WO2000064866A1

WO2000064866A1 - Diphenylurea derivatives

Info

Publication number: WO2000064866A1
Application number: PCT/GB2000/001542
Authority: WO
Inventors: Keith Oldham
Original assignee: AstraZeneca AB
Current assignee: AstraZeneca AB
Priority date: 1999-04-24
Filing date: 2000-04-19
Publication date: 2000-11-02
Anticipated expiration: 2001-10-24
Also published as: GB9909409D0; EP1173415A1; AU4132000A; JP2002543060A

Abstract

A compound of formula (I) wherein A' is NH or NR6, where R6 is C¿1-6?alkyl, C2-6alkenyl, C2-6alkanoyl or C1-6alkoxycarbonyl; B is oxygen or sulphur; Y' is a specified linker group; m is from 0 to 5; n is from 0 to 4; R?1 and R4¿ are each independently selected from specified organic groups; and R5 is an acidic functional group; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof. These compounds are useful in the treatment of a disease or medical condition mediated by the interaction between fibronectin and/or VCAM-1 (especially VCAM-1) and the integrin receptor α¿4?β1.

Description

DEPHENYLUREA DERIVAΗVES

This invention relates to compounds which are inhibitors of the interaction between the integrin α₄β,, also known as Very Late Antigen-4 (VLA-4) or CD49d/CD29, and its protein ligands, for example Vascular Cell Adhesion Molecule- 1 (VCAM-1) and fibronectin. This invention further relates to processes for preparing such compounds, to pharmaceutical compositions containing them and to their use in methods of therapeutic application. α₄β, is a member of the integrin family of heterodimeric cell surface receptors that are composed of noncovalently associated glycoprotein subunits (α and β) and are involved in cell adhesion to other cells or to extracellular matrix. There are at least 14 different human integrin α subunits and at least 8 different β subunits and each β subunit can form a heterodimer with one or more α subunits. Integrins can be subdivided based on their β subunit composition. α₄β, is one of several β, integrins, also known as Very Late Antigens (VLA).

The interactions between integrins and their protein ligands are fundamental for maintaining cell function, for example by tethering cells at a particular location, facilitating cell migration, or providing survival signals to cells from their environment. Ligands recognised by integrins include extracellular matrix proteins, such as collagen and fibronectin; plasma proteins, such as fibrinogen; and cell surface molecules, such as transmembrane proteins of the immunoglobulin superfamily and cell-bound complement. The specificity of the interaction between integrin and ligand is governed by the α and β subunit composition.

Integrin α₄β, is expressed on numerous hematopoietic cells and established cell lines, including hematopoietic precursors, peripheral and cytotoxic T lymphocytes, B lymphocytes, monocytes, thymocytes and eosinophils [Hemler, M.E. et al (1987), J. Biol. Chem., 262, 11478-11485; Bochner, B.S. et al (1991), J. Exp. Med., 173, 1553-1556]. Unlike other β, integrins that bind only to cell-extracellular matrix proteins, α₄β, binds to VCAM-1, an immunoglobulin superfamily member expressed on the cell surface, for example on vascular endothelial cells, and to fibronectin containing the alternatively spliced type III connecting segment (CS-1 fibronectin) [Elices, M.J. et al (1990), Cell, 60, 577-584; Wayner, E.A. et al (1989). J. Cell Biol, 109, 1321-1330].

The activation and extravasation of blood leukocytes plays a major role in the development and progression of inflammatory diseases. Cell adhesion to the vascular endothelium is required before cells migrate from the blood into inflamed tissue and is mediated by specific interactions between cell adhesion molecules on the surface of vascular endothelial cells and circulating leukocytes [Sharar, S.R. et al (1995). Springer Semin. Immunopathol, 16, 359-378]. α₄β, is believed to have an important role in the recruitment of lymphocytes, monocytes and eosinophils during inflammation. α₄β, /ligand binding has also been implicated in T-cell proliferation, B-cell localisation to germinal centres, haemopoeitic progenitor cell localisation in the bone marrow, placental development, muscle development and tumour cell metastasis.

The affinity of α₄β, for its ligands is normally low but chemokines expressed by inflamed vascular endothelium act via receptors on the leukocyte surface to upregulate α₄β, function [Weber, C. et al (1996), J. Cell Biol, 134, 1063-1073]. VCAM-1 expression is upregulated on endothelial cells in vitro by inflammatory cytokines [Osborn. L. et al (1989) Cell, 59, 1203-121 1] and in human inflammatory diseases such as rheumatoid arthritis [Morales-Ducret, J. et al (1992). J. Immunol., 149, 1424-1431], multiple sclerosis [Cannella, B. et al., (1995). Ann. Neural., 37, 424-435], allergic asthma [Fukuda, T. et al (1996), Am. J. Respir. Cell Mol. Biol., 14, 84-94] and atherosclerosis [O'Brien, K.D. et al (1993). J. Clin. Invest, 92, 945-951].

Monoclonal antibodies directed against the α₄ integrin subunit have been shown to be effective in a number of animal models of human inflammatory diseases including multiple sclerosis, rheumatoid arthritis, allergic asthma, contact dermatitis, transplant rejection, insulin- dependent diabetes, inflammatory bowel disease, and glomerulonephritis.

Integrins recognise short peptide motifs in their ligands The minimal α₄β, binding epitope in CS-1 is the tripeptide leucine-aspartic acid-valine (Leu-Asp-Val) [Komoriya, A., et al (1991). J. Biol. Chem., 266, 15075-15079] while VCAM-1 contains the similar sequence isoleucine-aspartic acid-serine [Clements, J.M., et al (1994). J. Cell Sci., 107, 2127-2135]. The 25-amino acid fibronectin fragment, CS-1 peptide, which contains the Leu Asp-Val motif, is a competitive inliibitor of ₄β, binding to VCAM-1 [Makarem, R., et al (1994). J. Biol. Chem., 269, 4005-4011]. Small molecule α₄β, inhibitors based on the Leu- Asp-Val sequence in CS-1 have been described, for example the linear molecule phenylacetic acid-Leu-Asp-Phe-D-Pro-amide [Molossi, S. et al (1995). J. Clin. Invest., 95, 2601-2610] and the disulphide cyclic peptide Cys-Trp-Leu-Asp-Val-Cys [Vanderslice, P., et al (1997). J. Immunol.. 158, 1710-1718]. More recently, non- and semi-peptidic compounds which inhibit α₄β,/VCAM binding and which can be orally administered have been reported in for example, WO96/22966 and WO98/04247.

There remains a continuing need for alternative compounds which inhibit the interaction between VCAM-1 and fibronectin with integrin α₄β, and, in particular, for compounds which can be administered by an oral route.

We have now found a group of compounds which contain a urethane or carbamosulfanyl linkage which inhibits this interaction.

Accordingly the present invention provides a compound of formula (I)

wherein

A^" is NH or NR⁶, where R⁶ is C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkanoyl or C,.₆alkoxycarbonyl;

B is oxygen or sulphur;

Y^" is a linker group comprising an optionally substituted hydrocarbyl chain which is optionally interposed by one or more heteroatoms independently selected from oxygen, nitrogen and sulphur and/or by a monocyclic or bicyclic ring system; or linker group Y and

A' can be taken together to form a 5 to 7 membered heterocyclic ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂_

₍₎alkenyloxy, C_2.6alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyl]amino and C₂.₆alkanoylamino; m is from 0 to 5; n is from 0 to 4;

R' and R⁴ are each independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,_₆alkyl, C₂.₆alkenyl,

C,_.6alkanoyl, C_2.6alkynyl, C,„₆ alkoxy, C₂.₆alkenyloxy, C,.₆alkynyloxy, C,_₆ alkylamino, di-[(C,.₆)alkyl]amino, C₂.₆alkanoylamino, N-C,.₆alkylcarbamoyl, Cι_₆alkoxylcarbonyl,

N,N-di-[(C,.₆alkyl]carbamoyl, C_MalkoxylC,.₆alkyl, (CH₂)_tOH where t is 1 or 2,

-CO₂R^a and CONR^aR° where R^a and R^b are independently hydrogen or C,_₆ alkyl or one of R⁴ can be taken together with A' to form a 5 to 7 membered heterocyclic ring optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy,

C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyljamino and C₂.₆alkanoylamino;

R² and R³ are each independently selected from hydrogen, C,.₆ alkyl, C,-₃ alkanoyl or

C|.₆ alkoxycarbonyl; and

R⁵ is an acidic functional group; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

As used herein, the term "hydrocarbyl chain" as used in relation to linker Y' refers to an alkylene, alkenylene and alkynylene group, for example of from 1 to 10 carbon atoms in the case of the alkylene group and from 2 to 10 carbon atoms in the case of alkenylene and alkynylene groups which may be optionally interposed with one or more of the groups listed above.

In particular, the linker group is an alkylene chain which is interposed by a monocyclic ring system and optionally also at least one heteroatom. The ring system is preferably a monocyclic ring system as defined hereinafter.

Suitable substitutents for the linker group Y' include any of the groups listed above for R¹ and R⁴ as well as aryl groups such as phenyl or naphthyl or aralkyl groups such as benzyl. In particular, the substituents include C,.₆alkyl, C₂.₆alkenyl, C_2.6alkynyl, aryl such as phenyl and aralkyl such as benzyl. Where the linker group Y' includes a nitrogen heteroatom, substitutents may be present on said atom.

Suitably, the linker group is such that the spacing of the group B and the group R⁵ by not more than 10 atoms.

In a preferred embodiment, a basic group, and in particular a moiety containing at least one nitrogen is present within linker Y' .

Preferably, the group A' is orientated in the meta or para position with respect to the ureido group in formula (I), and most preferably A' is orientated para- to the ureido group in formula (I).

