WO1994025435A1 - Peptidyl derivatives and their use as metalloproteinase inhibitors - Google Patents

Abstract

Compounds of formula (1), wherein R represents a -CONHOR6 [where R6 is a hydrogen atom or an acyl group], carboxyl (-CO¿2?H), esterified carboxyl, -SR?6¿ or -P(O)(X?1R7)-X2R8¿ group, where X?1 and X2¿, which may be the same or different, is each an oxygen or sulphur atom and R?7 and R8¿, which may be the same or different each represents a hydrogen atom or an optionally substituted alkyl, aryl, or aralkyl group; R1 represents a hydrogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group; R2 represents an optionally substituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, amino (-NH¿2?), substituted amino, carboxyl (-CO2H), or esterified carboxyl group; R?3¿ represents a hydrogen atom or an alkyl group; R4 represents a hydrogen atom or an alkyl group; R5 represents a group -C(R?9)(R10)Het-R11¿, wherein R?9 and R10¿ which may be the same or different is each an optionally substituted alkyl or alkenyl group optionally interrupted by one or more -O- or -S- atoms or -N(R12)- groups (where R12 is a hydrogen atom or an optionally substituted alkyl group), or an optionally substituted cycloalkyl, cycloalkenyl, aryl or heteroaryl group, or R?9 and R10¿ together with the carbon atom to which they are attached are linked together to form an optionally substituted cycloalkyl or cycloalkenyl group, Het is -O-, -S(O)¿p?- [where p is zero, or an integer 1 or 2] or -N(R?12¿)-, and R11 is a hydrogen atom or an aliphatic, cycloaliphatic, heterocycloaliphatic, aromatic, or heteroaromatic group; X is an amino (-NH¿2?), substituted amino, hydroxyl or substituted hydroxyl group, or is linked to the atom or group Het in R?5¿ to form a chain -X-Alk-R5- where X is -N(R12)-, Alk is an optionally substituted alkylene chain and R5 is -Het-C(R9)(R10)-; and the salts, solvates, hydrates and prodrugs thereof. The compounds are orally active metalloproteinase inhibitors, with a good duration of action and may be of use in the prophylaxis or treatment of diseases or disorders in which stromelysin, collagenase and gelatinase have a role, for example in the treatment of cancer to control the development of tumor metastases.

Description

PEPTIDYL DERIVATIVES AND THEIR USE AS METALLOPROTEINASE INHIBITORS

FIELD OF THE INVENTION

This invention relates to a novel class of peptidyl derivatives, to processes for their preparation and to their use in medicine.

BACKGROUND TO THE INVENTION In normal tissues, cellular connective tissue synthesis is offset by extracellular matrix degradation, the two opposing effects existing in dynamic equilibrium. Degradation of the matrix is brought about by the action of proteinases released from resident connective tissue cells and invading inflammatory cells, and is due, in part, to the activity of at least three groups of metalloproteinases. These are the collagenases, the gelatinases (or type-IV collagenases) and the stromelysins. Normally these catabolic enzymes are tightly regulated at the level of their synthesis and secretion and also at the level of their extracellular activity, the latter through the action of specific inhibitors, such as α - macroglobulins and TIMP (tissue inhibitor of metalloproteinase), which form inactive complexes with metalloproteinases.

The accelerated, uncontrolled breakdown of connective tissues by metalloproteinase catalysed resorption of the extracellular matrix is a feature of many pathological conditions, such as rheumatoid arthritis, corneal, epidermal or gastric ulceration; tumour metastasis or invasion; periodontal disease and bone disease. It can be expected that the pathogenesis of such diseases is likely to be modified in a beneficial manner by the administration of metalloproteinase inhibitors and numerous compounds have been suggested for this purpose [for a general review see Wahl, R.C. si a! Ann. Rep. Med. Chem. 25, 175-184, Academic Press Inc., San Diego (1990)].

Although numerous metalloproteinase inhibitors have been described, many have not been suitable for further development as medicines since they have lacked any useful activity when administered orally at pharmaceutically acceptable doses. What is therefore needed is a potent orally active compound with a good duration of action.

SUMMARY OF THE INVENTION We have now found a new class of peptidyl derivatives, members of which are metalloproteinase inhibitors. The compounds according to the invention have surprisingly good oral bioavailability, and after oral administration have an advantageously longer duration of action than related known compounds, such as those described in International Patent Specification No. WO92/09564.

Thus according to one aspect of the invention we provide a compound of formula (1)

wherein R represents a -CONHOR⁶ [where R^β is a hydrogen atom or an acyl group], carboxyl (-CO2H), esterified carboxyl, -SR^β or -P(0)(X¹R⁷)- -X²R⁸ group, where X¹ and X², which may be the same or different, is each an oxygen or sulphur atom and R⁷ and R⁸, which may be the same or different each represents a hydrogen atom or an optionally substitued alkyl, aryl, or aralkyl group;

R¹ represents a hydrogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group;

Ft² represents an optionally substituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, amino (-NH2), substituted amino, carboxyl (-CO2H), or esterified carboxyl group;

R³ represents a hydrogen atom or an alkyl group;

R⁴ represents a hydrogen atom or an alkyl group; R5 represents a group -C(R⁹)(R¹⁰)Het-R¹¹, wherein R⁹ and R¹⁰ which may be the same or different is each an optionally substituted alkyl or alkenyl group optionally interrupted by one or more -O- or -S- atoms or -N(R¹²)- groups (where R¹² is a hydrogen atom or an optionally substituted alkyl group), or an optionally substituted cycloalkyl, cyclo¬ alkenyl, aryl or heteroaryl group, or R⁹ and R¹⁰ together with the carbon atom to which they are attached are linked together to form an optionally substituted cycloalkyl or cycloalkenyl group, Het is -0-, -S(0)_p- [where p is zero, or an integer 1 or 2] or -N(R¹²)-, and R¹¹ is a hydrogen atom or an aliphatic, cycloaliphatic, heterocycloaliphatic, aromatic, or hetero- aromatic group;

X is an amino (-NH₂), substituted amino, hydroxyl or substituted hydroxyl group, or is linked to the atom or group Het in R⁵ to form a chain -X-Alk- R⁵- where X is -N(R¹²)-, Alk is an optionally substituted alkylene chain and R5 is -Het-C(R⁹)(R¹⁰)-;

and the salts, solvates, hydrates and prodrugs thereof.

It will be appreciated that the compounds according to the invention can contain one or more asymmetrically substituted carbon atoms, for example those marked with an asterisk in formula (1). The presence of one or more of these aysmmetric centres in a compound of formula (1) can give rise to stereoisomers, and in each case the invention is to be understood to extend to all such stereoisomers, including enantiomers and diastereoisomers, and mixtures, including racemic mixtures, thereof.

In the formulae herein, the -line is used at a potential asymmetric centre to represent the possibility of R- and S- configurations, the — m line and the line to represent an unique configuration at an asymmetric centre.

In the compounds according to the invention, when the group R represents an esterified carboxyl group, it may be for example a group of formula -C0₂R¹³ where R¹³ is a straight or branched, optionally substituted C-_|.₈alkyl group such as a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl or t-butyl group; a C₆.₁₂arylC₁.₈alkyl group suc as an optionally substituted benzyl, phenylethyl, phenylpropyl, α-naphthylmethyl or β-naphthylmethyl group; a C₆.ι₂aryl group such as an optionally substituted phenyi, α-naphthyl or β-naphthyl group; a C₆.₁₂aryloxyCι.₈alkyl group such as an optionally substituted phenyl- oxymethyl, phenyloxyethyl, α-naphthyloxymethyl or β-naphthyloxy- ^' methyl group; an optionally substituted C₁.₈alkanoyloxyC₁.₈alkyl group, such as a pivaloyloxymethyl, propionyloxyethyl or propionyloxypropyl group; or a C₆.₁₂aroyloxyCι.₈alkyl group such as an optionally substituted benzoyloxyethyl or benzoyloxypropyl group. Optional substituents present on the groups R¹³ include for example one or more halogen atoms such as fluorine, chlorine, bromine or iodine atoms, or C-^alkyl, e.g. methyl or ethyl, or C-^alkoxy, e.g. methoxy or ethoxy, groups.

