CA2139129A1 - 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 (-CO2H), esterified carboxyl, -SR6 or -P(O)(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 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 (-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 cycloalkyl or cycloalkenyl group, Het is -O-, -S(O)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 heteroaromatic 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. 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

wo 94/2543~ 213 91~ M PCT/GB94/00896 PEPTIDYL DERIYATIVES AND THE~R USE AS METALLOPROTEINASE INHIBITORS

5 FIELD QF THE ~NVENTIC)N
This invention relates to a novel class of peptidyl derivatives, to processes for their preparation and to their use in medicine.

BAC:KGROUND TO THE ItlVENTlON
10 In normal tissues, cellular connective tissue synthesis is offset by extracellular matrix degradation, the two opposing effects exis~ing in dynamic equilibrium. Degradation ot the ma~rix 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 15 three groups of metalloproteinases. These are the collagenases, the gelatinases (or type-lV collagenases~ and the stromelysins. Nom~ally these catabolic enzyrnes a-e tightly regulated at the level of their synthQsis and secretion and also at the level of their e~nracellular activity, the latter through the action of specific inhibitors, such as a2-20 macroglobulins and TIMP (tissue inhibitor o~ metalloproteinase), whichforrn inactive complexes with metalloproteinases.

The accelerated, uncontrolled breakdown of connective tissues by - metalloproteinase catalysed resorption of the extracellular matrix is a 2~ feature of msny pathological conditions, such as rheum~toid arthritis, comeal, epidennal or gastric ulceration; tumour metastasis or invasion;
periodontal disease and bone- disaase. ~ It can be expected tha~ the pathogenesis of such diseases is likely to be modifiad in a beneficial manner by the administration of metalloproteinase inhibitors and 30 numsrous compounds have been suggested for this purpose [for a general review see Wahl, R.C. ~al A~m. Rep. Med. Chem. 25, 175-184, Academic Press Inc., San DieQo (1990)l.

. .
Although numerous metalloproteinase inhibitors have been described, 35 many have not been suitable for further development as medicines since they have lacked any useful actiYity when administered orally at WO 94/2~435 PCT/GB94/00896 ?.,~39~9 2 pharmaceutically acceptable doses. What is therefore needed is a potent orally active compound with a good duration of action.

SUMMARY OF THE INVENT!ON
5 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 atter oral administr~tion have an advantageously longer duration of action than related known compounds, such as those described in International 10 Patent Specification No. W092/09564.

Thus according to one aspect of the invention we provide a compound of formula (1) R2 R3 o ~R~RsX
15 wherein R represents a -CONHOR~ lwhere R3 is a hydrogen atom or an acyl groupl, carboxyl (-C02H), esterified carboxyl, -SR~ or -P(o)(X1R7)--X2R8 group, where X1 and X2, which may be the same or dfflerent, is each an oxygen or sulphur atom and R7 and ~, which may be the same or different each represents a hydrogen atom or an optionally substitued 20 alkyl, aryl, or aralkyl group;
.
R1 represents a hydrogen atom or an optionally substituted alkyl, alkeny!, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group;

25 R2 represents an optionally substituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, amino (-NH2), substituted amino, carboxyl (-CO2H), or esterified carboxyl group;
, R3 !epresents a hydrogen atom or an alkyl group;
R4 represents a hydrogen atom or an alkyl group;

WO 94t25435 PCT/GB94/00896 213912``~

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 a~kenyl 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 aikyl group), or an optionally substituted cyc~oalkyl, cyclo-alkenyl, aryl or heteroaryl group, or Rg 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 intager 1 or 2l or -N(R12)-, and R11 is a hydrogen atom or an aliphatic, cycloaliphatic, heterocycloaliphatic, aromatic, or hetero-aromatic group;

X is an amino ( Ntl2), 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)(R1o)-;

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 diastewisomers~ and mktures, inc!uding racemic miktures, thereof.

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

.
In the compounds according to the in~ention, when the group R
represents an esterified carboxyl group, it may be for example a group of - 35 formula -CO2R13 where R13 is a straight or branched, optionally substituted C1.8alkyl group such as a methyl, ethyl, n~propyl, i-propyl, ~39~29 4 n-butyl, i-butyl, s-butyl or t-butyl group; a C6 12arylCl.8alkyl group such as an optionally substituted benzyl, phenylethyl, phenylpropyl, a-naphthylmethyl or ~-naphthylmethyl group; a C6.~2aryl group such as an optionally substituted phenyl, -naphthyl or ~-naphthyl group; a 5 C6.12aryloxyC1.~alkyl group such as an optionally substituted phenyl-oxymethyl, phenyloxyethyl, a-naphthyloxymethyl or ~-naphthyloxy-methyl group; an optionally substituted Cl.8alkanoyloxyCl.8alkyl group, such as a pivaloyloxymethyl, propionyloxyethyl or propionyloxypropyl group; or a C6.12aroyloxyC1.8alkyl group such as an optionally 10 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 Cl.4alkyl, e.g. methyl or ethyl, or C1~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 R~ in compounds of forrnula (1~ represents an acyl 20 group, it may b: for exarnple a group of formula R14C=X3 where X3 iS an oxygen or~sulphur atom and F~14 represents a hydrogen atom or a group selected from~ amine~ (-NH2), substituted amino (for example a group -NR17P~18 as described below in relation to the group X), or an optionally - - substituted G~allq~l, Cl~alkoxy,C1 ~Salkythio, C2~alkenyl, C2 ~Salkynyl, 25 C~.lyaryl,~ C~12aralkyl, C3.6cycloalkyl, C3.6heteroaryl or C3.6hetero-aralkyl group. ~Particular groups of these types include optionally - substi~utod~-methyl, e~hyl, n-propyl, i-propyl, methoxy, ethoxy, methylthio, ethyl~io, e~nyl, 1-propenyl, ethynyl, 1-propynyl, phenyl, 1-naphthyl, 2-naphthyl, benzyl, phenethyl, 1-naphthylmethyl, 2-naphthylmethyl, 30 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-s~stituents selected from those described below in relation to the group Rtor p~2 when such groups represent substitued alkyl, aryl or 35 heteroaryl groups.
, WO 94n543s 21 3 9 2 2 ~ PCT/G11194/00896 The group R7 and/or F~8 in compounds of forrnula (1) may each be a hydrogen atom or an optionally substituted straight or branched Cl.6alkyl, e.g. methyl, ethyl, n-propyl, i-propyl, n-butyl or i-butyl, C6.12aryl, e.g. phenyl, or C6 12arylC1.6alkyl, e.g. benzyl, phenylethyl or 5 phenylpropyl group. Optional substituents present on alkyl groups of this type include one ormore C1~alkoxy, e.g. methoxyor ethoxy, or C1.6alkylthio, e.g. methylthio or ethylthio groups or an optionally substituted C6.12aryloxy e.g. phenyloxy, C6.12arylthio e.g. pheny~thio, C6.12arylC1.6alkoxy e.g. benzyloxy or C6 l2arylC1.6alkylthio e.g.
10 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 15 optionally substituted alkyl or alkenyl group, it may be, for example, a straight or branched C1~ alkyl or C2.Balkenyl group, such as a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pen~yl, i-pentyl, n-hexyl, ethenyl, 1-propenyl, 1-butenyl or 2-butenyl group optionally substituted by one or more Cl~alkoxy, e.g. methoxy, ethoxy, propoxy, 20 C1~alkylthie, e.g. methylthio, ethylthio, propylthio, C~12arylCl~alkoxy, e.g. phenylC1~alkoxy such as ber~yloxy, aralkylthio, e.g. phenyl-C1.6alkylthio such as benzylthio, amino (-NH2), substttuted amino, lsuch as -NHR1~, where R15 is a C1~alkyl e.g~ methyl or ethyl, C~12aryl-C1~alkyl, e.g. phenylC1~alkyl, such as benzgl, Cff.12aryl, e.g. phenyl, 25 C3.8cyc!oalkyl, e.~. cyclohexyl,~ or Cj.8cycloalkylCl.6alkyl, ~.9.
cyclohexylmethyl groupl, carboxyl (-C02H) or -CO2~13 twhere R13 is as defined above~ groups.