In a particular embodiment, the invention comprises a compound of formula (IA)

wherein

A is nitrogen or NR⁶, where R⁶ is C,.₆alkyl, C₂.₆alkanoyl or C^alkoxycarbonyl;

B is oxygen or sulphur;

Y is a linker group connecting group B to group R⁵ and containing up to 10 atoms where each atom is independently selected from carbon, oxygen, nitrogen and sulphur and may optionally comprise a monocyclic or bicyclic ring system or linker group Y and A can be taken together to form a 5 to 7 membered heterocyclic ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.

₆alkynyloxy, C,.₆alkylamino, di-[C|_₆ alkyljamino and C₂.₆alkanoylamino; m is from 0 to 5; n is from 0 to 4;

R¹ and R⁴ are each independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl,

C,.₆alkanoyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C ₆ alkylamino, di-[(C,.₆)alkyl]amino, C₂.₆alkanoylamino, N-C_:.₆alkylcarbamoyl, C,.₆alkoxylcarbonyl,

N,N-di-[(C_l.6alkyl]carbamoyl, C,.₄alkoxylC,.₆alkyl, (CH₂)_tOH where t is 1 or 2,

-CO,R^a and CONR^aR^b where R^a and R^b are independently hydrogen or C,.₆ alkyl or one of R⁴ can be taken together with A to form a 5 to 7 membered heterocyclic ring optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₅alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,_₆alkoxy,

C_2.6alkenyloxy, C_2.6alkynyloxy, C ^alkylamino, di-[C,.₆ alkyl]amino and C₂.₆alkanoylamino;

C|.₆ alkoxycarbonyl; and R⁵ is an acidic functional group; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

It will be understood that Y or Y' preferably excludes those linker groups which are unstable in acid conditions such as those found in the stomach of a human or animal body, for example

With reference to the compounds of formula (I), the 5 to 7 heterocyclic can be an, optionally substituted, saturated or unsaturated ring with up to three heteroatoms independently selected from nitrogen, oxygen and sulphur. An example of such a ring is an oxazolidinone, oxazolone and thiazolone.

Linker group Y or Y'can comprise a monocyclic ring or a bicyclic ring. With reference to the compounds of formula (I), 'monocyclic ring' means a 5 to 7 membered ring. It can be an, optionally substituted, preferably aromatic ring with up to three heteroatoms independently selected from nitrogen, oxygen and sulphur. Examples of such rings include phenyl, pyrimidinyl, pyridyl, imidazolyl, thienyl, thiazolyl, pyridazinyl and pyrrolyl. Other examples include morpholino, tetrahydropyridyl or tetrahydropyrazolyl.

With reference to the compounds of formula (I), 'bicyclic ring system' means an 8 to 10 membered fused ring system wherein one or both rings may contain ring heteroatoms. The ring system may be totally or partially saturated, optionally substituted and contain up to five heteroatoms independently selected from oxygen, nitrogen or sulphur. Examples of suitable ring systems include naphthalene, quinazoline, benzothiophene, benzoxazole, benzothiazole, and benzofuran and the corresponding hydro derivatives, e.g. tetrahydronaphthalene and dihydroquinazoline .

The term 'acidic functional group' means a group which incorporates an acidic hydrogen and includes carboxylic acids, tetrazoles, acyl sulphonamides, sulphonic and sulphinic acids.

In one aspect of the invention, the group Y' or Y is the group

- D - (E)_p - (F)_q - G - where D, E, F, G, p and q are as defined below. Thus suitable compounds of the invention are compounds of formula (II)

wherein

A, B, R¹ to R⁵, m and n are as hereinbefore defined;

D and G are each independently C,_₄ alkyl or C₂.₄ alkenyl and each carbon is optionally substituted with halogeno, hydroxy, amino, nitro, phenyl, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl, C,.₆alkanoyl, C₂.₆alkynyl, C,.₅alkoxy,

C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆ alkylamino, di-[(C,.₆)alkyl]amino, C₂.₆alkanoylamino

N-C,.₆alkylcarbamoyl, C,.₆alkoxylcarbonyl, J^,N-di-[(C₁.₆alkyl]carbamoyl, C alkoxylC,.

₆alkyl, (CH₂),.OH where r is 1 or 2, -CO₂R⁷ and CONR⁷R⁸ where R⁷ and R⁸ are independently hydrogen or C,.₆ alkyl, or D and A can be taken together to form a 5 to 7 membered ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl,

C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C|.₆ alkyl]amino and C₂.₆alkanoylamino;

E is phenyl or a monocyclic heterocycle both optionally substituted with up to 3 substituents selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,_.6alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy,

C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆alkyl] amino, C₂.₆alkanoylamino, C₃.₆ alkenoyl- amino,

C₃.₆alkynoylamino, C,.₆alkoxycarbonyl, N-C,.₆alkylcarbamoyl, N,N-di-[C,.₆ alkyl]carbamoyl,

C,.₆alkanoyl, C,_.4alkoxylC,_.6alkyl, (CH₂)_sOH where s is 1 or 2, -CO₂R⁹ and CONR⁹R¹⁰ where

R⁹ and R¹⁰ are independently hydrogen or C,.₆ alkyl or E and D can be taken together to form a bicyclic ring system, optionally substituted with up to 3 substituents selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂_

₆alkynyl, C,_.6alkoxy, C_2.6alkenyloxy, C₂.₆alkynyloxy, C_Lgalkylamino, di-[C,.₆ alkyl]amino and

C₂.₆alkanoylamino;

F is selected from oxygen, sulphur, amino or CRⁿR¹²; p and q are each independently 0 or 1 ; R" and R¹² are each independently selected from hydrogen, halogeno, hydroxy, amino, nitro, phenyl, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C_2.6alkenyl, C,.₆alkanoyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,_₆ alkylamino, di-[(C,.₆)alkyl]amino, C,.₆alkanoylamino, N-C,.₆alkylcarbamoyl, C,_.6alkoxylcarbonyl, N,N-di-[(C,.₆alkyl]carbamoyl, C,.₄alkoxylC,.₆alkyl, (CH₂)_uOH where u is 1 or 2, -CO₂R¹⁵ and CNNR^,5R¹⁶ where R¹⁵ and R¹⁶ are independently hydrogen or C,_.6 alkyl with the proviso that not more than one ring system can be formed from taking together two groups from R⁴, A, D and E; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

With reference to the compounds of formula (II), the 5 to 7 membered ring that can be formed by taking together A and D is a heterocycle ring. It can be an, optionally substituted, saturated or unsaturated ring with up to three heteroatoms independently selected from nitrogen, oxygen and sulphur. An example of such a ring is an oxazolidinone, oxazolone and thiazolone.

With reference to the compounds of formula (II), 'monocyclic heterocycle' means a 5 to 7 membered ring. It can be an, optionally substituted, preferably aromatic ring with up to three heteroatoms independently selected from nitrogen, oxygen and sulphur. Examples of such rings include pyrimidinyl, pyridyl, imidazolyl, thienyl, thiazolyl, pyridazinyl and pyrrolyl.

In a further preferred embodiment, the "monocyclic heterocycle" is a non-aromatic heterocyclic ring. Preferably, the rings are linked either to the groups D and/or F by means of a nitrogen atom. Such groups include morpholino, tetrahydropyridyl or tetrahydropyrazolyl.

With reference to the compounds of formula (II), 'bicyclic ring system' means an 8 to 10 membered fused ring system wherein one or both rings may contain ring heteroatoms. The ring system may be totally or partially saturated, optionally substituted and contain up to five heteroatoms. independently selected from oxygen, nitrogen or sulphur. Examples of suitable ring systems include naphthalene, quinazoline, benzothiophene, benzoxazole, benzothiazole, and benzofuran and the corresponding hydro derivatives, e.g. tetrahydronaphthalene and dihydroquinazoline.

The term 'acidic functional group' means a group which incorporates an acidic hydrogen and includes carboxylic acids, tetrazoles, acyl sulphonamides, sulphonic and sulphinic acids. Particularly preferred values for E in formula (II) is phenyl. Groups D and F are preferably arranged in the ortho- or meta- positions on phenyl ring E, and most preferably are orientated meta- to each other.

Alternatively, E is a monocyclic heterocycle as defined above, and in particular is an N-linked 6 membered non-aromatic heterocycle.

Preferably A' in formula (I) is NH or NR⁶ where R⁶ is C,.₆alkyl such as methyl or C₂_ ₆alkenyl such as ethen-2-yl. More preferably A' (or A in Formula (IA) and (II)) is NH or NCH₃ and most preferably A or A' is NH.

Preferably, R¹ is a C,.₆alkyl group such as methyl, which is in the ortho position on the phenyl ring relative to the ureido group. Preferably m is 1.

Preferably R² and R³ are hydrogen or C,_₄ alkyl and most preferably are both hydrogen.

A particular preferred value for B is oxygen.

Particular values for R⁴ include C^alkyl such as methyl, or C,_₆ alkoxy such as methoxy. In this case, n is suitably 0, 1 or 2. A particularly preferred group of compounds of formula (I) are those of formula (III)

where R', R², R³, A, B, Y, R⁵ and m are as defined above and R^4a is either hydrogen or C,_ ₆alkoxy such as methoxy, and R^4b is either hydrogen or C,.₆alkyl such as methyl.

A preferred value for F in formula (II) is oxygen.

Alternatively, q is 0.

Most preferably R⁵ is carboxy.

