PT100558A - Peptidyl derivatives, pharmaceutical compositions that contain them, and process for preparation of these derivatives - Google Patents

Abstract

The invention relates to new peptidyl derivatives, with the formula I: <IMAGE> in which R represents a -CONHOH, carboxyl (-CO2H) or esterified carboxyl group; R1 represents a hydrogen atom or an alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group, which may possibly be substituted; R2 represents a phenylethyl, phenylpropyl or phenylbutyl group, which may possibly be substituted; R3 represents a hydrogen atom or an alkyl group; R4 represents a hydrogen atom or an alkyl group; R5 represents an alkyl or alkenyl group which may possibly be substituted, and which may possibly be interrupted by one or more -O- or -S- atoms or -N(R7)- groups [in which R7 represents a hydrogen atom or an alkyl group with 1 to 6 carbon atoms]; X represents an amino (-NH2), substituted amino, hydroxyl or substituted hydroxyl group; and the salts, solvates and hydrates thereof. The compounds are metalloproteinase inhibitors and in particular have a selective action against gelatinases, and can be used in the treatment of cancer in order to control the development of metastases of tumours.

Description

Description of the invention of OELLTECH LIMITED, British, industrial and commercial, established at 216 Bath Road, Slough, Berkshire SL 1 4M, UK (inventors: John Ro-bert Porter, Thomas Andrew Milican, John Richard Morphy and Higel Robert Amold Beeley, Residents in the United Kingdom) for &quot; PEPTIDILO DERIVATIVES: PHARMACEUTICAL COMPOSITIONS THAT THE CONTAINS AND PREPARATION PROCESS FOR THESE DERIVATIVES &quot;.

DESCRIPTION

Field of the Invention The present invention relates to a particular class of peptidyl derivatives, to processes for their preparation and to their use in medicine.

Background of the invention

In normal tissues the synthesis of cellular connective tissue is counterbalanced by the degradation of the extracellular matrix, with the two opposing effects in a dynamic equilibrium. The degradation of the matrix is caused by the action of M.A. · 1 -

released from resident connective tissue cells and by invasive inflammatory cells, and is due in part to the activity of at least three groups of metalloproteinases. These are collagenases, gelatinases (or type IV collagenases) and stromelysins. Usually these catabolic enzymes are tightly regulated at the level of their synthesis and secretion and also at the level of their extracellular activity, it is last through the action of specific inhibitors, such as O -macroglobulins and IIMP (metalloprotein inhibitor tissue if), which form inactive complexes with the metalloproteinases. Accelerated, uncontrolled breakage of junctional tissues by resorption, catalyzed by metalloproteinases, of the cell matrix is a feature of many pathological states, such as rheumatoid arthritis, corneal, epidermal or gastric ulceration, metastases or tumor invasion; periodontal diseases and bone diseases. It is expected that the pathogenesis of these diseases is likely to be beneficially modified by the administration of metalloproteinase inhibitors and numerous compounds have been suggested for this purpose for a general review see Wahl, RC et al. Ann. Rep. Med. Chem. 25, 175-184, Academic Press Inc., San Bi ego (1990) J,

Certain hydroxamic acid peptidyl derivatives have been described as collagenase and / or stromelysin inhibitors. See for example European Patent Specification Nos. 214,639, 231,081, 236,872 and 274,453, and International Patent Specifications nes ¥ 090 / 05716 and WG9Q / G5719__7.

SUMMARY OF THE INVENTION

We have now discovered a particular class of peptidyl derivatives, which members advantageously have a potent inhibitory action against gelatinase. - 2 -

There is now strong evidence that metalloproteinases are important in invasion of tumors and metastases. Tumor cell gelatinase, in particular, is associated with the potential of tumor cells to invade and form metastases. Invasion of tumors and formation of metastases are the major cause of failure of treatments for cancer patients, and the use of a selective inhibitor of gelatinase, such as a compound of the present invention, which is capable of inhibiting cell invasion of tumor, may prove to be improving the treatment of this disease.

Thus, according to one aspect, we disclose a compound of formula (I) of the present invention R R &quot; 0

THE.