Aryl groups represented by R1 and/or R2-in compounds of formula (1) 30 include optionally substituted mono or bicyclic C~;.t2 aryl groups such as phenyl or 1- or 2-naphthyl groups..- ~~ ~

Aralkyl ~roups represented by R1=nclu~e optionally substituted mon~
or bicyclic C6.12arylC1~alkyl groups-such as phenylCl~alkyl, or 1- or 2-naphthylC1.6alkyl, for example benzyl, phenylethyl, phenylpropyl~ I
phenylbutyl, phenylpentyl, 1- or 2- naphthylmethyl, naphthylethyl, wo 94/2s43s PCT/GBg4/00896 ~3~S 6 naphthylpropyl, naphthylbutyl 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 C3.9heteroarylC1.6alkyl group, such as an optionally substituted pyrrolylmethyl, furanylmethyl, thienylmethyl, imidazolylmethyl, t oxazolylmethyl, thiazolylmethyl, pyrazolylmethyl, pyridinylmethyl, or pyrimidinylmethyl group.

Heteroarylthioalkyl groups represented by Rl include optionally substituted mon~ or bicyclic C3 9heteroarylthioC ~.6alkyl groups such as optionally substituted pyrrolylthiomethyl, turanylthiomethyl, oxazolyl-thiomethyl, thiazolynhiomethyl, pyrazolylthiomethyl, pyridinylthiomethyl, or pyrimidinylthiomethyl groups.
When ~he group R2 in compounds of fom ula (1) represents an alkyl or alkenyl group it may be for example a straight or branched C1~alkyl, or C2~alkenyl group, such as a me*yl, ethyl, n-propyl, i-propyl, n~butyl, i-butyl, s-but~l, t-butyl, n-pentyl, i-pentyl, n-hexyl, e~henyl, 1-propenyl, 1-butenyl, or 2 bu~ny1 group. Optional substituents whic~h may be present on such groups include one or more C1~alkoxy, e.g. methoxy, ethoxy, 1;
propoxy, C1~alkylthio, e.g. methylthio, ethylthio, propy~thio, amino, substhuted amino~such as NHR15 where~R1s is as defined abovel, car~xyl or CO2F'~l3 Iwhere R13 is as defined above] groups.
- Cycloalkyl groups represen~d by the group R2 in compounds according to th-e; invention inc!ude 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-alky!C1~alkyl group such as a cyclopentylC1.6alkyl or cyclohexyl- !
C1.6alky! group, for example a cyclopentylmethyl, cyclopentylethyl, WO 94/;!5435 21 ? 9 ~ 2 f! PCT/GB94/00896 cyclopentylpropyl, cyclopentylbutyl, cyclohexylmethyl, cyclohexylethyl, cyclohexylpropy~, or cyclohexylbutyl group.

Optional substituents which may be present on aryl, aralkyl, 5 hetsroaralkyl 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 10 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 Cl.6alkyl, e.g. methyl or ethyl, C 1.~alkoxy e.g. methoxy or ethoxy, C2.6alkylenedioxy, e.g.
ethylenedioxy, haloC1.6alkyl, e.g. trifluoromethyl, C1.6alkylamino, e.g.
15 methylamino or ethylamino, C1.6dialkylamino, e.g. dimethylamino or diethylarnino, arnino (-NH2), nitro, c~sno, hydroxyl (-9H), carboxyl (-C02H), -CO2Rl3, where R13 is as defined above, Cl~alkylcarbonyl, e.g. acetyl, sulphonyl (-SO3H), Cl~alkylsulphonyl, e.g. methylsulphonyl, aminosulphonyl (-:SO2NH2), Cl.8 alkylaminosulphonyl, e.g. methyl-20 aminosulphonyl or ethyiamino~ulphonyl, Cl.~dialkylamino-sulphonyl e.g. dimethylaminosulphonyl or diethylaminosulphonyl, c~rboxamido (-CONH2), C1.6alkylaminocarbonyl, e.g. methylaminocarbonyl or ethylaminocarbonyl, Cl.6dialkylaminocarbonyl, e.g. dimethylamino-- carbonyl or diethylaminocarbon~l, sulphonylamino (-NHS02H), 25 C1~alkylsulphonylamino, e.g. methylsulphonylamino or ethylsulphonyl-amino, or C1~6dialkylsulphonylamino, e.g. dimethylsulphonylamino or diethylsulphonylamino groups. It will be appreciafed that where two or more R16 substituents are present, these need not necessarily be the same atoms andlor groups. The R16 substituenSs may be present at any 30 ring carbon atom away from ~hat attached te the rest of the molecule ef formula (1). Thus, for example, in phenyl groups any substRuents may be present at the 2-, 3-, 4-, S- or 6- positions relative to the ring carbon atom attached to the remainder of the molecule.

35 When the groups P/3 and R4 in compounds of formula (1) are alkyl groups, they may be for example straight or branched C1~alkyl groups " ,.. .

WO 94/2S43~ - PCTIGB94/00896 ? ~9~ 8 such as methyl or ethyl groups.