Suitable values for R¹ to R\ R⁶, R¹ ' to R¹⁶ and for various substituents on Y(or Y'), D, E, F, G or on any ring formed between Yor Y' and A or A', A or A' and D, D and E, A or A' and R⁴ include :- or halogeno: fluoro, chloro, bromo and iodo or C,.₆alkyl: methyl, ethyl, propyl, isopropyl and tert-butyl; or C₂.₆alkenyl: vinyl, allyl and but-2-enyl; or C,.₆alkanoyl formyl, acetyl, propionyl or butyryl; or C₂.₆alkynyl: ethynyl, 2-propynyl and but-2-ynyl; or C,.₆alkoxy: methoxy, ethoxy, propoxy, isopropoxy and butoxy; or C₂.₆alkenyloxy: vinyloxy and allyloxy; or C₂.₆alkynyloxy: ethynyloxy and 2-propynyloxy; or C,.₆alkylamino: methylamino, ethylamino, propylamino isopropylamino and butylamino; for di-C,.₆alkylamino: dimethylamino, diethylamino; for C₂.₆alkanoylamino: acetamido, propionamido and butyramido; for N-C,.₆alkylcarbamoyl: N-methylcarbamoyl, N-ethylcarbamoyl and N-propylcarbamoyl; for N,N-di-C alkylcarbamoyl: N,N-dimethylcarbamoyl,

N-ethyl-N-methylcarbamoyl and

N,N-diethylcarbamoyl; for C i ._ήalkoxy carbony 1 : methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and tert-butoxycarbony 1 ; for C|_₄alkoxyC alkyl: methoxymethyl, ethoxymethyl,

1 -methoxymethyl, 2 -methoxy ethyl, Particularly suitable compounds of formula (II) or pharmaceutically acceptable salts thereof include are those wherein, unless otherwise stated each of R¹ to R¹⁶, A, B, D to G, m, p to u has any of the meanings defined hereinbefore or below in sections a to h a) B is oxygen, D and G are each independently a C alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen and R⁵ is carboxy. b) B is oxygen, D and G are each independently a C alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen and is in the meta position with respect to E and R⁵ is carboxy. c) B is oxygen, D and G are each independently a C alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen, R⁵ is carboxy and R¹ and R⁴ are each independently C,.₆ alkyl or C,.₆ alkoxy. d) B is oxygen, D and G are each independently a C_M alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen, R⁵ is carboxy, R¹ and R⁴ are each independently C,.₆ alkyl or C,.₆ alkoxy and m and n are both 0 orl. e) B is oxygen, D and G are each independently a C,.₄ alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen, R⁵ is carboxy and R² and R³ are each independently C ₆ alkyl, preferably methyl. f) B is oxygen, D and G are each independently a C_M alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen, R⁵ is carboxy and R⁶ is C,_₆ alkyl, preferably methyl g) B is oxygen, D is a C_M alkyl, optionally substituted as hereinbefore defined, E is phenyl, q is zero and G is a C alkyl or C alkenyl, optionally substituted as hereinbefore defined and R³ is carboxy. h) B is oxygen, D and G are each independently C,_₄ alkyl, optionally substituted as hereinbefore defined, E is phenyl, F is oxygen, A and one of R⁴ together form a five membered ring and R⁵ is carboxy.

Other particular groups are compounds where i) B is oxygen, D and G are each independently a C,.₄ alkyl, optionally substituted as hereinbefore defined, q is 0, E is morpholino which is N-linked to the group G and is linked to D at the 2 position on the morpholine ring, and R⁵ is carboxy. j) B is oxygen, D and G are each independently a C_M alkyl, optionally substituted as hereinbefore defined, q is 0, E is tetrahydropyridyl which is N-linked to the group G and is linked to D at the 4 position on the ring, and R⁵ is carboxy; k) B is oxygen, D and G are each independently a C,_₄ alkyl, optionally substituted as hereinbefore defined, q is 0, E is tetrahydropyridyl which is N-linked to the group D and is linked to G at the 4 position on the ring, and R⁵ is carboxy;

1) B is oxygen, D and G are each independently a C alkyl, optionally substituted as hereinbefore defined, q is 0, E is tetrahydropyridyl which is N-linked to the group G and is linked to D at the 2 position on the ring, and R⁵ is carboxy; m) B is oxygen, D and G are each independently a C,_₄ alkyl, optionally substituted as hereinbefore defined, q is 0, E is tetrahydropyrazinyl which is N-linked to the group D and G, and R⁵ is carboxy.

Pharmaceutically acceptable salts include acid addition salts such as salts formed with mineral acids, for example, hydrogen halides such as hydrogen chloride and hydrogen bromide, sulphonic and phosphonic acids; and salts formed with organic acids, especially citric, maleic, acetic, oxalic, tartaric, mandelic, p-toluenesulphonic, methanesulphonic acids and the like. In another aspect, suitable salts are base salts such as alkali metals salts, for example, sodium and potassium; alkaline earth metal salts such as magnesium and calcium; aluminium and ammonium salts; and salts with organic bases such as ethanolamine, methylamine, diethylamine, isopropylamine, trimethylamine and the like. Such salts may be prepared by any suitable method known in the art.

In vivo hydrolysable derivatives include, in particular, pharmaceutically acceptable esters that hydrolyse in the human body to produce the parent compound. Such esters can be identified by administering, for example, intravenously to the test animal, the compound under test and subsequently examining the test animal's body fluids. Suitable in vivo hydrolysable esters for hydroxy include acetyl and for carboxy include, for example, alkyl esters, dialkylaminoalkoxy esters and C,.₆alkoxy methyl esters for example methoxymethyl, C -alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidyl esters, C₃.₈ cycloalkoxycarbonyloxyC,.₆alkyl esters for example 1 -cyclohexylcarbonyloxy ethyl; l,3-dioxolan-2-ylmethyl esters for example 5-methyl-l,3-dioxolan-2-ylmethyl; and C,.₆alkoxycarbonyloxy ethyl esters for example 1-methoxycarbonyloxyethyl.

It is to be understood that insofar as certain of the compounds of formula (I) as hereinbefore defined may exist in optically active or racemic forms by virtue of one or more asymmetric carbon atoms, the invention includes in its definition any such optically active or racemic form which possesses the property of inhibiting the interaction between VCAM-1 and fibronectin with integrin α₄β,. The synthesis of optically active forms may be carried out by standard techniques of organic chemistry well known in the art. These include, for example, synthesis from optically active starting materials or by resolution of a racemic form.

Particular Examples of compound of formula (I) are given in Tables 1, 2 and 3.

Table 2

Table 3

The activities of the compounds of this invention to inliibit the interaction between

VCAM-1 and fibronectin with integrin α₄βi may be determined using a number of in vitro and in vivo screens, as hereinafter defined. For example, compounds of formula (I) preferably have an IC₅₀ of <10μM, more preferably <lμM in the MOLT-4 cell/Fibronectin assay hereinafter described.

In order for it to be used, a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof is typically formulated as a pharmaceutical composition in accordance with standard pharmaceutical practice.

Thus, according to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof and a pharmaceutically acceptable carrier. The pharmaceutical compositions of this invention may be in a form suitable for oral use, for example a tablet, capsule, aqueous or oily solution, suspension or emulsion; for nasal use, for example a snuff, nasal spray or nasal drops; for vaginal or rectal use, for example a suppository; for administration by inhalation, for example as a finely divided powder or a liquid aerosol; for sub-lingual or buccal use, for example a tablet or capsule; or for parenteral use (including intravenous, subcutaneous, intramuscular, intravascular or infusion), for example a sterile aqueous or oily solution or suspension, or a depot formulation with drug incorporated in a biodegradable polymer. The composition may be in a form suitable for topical administration such as for example creams, ointments and gels. Skin patches are also contemplated. For these purposes, the compositions of this invention may be formulated by means known in the art, such as for example, as described in general terms, in Chapter 25.2 of Comprehensive Medicinal Chemistry, Volume 5, Editor Hansch et al, Pergamon Press 1990. Furthermore, the pharmaceutical composition of the present invention may contain one or more additional pharmacological agents suitable for treating one or more disease conditions referred to hereinabove in addition to the compounds of the present invention. In a further aspect, the additional pharmacological agent or agents may be co-administered, either simultaneously or sequentially, with the pharmaceutical compositions of the invention.

The composition of the invention will normally be administered to humans such that the daily dose will be 0.01 to 75mg/kg body weight and preferably 0.1 to 15mg/kg body weight. A preferred composition of the invention is one suitable for oral administration in unit dosage form for example a tablet or capsule which contains from 1 to 1 OOOmg and preferably 10 to 500mg of a compound according to the present invention in each unit dose. Thus, according to yet another aspect of the invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof for use in a method of therapeutic treatment of the human or animal body.

In yet a further aspect of the invention the present invention provides a method of treating a disease mediated by the interaction between VCAM-1 and/or fibronectin and the integrin receptor α₄β, in need of such treatment which comprises administering to said warm-blooded mammals an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof.

The present invention also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof in the production of a medicament for use in the treatment of a disease or medical condition mediated by the interaction between fibronectin and/or VCAM-1 (especially VCAM-1) and the integrin receptor α₄β,

In a preferred embodiment the mammal in need of treatment is suffering from multiple sclerosis, rheumatoid arthritis, asthma, coronary artery disease, psoriasis, atherosclerosis, transplant rejection, inflammatory bowel disease, insulin-dependent diabetes and glomerulonephritis.

Preferred compounds of formula (I) for use in these various embodiments of the invention are as set out above and include compounds of formula (IA) or (II). In another aspect of the invention, there is provided a process for preparing a compound of formula (I), a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof which method comprises either (a) reacting a compound of formula (IV)

where A\ R¹, R², R\ R⁴, m and n are as defined in relation to formula (I), with a compound of formula (V)

where B and Y' are as defined in relation to formula (I), Z is a leaving group, such as halo, and R'⁷ is a group R³ as defined in relation to formula (I) or a protected form thereof: or

(b) reacting a compound of formula (VI)

where A\ B, Y' R³, R⁴ and n are as defined in relation to formula (I) and R¹⁷ is as defined in relation to formula (V) with a compound of formula (VII)

where R' and m are as defined in relation to formula (I); and thereafter if desired or necessary, i) removing any protecting groups; and ii) optionally forming a pharmaceutically acceptable salt or in vivo hydrolysable derivative.

Particular protected groups R¹⁷ are ester groups an in particular C,.₆alkyl esters. Starting materials for the process may be obtained by standard organic chemistry procedures. The preparation of such starting materials is described in the following representative processes in conjunction with the accompanying Examples. Alternative starting materials are obtaintable by procedures which are analogous to those desctibed below and within the ordinary skill of an organic chemist. One process (a) above, comprises coupling together a compound of formula (IV) and (V) above, via the formation of a urethane or a carbamosulfanyl. The compounds are suitably reacted together in the presence of an inert solvent and in basic conditions.