In general, when the group R represents an esterified carboxyl group, it may be a metabolically labile ester of a carboxylic acid.

When the group R⁶ in compounds of formula (1) represents an acyl group, it may be for example a group of formula R¹⁴C=X³ where X³ is an oxygen or sulphur atom and R¹⁴ represents a hydrogen atom or a group selected from amino (-NH₂), substituted amino (for example a group -NR¹⁷R¹⁸ as described below in relation to the group X), or an optionally substituted C-|-₆alkyl, C₁-_€alkoxy,C₁.₆alkyth.o, C .₆alkenyl, C^alkynyl, C_θ.₁₂aryl, C₆_ι₂aralkyl, C₃.₆cycloalkyl, C₃.₆heteroaryl or C₃.₆hetero- aralkyl group. Particular groups of these types include optionally substituted methyl, ethyl, n-propyl, i-propyl, methoxy, ethoxy, methylthio, ethylthio, ethenyl, 1-propenyl, ethynyl, 1-propynyl, phenyi, 1-naphthyl, 2-naphthyl, benzyl, phenethyl, 1-naphthylmethyl, 2-naphthylmethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, furanyl, pyrrolyl, thienyl, furanylmethyl, pyrrolylmethyl or thienylmethyl groups. Optional substituents which may be present on such R¹⁴ groups include one or more substituents selected from those described below in relation to the group R¹ or R² when such groups represent substitued alkyl, aryl or heteroaryl groups. The group R⁷ and/or R⁸ in compounds of formula (1) may each be a hydrogen atom or an optionally substituted straight or branched

C-_|.₆alkyl, e.g. methyl, ethyl, n-propyl, i-propyl, n-butyl or i-butyl,

C₆-i2aryl, e.g. phenyi, or C₆-i2arylCι-₆alkyl, e.g. benzyl, phenylethyl or phenylpropyl group. Optional substituents present on alkyl groups of this type include one or more

e.g. methoxy or ethoxy, or

' Ci-βalkylthio, e.g. ^• methylthio or ethylthio groups or an optionally substituted C₆-i2aryloxy e.g. phenyloxy, C6-12ary.tl.io e.g. phenylthio,

C₆-i2arylC-|-₆alkoxy e.g. benzyloxy or C₆-12arylC1.6alkyltl.io e.g. benzylthio. Optional substituents present on the group R⁷ or R⁸ when it is an aryl or aralkyl group or an alkyl group substituted by an aryloxy or arylthio group include R¹⁶ groups as defined below.

When the group R¹ in compounds of formula (1) represents an optionally substituted alkyl or alkenyl group, it may be, for example, a straight or branched C-_|-₆ alkyl or C₂^alkenyl group, such as a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, i-pentyl, n-hexyl, ethenyl, 1-propenyl, 1-butenyl or 2-butenyl group optionally substituted by one or more C^alkoxy, e.g. methoxy, ethoxy, propoxy, C^alkylthio, e.g. methylthio, ethylthio, propylthio, C_β.ι₂arylC₁.₆alkoxy, e.g. phenylC₁.₆alkoxy such as benzyloxy, araikylthio, e.g. phenyl-

such as benzylthio, amino (-NH₂), substituted amino, [such as -NHR¹⁵, where R¹⁵ is a

Ci^al yl, e.g. phenylCι.₆alkyl, such as benzyl, C₆.ι₂aryl, e.g. phenyi, C₃.₈cycloalkyl, e.g. cyclohexyl, or C₃.₈cycloalkylCι.₆alkyl, e.g. cyclohexylmethyl group], carboxyl (-C0 H) or -C0₂R¹³ [where R¹³ is as defined above] groups.

Aryl groups represented by R¹ and/or R² in compounds of formula (1) include optionally substituted mono- or bicyclic C₆.-|₂ aryl groups such as phenyi or 1- or 2-naphthyl groups.

Aralkyl groups represented by R¹ include optionally substituted mono- or bicyclic C₆-ι₂aryiC-|^alkyl groups such as phenylC₁_₆alkyl, or 1- or 2- naphthylC-_|.₆alkyl, for example benzyl, phenylethyl, phenylpropyl, phenylbutyl, phenylpentyl, 1- or 2- naphthylmethyl, naphthylethyl, naphthyipropyl, naphthyibutyl or naphthylpentyl groups.

When the group R¹ in compounds of formula (1) is a heteroaralkyl group, it may be for example an optionally substituted mono-or bicyclic C₃-₉heteroarylCi-₆alkyl group, such as an optionally substituted pyrrolylmethyl, furanylmethyl, thienylmethyl, imidazolylmethyl, oxazolylmethyl, thiazolylmethyl, pyrazolylmethyl, pyridinylmethyl, or pyrimidinylmethyl group.

Heteroarylthioalkyl groups represented by R¹ include optionally substituted mono- or bicyclic C₃.gheteroarylthioC₁.₆alkyl groups such as optionally substituted pyrrolylthiomethyl, furanylthiomethyl, oxazolyl- thiomethyl, thiazolylthiomethyl, pyrazolylthiomethyl, pyridinylthiomethyl, or pyrimidinylthiomethyl groups.

When the group R² in compounds of formula (1) represents an alkyl or alkenyl group it may be for example a straight or branched C_halky. , or C2-6alkenyl group, such as a methyl, ethyl, n-propyl, i-propyl, n-butyl, i- butyl, s-butyl, t-butyl, n-pentyl, i-pentyl, n-hexyl, ethenyl, 1-propenyl, 1- butenyl, or 2-butenyl group. Optional substituents which may be present on such groups include one or more C-i-βalkoxy, e.g. methoxy, ethoxy, propoxy, C-i-βalkylthio, e.g. methylthio, ethylthio, propylthio, amino, substituted amino [such as -NHR¹⁵ where R¹⁵ is as defined above], carboxyl or -CO2R¹³ [where R¹³ is as defined above] groups.

Cycloalkyl groups represented by the group R² in compounds according to the invention include C₃.₈cycloalkyl groups such as cyclopentyl or cyclohexyl groups.

When the group R² in compounds of formula (1) is a substituted amino group, this may be for example a group -NHR¹⁵ where R¹⁵ is as defined above.

When R² is a cycloalkylalkyl group it may be for example a C₃.₈cyclo- alkylCi-βaikyl group such as a cyclopentylCi-βalkyl or cyclohexyl-

C-|-6alkyl group, for example a cyclopentylmethyl, cyclopenty (ethyl, cyclopentylpropyl, cyclopentylbutyl, cyclohexylmethyl, cyclohexylethyl, cyclohexyipropyl, or cyclohexylbutyl group.

Optional substituents which may be present on aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl groups represented by R¹ or R² include those R¹⁶ substituents discussed below.