When the group R9 or R10 in compounds of forrnula (1) is an optionally substRuted alkyl or alkenyl group it may be a straight or branched 5 C1~alkyl, 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(Rl2)- groups where Rl2 is a hydrogen atom or an optionally substituted C1~alky group such as a methyl, ethyl or propyl group.
: ~ ~
Optional substituents which may be present on such groups include one Of mor* C1.6alkoxy, Cl.6alkylthio, C6.l2arylCl.6alkoxy, aralkylthio, amino,-substituted amino, carboxyl, -C02Q13, aryl or heteroaryl groups as defined above in connection with the group Rl, or an optionally 15 subst~tuted ~cyc!oalkyl or~ cyoloalkeny~ group as defined below in connecbon wi~h~the~groups R9~and R10.

When-thé group R9, R10 or~ R9~;and R'0 togeUler with the~carbon atom to ch they~are attached, is~ an op~ionally substituted cycloalkyl or 20 ~cycloalkenyl~ group, ~it may~ be tor ~example~ a C3.8cycloalkyl, e.g.
cycbp~ cycbp~yl or~cyclohe~yl, or ~C3.8cycloalkenyl e,g. cyclo prope~l,~cyolop~or~cldlexenyl,gtoupoptionallysubstltuted by one,~ two~or ~ more~ C1~alkyl,~ e.g. ~ melhyl or eth~, Cl~alkoxy, e.g.
m~qi or ~rorJ, C~alkyl~b, ~e.g. me~io, or hydro~yl groups.

The tenn Hét in ~ pounds ot tomwh~ (1 j may-rep~esent ~, -S-, -S(O)-, S(O)~ or -N(~ ~*rè~Rl2~ b a hydrogen àtom or~a C1.6alkyl group as defined~ at~e.

30 When R1 l-in- compounds ~ot formula (1 ) is an aliphatic group it may be tor example an optionally substituted saturated or unsaturated straight or branct~ 61~alkyl chain optionally interrupted by one or more -O- or atoms or groups~ sel-cted from -N(R1~2)-, -CO-, -CON(R12)-, or -N(Rl2)CO-. Particular groups include optionally substituted methyl, 3 5~ ethy', n~propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, 1-propenyl, 1-buten~l or 2-butenyl groups. Optional substauents which 21~91~

may be present on groups of these types include one or more amino (-NH2), substituted am~ro lfor example a group -NRl7R18 as described below in relation to the group Xl, C6.l2aryl, e.g. optionally substituted phenyl, C6 12aryloxy e.g. optionally substituted phenoxy, lthe 5 optional substituents in each case being R~6 groups as defined abovel C3.8cycloalkyl, e.g. cyclopentyl or cyclohexyl, C3.8cycloalkoxy, e.g.
cyclopentyloxy or cyclohexyloxy, carboxyl (-CO2H) or -CO2P(l3 groups.

Cycloaliphatic groups represented by Rl1 in compounds of forrnula (1) 10 include optionally substituted C3.8cycloalkyl and C3.8cycloalkenyl groups, for example optionally substituted cyclopropyi, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl ~nd cyclohexenyl groups.
Optional substituents include those groups R16 described above.

1~ Heterocycloaliphatic groups represented by Rl1 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-20 methy!piperidinyl groups. Optional substituents include those groups R16 described above. The heterocycloalkyl groups represented by Rl1 may be attached to the remainder of the molecule through any ring carbon atom.

25 When the group Rl1 in compounds of torrbula ~1) is an aromatic group R
may be for exasnple an optionally substituted mono- or bicyclic C~l2aryl group, for example an optionally substitute~-phenyl or 1- or 2-naphthyl group. Optional substituents which rnay be present on groups of this type include those R16 substituents described above.
Heteroaromatic groups represented by'the group R11 include mono-or bicyclic C5.9heteroaromatic groups containing one, two or three heteroatoms selected from -O-~or--~, or-N(Q12) groups. Particular examples include pyrrolyl, furanyl, thl'ënyl,' imidazolyl, oxazolyl, thiazolyl, 35 pyrazolyl, 1-indolyl, 2-indolyl, 1-quinolinyl or 2-quinolinyl groups. Such groups may be optionally substituted, for example by one or more R16 ,~ 9 ~ 1 o substituents. The heteroaromatic group may be connected to the remainder of the compound of formula (1) through any nng carbon atom, or where appropriate through a heteroa~om or group -N(Rl2)-.

5 When X in the compounds of fonnula (1) represents a substituted amino group it may be for example a group of formula -NR~7R18, where R17 and Rl8, 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, 10 optionally interrup~ed by one or more O- or -S- atoms or -N(R12)- or aminocarbonyloxy ~-NHC(O)O-] groups or Rl7 and Rl8, together with the nitrogen atom to wtiich they are attached, may form an optionally substituted C3~cyclic amino group optionally possessing one or more other heteroatoms sebcted from -~ or -S-, or -N(R12)- groups.
Whén R17 and/or R18 is an alkyl group it ma~ be for example a C1~alkyl 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 -O- or S- atoms, or -N(R13)- or aminocarbonyloxy groups and may be for example a 20 methoxyrnethyl, ethoxymethyl, ethoxyrnethyl, ethoxyethyl or ethylamino-carbonyloxymethyl group. The optional substituents which may be present on such groups include hydroxyl (-OH), carboxyl (-C02H), esterified carboxyl (-CO2R13), carboxamido (-CONH2), substituted carboxamido, e.g. a group -CONR17R18 where NR17R18 is as defined`
25 hereinj- ammo (-NH2), substituted amino, for example a group of formula -NR17R18, aminosulphonylamino, for example -N(R12)SO2NH2 or -N(Rt2~SO2NRt7R18 or aryl? e.g. C~12 aryl such as phenyl, optionally substituted by onè, two or more R16 substituents selected from those listed above.
- . -Partic.ular examples of cyclic amino groups represented by ~NR17F~l8 include morpholinyl, imidazolyl, piperazinyl, pyrrolyl, oxazolyl, thiazolyl, pyra~I,~pyrrolidinyl, pyridinyl and pyrimidinyl groups.

3~ When the group X is a ~ubstituted hydroxyl group it may be for example a group -ORl1 where p/11 is as defined above, other than a hydrogen WO 94/25435 2 1 ~ ~ I Z ~ PCT/GB94/~0896 atom.

When X is linked to the atom or group Het in R5 to forrn a chain -X-Alk-R5, the optionally substituted alkylene chain represented by Alk may be 5 an optionally substituted straight or branched C2.g alkyl~ne chain, for example an ethylene, propylene or butylene chain. Optional substituents present on the alkylene chain inciude those described above in relation to the alkyl group represented by p~2. In compounds of this type, the group X is -N(R12)-, where R12 is as defined above. The 10 group P~5 iS -Het-C(R9)(R10)- where Het, R9 and R10 are as defined above.