(D) in which R represents a group -GNHGH, carboxyl (-CG2 H) or esterified carboxyl; 1, R3 represents a hydrogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl group R 2 represents a hydrogen atom or an alkyl group; R 4 represents a hydrogen atom or an alkyl group; R 5 represents an optionally substituted alkyl or alkenyl group; optionally substituted by one or more -G- or -S- or -NH- (R ') - groups wherein R represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms,

wherein X represents an amino or substituted amino group,

hydroxyl or substituted hydroxyl; and their salts, solvates and hydrates.

Note that the compounds according to the invention may contain one or more asymmetrically substituted carbon atoms, for example those which are marked with an asterisk in formula (I). The presence of one or more of these asymmetric centers in a compound of formula (I) may give this stereoisomers, and in each case the invention should be taken as extending to all such stereoisomers, including enantiomers and diastereoisomers, and their mixtures, including racemic mixtures.

In the formulas given herein the line I used in a potentially asymmetric center to represent the possiblity of the R and S configurations, and the lines w and ----- present a single configuration in an asymmetric center.

In the compounds according to the invention, when the group R represents a carboxy group which is esterified, it may for example be a group of the formula - in which R represents a straight or branched chain alkyl group with 1 to Optionally substituted carbon atoms, such as a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl or t-butyl group; an aryl-alkyl group having 6 to 12 carbon atoms in the aryl part and 1 to 8 carbon atoms in the alkyl part, such as an optionally substituted group selected from benzyl, phenylethyl, phenylpropyl, or naphthylmethyl or--naphthylmethyl; an aryl group having 6 to 12 carbon atoms such as a phenyl, o-naphthyl or optionally substituted-naphthyl group; an aryloxy-alkyl group having 6 to 12 carbon atoms in the aryl part and 1 to 8 carbon atoms in the alkyl part, such as an optionally substituted group is selected from phenyloxymethyl, phenyloxyethyl, (X-naphthyloxymethyl or α-naphthyloxymethyl group; alkanoyloxy-alkyl with 1 to 8-4-

carbon atoms in each of the optionally substituted alkyl parts, such as a pivaloyloxymethyl, propionyloxyethyl or propionyloxypropyl group; or an aroyloxycarbonyl group having 6 to 12 carbon atoms in the aryl part and 1 to 8 carbon atoms in the alkyl part such as an optionally substituted group selected from benzoyloxyethyl or benzoyloxypropyl. 8

Possible substituents present on the R groups include for example one or more halogen atoms, such as fluorine, chlorine, bromine or iodine atoms, or alkyl groups having 1 to 4 carbon atoms, for example methyl or ethyl groups , or alkoxy groups having 1 to 4 carbon atoms, for example methoxy or ethoxy groups.

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

When the group in the compounds of formula (I) has an optionally substituted alkenyl or alkenyl group, it may for example be an alkyl group having 1 to 6 carbon atoms or an alkenyl group having 2 to 6 carbon atoms, carbon, of straight or branched chains, such as a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, n-hexyl, ethenyl, 1- propenyl, 1-butenyl or 2-butenyl groups optionally substituted by one or more alkoxy groups having 1 to 6 carbon atoms, for example methoxy, ethoxy, propoxy, C 1 -C 6 -alkylthio groups such as methylthio, ethyl - thio, propylthio, aryl-alkoxy groups having 6 to 12 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkoxy part, for example phenyl alkoxy having 1 to 6 carbon atoms in the alkoxy part such as benzyloxy , aralkylthio, for example phenylalkylthio of 1 to 6 carbon atoms in the alkyl part, such as benzylthio, g amino groups (-M0), amino substituted / such such as -RHR, wherein R6 represents a C1-6 alkyl group,

carbon atoms, for example methyl or ethyl, aryl-C 6 -C 12 -alkyl in the aryl part and 1 to 6 carbon atoms in the alkyl part, for example phenyl-C 1 -C 6 -alkyl in the alkyl part, such as benzyl, aryl of 6 to 12 carbon atoms such as phenyl, cycloalkyl of 3 to 8 carbon atoms such as for example cyclohexyl, or cycloalkylalkyl of 3 to 8 carbon atoms in the cycloalkyl and 1 to 6-carbon atoms in the alkyl part, such as for example the cyclohexylmethyl 7 '-carboxyl group (-O-H) o O-2 or -GyGOi wherein R ° is defined as above

Aryl groups represented by R1 in the compounds of formula (I) include aryl groups having 6 to 12 carbon atoms, such as phenyl or o- or n-naphthyl groups.