The aryl, aralkyl, heteroaryl, heteroaralkyl or heteroarylthioalkyl groups represented by R¹ and/or R² in compounds of formula (1 ) may each optionally be substituted in the cyclic part of the group by one, two or more substituents [R¹⁶] selected from halogen atoms, e.g. fluorine, chlorine, bromine or iodine atoms, or C^ alkyl, e.g. methyl or ethyl, C -| .6^alkoχy ^θ-9- rnethoxy or ethoxy, C₂-6alkylenedioxy, e.g. ethylenedioxy, haloC-_|.₆alkyl, e.g. trifluoromethyl, Cι.₆alkylamino, e.g. methylamino or ethylamino, Ct.5dialkylan-.in0, e.g. dimethylamino or diethylamino, amino (-NH₂), nitro, cyano, hydroxyl (-OH), carboxyl (-C0₂H), -C0₂R¹³, where R¹³ is as defined above, C-|._ealkylcarbonyl, e.g. acetyl, sulphonyl (-SO₃H), C-,.₆alkylsulphonyl, e.g. methylsulphonyl, aminosulphonyl (-S0₂NH₂), Cι.₆ alkylaminosulphonyl, e.g. methyl- aminosulphonyl or ethylaminosulphonyl, Cι.₆dialkylamino-sulphonyl e.g. dimethyiaminosulphonyl or diethylaminosuiphonyl, carboxamido (-CONH₂), C-j.ealkylaminocarbonyl, e.g. methylaminocarbonyl or ethylaminocarbonyl, C-).₆dialkylaminocarbonyl, e.g. dimethyiamino- carbonyl or diethylaminocarbonyl, sulphonylamino (-NHS0 H), C-_|^alkylsulphonylamino, e.g. methylsulphonylamino or ethylsulphonyl- amino, or C^dialkylsulphonylamino, e.g. dimethylsulphonylamino or diethylsulphonylamino groups. It will be appreciated that where two or more R¹⁶ substituents are present, these need not necessarily be the same atoms and/or groups. The R¹⁶ substituents may be present at any ring carbon atom away from that attached to the rest of the molecule of formula (1). Thus, for example, in phenyi groups any substituents may be present at the 2-, 3-, 4-, 5- or 6- positions relative to the ring carbon atom attached to the remainder of the molecule.

When the groups R³ and R⁴ in compounds of formula (1) are alkyl groups, they may be for example straight or branched C-_|.₆alkyl groups such as methyl or ethyl groups.

When the group R⁹ or R¹⁰ in compounds of formula (1) is an optionally substituted alkyl or alkenyl group it may be a straight or branched

e.g. methyl, ethyl, n-propyl i-propyl, n-butyl, i-butyl, n-pentyl or n-hexyl or C₂.₆alkenyl e.g. ethenyl or 1-propenyl group optionally interrupted by one or more -O- or -S- atoms or -N(R¹²)- groups where R¹² is a hydrogen atom or an optionally substituted Cι.₆alkyl group such as a methyl, ethyl or propyl group.

Optional substituents which may be present on such groups include one or more C-i-βalkoxy, C-i-βalkylthio,

aralkylthio, amino, substituted amino, carboxyl, -CO2R¹³, aryl or heteroaryl groups as defined above in connection with the group R¹, or an optionally substituted cycloalkyl or cycloalkenyl group as defined below in connection with the groups R⁹ and R¹⁰.

When the group R⁹, R¹⁰ or R⁹ and R¹⁰ together with the carbon atom to which they are attached, is an optionally substituted cycloalkyl or cycloalkenyl group, it may be for example a C₃.₈cycloalkyl, e.g. cyclopropyl, cyclopentyl or cyclohexyl, or C₃.₈cycloaikenyl e.g. cyclo- propenyl, cyclopentenyl or cyclohexenyl, group optionally substituted by one, two or more C_halky!, e.g. methyl or ethyl,

e.g. methoxy or ethoxy, C-^alkylthio, e.g. methylthio, or hydroxyl groups.

The term Het in compounds of formula (1) may represent -0-, -S-, -S(O)-, -S(0)2- or -N(R¹²)- where R¹² is a hydrogen atom or a C βalkyl group as defined above.

When R¹¹ in compounds of formula (1) is an aliphatic group it may be for example an optionally substituted saturated or unsaturated straight or branched C_halky I chain optionally interrupted by one or more -O- or -S- atoms or groups selected from -N(R1²)-, -CO-, -CON(R¹²)-, or -N(R¹²)CO-. Particular groups include optionally substituted methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, 1-propenyl, 1 -butenyl or 2-butenyl groups. Optional substituents which may be present on groups of these types include one or more amino (-NH2), substituted amino [for example a group -NR¹⁷R¹⁸ as described below in relation to the group X], C₆-ι₂aryl, e.g. optionally substituted phenyi,

e.g. optionally substituted phenoxy, [the optional substituents in each case being R¹⁶ groups as defined above] C₃.₈cycloalkyl, e.g. cyclopentyl or cyclohexyl, C₃.₈cycloalkoxy, e.g. cyclopentyloxy or cyclohexyloxy, carboxyl (-CO2H) or -CO2R¹³ groups.

Cycloaliphatic groups represented by R¹¹ in compounds of formula (1) include optionally substituted C₃.₈cycloalkyl and C₃.₈cycloalkenyl groups, for example optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl and cyclohexenyl groups. Optional substituents include those groups R¹⁶ described above.

Heterocycloaliphatic groups represented by R¹¹ in the compounds of formula (1) include optionally substituted Cs.7heterocycloalkyl groups containing one or two heteroatoms selected from -O- or -S-, or a group -N(R¹²)-, for example optionally substituted piperazinyl, morpholinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, or N- methylpipendinyl groups. Optional substituents include those groups R16 described above. The heterocycloalkyl groups represented by R¹¹ may be attached to the remainder of the molecule through any ring carbon atom.

When the group R¹¹ in compounds of formula (1) is an aromatic group it may be for example an optionally substituted mono- or bicyclic C_-i2aryl group, for example an optionally substituted phenyi or 1- or 2-naphthyl group. Optional substituents which may be present on groups of this type include those R¹⁶ substituents described above.

Heteroaromatic groups represented by the group R¹¹ include mono-or bicyclic Cs-gheteroaromatic groups containing one, two or three heteroatoms selected from -O- or -S-, or -N(R¹²) groups. Particular examples include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, 1-indolyl, 2-indolyl, 1-quinolinyl or 2-quinolinyl groups. Such groups may be optionally substituted, for example by one or more R¹⁶ substituents. The heteroaromatic group may be connected to the remainder of the compound of formula (1) through any ring carbon atom, or where appropriate through a heteroatom or group -N(R¹²)-.

When X in the compounds of formula (1) represents a substituted amino group it may be for example a group of formula -NR¹⁷R¹⁸, where R¹⁷ and R¹⁸, which may be the same or different, is each a hydrogen atom (with the proviso that when one of R¹⁷ or R¹⁸ is a hydrogen atom, the other is not) or an optionally substituted straight or branched alkyl group, optionally interrupted by one or more -O- or -S- atoms or -N(R¹²)- or aminocarbonyloxy [-NHC(0)0-] groups or R¹⁷ and R¹⁸, together with the nitrogen atom to which they are attached, may form an optionally substituted C₃.₆cyclic amino group optionally possessing one or more other heteroatoms selected from -O- or -S-, or -N(R¹²)- groups.

When R¹⁷ and/or R¹⁸ is an alkyl group it may be for example a Chalky! group such as a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, or t-butyl group, optionally interrupted by one or more -0- or -S- atoms, or -N(R¹³)- or aminocarbonyloxy groups and may be for example a methoxymethyl, ethoxymethyl, ethoxymethyl, ethoxyethyl or et y lam i no- carbon y I oxy methyl group. The optional substituents which may be present on such groups include hydroxyl (-OH), carboxyl (-C0₂H), esterified carboxyl (-C0₂R¹³), carboxamido (-CONH₂), substituted carboxamido, e.g. a group -CONR¹⁷R¹⁸ where NR¹⁷R¹⁸ is as defined herein, amino (-NH₂), substituted amino, for example a group of formula -NR¹⁷R¹⁸, aminosulphonylamino, for example -N(R¹²)S0₂NH₂ or -N(R¹²)Sθ2NR¹⁷R¹⁸ or aryl, e.g. C₆.ι₂ aryl such as phenyi, optionally substituted by one, two or more R¹⁶ substituents selected from those listed above.

Particular examples of cyclic amino groups represented by -NR¹⁷R¹⁸ include morpholinyl, imidazolyl, piperazinyl, pyrrolyl, oxazolyl, thiazolyl, pyrazolyl, pyrrolidinyl, pyridinyl and pyrimidinyl groups.

When the group X is a substituted hydroxyl group it may be for example a group -OR¹¹ where R¹¹ is as defined above, other than a hydrogen atom.