A suitable solvent system for the aforementioned process is dichloromethane or tetrahydrofuran (THF) in the presence of pyridine.

A particular compound of formula (V) is a compound of formula (VIII)

where Y is the group - D - (E)_p - (F)_q - G - and where B, D, E, F, G, p and q are as hereinbefore defined and R¹⁷ is a COOC,_.6 alkyl. A particular example of a compound of formula (IV) is a compound of formula (IX)

where R¹, R², R³, R⁴, n and m are as defined in relation to formula (I).

An example of a compound of formula (VIII) where B is oxygen, D is CH₂ ,E is phenyl, p is 1, q is 1, F is oxygen, G is C₃ alkyl and R¹⁷ is COOMe

Compounds of formula (V) can be prepared by reacting a compound of formula (X)

with phosgene in the presence of toluene/dichloromethane. Where B is oxygen, this compound is the corresponding alcohol. These alcohols can be prepared by reaching 3-hydroxybenzyl alcohol and methyl 4- bromobutyrate under basic conditions. It will be appreciated that different alcohols can be prepared by substituting, for example, methyl 4-bromobutyrate by t-butylbromoacetate.

An example of a compound of formula (IX) where m and n are both 1, R¹ is methyl, R² and R³ are both hydrogen and R⁴ is methoxy is

Such a compound can be prepared from the corresponding 3-methoxy-4(N'-(2- methylphenyl)urea)nitrobenzene. The latter can be prepared from the reaction of the corresponding methoxy-nitroaniline together with methylphenylisocyanate.

It will appreciated by the skilled person that in some of the reactions it may be necessary to protect sensitive groups in the molecule. Protecting groups may typically be chosen from any of the groups described in literature or known to the skilled person.

Suitable protecting groups for an amino group include for example, formyl, and alkoxycarbonyl groups, for example tert-butoxycarbonyl.

Suitable protecting groups for a hydroxy group include lower alkyl groups, for example tert-butyl, and acyl groups, for example an alkanoyl group such as acetyl.

Suitable protecting groups for a carboxy group include, an esterifying group, for example a methyl or ethyl group. These groups may be removed by any convenient method described in the literature or known to the skilled person for removal of the specific protection group in question. The method of removal is chosen so as to minimise disturbance on the other groups in the molecule. For example when the protecting group is an acyl group or a C,_₂ alkyl group it may be removed by hydrolysis with a suitable base such as, for example, sodium hydroxide. A tert-butyl protecting group may be removed with, for example, an organic acid such as trifluoroacetic acid .

An alternative method of preparing a compound of formula (I), a pharmaceutically acceptable salt or an in vivo hydrolysable derivative comprises reacting together a compound of formula (VI) and (VII), to form a urea linkage. The reaction is suitably effected in the presence of an inert solvent. Suitable inerts solvents for the aforementioned process include dichloromethane, THF or ethyl acetate. As before, any functional group is protected if necessary and i) removing any protecting groups; and ii) optionally forming a pharmaceutically acceptable salt or in vivo hydrolysable derivative.

A particular example of a compound of formula (VI) is a compound of formula (XI)

where Y is the group - D - (E)_p - (F)_q - G - where R⁴, R'⁷' B, D, E, F, G, n, p and q are as hereinbefore defined. A particular example of a compound of formula (VII) is a compound of formula (XII)

where R' and m are as hereinbefore defined. An example of a compound of formula (V) is

(XIII) Such a compound can be prepared by reducing the corresponding nitroaniline derivatives. Thus compounds of formula (VI) can be prepared by reduction of a compound of fomula (XVI)

where A', Y', B, R¹⁷, R⁴ and n are as defined above.

A suitable reduction regime would be iron powder in the presence of ammonium chloride/water/methanol .

The nitroaniline derivative of formula (XVI) where A' is NH, can be prepared by reacting an appropriate compound of formula (XIV)

where B, Y' and R¹⁷ are as defined above, and B in particular is oxygen, with an isocyanate of formula (XV)

The reaction is suitably effected in an organic solvent such as dichloromethane of tetrahydrofuran (THF) in the present of a base such as triethylamine. Thus an exemplary scheme is as follows :

Inert solvent e.g dichloromethane or THF + small amt. base e,g triethylamine

The invention is further limited by the following biological test methods, data and non-limiting examples. Example 1 - Preparation of 4-(3-([({4-[(2-toIuidinocarbonyι)amino]anilino} carbonyl)oxy]methvUphenoxy)butanoic acid (Compound No. 1 in Table 1)

To methy l-4-(3 - { [( { 4- [(2-toluidinocarbonyl)amino]anilino } carbonyl)oxy]methyl } phenoxy)butanoate (0.085g) in dimethyl sulphoxide (DMSO) (1ml) was added 2N aqueous sodium hydroxide (0.2ml). The mixture was stirred for lhr at ambient temperature and then water (5ml) added. The pH was adjusted to pH 2 with concentrated hydrochloric acid. The precipitated solid product was filtered and washed with water and then dried to give the acid (0.055g) as a dark brown solid. (68%) Mass spectrum M-H 476.8

1H nmr (DMSO-d6) : 1.9 (m) 2H; 2.2 (s), 3H; 2.4 (t), 2H; 4.0 (t), 2H; 5.1 (s) 2H; 6.9 (m), 4H: 7.1 (m), 2H; 7.3 (t), 1H; 7.4 (s), 4H; 7.8 (m), 2H; 8.9 (s), 1H; 9.6 (s), 1H. a) Preparation of methyl 4-(3-hydroxymethyl)phenoxybutyrate

To a mixture of potassium carbonate (5.6g), 3-hydroxybenzyl alcohol (2.5g) in N- methylpyrollidinone (5ml) and methyl 4-bromobutyrate (2.2ml) were added followed by potassium iodide (2g). The mixture was stirred at room temperature for 48 hrs., added to water (30ml) and then extracted with diethyl ether (100ml). The organic layer was washed with dilute sodium hydroxide solution and brine. It was then dried over magnesium sulphate and evaporated to dryness. The residue was then purified by chromatography on silica using an increasingly polar mixture of ethyl acetate/ hexane (20% changing to 90%) to give methyl 4-(3-hydroxymethyl)phenoxybutyrate. (3.1g). 1H nmr (CDC1,) included the following resonances :

2.1 (m), 2H: 2.5 (t), 2H; 3.7 (s), 3H; 4.0 (t), 2H; 4.6 (s), 2H; 6.8 (q), 1H; 6.9 (m), 2H; 7.2 (m), 1H. b) Preparation of methyl-4-(3-{[({4-[(2-toluidinocarbonyl)amino] anilino}carbonyl)oxy]methyl}phenoxy)butanoate

To methyl 4-(3-hydroxymethyl)phenoxybutyrate (0.22g) in dichloromethane (3ml) was added 20% phosgene in toluene (3ml) and the mixture allowed to stand for 18hrs, evaporated and part of the residue (0.15g) in dichloromethane (2ml) added to 4-(2- tolylureido)aniline (CAS Registry Number 202874-38-2 and as described in WO 98/04247) (0.12g) in dichloromethane (3ml) containing pyridine (O.lg). The mixture was stirred for 2hrs and then evaporated and the residue purified by chromatography on silica using an increasingly polar mixture of methanol/ chloroform (0% changing to 10%) to give, after evaporation and trituration in diethyl ether the above material (0.15g) Mass spectrum: M+H 492 1H nmr (DMSO-d6) included the following resonances :

2.0 (m), 2H; 2.2 (s), 3H; 2.5 (m), 2H; 3.6 (s), 3H; 4.0 (t), 2H; 5.1 (s), 2H; 6.9 (m), 4H;

7.1 (m), 2H; 7.3 (t), 1H; 7.4 (s), 4H; 7.8 (m), 2H; 8.9 (s), 1H; 9.6 (s), 1H.

Example 2 - Preparation of 4-(3- ({methyl-4-[(2-toluidinocarbonvDaminolanilino] carbonyl)oxylmethy phenoxy^")butanoic acid (Compound No 5 in Table 1)

This was prepared by hydrolysis of methyl-4-(3-{[({methyl-4-[(2- toluidinocarbonyl)amino]anilino} carbonyl)oxy]methyl}phenoxy)butanoate using the process described in example 1

Mass spectrum M+H 492

1H nmr (DMSO-d6) included the following resonances :

1.9 (m), 2H; 2.2 (s), 3H; 2.4 (t), 2H; 3.2 (s), 3H; 4.0 (t), 2H; 5.1 (s), 2H; 6.8 (m), 3H; 6.9 (t), 1H; 7.2 (m), 6H; 7.4 (d), 2H; 7.8 (d), 2H; 7.9 (s), 1H; 9.0 (s), 1H. a) Preparation of methyl 4-[3-({[(4-nitroanilino)carbonyl]oxy} methyI)phenoxy]butanoate

To methyl 4-(3-hydroxymethyl)phenoxybutyrate (2.24g) in dichloromethane (10ml) was added 4-nitrophenylisocyanate (2g) and triethylamine (3 drops). The mixture was stirred for 2 hrs., evaporated to dryness and the residue triturated with diethyl ether to give methyl 4-[3-({[(4-nitroanilino)carbonyl]oxy}methyl)phenoxy]butanoate as a yellow powder (3.5g) 1H nmr (CDC1₃) included the following resonances :

2.1 (m), 2H; 2.5 (t), 2H; 3.7 (s), 3H; 4.0 (t), 2H; 5.2 (s), 2H; 6.9 (m), 3H; 7.1 (s), 1H; 7.3 (d), lH; 7.6 (d), 2H; 8.2 (d), 2H. b) Preparation of methyl 4-[3-({[(methyl-4-nitroanilino)carbonyl]oxy} methyl)phenoxy]butanoate

To methyl 4-[3-({[(4-nitroanilino)carbonyl]oxy}methyl)phenoxy]butanoate(lg) in N- methylpyrollidinone (5ml) added sodium hydride (1.1 equ.) and methyl iodide (1.3 equ.) and the mixture stirred for 4hrs, added to water (20ml) and extracted with diethyl ether (50ml), this washed with brine, dried and evaporated to dryness. The residue was purified by chromatography on silica using an increasingly polar mixture of ethyl acetate/ hexane (10% changing to 30%) to give product as a gum (0.8)g which crystallised on standing.