Aralkyl groups represented by R &quot; &quot; include aryl-alkyl groups having 6 to 12 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkyl part, such as phenyl-alkyl having 1 to 6 carbon atoms in the alkyl part, or a- or -naphthyl-C 1 -C 6 -alkyl in the alkyl part, for example benzyl, phenylethyl, phenylpropyl, phenylbutyl, phenylpentyl, &lt; RTI ID = 0.0 &gt; -naphthylmethyl, naphthylthio, naphthylpropyl, naphthylbutyl or naphthylpentyl .

When the group R1 in the compounds of formula (I) and a heteroaralkyl group may be for example a heteroaryl-alkyl group having 3 to 6 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkyl part, such as an optionally substituted group selected from pyrrolylmethyl, furanylmethyl, thienylmethyl, imidazolylmethyl, oxazolylmethyl, thiazolylmethyl, pyrazolylmethyl, pyrrolidinylmethyl, pyridinylmethyl, pyrimidinylmethyl, morpholinylmethyl, or piperazinylmethyl.

Heteroarylthioalkyl groups represented by R 1 include heteroarylthioalkyl groups having 3 to 6 ring atoms.

carbon atom in the heteroaryl part and 1 to 6 carbon atoms in the alkyl group, such as optionally substituted groups selected from pyrrolylthiomethyl, furanylthiomethyl, oxazolylthiomethyl, thiazolylthiomethyl, pyrazolylthiomethyl, pyrrolidinylthiomethyl, pyridinylthiomethyl, pyrimidinylthiomethyl, morpholinylmethyl, or piperazinylthiomethyl.

The aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl groups are represented by S, and the groups represent &quot; by &amp; in the compounds of formula (I) may each be optionally substituted in the cyclic part of the group by a factor of 1, 2 or more substituents selected from halogen atoms, for example fluorine atoms , chlorine, bromine or iodine, or alkyl groups having 1 to 6 carbon atoms, for example methyl or ethyl, alkoxy groups having 1 to 6 carbon atoms for example methoxy or ethoxy, alkylenedioxy groups having from 2 to 6 carbon atoms, carbon, for example ethylenedioxy, haloalkyl of 1 to 6 carbon atoms, for example trifluoromethyl, alkylamino of 1 to 6 carbon atoms, for example methylamino or ethylamino, dialkylamino of 1 to 6 carbon atoms in each of the parts alkyl, for example dimethylamino or diethylamino, amino (-NEL), nitro, cyano, hydroxyl (-OH), carboxyl (-CO2 H), -OR g, wherein R4 is defined as above, alkylcarbonyl with 1 to 6 carbon atoms in the alkyl part, for example acetyl, sulfonyl (-SQgH), alkylsulfonyl with 1 to 6 carbon atoms carbon atoms in the alkyl part, for example methylsulfonyl, aminosulfonyl, (G2 M2), alkylaminosulfonyl having 1 to 6 carbon atoms in the alkyl part, for example methylamino sulfonyl or ethylaminosulfonyl, dialkylaminosulfonyl having 1 to 6 carbon atoms in each of the , for example dimethylaminosulfonyl or diethylaminosulfonyl, carboxamido (-OMO 2), alkylaminocarbonyl having 1 to 6 carbon atoms in the alkyl part, for example methylaminocarbonyl or ethylaminocarbonyl, dialkylaminocarbonyl having 1 to 6 carbon atoms in each of the alkyl parts, for example - 7 -

(-NHSO2 H), alkylsulphonylamino having 1 to 6 carbon atoms in the alkyl part, for example methylsulphonylamino or ethylsulphonylamino, dialkylsulphonylamino having 1 to 6 carbon atoms in each of the parts alkyl, for example dimethylsulphonylamino or diethylsulfonylamino. Note that whenever two or more R substituents are present, they do not necessarily have to be identical atoms and / or groups. The substituents R * & &quot; 0 may be present on any ring carbon atom, other than that which is attached to the remainder of the molecule of formula (I). Thus, for example, phenyl groups having substituents may be present at the 2, 3, 4, 5, or 6 positions relative to the carbon atom of the ring linking the remaining molecule. 3 4 *

