2, 3 eq. DIPEA 82 white 0. 80 (CH2CI2/MeOH 9 : 1) 188-613. 3 n. d. i N w solid 189 +H + I I H I CM MHZ 38 GP C2 n. d. n. d. g,, n " ..

Step D General procedure DI (GP D1) : ester saponification A solution or suspension of the ester and 1. 1 eq. KOH in water/ethanol, methanol and/or dioxane was stirred at 25-50°C for 2-24 h. After washing with methyl-tert- butylether (80 ml) the volume of the reaction mixture was reduced until a slight turbidity was observed. The solution was acidified to pH 2 by the addition of 1 N aqueous HCI. The precipitate was collected by filtration, washed with cold water and dried in vacuum.

General procedure D2 (GP D2) : deprotection of benzyl esters/benzyl carbamate A solution or suspension of the ester and 10% Pd-C (10%) in dimethylformamide was hydrogenated for 12 h at r. t. and 50 bar hydrogen pressure. The reaction mixture was filtered through celite. Evaporation of the filtrate and purification of the crude product by preparative HPLC (LiChrospher RP-18, 12 M, 250x25 mm ; flow rate 40ml/min ; eluent : acetonitrile/water mixture with 0. 1% trifluoroacetic acid (vol./vol.), linear gradient of : 0 min. = 40% acetonitrile, 20 min. = 80% acetonitrile) afforded the product.

General procedure D3 (GP D3) : deprotection of benzyl esters A solution or suspension of the ester and 10% Pd-C (10%) in tetrahydrofurane was hydrogenated for 18 h at r. t. under atmospheric hydrogen pressure. The reaction mixture was filtered through celite. Evaporation of the filtrate afforded the product.

Table 3: following examples were prepared according to the general procedures D1 - D3:<BR> Example Structure Procedure Yicld Product Rf M.p. [°C] ESI-MS HPLC tR[min]<BR> No. [%] 39,, GPD1 ; 10---190-n. d. 9 eq. KOH H Who 0 I 40, o, GPD1 ; 10---220-n. d. HNJYH f wNe eq. KOH 41 GP D1 ; 10 90 white 0. 24 219-223 517. 0 21. 3 Method A eq. H eq. KOH solid (CHzCl2/MeOH/AcO [M+H] + H 9 : 1 : 0. 1) 42 HNS y GPD1 ; 10 87 yellow 0. 06 186-190 531 26. 6 Method A w in-H jO''Tj eq. KOH solid (CH2Cl2/MeOH/AcO [M+H]" ""'o H9 : 1 : 0. 1)) 43 GPDI 'OH H eq. KOH solid (CH2Cl2/MeOH/AcO [M+H] + 9 H9 : 1 : 0. 1) I I NH H 9 : 1 : 0. 1) H H Example Structure Procedure Yicld Product Rf M.p. [°C] ESI-MS HPLC tR[min]<BR> No. [%] 44 H GP DI ; 10 206 n. d. JN eq. KOH N I Y eq. KOH H H 45 GP D1 ; 1. 5 64 pale 0. 34 154-158 551. 3 22. 2 Method B eq. eq. KOH brown (CH2Cl2/MeOH/AcO [M+H] + 1 I -H oh H solid H9 : 1 : 0. 1) 46 GP D1 ; 1. 6 6 16 white 0. 28 138-142 546. 8 23. 1 Method B Njlt eq. NaOH solid (CHzCl/MeOH AcO >+H] + Nu, H H H 9 : 1 : 0. 1) 47 GP D1 ; 1. 1 36 pale red 0. 36 178-180 551. 1 21. 6 Method A OU eq. KOH solid (CH2Cl2/MeOH/AcO [M+H] + = H H H --H 9 : 1 : 0. 1) 48 GP D1 ; 3. 5 68 pale 0. 30 164-169 608. 0 23. 7 Method A eq. KOH brown (CH2Cl2/MeOHlAcO [M+H] solid H 9 : 1 : 0. 1) Example Structure Procedure Yicld Product Rf M.p. [°C] ESI-MS HPLC tR[min]<BR> No. [%] 49 GP D1 ; 1. 1 18 pale 0. 62 158-160 609. 2 23. 1 Method A eq. KOH brown (CH2ClJMeOH/AcO [M+H] + NIN, [ : : Ir = H solid H 9 : 1 : 0. 1) 50 g GPD1 oCJk eq. KOH solid (CH2ClJMeOH/AcO [M+H] + y H H Y H 9 : 1 : 0. 1) 51 OqzOH GPD1 ; 1. 5 45 yellow O. lO (CH2CI2/MeOH 188-190 562. 0 23. 5MethodA ,, i N eq. NaOI4 solid 9 1) [M+H] HAN H H 52 GP Dl 1. 1 76 white n. d. n. d. 557 22. 8 eq. KOH foam [M+H] + Method A MUS MS 53 GP Dl ; 1. 1 26 white 0. 74 206-208 599. 4 19. 2 23. 5 Method eq. KOH solid (CH2ClJMeOH/AcO [M+H] + B ; 2 dia- i AH 9 : 1 : 0. 1) steromeres Example Structure Procedure Yicld Product Rf M.p. [°C] ESI-MS HPLC tR[min]<BR> No. [%] 54 s° GPD1 ; 1. 5 26 pale 0. 30 178-180 538. 2 29. 4 Method A OH KOH brown (CH2CI2/MeOH/AcO [M+H] + solid H 9 : 1 : 0. 1) 55 H °, qN~NoO GPD1 ; 1. 5 33 pale 0. 18 187-189 539. 2 28. 7MethodA w i Nr. soH o Y eq. KOH brown (CH2Cl2/MeOH/AcO [M+H] + solid H 9 : 1 : 0. 1) 56 GP Dl ; 1. 1 2 pale 0. 26 172-174 552. 07 21. 3 Method A /bH eq. KOH brown (CH2Cl2/MeOH/AcO [M+H] + solid H 9 : 1 : 0. 1) HN' '57'a. H C GP D1 ; 1. 5 22 pale 0. 08 240 de-556. 2 29. 4 Method A w i/ eq. KOH brown (CH2ClJMeOH/AcO composi- [M+H] + Sr"-Y solid H 9 : 1 : 0. 1) tion 58 GP D2 3 white 0. 05 168-169 532. 1 n. d. g solid (CH2Cl2/MeOH/AcO [M+H] + W Jl W o 1" H X eO H 9 : 1 : 0. 1) LC-MS NH ; "Ha Example Structure Procedure Yicld Product Rf M.p. [°C] ESI-MS HPLC tR[min]<BR> No. [%] 59 GP D1 ; 1. 1 76 white 0. 48 210-218 570. 8 20. 0 Method A OH eq. KOH solid (CH2CI2/MeOI-I/AcO [M+Kl+ --y H H 9 : 1 : 0. 1) 60 GP D3 95 white 0. 12 175 523. 2 20. 2 Method B FI Ç < solid (CH2CI/MeOH AcO decompo- [M+H] + H 9. 5 : 0. 5 : 0. 1) sition 61 GPD1, 1. 5 1 pale 0. 10 n. d. 523. 2 20. 0 Method A H eq. KOH brown (CH2CI2/MeOH/AcO [M+H] + 0 OH solid H 9. 5 : 0. 5 : 0. 1)