1H nmr (CDC1,) included the following resonances :

2.1 (m), 2H; 2.5 (t), 2H; 3.4 (s), 3H; 3.7 (s), 3H; 4.0 (t), 2H; 5.2 (s), 2H; 6.9 (m), 3H; 7.3 (m), lH; 7.6 (d), 2H; 8.2 (d), 2H. c) Preparation of methyI-4-(3-{[({methyl-4-[(2-toluidinocarbonyl)amino] ani!ino}carbonyl)oxy]methyl}phenoxy)butanoate

To methyl 4-[3-({ [(methyl-4-nitroanilino)carbonyl]oxy}methyl)phenoxy] butanoate(0.7g) in methanol (10ml) was added iron powder (0.7g) and a solution of ammonium chloride (0.13g) in water (4ml). The mixture was refluxed for lhr., ethyl acetate (30ml) added and the mixture filtered to remove metal salts, and evaporated to dryness. The residue in dichloromethane was dried with magnesium sulphate, filtered and evaporated to dryness. The residue in dichloromethane (5ml) was treated with o-tolylisocyanate (1.1 equ.) and allowed to stand for 18hrs., evaporated to dryness and triturated with diethyl ether to give methyl-4-(3-{[({methyl-4-[(2-toluidinocarbonyl)amino] anilino} carbonyl)oxy]methyl}phenoxy]butanoate (0.6g) as an off-white solid.

1 H nmr (DMSO-d6) included the following resonances :

2.0 (m), 2H; 2.2 (s), 3H; 2.45 (m), 2H; 3.2 (s), 3H; 3.6 (s), 3H; 3.9 (t), 2H; 5.0 (s), 2H; 6.8

(m), 3H; 6.9 (m), 1H; 7.2 (m), 5H; 7.4 (d), 2H; 7.8 (d), 1H; 7.9 (s), 1H; 9.0 (s), 1H. „„

- 29 -

Example 3 - Preparation of 4-(3-{[({4-[(2-toluidinocarbonvDamino]anilino) carbonvDoxy] methyl} phenoxy)ethanoic acid (Compound No. 6 in Table 1)

Tertiary-butyl-4-(3 - { [( { 4- [(2-toluidinocarbonyl)amino]anilino } carbonyl) oxy]methyl}phenoxy)ethanoate (0.6g) was stirred in a mixture of dichloromethane (4ml) and trifluoroacetic acid (4ml) for 2hrs. and then evaporated to dryness. The residue was triturated in diethyl ether to give 4-(3-{[({4-[(2- toluidinocarbonyl)amino]anilino}carbonyl)oxy]methyl}phenoxy)ethanoic acid as a white powder (0.48g, 87% ) Mass spectrum M-H 448

1H nmr (DMSO-d6) included the following resonances :

2.2 (s), 3H; 4.6 (s), 2H; 5.1 (s), 2H; 6.9 (m), 4H; 7.1 (m), 2H; 7.3 (t), 1H; 7.35 (s), 4H; 7.8

(m), 2H; 8.9 (s), 1H; 9.6 (s), 1H. a) Preparation of t-butyl 4-(3-hydroxymethyl)phenoxyacetate To 3-hydroxybenzyl alcohol (95 g) in acetone (10ml) was added potassium carbonate

(1 lg ) and 1 spoonful of potassium iodide and the mixture stirred whilst t-butylbromo acetate (6.5ml) was added. The mixture was then stirred for a further 18 hrs. The acetone was then evaporated and the residue partitioned between water (60ml) and ethyl acetate (60ml). The organic layer was dried and evaporated and the residue purified by chromatography on silica using an increasingly polar mixture of ethyl acetate in hexane. The appropriate fractions evaporated to dryness gave 7g of white crystalline product. 1 H nmr (CDC1₃) contained the following resonances : 1.5 (s), 9H; 4.5 (s), 2H; 4.65 (d), 2H; 6.8 (q), 1H; 7.0 (m), 2H; 7.3 (t), 1H. b) Preparation of t-butyl-4-[3-({[(4-nitroanilino)carbonyl]oxy} methyl)phenoxy]ethanoate

To t-butyl 4-(3-hydroxymethyl)phenoxyacetate (1.2g) in dichloromethane (10ml) was added 4-nitrophenylisocyanate (1.1 equ.) and triethylamine (3 drops). The mixture was stirred for 2 hrs., evaporated to dryness and the residue triturated with diethyl ether to give t-butyl-4- [3-({[(4-nitroanilino)carbonyl]oxy}methyl)phenoxy]ethanoate as a yellow powder (1.7g) lH nmr (DMSO-d6) included the following resonances :

1.4 (s), 9H; 4.6 (s), 2H; 5.15 (s), 2H; 6.85 (q), 1H; 6.95 (s), 1H; 7.0 (d), 1H; 7.3 (t), 1H; 7.7 (d), 2H; 8.2 (d), 2H. c) Preparation of t-butyl-4-(3-{[{(4-[(2-toluidinocarbonyl)amino] anilino}carbonyl)oxy]phenoxy)ethanoate

T-buty 1-4- [3 -( { [(4-nitroanilino)carbony 1] oxy } methy l)phenoxy] ethanoate(0.7g) was reduced and reacted with o-tolylisocyanate by the method described in example 2 to give t- butyl-4-(3-{[({4-[(2-toluidinocarbonyl)amino]anilino}carbonyl)oxy]methyl} phenoxy)ethanoate (0.8g) as an off-white powder. IH nmr (DMSO-d6) included the following resonances :

1.4 (s), 9H; 2.2 (s), 3H; 4.6 (s), 2H; 5.1 (s), 2H; 6.9 (m), 4H; 7.1 (m), 2H; 7.3 (t), IH; 7.4 (s), 4H; 7.8 (m), 2H; 8.85 (s), IH; 9.6 (s), IH.

Example 4 - Preparation of 4-(3-{K(3-methoxy-4-[(2-toluidinocarbonyl)amino] anilino}carbonyl)oxy]methyl)phenoxy)butanoic acid (Compound No. 2 in Table 1)

Methyl 4-(3 - { [( { 3 -methoxy -4- [(2-toluidinocarbonyl)amino]anilino } carbonyl) oxy]methyl}phenoxy)butanoate(0.15 g) was hydrolysed by the method described in example 1 to give 0.07g of product as a dark powder. Mass spectrum M+H 508

IH nmr (DMSO-d6) included the following resonances:

1.9 (m), 2H; 2.2 (s), 3H; 2.4 (t), 2H; 3.8 (s), 3H; 4.0 (t), 2H; 5.1 (s), 2H; 6.9 (m), 5H; 7.1 (m), 2H; 7.3 (m), 2H; 7.8 (d), IH; 8.0 (d), IH; 8.4 (s), IH; 8.5 (s), IH; 9.6 (s), IH. a) Preparation of 3-methoxy-4(N'-(2-methylphenyl)urea) nitrobenzene

2-methoxy-4-nitroaniline(16.8g) in ethyl acetate(90ml) was treated with stirring with 2-methylphenylisocyanate (16.1ml). The solution was heated to 60°C for 12 hours. The solution was then chilled to 0°C and the precipitate filtered and washed with cold ethyl acetate. This gave 3-methoxy-4(N'-(2-methylphenyl)urea)nitrobenzene (lOg). lH nmr (DMSO-d6) included the following resonances:

2.2 (s), 3H; 4.0 (s) 3H; 6.9 (t) IH; 7.2 (m) 2H; 7.7 (d) IH; 7.8 (s) IH; 7.9 (d) IH; 8.4 (d) IH; 8.8 (s) IH; 9.2 (s) IH. m/Z 302(M+H), 300(m-H). b) Preparation of N-(4-amino-2-methoxyphenyl)-N'-(2-methylphenyl)urea) At ambient temperature a rapidly stirred solution of 3-methoxy-4(N'-(2- methylphenyl)urea)nitrobenzene(5g) in DMF (50ml) containing 10% palladium on carbon (500mg) was exposed to an atmosphere of hydrogen. When uptake of hydrogen had ceased the solution was filtered and the filter cake washed with dichloromethane. The combined filtrates were evaporated to dryness under reduced pressure, triturated with diethylether and filtered to give product (4.14g) as a off- white solid. IH nmr (DMSO-d6) included the following resonances:

2.2 (s) 3H; 3.8 (s) 3H; 4.8 (s) 2H; 6.1 (d) IH; 6.3 (s) IH; 6.8 (t) IH; 7.1 (m) 2H; 7.5 (d) IH; 7.8 (d) IH; 8.05 (s) IH; 8.15 (s) IH. m/Z 272(M+H) c) Preparation of methyl 4-(3-{[({3-methoxy-4-[(2-toluidinocarbonyl)amino] anilino}carbonyl)oxy] methyl} phenoxy)butanoate

N-(4-amino-2-methoxyphenyl)-N'-(2-methylphenyl)urea)(0.27g) was treated with the chloroformate of methyl 4-(3-hydroxymethyl)phenoxybutyrate as described in example 1 to give product (0.16g).

IH nmr (DMSO-d6) included the following resonances : 2.0 (m), 2H; 2.2 (s), 3H; 2.5 (m), 2H; 3.6 (s), 3H; 3.8 (s), 3H; 4.0 (t), 2H; 5.1 (s), 2H;

6.9 (m), 5H; 7.1 (m), 2H; 7.3 (m), 2H; 7.8 (d), IH; 8.0 (d), IH; 8.4 (s), IH; 8.5 (s), IH; 9.6 (s), IH.