When the groups R and R in the compounds of formula (I) are alkyl groups, they may be for example alkyl groups having 1 to 6 carbon atoms, such as methyl or ethyl groups. Group B? in the compounds of formula (I) may be an optionally substituted straight-chain or branched alkyl group of 1 to 6 carbon atoms, for example a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n or an alkenyl group having 2 to 6 carbon atoms, for example ethenyl or 1-propenyl, optionally interrupted by one or more -O- or -S- atoms, or -N (R 2) wherein R3 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, such as a methyl group.

The possible substituents which may be present on alkyl or R 2 -alkenyl groups include aryl-alkyl groups having 6 to 12 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkyl part, such as phenyl 1 to 6 carbon atoms in the alkyl part, optionally -

for example benzyl groups, aryl-alkoxy groups having 6 to 12 carbon atoms in the aryl part and 1 to 6 carbon atoms in the alkoxy part, such as phenyl-alkoxy groups having 1 to 6 carbon atoms in the alkoxy part, optionally substituted aryl groups, for example benzyloxy groups, aryl groups having 6 to 12 carbon atoms, for example optionally substituted phenyl groups, heteroaryl groups having 3 to 8 carbon atoms, for example optionally substituted groups selected from indole, imidazole or quinoline, alkoxy-aryl having 6 to 12 carbon atoms in each of the aryl parts and 1 to 6 carbon atoms in the alkoxy part, for example benzyloxy-phenyl, -OH, -SH, alkylthio having 1 to 6 carbon atoms , for example methylthio or ethylthio, carboxy (-CO 2 H), amino carboxamido (-OOIH 2) or guanido groups -MC (NH 2) = HH. The possible substituents present in these groups may be substituents R as mentioned above.

When X in the compounds of formula (I) represents a substituted amino group, this may be for example a group of the formula -HRXXRX wherein Rx and Rx, which may be the same or different, each represent a hydrogen atom (with the proviso that when one of the substituents R or R1 is a hydrogen atom, the other is not) or a straight-chain or branched alkyl group optionally substituted, e-terminally interrupted by one or more -O- or -S - or NH (O) O-groups or R 1 and R 2 together with the nitrogen atom to which they are attached may form a cyclic amino group having from 3 to 6 ring atoms optionally substituted with one or more other heteroatoms selected from -O- or n-S-, or -U (R) groups. 11 12

When R and / or R represent an alkyl group, this may be for example an alkyl group having 1 to 6 carbon atoms, such as a methyl, ethyl, n-propyl, iso-

propyl, n-butyl, isobutyl, s-butyl or t-butyl optionally interrupted by one or more -O- or -S-, or group -R (R ') - or aminocarbonyloxy and may be for example methoxymethyl, ethoxymethyl, ethoxyethyl or ethylaminoearbonyloxymethyl group. Possible substituents which may be present in these groups include hydroxyl (-OH), carboxyl (-ΟΟΗ), esterified carboxy (-CONO), carboxamido (-OONH0), substituted carbamate, for example a -COHR group in which 1 → 1 → 12 is defined as above, amino (-HEL), substituted amino, for example a group of formula -NR1; , or aryl, for example aryl of 6 to 12 carbon atoms, such as phenyl, optionally substituted by 1, 2 or more H10 substituents chosen from those already enumerated above in relation to the R group.

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

When the group Σ is a substituted hydroxyl group it may be for example a group -O RXJ- wherein RXJ &quot; and defined as above, but different from a hydrogen atom.

The salts of the compounds of formula (I) include pharmaceutically acceptable salts, for example acid addition salts derived from organic or inorganic acids, such as the hydrochlorides, hydrobromides, hydroiodides, p-toluenesulfonates, phosphates, sulfates, perchlorates, acetates, trifluoroacetates , propionates, citrates, malonates, succinates, lactates, oxalates, tartrates and benzoates.