Salts of compounds of formula (1) include pharmaceutically accsptable salts, for example acid addition salts derived from inorganic or organic 15 acids, sush as hydrochlorides, hydrobromides, hydroiodides, p-toluene sulphonates, phosphates, sulphates, perchlorates, acetates, trifluoro-acetates, propionates, citrates, malonates, succinates, lactates, oxalates, tartarates and benzoates.

- 20 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 morpholine, piperidine, dimethylarniae- or diethylarnine salts.
Prodrugs of compounds of formula (1) include those compounds, for example esters, alcohols or amines,~which-are convertible, In viv~. by metabolic means, e.g. by hydrolysist reduction, oxidation or transesterification, to compounds o~ formula (1).
When the group R in compounds of- the invention is an esterified carboxyl group, it may be a metabolicatly labile ester of formula -CO2Rl3 where Rl3 may be an ethyl, benzy!, 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,~trimethylbenzoyloxymethyl or pivaloyloxymethyl group.

~3~ ,~ PCTIGB94/00896 Wl~en the group R in compounds of formula (1) is a -P(O)(XlR7)X2R8 group it may in particular be a P(O)(OR7)0R8, e.g. a -P(O)(OH)OR8 group, or a ~P(O)(SH)OR8 or -P(O)(OH)SR~ group. Example-c of such groups include -P(O)(OCH3)0CH3, -P(O)(OCH2CH3)0CH2CH3, -P(O~(OH)OH, P(O)(OH)SH, -P(O)(SH)OH, -P(O)(OH)OCH3, -P(O)(OH)SCH3,-P(O)(OH)OCH2CH3,-P(O)(OH~OPh,-P(O)(OH)SPh, -P~O)(OH)OCH2Ph or -P(O)(OH)SCH2Ph, where Ph is a phenyl group optionally substitued by one or more substituents R16.
In the compounds of formula (1) the group R1 may in particular be a C1.6alkyl 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 thienyUhiomethyl, pyridinyl~
thiomethyl or pynmidinylthiomethyl group or is especially a hydrogen atom.

The group R2 in compounds of fo~ula (1) may be in particular an optionally substituted C1.6alkyl, C~cycloalkyl, or C6.12aryl 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; phenyl; 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 -rnethyl group, or, especially, a hydrogen atom.
I
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 Rl 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 t1) the group R5 may be a group -C(R9)(R10)Het-R~1 where Rl1 is an aliphatic, cycloaliphatic, WO 9412~;435 PCT/GB94/00896 213g12 ~rl heterocycloaliphatic, 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 5 (-NH2) or-NR17R18 group. Particular-NR17R18 groups are -NHR18 groups. Groups of this type include those where R18 is a C1.6alkyl 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)-l or aminocarbonyloxy groups and optionally substituted by a 10 hydroxyl, carboxyl, carboxyalkyl, e.g. carboxymethyl, carboxamido, amino, -NR1 7R 18, lfor example di-C1 -6alkylamino such as dimethylamino, C1.6alkylamino such as methylamino, or C3.6 cyclic amino such as morpholinyl, pyrrolidinyl or pyridinyll or phenyl optisnally substituted by one, two or more Rl~ substituents.
- -A particularly useful group of compounds according to the invention is that of formula (1) wherein R5 is a group -C(R9)(R~0)Het-R11 where Het is -S(O)p and R9, R10 and R11 are as defined for formula (1) Compounds of this type wherein Het is -S- are particularly useful.
A further particularq~ 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, In general, in compounds of forrnula (1) the groups Rl, R3 and R4 is each preferably a hydrogen atom. ~ ~

In a further preference, the group R in compounds according to the 30 invention is a -CONHOH or a -CO2H Qroup or a metabolically labile ester thereof, or a group P~O)(OH)OR~. In a particular preference, however, R is a -CO2H, -P(O)(OH)2 or,~ especially, a -CONHOH group.

, .
An especially useful group of compounds according to the invention has 35 the fonnula (1a) 9~ 4 X
o Rs (1a) wherein R, R2, R5 and X are as defined for forrnula tl ); and the salts, solvatss, hydrates and prodrugs thereof.

A particularly useful group of compounds of ~orrnula (1a) are those wherein R r~present a -CONHOH, -CQ2H or ~P(O)(OIt)2 group; p~2 and R5 are as defined for formula ~1~; X is an amino (-NH2) or substitu~ed amino group; and the satts, solvates, hydrates and prodrugs thereof.

Particularly useful compounds of forrnula (1a3 ars those wherein R5 is a - group -C(R~)tRl0)S(O)pR1l. Compounds of this type in which R5 is a -C(P~9~(R10)SR11 group are especially useful.

Other useful compounds of ~onnula ~1a) inelude those wherein R2 1~ represents a C3.~3alkyl group, partieularly an isobutyl or n-peniyl group, or a cycloalkylC3 6alkyl ~roup, partieularly a cyelohQxylpropyl, cyclo hea~lbutyl or cyclohexyipentyl ~roup.

In the compounds of formula (1a) X may be a -NH2 group or a group -NR17Ftl8 as defined for compounds of formula (13, particularly a -NHR18 group.

An especially us~ful group of compounds ac~ording to the invention has the formul~ (1a) wherein R7 is a C3.~3alkyl group, R5 is a group C(R9~(R10)SR~1 where R9 and Rl is each the same and is each an optionally substituted Cl~ alkyl group, and Rll is as defined for formula ; and X is an amino ( NH2) or NHR18 group, particularly where R18 is an optionally substituted C1.6 alkyl group. Compounds of this type whe^roin- Q5 is a group -C(CH3)2SR11 are particularly useful, especially where the group R11 is a hydrogen atom or an optionally substituted saturatad Cl.6 alkyl chain. In compounds of this last type X is preferably an amino (-NH2) group or a -NHCH3 group.