When X is linked to the atom or group Het in R⁵ to form a chain -X-Alk- R⁵, the optionally substituted alkylene chain represented by Alk may be an optionally substituted straight or branched C₂.g alkylene chain, for example an ethylene, propylene or butylene chain. Optional substituents present on the alkylene chain include those described above in relation to the alkyl group represented by R². In compounds of this type, the group X is -N(R¹²)-, where R¹² is as defined above. The group R5 is -Het-C(R⁹)(R¹⁰)- where Het, R⁹ and R are as defined above.

Salts of compounds of formula (1) include pharmaceutically acceptable salts, for example acid addition salts derived from inorganic or organic acids, such as hydroch rides, hydrobromides, hydroiodides, p-toluene sulphonates, phosphates, sulphates, perchlorates, acetates, trifluoro- acetates, propionates, citrates, mabnates, succinates, iactates, oxalates, tartarates and benzoates.

Salts may also be formed with bases. Such salts include salts derived from inorganic or organic bases, for example alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as magnesium or calcium salts, and organic amine salts such as moφholine, piperidine, dimethylamine or diethyl amine salts.

Prodrugs of compounds of formula (1) include those compounds, for example esters, alcohols or amines, which are convertible, in vivo, by metabolic means, e.g. by hydrolysis, reduction, oxidation or transesterification, to compounds of formula (1).

When the group R in compounds of the invention is an esterified carboxyl group, it may be a metabolically labile ester of formula -C0₂R¹³ where R¹³ may be an ethyl, benzyl, phenylethyl, phenylpropyl, 1- or 2- naphthyl, 2,4-dimethylphenyl, 4-t-butylphenyl, 2,2,2-trifluoroethyl, 1- (benzyloxy)benzyl, 1-(benzyloxy)ethyl, 2-methyl-1-propionyloxypropyl, 2,4,6-trimθthylbenzoyloxymethyl or pivaloyloxymethyl group. When the group R in compounds of formula (1) is a -P(0)(X¹R⁷)X²R⁸ group it may in particular be a P(0)(OR⁷)OR⁸, e.g. a -P(0)(OH)OR⁸ group, or a -P(0)(SH)OR⁸ or -P(0)(OH)SR⁸ group. Examples of such groups include -P(0)(OCH₃)OCH₃, -P(0)(OCH₂CH₃)OCH₂CH₃, -P(0)(OH)OH, -P(0)(OH)SH, -P(0)(SH)OH, -P(0)(OH)OCH₃, -P(0)(OH)SCH₃, -P(0)(OH)OCH₂CH₃, -P(0)(OH)OPh, -P(0)(OH)SPh, -P(0)(OH)OCH₂Ph or -P(0)(OH)SCH₂Ph, where Ph is a phenyi group optionally substitued by one or more substituents R¹⁶.

In the compounds of formula (1) the group R¹ may in particular be a C-,.₆alkyl group such as a methyl group, an aralkyl group such as benzyl group, an arylthioalkyl group such as a phenythiomethyl group or a heteroarylthioalkyl group such as thienylthiomethyl, pyridinyl- thiomethyl or pyrimidinylthiomethyl group or is especially a hydrogen atom.

The group R² in compounds of formula (1) may be in particular an optionally substituted d-βalkyl, C₃.8cycloalkyl, or Cβ-i2 ryl group. Particular types of these groups are optionally substituted C₃.₆alkyl, such as n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl or i- pentyl; cyclopentyl; cyclohexyl; phenyi; 1- or 2-naphthyl. Each of these cycloalkyl or aryl groups may be substituted, by one, two or more substituents R¹⁶ described above.

The groups R³ and R⁴ in compounds of formula (1) may each in particular be a methyl group, or, especially, a hydrogen atom.

The group R⁵ in compounds of formula (1) may in particular be a group -C(R⁹)(R¹⁰)Het-R¹¹ where R⁹ and R¹⁰ are the same. Particular compounds of this type are those wherein R⁹ and R¹⁰ is each the same and is each an optionally substituted alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heteroaryl group.

In another group of compounds of formula (1) the group R⁵ may be a group -C(R⁹)(R¹⁰)Het-R^{1 1} where R¹¹ is an aliphatic, cycloaliphatic, heterocycioaiiphatic, aromatic or heteroaromatic group as described above for compounds of formula (1).

The group X in compounds of formula (1) may be in particular an amino (-NH₂) or -NR ⁷R¹⁸ group. Particular -NR¹ R¹⁸ groups are -NHR^{1 8} groups. Groups of this type include those where R¹⁸ is a C-|.₆alkyl group, for example a methyl, ethyl, or n-propyl group, optionally interrupted by one or more -O- or -S- atoms or -N(R¹²) [e.g. -NH- or -N(CH₃)-] or aminocarbonyloxy groups and optionally substituted by a hydroxyl, carboxyl, carboxyalkyl, e.g. carboxymethyl, carboxamido, amino, -NR^{1 7}R ^{1 8}, [for example di-C-j .salkylamino such as dimethylamino, C-_|.₆alkylamino such as methylamino, or C₃.₆ cyclic amino such as morpholinyl, pyrrolidinyl or pyridinyl] or phenyi optionally substituted by one, two or more R¹⁶ substituents.

A particularly useful group of compounds according to the invention is that of formula (1) wherein R⁵ is a group -C(R⁹)(R¹⁰)Het-R¹¹ where Het is -S(0)p and R⁹, R¹⁰ and R^{1 1} are as defined for formula (1). Compounds of this type wherein Het is -S- are particularly useful.

A further particularly useful group of compounds of formula (1) are those wherein X is an amino or substituted amino group. Particularly useful compounds of this type are those wherein X is -NHCH₃ or, especially, -NH₂-.

In general, in compounds of formula (1) the groups R¹, R³ and R⁴ is each preferably a hydrogen atom.

In a further preference, the group R in compounds according to the invention is a -CONHOH or a -C0₂H group or a metabolically labile ester thereof, or a group P(0)(OH)OR⁸. In a particular preference, however, R is a -C0₂H, -P(0)(OH)2 or, especially, a -CONHOH group.

An especially useful group of compounds according to the invention has the formula (1a)

wherein R, R², R⁵ and X are as defined for formula (1 ); and the salts, ^' solvates, hydrates and prodrugs thereof.

A particularly useful group of compounds of formula (1a) are those wherein R represents a -CONHOH, -C0₂H or -P(0)(OH)2 group; R² and R⁵ are as defined for formula (1 ); X is an amino (-NH₂) or substituted amino group; and the salts, solvates, hydrates and prodrugs thereof.

Particularly useful compounds of formula (1a) are those wherein R⁵ is a group -C(R⁹)(R¹⁰)S(O)pR¹¹. Compounds of this type in which R⁵ is a -C(R⁹)(R¹⁰)SR¹¹ group are especially useful.

Other useful compounds of formula (1a) include those wherein R² represents a C₃.₆alkyl group, particularly an isobutyl or n-pentyl group, or a cycloalkylC₃-₆alkyl group, particularly a cyclohexylpropyl, cyclo- hexylbutyl or cyclohexylpentyl group.

In the compounds of formula (1a) X may be a -NH₂ group or a group -NR¹ R¹⁸ as defined for compounds of formula (1), particularly a -NHR¹⁸ group.

An especially useful group of compounds according to the invention has the formula (1a) wherein R² is a C₃-6alkyl group, R⁵ is a group -C(R⁹)(R¹⁰)SR¹¹ where R⁹ and R¹⁰ is each the same and is each an optionally substituted C-i-e alkyl group, and R ¹ is as defined for formula (1); and X is an amino (-NH2) or -NHR¹⁸ group, particularly where R¹⁸ is an optionally substituted C1-₆ alkyl group. Compounds of this type wherein R⁵ is a group -C(CH₃) SR¹¹ are particularly useful, especially where the group R¹¹ is a hydrogen atom or an optionally substituted saturated C-ι-₆ alkyl chain. In compounds of this last type X is preferably an amino (-NH2) group or a -NHCH₃ group. In a still further useful group of compounds of formula (1a), R is a -CONHOH, -C0₂H or -P(0)(OH)₂ group, R² is an isobutyl group, R⁵ is a group -C(CH₃)₂SR¹¹ where R¹¹ is a hydrogen atom or an optionally substitued C-i-β alkyl group, and X is an amino (-NH₂) or -NHR¹⁸ group where R¹⁸ is an optionally substituted C-ι-₆ alkyl group. Compounds of this type wherein R is a -CONHOH group are particularly useful; as are those compounds wherein R¹¹ is a hydrogen atom or a methyl group; and those compounds wherein R¹⁸ is a hydrogen atom or a methyl group.