Example 5 - Preparation of 2-(3-{2[((4[2-toIuidinocarbonyl)amino]anilino} carbonyl)oxylethyl}phenoxy)acetic acid (Compound No. 7 in Table 1)

Tertiary -butyl-2-(3 - { 2 [( { 4 [2-toluidinocarbonyl)amino]anilino } carbonyl)oxy] ethyl } phenoxy)ethanoate (0.5g) was hydrolysed by the method described in example 3 (0.41 g) Mass spectrum M-H 462 IH nmr (DMSO-d6) included the following resonances :

2.2 (s), 3H; 2.9 (t), 2H; 4.3 (t), 2H; 4.7 (s), 2H; 6.7 (q), IH; 6.9 (m), 3H; 7.1 (m), 3H; 7.3 (s), 4H; 7.8 (m), 2H; 8.8 (s), IH; 9.5 (s), IH. a) Preparation of t-butyl 3-(2-hydroxyethyl)phenoxyacetate

To 3-(2-hydroxyethyl)phenol (2.8g) in N-methylpyrollidinone (8ml) was added potassium carbonate (5.6g) and t-butylbromoacetate (4.4g). The mixture was stirred for 18hrs, added to water (40ml) and extracted into diethyl ether. The organic layer was washed with brine, dried and evaporated to dryness to give t-butyl 3-(2-hydroxyethyl)phenoxyacetate (5.3g) used without further purification

1 H nmr (CDC1₃) included the following resonances :

1.5 (s), 9H; 2.8 (t), 2H; 3.8 (q), 2H; 4.5 (s), 2H; 6.8 (m), 3H; 7.2 (t), IH. b) Preparation of t-butyl 2-[3-(2-{[(4-nitroanilino)carbonyl]oxy} ethyl)phenoxy] ethanoate

Tertiary-butyl 3-(2-hydroxyethyl)phenoxyacetate (2.5g) was reacted with 4- nitrophenylisocyanate (1.6g) by the method described in example 3 to give the product(2.9g) IH nmr (CDC1,) included the following resonances : 1.5 (s), 9H; 3.0 (t), 2H; 4.4 (t), 2H; 4.6 (s), 2H; 6.8 (m), 3H; 7.2 (m), 2H; 7.6 (d), 2H;

8.2 (d), 2H. c) Preparation of t-butyl-2-(3-{2[({4[2-toluidinocarbonyl)amino]anilino} carbonyl)oxy]ethyl}phenoxy)ethanoate t-butyl 2-[3-(2-{[(4-nitroanilino)carbonyl]oxy}ethyl)phenoxy]ethanoate (0.8g) was reduced and reacted with o-tolylisocyanate by the method described in example 2 to give the product(0.7g) as a white powder.

1 H nmr (DMSO-d6) included the following resonances :

1.4 (s), 9H; 2.2 (s), 3H; 2.9 (t), 2H; 4.4 (t), 2H; 4.6 (s), 2H; 6.9 (m), 4H; 7.1 (m), 3H; 7.8 (m), 2H; 8.8 (s), IH; 9.5 (s), IH.

Example 6

Preparation of Compound No. 31 in Table 2

To the methyl ester of the Compound 31 (0.099g) in MeOH (1ml) was added 2N aq NaOH solution (0.08ml), and the mixture stirred at ambient temperature for 16 hrs. This was then evaporated to dryness, the residue dissolved in THF (5ml) and acidified with cone, hydrochloric acid. This solution was then adsorbed onto silica, and purified by chromatography on silica, using increasingly polar mixtures of CH₂Cl₂/MeOH (5%-20%) and the appropriate fractions evaporated to dryness to give the Compound 31 (0.054g) as an off- white solid (56%). MS M+H 515

NMR spectrum (in DMSO-d₆) included the following resonances: 1.62(m) 2H, 1.80-2.04 (m) 2H, 2.05 (s) 3H, 2.20-2.30 (m) 7H, 2.62 (d) 2H, 2.68 (d) 2H, 3.82 (s) 3H, 4.04 (m) 3H, 6.95 (m) 2H, 7.14 (m) 2H, 7.76 (d) IH, 7.90 (s) IH, 8.52 (s) IH, 8.54 (s) IH, 8.80 (s) 1 a) Preparation of alcohol intermediate

2-(Hydroxymethyl) morpholine (0.8g) was dissolved in acetonitrile (15 ml), and to this solution were added anhydrous potassium carbonate (2.85g; 3 equivalents), potassium iodide (0.025g; catalytic amount) and methyl-4-bromobutyrate (0.95 ml; 1.05 equivalents). This reaction mixture was then stirred at 60° C for 16 hours. The cooled suspension was then filtered, washing with more acetonitrile, and the filtrate concentrated under reduced pressure to give the desired alcohol as a yellow oil (1.48g; 100% yield not purified).

MS Data: M+H 218 b) Preparation of nitrophenyl intermediate The alcohol intermediate of step 1 was reacted with 4-nitrophenylisocyanate as described in Example 2a to give the corresponding nitro intermediate of formula

MS M+H 426 NMR spectrum (in DMSO-d₆) included the following resonances:

1.64-1.72 (m) 2H, 1.80-1.86 (m) IH, 1.92-2.04 (m) IH, 2.20 (s) 3H, 2.26-2.36 (m) 4H, 2.60- 2.64 (m) IH, 2.72-2.78 (m) IH, 3.42-3.52 (m) IH, 3.58 (s) 3H, 3.66 (m) IH, 3.80 (m) IH, 3.84 (s) 3H, 4.10 (d) 2H, 7.60 (s) IH, 7.78 (s) IH, 9.28 (s) IH c)Preparation of aniline intermediate

The nitro compound from (b) (0.47g) was dissolved in ethanol (10 ml) and ammonium formate (0.625g; 10 equivalents) was added. The reaction flask was flushed with argon, and the catalyst (10% Pd/C, 0.047mg; 0.1 equivalents by weight) was then added. The reaction was heated to reflux and stirred at this temperature for 1 hour. The reaction mixture was then cooled and filtered through a bed of celite, washing through with more ethanol. The solvent was removed under reduced pressure, and the residue dissolved in ethyl acetate (50ml). This solution was then washed with water, brine, dried over anhydrous magnesium sulphate and concentrated under reduced pressure to give the above aniline as a brown oil (0.43g; 98% yield).

MS Data: M+H 438 d) Preparation of Methyl Ester precursor to Compound 31 The aniline intermediate of step c was reacted with o-tolylisocyanate as described in Example 2c to yield the required methyl ester.

MS Data: M+H 529

Example 7 Preparation of Compound No. 29 in Table 2

To the methyl ester of the above (0.075g) in THF (2ml) was added a solution of LiOH.H₂0 (0.04g) in H₂0 (2ml), and the mixture stirred at ambient temperature for 16 hrs. This was acidified with cone, hydrochloric acid and evaporated to dryness. Purified by chromatography on silica, using increasingly polar mixtures of CH₂Cl₂/MeOH (5%-20%) and the appropriate fractions evaporated to dryness to give the above material (0.064g) as an off- white solid (88%).

MS M+H 455

Example 8

Using the methods of the Examples as described above, where Method A is as described in Example 1 , Method B is as described in Example 2, Method C is as described in Example 7, Method D is as described in Example 7, the following compounds whose structures are listed in Tables 1 , 2 and 3 were prepared:

Table 4

a) Alkyl (usually methyl or ethyl) ester intermediates (i.e. compounds of formula (I) where R⁵ is an methyl or ethyl carboxylic ester) used in the process were generally obtained as described in Example 2c. The characterising data where available of the esters are set out in Table 5.

Table 5

In the above Table, the compound nos refer to the acids which were ultimately derived from the methyl esters. Examples with the note 'alkylation' had the appropriate nitro intermediate alkylated as described in Example 2b. ai) The methyl ester of Compound 3 in Table 1 was obtained from a urea carbamate ester intermediate -ureidoaniline + chloroformate using a method analogous to that described in Example 1

MS M+H 462

NMR spectrum (in DMSO-d₆) included the following resonances: 2.2 (s) 3H, 2.7 (t) 2H, 2.9 (t) 2H, 3.6 (s) 3H, 5.1 (s) 2H, 6.9 (m) IH, 7-7.4 (m) 8H, 7.8 (m) 2H, 8.8 (s) IH, 9.4 s) lH b) Details of the intermediate anilines of formula (VI) are set out in Table 6. In this Table, the method indicates the following: Y¹ Fe/NH₄Cl/EtOH/H₂O As Ex 2c

Y² HCO₂NH₄/Pd/C/EtOH As Ex 6c

Y³ H₂/Pd/C/EtOH As Ex 4b

The compound No. refers to the Compound of formula (I) which was ultimately derived from the intermediate.

Table 6

c) The above described anilines were obtained from the corresponding nitro compounds of formula (XIV) as set out above using methods analogous to that of Example 2a. Data for the nitro compounds is set out in Table 7. Table 7

Again, the compound No. column refers to the compound of formula (I) ultimately obtained from this intermediate.

ci) In the case of the nitro compound used in the preparation of compound 54 in Table 3, (i.e. the compound of formula

a modified route was used.