WO 94125435 213 9 ~ 2 `~ PCTIGB94/00896 In a still turther useful group of compounds of formula (1a), R is a -CONHOH, -C02H or -P(0)(01~)2 group, R2 is an isobutyt group, R5 is a group -C(CH3)2SR11 where R11 is a hydrogen atom or an optionally substitued C1~ alkyl group, and X is an amino (-NH2) or -NHR18 group 5 where R18 is an optionally substituted C1 6 alkyl group Compounds of this lype 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 hrther group of compounds according te the invention has the formula~ (1a) wh~ rein R and R2 are as defined~ for formula (1), Rs is a ~group~-C~CH3)25H o r -C(CH3)2SCH3 and X is -NH2 or -NHCH3 Particularly useful compounds of this type are those wherein R is a ~5 - group -CONHOH, -CO2H or -P(O)(OH)2 and R2 is a Cwalkyl group P~articular~y useful compounds are~those wh-re R2 is an isobutyl group . ~ -In~compoun:ds~of~the above~described types, R5 is preferably a group C(CH~3)2S~CH3 ~ 1n~;th-se ~compounds,~R is preferably -CO2H or Z0 ~ P(O)(OH2) or~is ~blly -CONHOH X is prderably -NH2 or -NHCH3 *~ -impon~nt~ cornpoùnd according to th inv~nL;on is 4-(N-Hyd~nmo)-2(R~(2-methylpropyl)succinyl]-L IS-(methyl) 25 ~ ~nlbillaminel ~N-methylami~; and:. the~ salts,: solvat s, hydrates and prodrugs~the~

me Gompounds~ ac~c~rdhg to the invention -may be ~prepared by the following gen-ràl~ processes, more specifically described in the 3 0 ~ l~xamples hereinafter In the d scriptbn and formulae below the groups R, R1, R2, R3, R4, R5 and X are as- defined above, except where ;otherwis indicated It will be appreciated that functional groups, such as~ amino, hydroxyl or carb~xy~-groups, present in the various - ~ - compounds described ~bebw, a~d ~ ch it is desired to retain, may need : 35: to :be: in protect-d~ form ~before~ any reaction is initiated In such instances, removal of the protecting group may be the final step in a ~':
~' : .

?~ ~ PCTIGB94/00896 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 5 example palladium on a suppon 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-10 butyl group which may be removed from a protected derivative bytreatment 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, hydro~l or carboxyl group may be achieved using standard alkylation or esterification procedures, for 15 example as described bebw.
: l Thus according to a further aspect of the invention a compound of formulà~(1) may be prepared by coupling an acid of formula (2) R~

~1 o (2) 20 or an active derivative thereof, with an amine of formula (3) - ' ~ ~ X
R4 (3) followed by removal of any protecting groups.
: - .
.
~-25- Active derivatives of acids for formula (2) include for examp~e acid ;l anhydrides, or acid halides, such as acid chlorides. ~i~
. _ = .
The coupling reaction may be performed using standard conditions for amination readions of this type. mus, for example the reaction may be 30 achieved in a solvent, for example an inert organic solv~nt such as an I

WO 9412543~ PCT/GB94/00896 21~912~

ether, e.g. a cyclic ether such as tetrahydrofuran, an amide e.g. a substituted amide such as dimethylformamide, or a halogenated hydrocarbon such as dichloromethane at a low temperature, e.g. -30C
to ambient temperature, such as -20C to 0C, optionally in the 5 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 forrnula (2) is used, the reaction may additionall~ be performed in the presence of a condensing agent, for example a diimide such as N,N'-dicyclohexylcarbodiimide, or 1-(3-dimethylaminopropyl)-3-ethyl-10 car~odiimide, advantageously in the presence of a triazole such as2-hydroxybenzotriazole. Altematively, the acid may be reacted with a chloroformate for example ethylchlorofonnate, prior to reaction with the amine of formula (3).

15 Fr~e hydro~l or carboxy~ groups in the starting materials of formulae (2) - lwhere R is -CONHOH or CO2Hl and (3) may need to b~ protected - during the coupling reaction. Suitable proteoting groups and methods for their removal may be those mentioned above. Where R in the interrnediates ol fonnula (2) is a -P(O)(XlR7)X2R8 group, at least one of 20 P~7 or R8 is other than a hydrogen atom. Conveniently, each of R7 and RB 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 Q7 and~or R8 is each a hydrogen at-om.
It will be appreciated that where a particular stereoisomer of forrnula (1) is required, this may be obtained by resolution of a mixture of isomers following the coupling reaction of an acid of forrnula (2) and an amine of formula (3). Conventional resolution techniques may be used, for 30 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 preparad by 35 reaction of a compound of formula (2a) WO 94125435 ~ PCT/GB94/00896 .
R~

o (2a) with an amine of formula (3a) ~x Rs (3a) as described above Intermediate acids of formula (2) wh~rein P~ is a carboxyl or esterified carboxyl group or a group -P(O)(X~R7)X2R8 or-SP~8 may be prepared 10 from a corrsspanding ester of forrnula (4) f~2 R~

R1 0 ~4) where Rl9 is an alkyl group, for example a meth~ or t-buty~ group, using for example tritluoroace~ic acid, or, when Rlg is an aralkyl group, such - as a benzyl group, by hyqrogenolysis~ for example by r~action with 15 hydrogen in the presence of a metai catalyst, e.g. palladium, on a support such as carbon in a solv~nt such as an alcohol, e.g. methanol optionally at an eleYated pressllre and temperature. ~
.
Ar ester of fonnula (4) whQre R is a carboxyl or esterified carboxyl group 20 may be preparad by es~erification ot the corresponding~ acid of formula (5) R - (S) ~8~

WO 94/2~435 PCT/GB94/00896 2139~2.i, using an appropriate acyl halide, for example an acyl chloride in a solvent such as ~n alcohol, e.g. methanol at a low temp~rature, e.g.
around OC.

5 Acids of fsrmula (5) may be prepared by alkylation of a compound of ~orrnula (6) O~ 12a~

Q1 0 (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 pres~nce of a 10 base, for example an alkoxide su~h as sodium ethoxide in a solve~t 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.

15 Int~rrnediates of farmuia (6) are either known ccmpounds or may be prepared by rnethods analogous to those used for the pr0paration of the known compounds.

Intermediate esters of formula (4) where P( is a -P(O)(XlR7)X2R~ group 2û may be prepared by reaction ot an acrylate RlCHC(R2)CORl9 with a phosphite:P(OR20~(X1R7)X2P48 lwhere R20 is a leaving group, for ex~mple a sily~ group such as a trialkylsilyl group e.g. a trimethylsilyl group~ at an elevated temperature. - -25 Ac~ylatee of ~ormula RlCHC(R2)CORl9 may be prepared ~y 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 30 elevated tempe~ature.

Mono-esters of formula HOOCCH(R2)COOR19 may b~ pr~par3d by WO 94/25435 PCT/GB94/û0896 9~'~`3 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 0C. The di-esters for use in this reaction may be prepared by alkylation of the corresponding 5 malonates of formula R19OOCCH2COOR19 with a halide R2Hal [where Hal is a halogen atom such as a chlorine or bromine atoml 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 R19OOCCH2COOR19 are either known compounds or may be prepared 10 by methods analogous to those used for the preparation of the known compounds.
i Intermediate phosphites of formula P(OR20)(X1R7)X2R8 may be prepared by reaction of a phosphite HP(O)(X1R7)X2R8 with an 15 appropriate amine (R2)2NH e.g. a silazane, at an elevated temperature, e.g. the reflux temperature. Phosphites of formula HP(O)(X1R7)X2R8 are either known compounds or may be prepared by methods analogous to those used for the preparation of the known compounds.