One further group of compounds according to the invention has the formula (1a) wherein R and R² are as defined for formula (1), R⁵ is a group -C(CH₃)₂SH or -C(CH₃)₂SCH₃ and X is -NH₂ or -NHCH₃. Particularly useful compounds of this type are those wherein R is a group -CONHOH, -C0₂H or -P(0)(OH)2 and R² is a C₃-6alkyl group. Particularly useful compounds are those where R² is an isobutyl group.

in compounds of the above described types, R⁵ is preferably a group

-C(CH3)2SCH₃. In these compounds, R is preferably -CO2H or -P(0)(OH₂) or is especially -CONHOH. X is preferably -NH₂ or -NHCH3.

An important compound according to the invention is:

[4-(N-Hydroxyamino)-2(R)-3-(2-methylpropyl)succinyl]-L-[S-(methyl) penicillamine] N-methylamide; and the salts, solvates, hydrates and prodrugs thereof.

The compounds according to the invention may be prepared by the following general processes, more specifically described in the Examples hereinafter. In the description and formulae below the groups R, R¹ , R², R³, R⁴, R⁵ and X are as defined above, except where otherwise indicated. It will be appreciated that functional groups, such as amino, hydroxyl or carboxyl groups, present in the various compounds described below, and which it is desired to retain, may need to be in protected form before any reaction is initiated. In such instances, removal of the protecting group may be the final step in a particular reaction. Suitable amino or hydroxyl protecting groups include benzyl, benzyloxycarbonyl or t-butyloxycarbonyl groups. These may be removed from a protected derivative by catalytic hydrogenation using for example hydrogen in the presence of a metal catalyst, for example palladium on a support such as carbon in a solvent such as an alcohol e.g. methanol, or by treatment with trimethylsilyl iodide or trifluoroacetic acid in an aqueous solvent. Suitable carboxyl protecting groups include benzyl groups, which may be removed from a protected derivative by the methods just discussed, or alkyl groups, such as a t- butyl group which may be removed from a protected derivative by treatment with trifluoroacetic acid in an aqueous solvent. Other suitable protecting groups and methods for their use will be readily apparent. The formation of the protected amino, hydroxyl or carboxyl group may be achieved using standard alkylation or esterification procedures, for example as described below.

Thus according to a further aspect of the invention a compound of formula (1) may be prepared by coupling an acid of formula (2)

or an active derivative thereof, with an amine of formula (3)

followed by removal of any protecting groups.

Active derivatives of acids for formula (2) include for example acid anhydrides, or acid halides, such as acid chlorides.

The coupling reaction may be performed using standard conditions for amination reactions of this type. Thus, for example the reaction may be achieved in a solvent, for example an inert organic solvent such as an ether, e.g. a cyclic ether such as tetrahydrofuran, an amide e.g. a substituted amide such as d im ethy If orm amide, or a halogenated hydrocarbon such as dichloromethane at a low temperature, e.g. -30°C to ambient temperature, such as -20°C to 0°C, optionally in the presence of a base, e.g. an organic base such as an amine, e.g. triethylamine or a cyclic amine such as N-methylmorpholine. Where an acid of formula (2) is used, the reaction may additionally be performed in the presence of a condensing agent, for example a diimide such as N.N'-dicyclohexylcarbodiimide, or 1 -(3-dimethylaminopropyl)-3-ethyl- carbodiimide, advantageously in the presence of a triazole such as 2-hydroxybenzotriazole. Alternatively, the acid may be reacted with a chloroformate for example ethylchloroformate, prior to reaction with the amine of formula (3).

Free hydroxyl or carboxyl groups in the starting materials of formulae (2) [where R is -CONHOH or C0₂H] and (3) may need to be protected during the coupling reaction. Suitable protecting groups and methods for their removal may be those mentioned above. Where R in the intermediates of formula (2) is a -P(0)(X¹R⁷)X²R⁸ group, at least one of R⁷ or R⁸ is other than a hydrogen atom. Conveniently, each of R⁷ and R⁸ is an optionally substituted alkyl, aryl or aralkyl group. Such groups, when present in compounds of the invention may be cleaved as described below to yield other compounds of the invention wherein R⁷ and/or R⁸ is each a hydrogen atom.

It will be appreciated that where a particular stereoisomer of formula (1) is required, this may be obtained by resolution of a mixture of isomers following the coupling reaction of an acid of formula (2) and an amine of formula (3). Conventional resolution techniques may be used, for example separation of isomers by chromatography e.g. by use of high performance liquid chrormatography. Where desired, however, appropriate homochiral starting materials may be used in the coupling reaction to yield a particular stereoisomer of formula (1). Thus, in particular process a compound of formula (1a) may be prepared by reaction of a compound of formula (2a)

(2a)

with an amine of formula (3a)

as described above

Intermediate acids of formula (2) wherein R is a carboxyl or esterified carboxyl group or a group -P(0)(X¹R⁷)X²R⁸ or -SR^β may be prepared from a corresponding ester of formula (4)

where R¹⁹ is an alkyl group, for example a methyl or t-butyl group, using for example trifluoroacetic acid, or, when R¹⁹ is an aralkyl group, such as a benzyl group, by hydrogenolysis, for example by reaction with hydrogen in the presence of a metal catalyst, e.g. palladium, on a support such as carbon in a solvent such as an alcohol, e.g. methanol optionally at an elevated pressure and temperature.

An ester of formula (4) where R is a carboxyl or esterified carboxyl group may be prepared by esterification of the corresponding acid of formula (5)

(5) using an appropriate acyl halide, for example an acyl chloride in a solvent such as an alcohol, e.g. methanol at a low temperature, e.g. around 0°C.

Acids of formula (5) may be prepared by alkylation of a compound of formula (6)

with an appropriate halide, e.g. a compound R²Hal, where Hal is a halogen atom such as a chlorine or bromine atom in the presence of a base, for example an alkoxide such as sodium ethoxide in a solvent such as an alcohol, e.g. ethanol at ambient temperature, followed by decarboxylation using for example concentrated hydrochloric acid at an elevated temperature.e.g. the reflux temperature.

Intermediates of formula (6) are either known compounds or may be prepared by methods analogous to those used for the preparation of the known compounds.

Intermediate esters of formula (4) where R is a -P(0)(X¹R⁷)X²R⁸ group may be prepared by reaction of an acrylate R¹CHC(R²)COR¹⁹ with a phosphite:P(OR²⁰)(X¹R )X R⁸ [where R ° is a leaving group, for example a silyl group such as a trialkylsilyl group e.g. a trimethylsilyl group] at an elevated temperature.

Acrylates of formula R¹CHC(R²)COR¹⁹ may be prepared by reaction of a mono-ester HOOCCH(R²)COOR¹⁹ with an aldehyde R¹CHO or a polymer thereof e.g. paraformaldehyde or paraldehyde in the presence of a base, for example an organic base such as piperidine. The reaction may be performed in a solvent, such as pyridine, optionally at an elevated temperature.

Mono-esters of formula HOOCCH(R²)COOR¹⁹ may be prepared by hydrolysis of the corresponding di-ester R¹⁹OOCCH(R²)COOR¹⁹ using a base, for example an alkali hydroxide, in an inert solvent such as dioxane at a low temperature e.g. around 0°C. The di-esters for use in this reaction may be prepared by alkylation of the corresponding malonates of formula R¹⁹OOCCH2COOR¹⁹ with a halide R²Hal [where Hal is a halogen atom such as a chlorine or bromine atom] in the presence of a base, e.g. a hydride such as sodium hydride in a solvent such as tetrahydrofuran at ambient temperature. Malonates of formula R¹⁹0OCCH2COOR¹⁹ are either known compounds or may be prepared by methods analogous to those used for the preparation of the known compounds.