Salts with bases may also be formed. These salts include salts derived from organic or inorganic bases, for example alkali metal salts such as sodium or potassium salts,

alkaline earth metal salts, such as magnesium or calcium salts, and salts of organic amines, such as morpholine, piperidine, dimethylamine or diethylamine salts.

When the group R in the compounds of the present invention is a esterified carboxyl group, it may be a metabolically dissociable ester of the formula -CH 2 CH 2 wherein R 2 may be an ethyl, benzyl, phenylethyl, phenylpropyl, - or β-naphthyl, 2,4-dimethylphenyl, 4-t-butylphenyl, 2,2,2-trifluoroethyl, 1- (benzyloxy) benzyl, 1- (benzyloxy) ethyl, 2-methyl- propionyloxypropyl, 2,4,6-trimethylbenzoyloxymethyl or pivaloyloxymethyl.

In the compounds of formula (I) the group R1 may in particular be an alkyl group having 1 to 6 carbon atoms, such as a methyl group, an aralkyl group such as the benzyl group, an aryl-thioalkyl group such as a phenylthio-methyl, or a heteroarylthioalkyl group such as a thienylthiomethyl, pyridinyl-thiomethyl or pyrimidinylthio-methyl group, or is in particular a hydrogen atom.

The groups R1 and R2 in the compounds of formula (I) may each be in particular a methyl group, or especially a hydrogen atom. The group 1 in the compounds of formula (I) may in particular be an alkyl group having 1 to 6 carbon atoms, for example an isopropyl group or an isobutyl group, or a benzyl, benzyloxybenzyl or indolylmethyl group optionally substituted. The group X in the compounds of formula (I) may be in particular an amino group (-NH 2) or a group. Particular -1¾¹½12 groups are -MK12 groups. Typical groups of the type include those in which R represents an al-

for example a methyl, ethyl or n-propyl group, optionally interrupted by one or more -O- or -S- or -N (R) -alkyl groups, for example -NH - or -H (OH) -J or aminocarbonyloxy and optionally substituted by a hydroxyl, carboxy, carboxyalkyl, for example carboxymethyl, carboxamido, amino, - for example dialkylamino group having 1 to 6 carbon atoms in each of such as dimethylamino, alkylamino having 1 to 6 carbon atoms, such as methylamino, or cyclic amino having 5 to 6 carbon atoms, such as morpholinyl, pyrrolidinyl or pyridinyl-J or phenyl optionally substituted by 1, 2 or

&quot; &quot; 1Q plus R substituents.

A particularly useful group of compounds according to the invention are compounds of formula (I) in which R 2 represents an optionally substituted phenylpropyl group.

Another particularly useful group of compounds of formula (I) are those in which X represents an α-amino or substituted amino group.

In general in the compounds of formula (I) the groups R1 and R2 are preferably each a hydrogen atom.

A further preference is the R group in the compounds according to the invention and a -CH2 OH group or a -CH2 H group or a metabolically dissociable ester thereof. In a particular preference, however, R represents a group -OQNHQH or a group -COgH. An especially useful group of compounds according to the invention has formula (Ia)

Wherein R, R 1, R 2 and X are as defined for formula (I); and the salts, solvates and hydrates thereof.

A particularly useful group of compounds of formula (la) are those wherein R 1 is -CQNHGH or 2-OH; R represents an optionally substituted phenylpropyl group; X represents an amino (-NH 2) or substituted amino; and the salts, solvates and hydrates thereof.

In the compounds of formula (la) X may be -NH- group or -ΝΕ 12 12 group as defined for the compounds of formula (I). The group R may in particular be an alkyl group having 1 to 6 carbon atoms, such as an isopropyl or isobutyl group, or a optionally substituted group selected from benzyl, benzyloxybenzyl or indolylmethyl.