20 Intermediates o f formula (4) where R is a -SR~ group are either knom compounds or may be prepared from known starting materials of formula R~SCH(R1)CH(CO2CH2CH3)2 by using a similar series of reactions to those just described for the preparation of compounds of - formula (4) whero R is a carboxyl group.
In another process, intermediate acids of formula (2) wherein R is a -P(O)(X1R7)X2R8 group may be prepared by re ction-:of-an acid - ~ R2CH2CO2H with a phosphonate P(O)(X1R7)(X2R8)CH2OR21 ~where ~21 iS a leaYing group, for example a trifluoromethylsulphonyloxy group 30 in the presence of a base such as n-butyllithium in a solvent such as tetrahydrofuran. Phosphonates fo~ use in this reaction rnay be prepared from the corresponding compound P(O)(X1R7)(X2R8)CH2oH by reaction with paraformaldehyde in the presence of a base such-~s- triethylamine at an elevated temperature followed by reaction with a halidè R21Hal in 35 the presence of a base such as sodium hydride in a so~ent such as an sth~r. Phosphonatos P(O)(X1R7)(X2R8)CH20H and acids R2CH2CO2H

WO 94/25435 213 ~J 1 2 '~ PCT/GB94/00896 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.

Interrnediate acids of fonnula (2) wherein R is a -CONHOR6 group or a protected derivative thereof may be prepared by reaction of an anhydride of formula (7) R2~ l~
,1 ~
o (7) with a hydroxylamine swch as O-benzylhydroxylamine or NH20R6 10 where R6 is an acyl group in a solvent such as tetrahydroturan at a low temperature, e.g. around -20C, followed where desired by removal ot 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 -CO2H with an acyl chloride such as acetyl chloride.
.
The homochiral~ acids of formyla 12a) may be prepared according to another feature of the invention by oxidation of an oxazolidinone of - -~; formula (8) :- ~ R2 R~ N~ ~0 - Ph (8) (where Ph is a phenyl group) ~' ` -1, - 25 using an oxidising agent such as peroxide, e.g. hydrogen peroxWe in a ~-~~
solvent such as an ether e.g.~ a cyclic ether such as tetrahydrofuran, a~ a low temperature, e.g. around 0C followed by treatment with a base, such as lithium hydroxide, at an elevated temperature.

WO 9~/25435 ~ PCT/GB94/00896 'l.~ 3~

The compounds of forrnula (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-5 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 -78C.

Acyl halides PsCH2CH(R2)COHal may be prepared by treatment of the 10 corresponding known acids RCH2CH(R2)CO2H with conventional halogenating agents for example thionyl halides under standard reaction conditions.

Intermediates of formula (3) are either known compounds or may be 15 prepared from known amino acid starting materials using standard methods, for example by employing a séries of substitution reactions to manipulate the groups R5 and X as described in the Examples hereinafter, or for example as described by Wessjohann .~L Chem.
Ber. 1992, ~2~, 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 forrnula (1) wherein R is ~ -CO2H
25 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 R60NH2 where P/~ is an acyl group.- The reaction may be performed using the reagents and conditions described above in the preparation of compounds ot fomlula (1) from the starting 30 materials of fonnulae (2) and (3).

In another interconversion process, compounds of fomluia (1) wherein R
is -C02H andlor X contains a -CO2H group may-b~ prepared by hydrolysis of the corresponding esterified compounds~(for example 35 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 methanol, 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 CO2P~13 group andlor X contains a -CO2R13 group may be prepared by reaction of the corresponding acids, where R is a -CO2H group and/or X contains a -CO2W group or an active derivative thereof, with an alcohol R13OH
10 using standard conditions.

In another interconversion process, a compound of fonnula (1) wherein R5 is a group -C(R9)(R10)S-R11 may be oxidised to a corresponding compound where F15 iS a group -C(R9)(Rl0)SOR11 or 15 -C(R9)(R10)SO2R11 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 pero~yacid in a halogenated hydrocarbon solvent such as dichloromethane at a low temperature, e.g. around -78G.
The compounds according to the invention are potent inhibitors of the metalloprote~inases collagenase, stromelysin and gelatinase and advantageously~have~ ~a long duration of action when administered ora!ly.; The activity of the com-pounds may be detemlined by the use of 25 appropriat- enz~me inhibi~ion tests for example as described in Exar:nple~ A~hereinafter or by oral administration to mice as described hereinafter in~Exarinple 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 30 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 35 the compounds of formula (1) may be of use in the prophylaxis or treatment of musculo-skeletal disorders, for example arthritic diseases .

WO 94/25435 ~;,9 24 PCT/G119-100896 such as rheumatoid arthritis, osteoarthritis and septic arthritis, and to be of usa to prevent tumour cell metastasis and inv~sion. The compounds may therefore be o~ use in the treatment of cancer, partieularly in conjunction with radiotherapy, chemotherapy or surgery, or in patients 5 presenting with primary tumours, to control the development of tumour metastasis. Particular caneers may inelude breast, melanoma, iung, head, neck or bladder caneers. Other uses to whieh the compounds of the invention may be put, include use for prevention of myelin degradation in the central and peripheral nervous system, for exampl~
10 in the treatment of multiple sclerosis, use for eontrolling peridontal diseases sueh as gingivitis, and use in tissue remodelling.

The eompounds aeeording to the invention ean also be expeeted to be of use in the prophylaxis or treatment of angiogenic diseases. Sueh 15 diseases may be characterised by the pathologieal growth or new eapillaries ~see, for-example Folkman, J. and Klagsbrun, M. Scienee ~, 442-447 (1987) and Moses, M. ~ and Langer, R. Biorrechnology ~, 630-634 (1991~1. Particular angiogenesis dependent diseases include solid tumours and arthritic diseases as described above, and, 20 additionally, psoriasis, eye diseases sueh as the proliferative reinopathies, neovascular glaucome and ocular tumours, angiofibromas, and hemangiomas.

- For use in the above applieations, the eompounds of formula (1) may be 25 formulated in a eonventional manner, optionally with one or more physiologically aeeeptable earriers, diluents or excipients.