To methyl 4-(3-hydroxymethyl)phenoxybutyrate (1.12g) in dichloromethane (6ml) was added 20%) phosgene in toluene (12ml) and the mixture allowed to stand for 18hrs, evaporated and the residue in dichloromethane (15ml) added to 4-nitroindoline (0.82g) in pyridine (2ml) and the mixture stirred overnight, evaporated to dryness and partitioned between aqueous acid and diethyl ether. A crystalline material precipitated and was filtered, washed with ether and dried to give the above (0.8g)

NMR

2. 0 (m) 2H, 2.4 (m) 2H, 3.2 (t) 2H, 3.6 (s) 3H, 4.0 (t) 2H, 4.1 9t) 2H, 5.2 (s) 2H, 6.9 (m) 3H,

7.3 (t) IH, 7.8 (m) IH, 8.1 (m) 2H Alcohol intermediates (of general formula (XIV) above) used in the preparation of the nitro compounds were prepared by various methods. These are outlined below or summarised in Table 8.

dl) Alcohol for use in the Preparation of Compound No. 28 in Table 1

Piperidine methanol (0.5g) was dissolved in dichloromethane (10 ml) and triethylamine (0.64 ml; 1.05 equivalents) was added, followed by methyl-4-bromobutyrate (0.6 ml; 1.05 equivalents). The resulting solution was stirred at room temperature for 16 hours, then diluted with a further 40 ml dichloromethane and washed with water (25 ml). The organic solution was dried using phase separating paper and concentrated under reduced pressure. Purification on a silica Bond Elut column using dichloromethane/methanol mixtures of increasing polarity as eluent gave the desired alcohol (Example 6 Id) as a yellow oil (0.309g; 33% yield).

MS Data: M+H 216

NMR Data: (CDC1₃) 1.20-1.34 (m) IH, 1.58-1.70 (m) 2H, 1.70-1.90 (m) 4H, 2.04 (m) IH, 2.14 (m) IH, 2.26 (m) IH, 2.36-2.44 (m) 4H, 2.66 (m) IH, 2.84 (m) IH, 3.56 (m) IH, 3.66 (m) lH, 3.68 (s) 3H,

d2) Alcohol for use in the Preparation of Compound No. 30 in Table 1

2-(Hydroxymethyl) morpholine (0.8g) was dissolved in acetonitrile (15 ml), and to this solution were added anhydrous potassium carbonate (2.85g; 3 equivalents), potassium iodide (0.025g; catalytic amount) and methyl-4-bromobutyrate (0.95 ml; 1.05 equivalents). This reaction mixture was then stirred at 60° C for 16 hours. The cooled suspension was then filtered, washing with more acetonitrile, and the filtrate concentrated under reduced pressure to give the desired alcohol (Example 63d) as a yellow oil (1.48g; 100%) yield not purified).

MS Data: M+H 218

d3) Alcohol for use in the Preparation of Compound No. 48 in Table 1

Method exactly in method d2, but using the alkyl chloride in place of the alkyl bromide. MS Data: (EI+) M+ 243

d4) Alcohol for use in the Preparation of Compound No. 4 in Table 1 To 3 -hydroxy acetophenone (7g) in NMP (70ml) was added potassium carbonate

(13.8g) and potassium iodide (2g), followed by methyl-4-bromobutyrate (6.2ml) and the mixture stirred for 18hrs, then added to water (200ml) and extracted with ethyl acetate. The organic extracts were wshed with brine and evaporated to dryness, purified by chromatography on silica (hexane/ethyl acetate) and the appropriate fractions evaporated to dryness. lg of this material was dissolved in ethanol and sodium borohydride (0.16g) was added and the mixture stirred for 18hrs, evaporated , and partitioned between dilute acid and diethyl ether. The organic layer was separated, dried and evaporated and the residue purified by chromatography on silica (10% ethyl acetate in hexane increasing to 90%). The aapropriate fractions evaporated to dryness gave an approx. 50:50 mixture of ethyl & methyl 4-(3-[l- hydroxy]ethyl)phenoxybutyrate. NMR

1.2 (t) 1.5H ethyl ester , 1.5 (d) 3H, 2.1 (m) 2H, 2.5 (m) 2H, 3.7 (s) 1.5H methyl ester, 4.0 (t) 2H, 4.1 (q) IH, ethyl ester, 4.9 (m) IH, 6.8 (d) IH, 6.9 (s) 2H, 7.2 (m) IH. d5) Alcohol for use in the Preparation of Compound No. 23

To 1,3 benzenedimethanol (1.4g) in THF (10ml) added t-butylacrylate (2g) followed by a 2 drops of benzyltrimethylammonium hydroxide solution. The mix stirred for lhr. , evaporated and purified by chromatography on silica using an increasingly polar mixture of hexane/ethyl aceate to give after evaporation of the appropriate fractions 0.55g of t-butyl (3- hydroxymethyl)benzyloxypropionate as an oil.

M+H 267

d61 Alcohol for use in the Preparation of Compound No. 35

To 4-hydroxypiperidine (2g) in DMf (35ml) was added potassium carbonate (8.2g) and methyl 4-bromobutyrate (2.9ml). The mix was stirred at 70C for 8hrs and then cooled, and partitioned between ethyl acetate and water, the organic layer washed with brine, dried and evaporated to dryness. The residue has a consistent M+H and was used without further purification or characterisation.

d7) Alcohol for use in the Preparation of Compound No. 41 To 4-(3-hydroxypropyl)pyridine (1.4g) in acetonitrile (10ml) was added t-butyl bromoacetate (2g) and the mixture allowed to stand for 24 hrs., and evaporated. The residue was dissolved in methanol and hydrogenated at atmospheric pressure in the presence of 10% palladium on carbon catalyst at 55C for 16 hrs. The mix was cooled, filtered and evaporated to give t-butyl 4-(3-hydroxypropyl)piperidinylacetate M+H 258

d8) Alcohol for use in the Preparation of Compound No. 43

To 4-(2-hydroxyethyl)piperidine (1.29g) in acetonitrile (5ml) was added t-butyl acrylate (1.28g) and the mixture allow to stand for 18hrs, evaporated to give t-butyl 4-(2- hydroxyethyl)piperidinylpropionate M+H 258

d9) Alcohol for use in the Preparation of Compound No. 25 To 3-(hydroxyethoxy)phenol (CAS49650-88-6) (1.54g) in N-methylpyrollidinone

(3ml) was added potassium carbonate (2.8g) and t-butylbromoacetate (1.95g) and the mix stirred for 18hrs, then added to a mixture of brine and diethyl ether. The organic layer was washed with brine, dried and evaporated to dryness and the residue purified by chromatography using an increasingly polar mixture of hexane / ethyl acetate to give t-butyl 3 -(hydroxy ethoxy)phenoxy acetate M+H 269

dlO) Alcohol for use in the Preparation of Compound No. 26 in Table 1

To methyl 4-iodophenylacetate (27g) in acetonitrile (300ml) were stirred and palladium chloride (0.17g), copper (I) iodide (0.37g), triphenylphosphine (0.5 lg) , 3-butyn-l - ol (8.85g ) and triethylamine (20ml) were added and the mixture stirred under an inert atmosphere in the absence of light for 18hrs. and then the solvent removed by evaporation and the residue partitoned between water and ethyl acetate, the organic layer removed, dried and evaporated to dryness. The residue was purified by chromatography on silica using an increasingly polar mixture of ethyl acetate/hexane. The appropriate fractions evaporated to dryness gave methyl 4-(4-hydroxybut-3-ynyl)phenylacetate (18.6g)

This was reduced in ethanol(250ml) using Pd/C catalyst (5%,2g) under an atmosphere of hydrogen gas at a pressure of 3 atmospheres for lhr, filtered and evaporated to dryness to give methyl 4-(4-hydroxybutyl)phenylacetate (18g) as a yellow oil used without further purification or characterisation.

Table 8

Example 9

The compounds of the invention or pharmaceutically acceptable salts thereof may be formulated into tablets together with, for example, lactose Ph.Eur, Croscarmellose sodium, maize starch paste (5% w/v paste) and magnesium stearate for therapeutic or prophylactic use in humans. The tablets may be prepared by conventional procedures well known in the pharmaceutical art and may be film coated with typical coating materials such as hydroxypropylmethylcellulose.

In Vitro and In Vivo Assays

The following abbreviations are used. Suitable sources of materials are listed below. MOLT-4 cells - human T-lymphoblastic leukaemia cells (European Collection of Animal Cell Cultures, Porton Down).

Fibronectin - purified from human plasma by gelatin-sepharose affinity chromatography according to the methods described in E.Nengvall, E.Ruoslahti, Int. J. Cancer, 1977, 20, pages 1-5 and J. Forsyth et al, Methods in Enzymology, 1992, 215, pages 31 1-316).

RPMI 1640 - cell culture medium. (Life technologies, Paisley UK). PBS - Dulbecco's phosphate buffered saline (Life Technologies). BSA - Bovine serum albumin, fraction V (ICN, Thame, UK). CFA - Complete Freund's Adjuvant (Life Technologies).

In the following assays and models references to compound(s) refers to the compounds of formula (I) according to the present invention. 1.1 In vitro assay : MOLT-4 cell/ Fibronectin adhesion assay.

The MOLT-4 cell /fibronectinadhesion assay was used to investigate the interaction of the integrin α₄-β, expressed on the MOLT-4 cell membrane with fibronectin. Polystyrene 96 well plates were coated overnight at 4°C with fibronectin, 100 μl of 10 μg/ml in PBS. Nonspecific adhesion sites were blocked by adding 100 μl BSA, 20 mg/ml. After incubating for 1 h at room temperature, the solutions were aspirated. MOLT-4 cells suspended in serum-free RPMI-1640 medium 2E6 cells/ml (50 μl) and solutions of compound diluted in the same medium (50 μl) were added to each well. After incubation for 2 h at 37°C in a humidified atmosphere of 5% (v/v) CO₂, non-adherent cells were removed by gentle shaking followed by vacuum aspiration. Adherent cells were quantified by a colorimetric acid phosphatase assay. To each well was added 100 μl p-nitrophenyl phosphate (6 mg/ml) in 50 mM sodium acetate buffer, pH 5.0, containing 1% Triton X-100. After incubation for 1 h at 37°C, 50 μl sodium hydroxide (1M) was added to each well and the absorbance 405 nm was measured on a microplate spectrophotometer. Compounds which inhibited adhesion gave a lower absorbance reading. Standard, control and test conditions were assayed in triplicate. Percentage inhibition was calculated with respect to total (no inhibitor) and non-specific (no fibronectin) standards on each plate.