Intermediate phosphites of formula P(OR²⁰)(X¹R )X²R⁸ may be prepared by reaction of a phosphite HP(0)(X¹ R⁷)X R⁸ with an appropriate amine (R²⁰)2NH e.g. a silazane, at an elevated temperature, e.g. the reflux temperature. Phosphites of formula HP(0)(X¹R⁷)X²R⁸ are either known compounds or may be prepared by methods analogous to those used for the preparation of the known compounds.

Intermediates of formula (4) where R is a -SR^β group are either known compounds or may be prepared from known starting materials of formula R^βSCH(R¹)CH(C0₂CH₂CH3)₂ by using a similar series of reactions to those just described for the preparation of compounds of formula (4) where R is a carboxyl group.

In another process, intermediate acids of formula (2) wherein R is a -P(0)(X¹R⁷)X²R⁸ group may be prepared by reaction of an acid R CH₂C0₂H with a phosphonate P(0)(X¹R⁷⁾(X R⁸)CH₂OR²¹ where R21 is a leaving group, for example a trifluoromethylsulphonyloxy group in the presence of a base such as n-butyllithium in a solvent such as tetrahydrofuran. Phosphonates for use in this reaction may be prepared from the corresponding compound P(0)(X¹R⁷)(X²R⁸)CH2θH by reaction with paraformaldehyde in the presence of a base such as triethylamine at an elevated temperature followed by reaction with a halide R²¹Hal in the presence of a base such as sodium hydride in a solvent such as an ether. Phosphonates P(0)(X¹R )(X²R⁸)CH₂OH and acids R CH₂C0 H for use in the above reactions are either known compounds or may be prepared by methods analogous to those used for the preparation of the known compounds.

Intermediate acids of formula (2) wherein R is a -CONHOR⁶ group or a protected derivative thereof may be prepared by reaction of an anhydride of formula (7)

with a hydroxylamine such as O-benzylhydroxyiamine or NH2OR⁶ where R⁶ is an acyl group in a solvent such as tetrahydrofuran at a low temperature, e.g. around -20°C, followed where desired by removal of the protecting group as described above.

The intermediate anhydrides of formula (7) may be prepared for example by heating for example at the reflux temperature, a diacid of formula (5) where R is -C0₂H with an acyl chloride such as acetyl chloride.

The homochiral acids of formula (2a) may be prepared according to another feature of the invention by oxidation of an oxazolidinone of formula (8)

(8)

(where Ph is a phenyi group)

using an oxidising agent such as peroxide, e.g. hydrogen peroxide in a solvent such as an ether e.g. a cyclic ether such as tetrahydrofuran, at a low temperature, e.g. around 0°C followed by treatment with a base, such as lithium hydroxide, at an elevated temperature. The compounds of formula (8) may be prepared by reaction of an acyl halide RCH₂CH(R²)COHal (where Hal is a halogen atom such as a chlorine, bromine or iodine atom) with a solution of (S)-4-(phenyl- methyl)-2-oxazolidinone in the presence of a base such as n-butyl¬ lithium in a solvent such as tetrahydrofuran at a low temperature, e.g. around -78°C.

Acyl halides RCH₂CH(R²)COHal may be prepared by treatment of the corresponding known acids RCH₂CH(R²)C0 H with conventional halogenating agents for example thionyl halides under standard reaction conditions.

Intermediates of formula (3) are either known compounds or may be prepared from known amino acid starting materials using standard methods, for example by employing a series of substitution reactions to manipulate the groups R⁵ and X as described in the Examples hereinafter, or for example as described by Wessjohann βt al. Chem. Ber. 1992. 125. 867-882.

Compounds of formula (1) may also be prepared by interconversion of other compounds of formula (1). Thus, for example, a compound of formula (1) wherein R is a -CONHOR⁶ group may be prepared by reaction of a corresponding acid of formula (1) wherein R is a -C0₂H group or an active derivate thereof (for example an acid chloride or an acid anhydride) with hydroxylamine or an O-protected derivative or a salt thereof or a reagent R⁶ONH₂ where R^β is an acyl group. The reaction may be performed using the reagents and conditions described above in the preparation of compounds of formula (1) from the starting materials of formulae (2) and (3).

In another interconversion process, compounds of formula (1) wherein R is -C0₂H and/or X contains a -C0₂H group may be prepared by hydrolysis of the corresponding esterified compounds (for example where R is a -C0₂R¹³group and/or X contains a similar group) using conventional procedures, for example by treatment with a base, e.g. an alkali metal hydroxide such as lithium hydroxide in a solvent such as an aqueous alcohol, e.g. aqueous methanoi, or by treatment with an acid such as a mineral acid, e.g. hydrochloric acid in the presence of a solvent, e.g. dioxane.

, Similarly esters of formula (1), for example where R is a C0 R¹³ group and/or X contains a -C0₂R¹³ group may be prepared by reaction of the corresponding acids, where R is a -C0₂H group and/or X contains a

-C0₂H group or an active derivative thereof, with an alcohol R¹³OH using standard conditions.

In another interconversion process, a compound of formula (1) wherein R5 is a group -C(R⁹)(R¹⁰)S-R¹¹ may be oxidised to a corresponding compound where R⁵ is a group -C(R⁹) (R^{1 0}) S O R ^{1 1} or -C(R⁹)(R^{1 0})SO₂R^{1 1} using an oxidising agent, for example a peroxymonosulphate such as potassium peroxymonosulphate, in a solvent such as an aqueous alcohol at ambient temperature or a peroxyacid in a haiogenated hydrocarbon solvent such as dichloromethane at a low temperature, e.g. around -78°C.

The compounds according to the invention are potent inhibitors of the metalloproteinases collagenase, stromelysin and gelatinase and advantageously have a long duration of action when administered orally. The activity of the compounds may be determined by the use of appropriate enzyme inhibition tests for example as described in Example A hereinafter or by oral administration to mice as described hereinafter in Example B. In our tests using this approach, compounds according to the invention have been shown to inhibit stromelysin, and, in particular, collagenase and gelatinase with Ki values in the nanomolar range.

The compounds according to the invention can be expected to be of use in the prophylaxis or treatment of diseases or disorders in which stromelysin, collagenase and gelatinase have a role. Thus for example the compounds of formula (1) may be of use in the prophylaxis or treatment of musculo-skeletal disorders, for example arthritic diseases such as rheumatoid arthritis, osteoarthritis and septic arthritis, and to be of use to prevent tumour cell metastasis and invasion. The compounds may therefore be of use in the treatment of cancer, particularly in conjunction with radiotherapy, chemotherapy or surgery, or in patients presenting with primary tumours, to control the development of tumour metastasis. Particular cancers may include breast, melanoma, lung, head, neck or bladder cancers. Other uses to which the compounds of the invention may be put, include use for prevention of myelin degradation in the central and peripheral nervous system, for example in the treatment of multiple sclerosis, use for controlling peridontal diseases such as gingivitis, and use in tissue remodelling.

The compounds according to the invention can also be expected to be of use in the prophylaxis or treatment of angiogenic diseases. Such diseases may be characterised by the pathological growth or new capillaries [see, for example Folkman, J. and Klagsbrun, M. Science 235. 442-447 (1987) and Moses, M. A. and Langer, R. Bio/Technology 2, 630-634 (1991)]. Particular angiogenesis dependent diseases include solid tumours and arthritic diseases as described above, and, additionally, psoriasis, eye diseases such as the proliferative reinopathies, neovascular glaucome and ocular tumours, angiofibromas, and hemangiomas.

For use in the above applications, the compounds of formula (1) may be formulated in a conventional manner, optionally with one or more physiologically acceptable carriers, diluents or excipients.