Thus according to a further aspeet of the invention-we provide a pharmaeeutieal composition comprising a eompound of formula (1) and 30 a pharmaeeutieally aeceptable diluent, earrier or exeipient.

In a still further aspeet the invention provides a proeess for the produetion of a pharmaeeutieal eomposition eom~rising bringing a eompound of formula (1) into assoeiaiion with a pharmaeeutieally 35 aeeeptable diluent, earrier or exeipient.

WO 94125435 2 1 ~ 9 1 2 ~. PCTIGB94 00896 Compounds for use according to the present invention may be forrnulated for oral, bucczl, parental or rectal administration or in a forrn suitable for nasal administration or administration by inhalation or insufflation.

For oral administra~ion, 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 10 hydroxypropl methylcel~lulose); fillers (e.g. Iactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc or silica); disintegrants (e.g. potato starch or sodium glycollate); or wetting agents (e.g. sodium lauryl sulphate). The tablets may be coated by methods well known in the art. Uquid preparations for 15 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, 20 emulsifying~agents, non-aqueous vehicles; and preservatives. The prepara~ions may also contain buffer salts, flavouring, colouring and sweetening agents as appropriate.

Preparations for oral administration may be suitably fomnulated to give 25 controlled r-base of the active compound.

For buccal administration the compositions may take the form of tablets or lozen~es forrnulated in conventional manner.

30 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 forrn. The compositions for injection may take such forms as suspensions, solutions or emulsions in oil~ or aqueous vehicles, and may contain 35 formulatory agents such as suspending, stabilising and/or dispersing agents. Altematively, the active ingredient may be in powder form for WO 94/Z543~i ~3 9 ~? 3 PCTIGB94/00896 constitution with a suitable vehicle, e.g. sterile pyrogen-~ree water, before use.

The compounds of formula ~1) may also be formulated in rectal compositions such as suppositories or reten~ion 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 inj~ction.

For nasal administration or administration by inhalation the compounds 1~ for use according to the present invention are conventiently delwered in the form of an aerosol spray presentation for pressurised packs or a nebuliser, with the use of suitable propellant, e.g. dichloro difluoramethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other sultable 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.
f The doses of compounds of formula (1) used in the above app!ications wili vary depending on the disease or disorder and condition of the patient to be troated but in general may be in the range~around 0.5mg to 100mgJkg 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 wit~ conventional practice. - ~
_ DESCRIPTIONS OF SPECIFIC EMBODIME~I~
The invention is further illustrated in the following noFl-lirniting Examples.

WO 94/t~435 PCTIGB94/00896 21391~9 In the Examples, the following abbreviations are used:

~r - room temperature DMF - dimethylformamide THF - tetrahydrofuran TFA - trifluoroacetic acid EXAMPI.E~

10 Th~ activity of the compounds of the invention may be deterrnined as described balow.

~11 enzyrne assays to determine Ki values were perforrned using the peptide substra~e Dnp~Pr~Leu-Gly-Leu-Trp-Ala-D-Arg-NH2. [M. Sharon 15 Stock and Rob0rt D. Gray. JBC ~ 4277-81, 1989l. The enzyrnes cleave ~t tha Gly-Leu bond which can be followed Sluorirnetrically by measuring ~he increase in Trp fluorescsnce emission assvciated with the removal of the quenching dinitrophenol ~Dnp) group.

20 Essentially, enzyrne (e.g. gelatinase, stromelysin, collagenase) at 0.08-2nM; a range of inhibitor concentrations (0.1-50 x Ki) and substrate (approx. 2011m~ are incubated ovemight in 0.1M Tris/HCI buffer, pH 7.~, containing û.1M NaCI, 10mM CaCi2 and 0.05%. Brij 35 at either roorn tempefature or 37C dependlng on the enzyme. ~he reaction is 25 stopped by adjusting the pH to 4 using 0.1M sodium acetate buffer and the fluorescence read at an excitation wave~ength of 2~0nm and emission wavelength of 346nm.

Kj val~Jes can be established using the equation for tight-binding 30 inhibition:-Vj = vO /
~ ,pp) + [llz + 2 (~ ID~ + ~ (4pr lll - Iq) where V0 is the initiai rate of reaction in the absence of inhibitor, Vj is ~he ~ 9~ 28 initial rate in the presence of inhibitor, EE] is the total enzyme concentration and ~I] the total~ inhibitor concentration in the reaction mixture.

5 For stromelysin and collagenase, Kj (app) was assumed to approximate to the true Kj as lS~ ~ Km for the substrate hydrolysis. For gelatinase the Kj was determined by performing the analyses at several substrate concentrations. A plot of Ki(app) ~/s. [S3 then gave the true Kj as the value of the y-axis intercept.
The following results were obtained with the compound of Example 1c):
Kl(nUI) Collageo~e Stromelysin-1 Ge6~tinas~72kD
2.9 90.0 1.55 EXAMPLE B
The oral activity of the compounds according to the imention may be deterrnined using the mouse pleural cavity assay described below. ~his assay measures the ability of compounds of the invention when 20 administered orally to inhibit a subsequent inoculation of ~elatinase into the mouse pbural cavity.

A 2ml solution of the test compound (for example around 25~1m/kg) in an appropriate sohent (e.g. 50% polyethylene glycol (PG~plus a variable 25 proportion of dimethyl sulphoxide (DMSO) (~ 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 foml at~a-c~centration of 20nM) and radiolabelled [1 4C]~gelatin (at an approximate concentration of 1011M i.e. 500 times molar excess) is injected into the 30 pleural cavity and maintained at 4C. A~ter 35 min mice are overdosed with anaesthetic, the contents of the pleural cavity aspirated and the aspirates cleared by centrifugation at 4C then diluted to t5% in trichloroacetic acid (TCA) and lett ovemight at 4C. Tfie resuiting TCA
precipitate is then separated by centrifugation and radioactivity in each 35 supernatant measured by scintillation counting. Results are exprsssed as a % inhibition of enzyme activity calculated by comparing the wo s4ns43s PCTIGB94/00896 21391 2~

radioactivity measured for each test compound with a control value obtained by performing the same assay in the absence of a gelatinase inhibitor.