1.2 In-vivo Inflammation Models

Activity of a compound can be tested in the following models. 1.2.1 Ovalbumin Delayed type Hypersensitivity in mice

Balb/c female mice (20-25g) are immunised on the flank with an 1 : 1 (v/v) emulsion of ovalbumin (2 mg/ml) with CFA. Seven days later the mice are challenged by subplantar injection of 1% heat aggregated ovalbumin in saline (30 μl) into the right hind foot pad. Swelling of the foot develops over a 24 hour period following which foot pad thickness is measured and compared with the thickness of the contralateral uninjected foot. The percentage increase in foot pad thickness is calculated. Compounds are dosed orally by gavage to groups of 5 mice at doses ranging from 0.001 mg/kg to 100 mg/kg. Inhibition of the inflammatory response is calculated comparing vehicle treated animals and compound treated groups. 1.2.2. Collagen-induced arthritis in mice

DBA/1 male mice are immunised with 0.1ml of an emulsion prepared from equal volumes of bovine collagen type II in 0.05M acetic acid (2 mg/ml) and CFA. This mixture is injected at the base of the tail. Twenty days later compounds are dosed orally by gavage at doses ranging from O.OOlmg/kg/day to 100 mg/kg/day. On the day following the first dose, each animal receives an intra-peritoneal booster injection of 0.1ml of collagen type II in acetic acid. The mice are assessed for the incidence and severity of arthritis in all four limbs for up to 28 days. Inhibition of arthritis is calculated by comparing vehicle treated and compound treated mice.

Claims

Claims:

A compound of formula (I)

wherein

A' is NH or NR⁶, where R⁶ is C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkanoyl or C^alkoxycarbonyl; B is oxygen or sulphur;

Y' is a linker group comprising an optionally substituted hydrocarbyl chain which is optionally interposed by one or more heteroatoms independently selected from oxygen, nitrogen and sulphur and/or by a monocyclic or bicyclic ring system; or linker group Y' and A' can be taken together to form a 5 to 7 membered heterocyclic ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂_₆alkenyl, C₂.₆alkynyl, C_|.₆alkoxy, C₂_ ₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyl]amino and C₂.₆alkanoylamino; m is from 0 to 5; n is from 0 to 4; R¹ and R⁴ are each independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl, C,_.6alkanoyl, C₂.₆alkynyl, C,_₆ alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆ alkylamino, di-[(C_!.6)alkyl]amino, C_2.6alkanoylamino, N-C,_₆alkylcarbamoyl, C_μ6alkoxylcarbonyl, N,N-di-[(C,_.6alkyl]carbamoyl, C,.₄alkoxylC,.₆alkyl, (CH₂)_tOH where t is 1 or 2, -CO₂R^a and CONR^aR^D where R^a and R^b are independently hydrogen or C,.₆ alkyl or one of R⁴ can be taken together with A to form a 5 to 7 membered heterocyclic ring optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C^alkyl, C₂.₆alkenyl, C_2.6alkynyl, C,_.6alkoxy, C_2.6alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyl]amino and C₂.₆alkanoylamino; R² and R³ are each independently selected from hydrogen, C,_₆ alkyl, C,-₃ alkanoyl or C,_.6 alkoxycarbonyl; and R⁵ is an acidic functional group; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

2. A compound according to claim 1 wherein Y' is an alkylene chain which is interposed by a monocyclic ring system and/or a heteroatom.

3. A compound according to claim 1 wherein the group A' is orientated in the meta or para position with respect to the ureido group in formula (I), and most preferably A' is orientated para- to the ureido group in formula (I).

4. A compound according to claim 1 of formula (IA)

wherein

A is nitrogen or NR⁶, where R⁶ is C,.₆alkyl, C₂.₆alkanoyl or C_Le-alkoxycarbonyl; B is oxygen or sulphur;

Y is a linker group connecting group B to group R⁵ and containing up to 10 atoms where each atom is independently selected from carbon, oxygen, nitrogen and sulphur and may optionally comprise a monocyclic or bicyclic ring system or linker group Y and A can be taken together to form a 5 to 7 membered heterocyclic ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂_ ₆alkynyloxy, C,.₆alkylamino, di-[C,_₆ alkyljamino and C₂.₆alkanoylamino; m is from 0 to 5; n is from 0 to 4;

R¹ and R⁴ are each independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C_2.6alkenyl, C_:.₆alkanoyl, C₂.₆alkynyl, C,.₆ alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,_₆ alkylamino, di-[(C,.₆)alkyl]amino, C₂.₆alkanoylamino, N-C^alkylcarbamoyl, C,.₆alkoxylcarbonyl, N,N-di-[(C,.₆alkyl]carbamoyl, C,.₄alkoxylC,.₆alkyl, (CH₂),OH where t is 1 or 2, -CO₂R^a and CONR^aR^b where R^a and R^b are independently hydrogen or C,.₆ alkyl or one of R⁴ can be taken together with A to form a 5 to 7 membered heterocyclic ring optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,_.6alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyl]amino and C₂.₆alkanoylamino; R² and R¹ are each independently selected from hydrogen, C,_₆ alkyl, C,-, alkanoyl or C,_.6 alkoxycarbonyl; and R⁵ is an acidic functional group; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

5. A compound according to any one of the preceding claims of formula (II)

wherein

A, B, R¹ to R⁵, m and n are as defined in claim 4,

D and G are each independently C alkyl or C₂.₄ alkenyl and each carbon is optionally substituted with halogeno, hydroxy, amino, nitro, phenyl, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl, C,.₆alkanoyl, C₂.₆alkynyl, C,_₆ alkoxy,

C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,_₆ alkylamino, di-[(C,.₆)alkyl]amino, C₂.₆alkanoylamino

N-C_|.6alkylcarbamoyl, C,.₆alkoxylcarbonyl, ^J,N-di-[(C,.₆alkyl]carbamoyl, C_MalkoxylC,.

₆alkyl, (CH₂),OH where r is 1 or 2, -CO₂R⁷ and CONR⁷R⁸ where R⁷ and R⁸ are independently hydrogen or C,_₆ alkyl, or D and A can be taken together to form a 5 to 7 membered ring, optionally substituted with up to 3 substituents independently selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₆alkenyl, C,.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,_₆ alkyl]amino and C₂.₆alkanoylamino; E is phenyl or a monocyclic heterocycle both optionally substituted with up to 3 substituents selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl, C₂.₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C|.₆alkyl]amino, C₂.₆alkanoylamino, C₃.₆ alkenoyl- amino, C₃.₆alkynoylamino, C,.₆alkoxycarbonyl, N-C,.₆alkylcarbamoyl, N,N-di-[C,.₆ alkyl]carbamoyl, C,.₆alkanoyl, C_MalkoxylC,.₆alkyl, (CH₂)_sOH where s is 1 or 2, -CO₂R⁹ and CONR⁹R¹⁰ where R⁹ and R¹⁰ are independently hydrogen or C,_₆ alkyl or E and D can be taken together to form a bicyclic ring system, optionally substituted with up to 3 substituents selected from halogeno, hydroxy, amino, nitro, trifluoromethyl, trifluoromethoxy, cyano, C,.₆alkyl, C₂.₅alkenyl, C_2. ₆alkynyl, C,.₆alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆alkylamino, di-[C,.₆ alkyljamino and C₂.₆alkanoy lamino ;

F is selected from oxygen, sulphur, amino or CR"R¹²; p and q are each independently 0 or 1 ; R" and R¹² are each independently selected from hydrogen, halogeno, hydroxy, amino, nitro, phenyl, trifluoromethyl, trifluoromethoxy, cyano, carboxy, carbamoyl, C,.₆alkyl, C₂.₆alkenyl, C,.₆alkanoyl, C₂.₆alkynyl, C,.₆ alkoxy, C₂.₆alkenyloxy, C₂.₆alkynyloxy, C,.₆ alkylamino, di-[(C|.₆)alkyl]amino, C₂.₆alkanoylamino, N-C,.₆alkylcarbamoyl, C^alkoxylcarbonyl, N,N-di-[(C_μ6alkyl]carbamoyl, C_l.₄alkoxylC,.₆alkyl, (CH₂)_uOH where u is 1 or 2, -CO₂R'⁵ and CNNR¹⁵R¹⁶ where R¹⁵ and R¹⁶ are independently hydrogen or C,.₆ alkyl with the proviso that not more than one ring system can be formed from taking together two groups from R⁴, A, D and E; or a pharmaceutically acceptable salt or in-vivo hydrolysable derivative thereof.

6. A compound according to claim 5 wherein E is phenyl or a monocyclic heterocycle.

7. A pharmaceutical composition which comprises a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof and a pharmaceutically acceptable carrier.

8. A compound according to any one of claims 1 to 6 for use in a method of therapeutic treatment of the human or animal body.

9. The use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof in the production of a medicament for use in the treatment of a disease or medical condition mediated by the interaction between fibronectin and/or VCAM-1 (especially VCAM-1) and the integrin receptor α₄β,

10. A process for preparing a compound of formula (I), a pharmaceutically acceptable salt or an in vivo hydrolysable derivative thereof which method comprises either

(a) reacting a compound of formula (IV)

where A', R', R², R³, R⁴, m and n are as defined in relation to formula (I), with a compound of formula (V)

where B and Y' are as defined in relation to formula (I), Z is a leaving group, such as halo, and R'⁷ is a group R⁵ as defined in relation to formula (I) or a protected form thereof: or

(b) reacting a compound of formula (VI)

where A', B, Y' R³, R⁴ and n are as defined in relation to formula (I) and R¹⁷ is as defined in relation to formula (V) with a compound of formula (VII)

where R¹ and m are as defined in relation to formula (I); and thereafter if desired or necessary, i) removing any protecting groups; and. ii) optionally forming a pharmaceutically acceptable salt or in vivo hydrolysable derivative.