The compounds according to the invention may be prepared by the following processes. In the description and formulas below the R, R, R, R, R, R, R, and X groups are as previously defined except where expressly indicated to the contrary. Note that functional groups such as amino, hydroxy or carboxy, present in the various compounds described below and intended to be maintained in the molecule, may need to be protected before any reaction is initiated. In these circumstances the removal of the nozzle groups may be the final step in a particular reaction. The 13-

Suitable amino or hydroxyl blocking groups include the benzyl, benzyloxycarbonyl or t-butyloxycarbonyl groups. These may be removed from a blocked derivative by catalytic hydrogenation, for example using hydrogen in the presence of a metal catalyst, for example palladium on a carrier, such as charcoal, in a solvent such as an alcohol, for example methanol, or by treatment with trimethylsilyl iodide or trifluoroacetic acid in an aqueous solvent. Suitable carboxyl blocking groups include benzyl groups, which may be removed from a blocking derivative by the methods discussed above, or alkyl groups, such as the t-butyl group, which may be removed from a blocked derivative by treatment with trifluoroacetic acid in an aqueous solvent. Other suitable blocking groups may be evident, as well as the methods for their use. The formation of a blocked amino, hydroxyl or carboxyl group can be achieved using conventional alkylation or esterification processes, for example those which are described below.

Thus, according to a further aspect of the present invention, a compound of formula (I) may be prepared by the coupling of an acid of formula (II).

OH (II) or an active derivative thereof, with an amine of formula (III)

R (III) followed by removal of any blocking group present.

The active derivatives of acids of formula (II) include, for example, acid anhydrides or acid halides, such as acyl chlorides. The coupling reaction may be carried out using standard conditions for such an amination reaction. Thus, for example, the reaction may be carried out in a solvent, for example an inert organic solvent such as an ether, for example a cyclic ether such as tetrahydrofuran, an amide for example a substituted amide, such as dimethylformamide, or a hydrocarbon halogenated such as dichloromethane, at a low temperature, for example from -30 ° C to ambient temperature, such as -20 ° C to 0 ° C, optionally in the presence of a base, for example an organic base such as amine, for example triethylamine or a cyclic amine, such as N-methylmorpholine. When using an acid of formula (II) the reaction may be further carried out in the presence of a condensing agent, for example a diamide such as N, N-diethylhexylcarbodiimide, preferably in the presence of a triazole such as 1-hydroxybenzotriazole. Alternatively the acid may be reacted with a chloroformate, for example ethyl chloroformate, prior to reaction with the amine of formula (III).

The free hydroxyl or carboxyl groups in the starting compounds of the formulas (II) in which R 2 represents -CORH 2 OH or CO 2 H 7 and (III) may need to be protected from

the coupling reaction. Suitable blocking groups, as well as the methods for their removal, may be those already described above.

It should be noted that where a particular stereoisomer of formula (I) is desired, it can be obtained by reacting a mixture of isomers after the coupling reaction of an acid of formula (II) and an amine of formula (III). Conventional resolution techniques may be used, for example, separation of the isomers by chromatography, for example by use of high performance liquid chromatography. Where desired, however, suitable homo-pharynical starting materials may be used in the coupling reaction to drive a particular stereoisomer of formula (1). Thus, in a particular process, a compound of formula (Ia) can be prepared by reacting a compound of formula (IIa)

with an amine of formula (IIIa)

as described above.

The intermediate acids of formula (II)

R represents a carboxyl group or a esterified carboxyl group, can be prepared by hydrolyzing a corresponding ester of formula (IY)

(IV) in which R represents an alkyl group, for example a methyl or t-butyl group, for example using trifluoroacetic acid, or when R3 represents a methyl group using the enzymatic hydrolysis, as for example with Ot In this reaction the enzymatic hydrolysis (for example as will be described more particularly in the examples below) provides particularly useful a method of selecting the isomers. The ester of formula (IY) may be prepared by esterifying the corresponding acid of formula (Y)

using an appropriate acyl halide, for example an acyl chloride in a solvent such as an alcohol, for example methanol, at a low temperature, for example on the order of 0 ° C.

The acids of formula (Y) may be prepared by alkylating a compound of formula (YI)

with a suitable halide, for example a compound of formula R 1 Hal, in which Hal represents a halogen atom such as a chlorine atom or &quot; bromine, in the presence of a base, for example an alkoxide, such as sodium in a solvent such as an alcohol, for example ethanol, at room temperature, followed by decarboxylation using for example concentrated hydrochloric acid at an elevated temperature, for example at reflux temperature.