5 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 10 evaluated in~ accordance wi~h standard~ procedures. In out tests, compounds~ according to the invention, when administered orally in a singb;,dose~at~ 10~kg to mice in the; above model have reduced lung tumour~ cobnisa,tion~ to~ negligable levels ~for periods of twelve hours duration or longer.
I n g éneral, ~comp~unds~ accolding to the inventbn are non-to~ac at ; pharmaceùtically u~ul~ doses. Thus, for example, when the I
~re;~admini~é~to mice at~the d~s d~ri~ above no i~nvl-l~D_~
~mmol) in 10% wh aqueous sodium~arbonàt,e ~olutbn~ was a~ d~-tert-butyl dicarbonate ',~':25~61mmol)~in~te~Pnol~(300inl).~Afterstirring~thereaction --mi~re tor'-18 hr~at ~ `the;vo~m~ reduced by appro~amately one half *~ rKi~t p~re an~t~the~ptl was~adjusted to 2 using 1N
H~ hydr~lo~ d.~ The~result~gs~urlywas~e~ra ed~seyeraltimeswith die~yl~ether,~;the;` l~ rs~beinq combined,~d~ (MgS04) and 30 evaporated~to'give the-title com~ound (36.79) as actear gum- ~H
(CDCb) 8.65 (1;H, ~br s); S.50 (1H, d), 4.35 (1 H, d); 2.00 (1 H, br s); 1.60 (3H, s): 1.50 (9H, s); 1.45~ (3H, s).
~, , 35 ~ INI E_~
t-i~ ~oll-c-rbonvl-L-D-nlolll-mlne~N~methv~'amlde A solution in anhydrous~ DMF (250ml) of Intermediate 1 (11.699;

~ ~ 9 ~ 30 33.5mmol), N-hydroxybenzotriazole (4.539; 33.5mmol), methylamine hydrochloride (11.39; 167.5 mmol); N-methylmorpholine (20.6ml;
1 84mmol), 1 -(3-dimethylaminopropyl)-3-ethyl carbodiimide hydro-chloride (7.19; 36.9mmol) and a trace of 4-dimethyiaminopyridine was 5 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 s6parated, washed with 10% w/v aq. NaHCO3 solution (500ml), dried (MgSO4) and evaporated. Following chromatography on silica, eluting with 20-50%
10 ethyl acetate in hexane, the title comDolmd was obtained as a clear glass (6.369). ~H (CDCI3) 6.~5 (1H, m); 5.75 (lH, d); 2.75 (3H, d); 2.50 (1 H, br s); 1.50 (3H, s); 1.45 (9H, s); 1.35 (3H, s).

15 N-tert-butvloxvcarbonvl~ S-(methyl)~enlclllamlne1-N-methvlamld~
To a solution of Inte~ediate 2 (19; 3.82mmol) in 2N aq. NaOH/CH30H
(lOmll30ml)was added iodomethane (1.18ml; 19mmol) in CH30H
(4ml). After stirring at RT for 2 hr, the reaction mixture was concentrated 20 to one quarter volume, then partitioned between diethyl ether and brine.
The organic la~er was washed with 10% w/v aq. citric acid, dried (MgSO4) and evaporated to give the title co~Qound (810mg) as a colourless glass. ~H (CDCI3) 6.80 (1H, m); 5.65 (1H, d); 4.20 (1H, d);
- 2.80 (3H, s); 2.10 ~3H, s); 1.45 (9H, s); 1.40 (3H, s); t.30 ~3H, s).

S~(M-thvl)D~nlcll!amlneJ-N~ thylamlde trl~oroa6et~te A solution ot Intermediate 3 (810mg; 2.93mmol) in TFA/dichtoromethane (10mU10ml) was stirred at RT for 2 hr. The solvent was then removed 30 under reduced pressure with the aid of a toluene/THF azeotrope. The title comDound (855mg) was obtained as a ye!!ow tinged glass in quantitative yield. ~H (CDCI3) 8.4 (3H, br s); 7.9 (1H, q); 4.20 (1 H, s);
2.80 (3H, d); 2.0 (3H, s); 1.45 (3H, s); 1.35 (3H, s~

35 EXAMPLE~ 1 a) [4-t-Butoxy-2(R) 3-(2-methvlprQeyl)succinvll L-~S

WO 94/25435 21 3 ~1 2 PCT/GB94100896 (methyl!~enicilLamioel-N-methyl~m~de 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~ (3-di-methylaminopropyl)~3-ethyl carbodiimide hydrochloride (3. t 9 mmol) and a trace amount of 4-dimeth~laminopyridine was stirred at RT under an atrnosphere of nitrogen ~or 18 hr. The reaction mixture was poured into 10% w/v aq.citric acid (100ml) and extracted into diethyl ether (100ml).
The organic layer was washed with 10% wlv aq. NaHCO3, separated, dried (MgSO4) and evaporated. The residue was chrornatographed on silica, eluting with 2-4% CH30H in CH2CI2, to give the titie comDound.
b) l4-Hygrox~-2(R)-3-(2-methylQro~vll~u~ç~ -L-~
(methvl)~enlclllamlnel-N methylaml~
A solution of the compound of Example 1a (0.933mmol) in a mixture of TFA (10ml) and wat~r (0.5ml) was lèft to stand at 4C
for 18 hr. The solvent was evaporated with the aid of a toluenelTHF azeotrope to obtain ~he ~l~omQ~nd.

c) ~4-(N-Hvdroxv~ nQ~ 2(R~-3-(2 meth~lptQ~yl1~ clnvl1-L~rs~(methyl)Den~ el~N~m~hv~ d~
To a solution in anhydrous THF of the compound of Example 1b (0.933 mmol) at -20C 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 ovemight. 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 (MgSO4) and evaporated The residue was purified (SiO2; 5-10% CH30H in CH2C12) to give the ~ H (CD30D) 4.50 (tH, s); 2.95 (lH, m);

? ~39~q*~ 32 2.7~ (3H, s); 2.40 (1H, dd); 2.15 (1H, dd); 1.50 (2H, m); 1.40 (3H;
s); 1.35 ~3H, s); 1.20 (1H, m); 0.90 (6H, 2d). -=

, _ .

,

Claims (12)

1. A compound of formula (1):

(1) wherein R represents a -CONHOR6 [where R6 is a hydrogen atom or an acyl group], carboxyl (-CO2H), esterified carboxyl, -SR6 or -P(O)(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, R2 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-(R10)-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 C3-6cycloalkyi or cycloalkenyl group, Het is -O-, -S(O)p- [where p is zero, or an integer 1 or 2] or -N(Rt2)-, 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 -NtR12)-, Alk is an optionally substituted alkylene chain and R5 is-Het-C(R9)(R10)-;

and the salts, solYates, 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 C1-6alkyl group.

5. A compound according to any of Claims 1 to 4 where R9 and R10 is each an optionally substituted C1-6 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 whers Het is a sulphur atom.

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

9. A compound accordins 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-methylpropyl)succinyl]-L-[S-(methyl)penicillarninelN-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) (2) or an active derivative thereof, with an amine of formula (3), (3) followed by removal of any protecting groups; or (b) interconverting a compound of formula (1), to another compound of formula (1).