In vitro assay : adhesion of Jurkat/Ramos cells to immobilized VCAM-1 (do- mains 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 purifi- cation of the PCR product, the fragment was digested with KpnI-AvrII. The digested 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. coli 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 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 (pHlO) was constructed by ligation of 0. 9 kb fragment from AvrII/Klenow/BclI digests of pH7 into SalIlKlenow/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- 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 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 microtiter plates Recombinant human VCAM-1 (extracellular domains 1-3) was dissolved at 1. 0 llg/ml in PBS. Each well of the microtiter plates (Nalge Nunc International, Fluoro- nunc Cert, 437958) was coated with 100 gl 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 u, l 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 mM KC1 and 2 mM MnCl2 just be- fore addition of the assay.

In vitro assay using Jurkat cells Preparation of fluorescence labeled Jurkat cells : Jurkat cells (American Type Culture Collection, Clone E6-1, ATCC TIB-152) were cultured in RPMI 1640 medium (Nikken Bio Medical Laboratory, CM1101) supple- mented with 10% fetal bovine serum (Hyclone, A-1119-L), 100 U/ml penicilin (Gibco BRL, 15140-122) and 100 Zg/ml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37 °C with 5% CO2.

Jurkat cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25 uM 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 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-HCl (pH 7. 4), 137 mM NaCl, 27 mM KC1, 4 mM glucose, 0. 1 % bovine serum albumin (BSA, Sigma, A9647) and 2 mM MnCl2.

Assay procedure (Jurkat cells) The assay solution containing each test compounds was transferred to the VCAM-1 coated plates. The final concentration of each test compounds was 5 JIM, 10 uM or various concentrations ranging from 0. 0001 uM to 10 I1M using a standard 5-point serial dilution. The assay solution containing the labeled Jurkat 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 meas- urement in a fluorometer (Wallac, ARVO 1420 multilabel counter).

The adhesion of Jurkat 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 fluo- rescent intensity from wells lacking VCAM-1 and FTS is the fluorescent intensity from wells containing the test compound of this invention.

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 Fg/ml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37 °C with 5% C02.

Ramos cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25, uM 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 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-HCl (pH 7. 4), 137 mM NaCl, 27 mM KC1, 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 ug/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 uM, 10 uM or various con- centrations ranging from 0. 0001 uM to 10 uM using a standard 5-point serial dilu- tion. 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 calcu- lated 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 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.

In vitro activity : In the Jurkat-VCAM-1 assay (indicated as Jurkat-VCAM-1) and the Ramos- VCAM-1 (indicated as Ramos-VCAM-1) the observed IC50 value ranges are indi- cated Table 4.

D > 10 µM # C > 2 µM # B > 0.5 µM # A Table 4.

No Structure IC50 Cell Type 27 Ramos H N tJ-'N, FI N H 39 i D Jurkat H H H o HO O 40 H D Jurkat \ H oh 41 A Jurkat OH AH S Jurkat NJ ec) rlo N 42 D Jurkat COR Y OH 43 B Jurkat 'OU / \ p N I , p H IH H 0 44 gj zu wOH C Jurkat n H H y H H 45 A-B Jurkat zou nu I/H'H \ I H I 46 D Ramos H \ N \ I H H No Structure IC50 Cell Type 47 A Ramos ou CL JE ) H H/- 48 A Ramos OH H Ji N) H-1 6 I H oh H H 49 t A Ramos cul gN) H 50 C Ramos ci H 49 t A Ramos gH < H 6 51 R C Ramos 0 H H N H C Ramos p_ NlN H H 52 C Ramos "Q I ON H H 53 Oq Jurkat I V H H y 54 ° C Jurkat OH N--,, zon No Structure IC50 Cell Type 55 H c Jurkat PiN, ls, OH H H Y OU 56 ° D Ramos KOH N jazz W )- OH H 57 D Jurkat 9>N, OH 58 A Ramos I OH \- MHZ NH3 59 A Ramos OU Jl I H H H 60 C Jurkat H 9 tY-OH "YH 61 H C Ramos I I' T 0 OH