Thus according to a further aspect of the invention we provide a pharmaceutical composition comprising a compound of formula (1) and a pharmaceutically acceptable diluent, carrier or excipient.

In a still further aspect the invention provides a process for the production of a pharmaceutical composition comprising bringing a compound of formula (1 ) into association with a pharmaceutically acceptable diluent, carrier or excipient. Compounds for use according to the present invention may be formulated for oral, buccal, parental or rectal administration or in a form suitable for nasal administration or administration by inhalation or insufflation.

For oral administration, the pharmaceutical compositions may take the form of, for example, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g. pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropl methyicellulose); fillers (e.g. lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc or silica); disintegrants (e.g. potato starch or sodium giycollate); or wetting agents (e.g. sodium lauryl sulphate). The tablets may be coated by methods well known in the art. Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents, emulsifying agents, non-aqueous vehicles; and preservatives. The preparations may also contain buffer salts, flavouring, colouring and sweetening agents as appropriate.

Preparations for oral administration may be suitably formulated to give controlled release of the active compound.

For buccal administration the compositions may take the form of tablets or lozenges formulated in conventional manner.

The compounds of formula (1) may be formulated for parental administration by injection e.g. by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form. The compositions for injection may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilising and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile pyrogen-free water, before use.

The compounds of formula (1) may also be formulated in rectal compositions such as suppositories or retention enemas, e.g. containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described above the compounds of formula (1) may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation or by intramuscular injection.

For nasal administration or administration by inhalation the compounds for use according to the present invention are conventiently delivered in the form of an aerosol spray presentation for pressurised packs or a nebuliser, with the use of suitable propellant, e.g. dichloro- difluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas or mixture of gases.

The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack or dispenser device may be accompanied by instructions for administration.

The doses of compounds of formula (1) used in the above applications will vary depending on the disease or disorder and condition of the patient to be treated but in general may be in the range around 0.5mg to 100mg/kg body weight, particularly from about 1mg to 50mg/kg body weight. Dosage units may be varied according to the route of administration of the compound in accordance with conventional practice.

DESCRIPTIONS OF SPECIFIC EMBODIMENTS The invention is further illustrated in the following non-limiting Examples. In the Examples, the following abbreviations are used:

RT - room temperature

DMF - dim ethy If orm am ide

THF - tetrahydrofuran

TFA - trifluoroacetic acid

EXAMPLE A -

The activity of the compounds of the invention may be determined as described below.

All enzyme assays to determine Ki values were performed using the peptide substrate Dnp-Pro-Leu-Gly-Leu-Trp-Ala-D-Arg-NH₂. [M. Sharon Stock and Robert D. Gray. JBC 264. 4277-81, 1989]. The enzymes cleave at the Gly-Leu bond which can be followed fluorimetrically by measuring the increase in Trp fluorescence emission associated with the removal of the quenching dinitrophenol (Dnp) group.

Essentially, enzyme (e.g. gelatinase, stromelysin, collagenase) at 0.08- 2nM; a range of inhibitor concentrations (0.1-50 x Ki) and substrate (approx. 20μm) are incubated overnight in 0.1 M Tris/HCI buffer, pH 7.5, containing 0.1 M NaCI, 10mM CaC-2 and 0.05%. Brij 35 at either room temperature or 37°C depending on the enzyme. The reaction is stopped by adjusting the pH to 4 using 0.1M sodium acetate buffer and the fluorescence read at an excitation wavelength of 280nm and emission wavelength of 346nm.

Kj values can be established using the equation for tight-binding inhibition:-

(K._(app)+ PI - IE]

where V₀ is the initial rate of reaction in the absence of inhibitor, V; is the initial rate in the presence of inhibitor, [E] is the total enzyme concentration and [I] the total inhibitor concentration in the reaction mixture.

For stromelysin and collagenase, Kj (app) was assumed to approximate to the true Kj as [S] « K_m for the substrate hydrolysis. For gelatinase the Kj was determined by performing the analyses at several substrate concentrations. A plot of Kj(app) vs. [S] then gave the true Kj as the value of the y-axis intercept.

The following results were obtained with the compound of Example 1c):

KI(nM) Collagenase Stromelvsin-1 Gelatinase-72kD

2.9 90.0 1.55

EXAMP E B

The oral activity of the compounds according to the invention may be determined using the mouse pleural cavity assay described below. This assay measures the ability of compounds of the invention when administered orally to inhibit a subsequent inoculation of gelatinase into the mouse pleural cavity.

A 2ml solution of the test compound (for example around 25μm/kg) in an appropriate solvent (e.g. 50% polyethylene glycol (PG)) plus a variable proportion of dimethyl sulphoxide (DMSO) (if required) is administered orally. After an interval of up to 24 hrs, 0.4ml of a mixture of an equal volume (2.2ml) of the enzyme gelatinase A (72K form at a concentration of 20nM) and radiolabelled [¹⁴C]-gelatin (at an approximate concentration of 10μM i.e. 500 times molar excess) is injected into the pleural cavity and maintained at 4°C. After 35 min mice are overdosed with anaesthetic, the contents of the pleural cavity aspirated and the aspirates cleared by centrifugation at 4°C then diluted to 15% in trichloroacetic acid (TCA) and left overnight at 4°C. The resulting TCA precipitate is then separated by centrifugation and radioactivity in each supernatant measured by scintillation counting. Results are expressed as a % inhibition of enzyme activity calculated by comparing the radioactivity measured for each test compound with a control value obtained by performing the same assay in the absence of a gelatinase inhibitor.

The ability of compounds of the invention to prevent tumour cell invasion may be demonstrated in a standard mouse model. Thus, briefly, nude mice may be inoculated with a tumour cell line showing gelatinase - dependent invasion and the ability of compounds according to the invention to reduce subsequent lung tumour colonisation may be evaluated in accordance with standard procedures. In out tests, compounds according to the invention, when administered orally in a single dose at 100mg/kg to mice in the above model have reduced lung tumour colonisation to negligabie levels for periods of twelve hours duration or longer.

In general, compounds according to the invention are non-toxic at pharmaceutically useful doses. Thus, for example, when the compounds were administered to mice at the doses described above no adverse effects were observed.

INTERMEDIATE 1 N-tert-butoxvcarbonvl-L-penlclllamlne

To a solution of L(+)- penicillamine (24g; 161mmol) in 10% w/v aqueous sodium carbonate solution (300ml) was added di-tert-butyl dicarbonate (35.1g; 161mmol) in tert-butanol (300ml) . After stirring the reaction mixture for 18 hr at RT, the volume was reduced by approximately one half under reduced pressure and the pH was adjusted to 2 using 1 N hydrochloric acid. The resulting slurry was extracted several times with diethyl ether, the ethereal layers being combined, dried (MgSθ4) and evaporated to give the title compound (36.7g) as a clear gum. 5H (CDCI₃) 8.65 (1H, br s); 5.50 (1H, d), 4.35 (1H, d); 2.00 (1H, br s); 1.60 (3H, s); 1.50 (9H, s); 1.45 (3H, s).

INTERMEDIATE 2

N-tert-butoxvcarbonvl-L-DβnicHlamlnβ-N-mβthvlamlde

A solution in anhydrous DMF (250ml) of Intermediate 1 (11.69g; 33.5mmoi), N-hydroxybenzotriazole (4.53g; 33.5mmol), methylamine hydrochioride (11.3g; 167.5 mmol); N-methylmorpholine (20.6ml; 184mmol), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydro- chloride (7.1g; 36.9mmol) and a trace of 4-dimethylaminopyridine was stirred at RT under an atmosphere of nitrogen for 18 hr. The reaction mixture was poured into 10% w/v aq.citric acid (600ml) and extracted into diethyl ether (600ml). The organic layer was separated, washed with 10% w/v aq. NaHC0₃ solution (500ml), dried (MgSθ4) and evaporated. Following chromatography on silica, eluting with 20-50% ethyl acetate in hexane, the title compound was obtained as a clear glass (6.36g). δ_H (CDCI₃) 6.85 (1 H, m); 5.75 (1 H, d); 2.75 (3H, d); 2.50 (1H, br s); 1.50 (3H, s); 1.45 (9H, s); 1.35 (3H, s).

INTE-RMEPIATE 3 N-tert-butyloxycarbonyl-L-rS- methvnpeniclllamlne1-N- m thvlami-te

To a solution of Intermediate 2 (1g; 3.82mmol) in 2N aq. NaOH/CH₃OH (10ml/ 30ml) was added iodomethane (1.18ml; 19mmol) in CH₃OH (4ml). After stirring at RT for 2 hr, the reaction mixture was concentrated to one quarter volume, then partitioned between diethyl ether and brine. The organic layer was washed with 10% w/v aq. citric acid, dried (MgS04) and evaporated to give the title compound (810mg) as a colourless glass, δμ (CDCI₃) 6.80 (1 H, m); 5.65 (1 H, d); 4.20 (1 H, d); 2.80 (3H, s); 2.10 (3H, s); 1.45 (9H, s); 1.40 (3H, s); 1.30 (3H, s).

INTERMEDIATE 4

L-rS-(MβthvhDenlclllaπHne1-N-mβt vlaιτιlde trlfluoroacetate A solution of Intermediate 3 (810mg; 2.93mmol) in TFA/dichloromethane (10ml/10ml) was stirred at RT for 2 hr. The solvent was then removed under reduced pressure with the aid of a toluene/THF azeotrope. The title compound (855mg) was obtained as a yellow tinged glass in quantitative yield. δ_H (CDCI₃) 8.4 (3H, br s); 7.9 (1H, q); 4.20 (1H, s); 2.80 (3H, d); 2.0 (3H, s); 1.45 (3H, s); 1.35 (3H, s).

EXAMPLE 1 a) r4-t-Butoxy-2,Ri-3-{2-methylpropyt-succlnvn-L-rS- rmethvhpenicillaminβl-N-methylamide

A solution in anhydrous DMF (30ml) of 2-(R)-(2- methylpropy succinic acid-4-t-butyl monoester [2.9mmol; prepared from t-butylbromoacetate, BuLi and (S)-4- (phenylmethyl)-2-oxazolidinone according to the procedure of

Intermediate 4 in W093/24475], Intermediate 4 (2.93mmol), N- hydroxybenzotriazole (2.93mmol); N-methylmorpholine (8.79mmol), 1-(3-di-methylaminopropyl)-3-ethyl carbodiimide hydrochloride (3.19 mmol) and a trace amount of 4- dimethylaminopyridine was stirred at RT under an atmosphere of nitrogen for 18 hr. The reaction mixture was poured into 10% w/v aqxitric acid (100ml) and extracted into diethyl ether (100ml). The organic layer was washed with 10% w/v aq. NaHC0₃, separated, dried (MgSθ4) and evaporated. The residue was chromatographed on silica, eiuting with 2-4% CH₃OH in CH₂CI_2> to give the title compound.

b) r4-Hvdroxv-2(R>-3-(2-mθthvlpropvπsucclnvn-L-rS- (mβthvnpβnlclHamlnβ--N-mothvlamtde A solution of the compound of Example 1a (0.933mmol) in a mixture of TFA (10ml) and water (0.5ml) was left to stand at 4°C for 18 hr. The solvent was evaporated with the aid of a toluene THF azeotrope to obtain the title compound.

c) r4-(N-Hvdroxva ino)-2(R 3-(2-mβt vlpropyl succlπvn- L-rS- methvnpenlclllamlne1-N-methvlamlde

To a solution in anhydrous THF of the compound of Example 1b (0.933 mmol) at -20°C was added N-methylmorpholine (1.87 mmol), and ethyl chloroformate (1.12 mmol). After 1 hr, 0-tri- methylsilylhydroxylamine (3.75mmol) was added and the reaction mixture was allowed to warm to RT overnight. The solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and 10% w/v aq. citric acid. The organic layer was separated, dried (MgS04) and evaporated. The residue was purified (Si0₂; 5-10% CH₃OH in CH₂CI₂) to give the title com^pound, δμ (CD₃OD) 4.50 (1H, s); 2.95 (1H, m); 2.75 (3H, s); 2.40 (1 H, dd); 2.15 (1 H, dd); 1.50 (2H, m); 1.40 (3H, s); 1.35 (3H, s); 1.20 (1H, m); 0.90 (6H, 2d).

Claims

A compound of formula (1):

wherein R represents a -CONHOR⁶ [where R^β is a hydrogen atom or an acyl group], carboxyl (-C0₂H), esterified carboxyl, -SR⁶ or -P(0)(X¹R⁷) X R⁸ group, where X¹ and X², which may be the same or different, is each an oxygen or sulphur atom and R⁷ and R⁸, which may be the same or different each represents a hydrogen atom or an optionally substitued alkyl, aryl, or aralkyl group;

R¹ represents a hydrogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group;

R² represents an optionally substituted alkyl, alkenyl, cycloalkyl, cycioalkylalkyl, aryl, amino (-NH₂), substituted amino, carboxyl (-CO₂H), or esterified carboxyl group;

R3 represents a hydrogen atom or an alkyl group;

R⁴ represents a hydrogen atom or an alkyl group;

R⁵ represents a group -C(R⁹)(R¹⁰)Het-R¹¹, wherein R⁹ and R¹⁰ which may be the same or different is each an optionally substituted alkyl or alkenyl group optionally interrupted by one or more -O- or -S- atoms or -N(R¹²)- groups (where R¹² is a hydrogen atom or an optionally substituted alkyl group), or an optionally substituted cycloalkyl, cycloalkenyl, aryl or heteroaryl group, or R⁹ and R¹⁰ together with the carbon atom to which they are attached are linked together to form an optionally substituted C₃-₆cycloalkyl or cycloalkenyl group, Het is -0-, -S(0)_p- [where p is zero, or an integer 1 or 2] or -N(R¹²)-, and R¹¹ is a hydrogen atom or an aliphatic, cycioaliphatic, heterocycloaliphatic, aromatic, or hetero-aromatic group;

X is an amino (-NH₂), substituted amino, hydroxyl or substituted hydroxyl group, or is linked to the atom or group Het in R⁵ to form a chain -X-Alk-R⁵- where X is -N(R¹²)-, Alk is an optionally substituted alkylene chain and R⁵ is -Het-C(R⁹)(R¹⁰)-;

and the salts, solvates, hydrates and prodrugs thereof.

2. A compound according to Claim 1 where R is a -CONHOH group.

3. A compound according to Claim 1 or 2 where R¹, R³ and R⁴ each represents a hydrogen atom.

4. A compound according to any of Claims 1 to 3 where R² represents a straight or branched d-βalkyl group.

5. A compound according to any of Claims 1 to 4 where R⁹ and R¹⁰ is each an optionally substituted Ci-β alkyl group.

6. A compound according to Claim 5 where R⁹ and R¹⁰ is each a methyl group.

7. A compound according to any of Claims 1 to 6 where Het is a sulphur atom.

8. A compound according to any of Claims 1 to 7 where R¹¹ is a hydrogen atom or a methyl group.

9. A compound according to any of Claims 1 to 8 where X is an amino or a N-methylamino group.

10. [4-(N-Hydroxyamino)-2(R)-3-(2-methylpropyi)succinyl]-L-[S- (methyl)penicillamine]N-methylamide; and the salts, solvates, hydrates and prodrugs thereof.

11. A pharmaceutical composition comprising a compound according to any of Claims 1 to 10 and a pharmaceutical diluent, carrier or excipient.

12. A process for preparing a compound of formula (1) as defined in Claim 1 , which comprises

(a) coupling an acid of formula (2)

or an active derivative thereof, with an amine of formula (3),

followed by removal of any protecting groups; or

(b) interconverting a compound of formula (1), to another compound of formula (1 ).