The intermediate compounds of formula (VI) are either known compounds, or may be prepared by methods analogous to those used for the preparation of the known compounds.

The intermediate acids of formula (IV) in which R represents a -CGNHOH group or a blocked derivative thereof may be prepared by reacting an anhydride of formula (VII)

with a hydroxylamine such as G-benzylhydroxylamine, in a solvent such as tetrahydrofuran, at a low temperature,

xa, for example of the order of -20 ° C, followed, where appropriate, by removal of the blocking group, as already described above.

Intermediate anhydrides of formula (7) may be prepared, for example, by heating, for example at reflux temperature, a diacid of formula (V) in which R 2 represents -GQ 2 H with an acyl chloride such as acetyl chloride.

Homophylic acids of formula (IIa) may be prepared according to another feature of the invention by oxidation of an oxazolidinone of formula (7111)

(in which Ph represents a phenyl group) using an oxidizing agent such as a peroxide, for example hydrogen peroxide, in a solvent such as an ether, for example a cyclic ether, such as tetrahydrofuran, at a low temperature, for example order of G ° O, followed by treatment with a base, such as lithium hydroxide, at an elevated temperature.

The compounds of formula (IIIII) are novel, are particularly useful intermediates for the preparation of stereoisomers of formula (Ia) and form a further aspect of the invention.

The compounds of formula (7III) may be prepared by reaction of an acyl halide of formula RGELCH

(Wherein Hal represents a halogen atom, such as a chlorine, bromine or iodine atom) with a solution of (s) -4- (phenylmethyl) -2-oxazolidinone in the presence of a solution of (R) base such as n-butyllithium in a solvent such as tetrahydrofuran at a low temperature, for example about -78 ° C.

The acyl halides of the formula RCH 2 CH (R 2) CO-Hal may be prepared by treating the corresponding known ethers of the formula RCH 2 H (R 2) OH with conventional halogenating agents, for example thionyl halides, under standard reaction conditions.

In another process according to the invention, a compound of formula (I) in which R represents a carboxyl group may be prepared by the decarboxylation of a corresponding compound of formula (IX). 2 v.

R O

This reaction may be carried out using standard conditions, for example by heating a compound of formula (IS) in an inert solvent, such as an aromatic hydrocarbon, for example xylene, at the reflux temperature.

The intermediate acids of formula (IX) may be prepared by reacting a blocked acid of formula (I)

in which S represents a blocked carboxyl group such as a benzyloxycarbonyl group and Z 'represents a blocking group such as a benzyl group with an amine of formula (III) using reagents and conditions as described above for the coupling of compounds of formulas (II) and (III), followed by removal of the blocking groups.

Intermediates of formula (X) may be prepared by treatment of an appropriate malonic Ister of the formula RCE-CO-Z1 with a halide of formula (XI)

(in which Hal represents a halogen atom, for example a chlorine or bromine atom) in the presence of a base, such as potassium t-butoxide, in a solvent such as dimethylformamide at room temperature.

The halides of formula (H) may be prepared by halogenation and subsequent decarboxylation of a diacid of formula (XII)

using for example a halogenating agent, such as bromine, in a solvent, such as diethyl ether, at room temperature, followed by heating the resulting halogenated intermediate in a solvent, such as an aromatic hydrocarbon, e.g. xylene, at the temperature of reflux.

The intermediate compounds of formula (XII) may be prepared by hydrolyzing the corresponding dhe-1-yl ester (for example dimethyl or diethyl ester) using a base such as sodium hydroxide or potassium hydroxide in a solvent such as an alcohol , for example methanol, at reflux temperature. The starting compounds, dialkyl esters, or are known compounds can be prepared by methods analogous to those used for the preparation of the known compounds, for example as described herein in the examples.

The compounds of formula (I) may also be prepared by interconversion of other compounds of formula (I). Thus, for example, a compound of formula (I) in which R represents an -OMOHOH group can be prepared by reacting a corresponding acid of formula (I) in which R represents a group -COOH, or an active derivative thereof for example an acid chloride or an acid anhydride) with hydroxylamine or an O-blocked derivative of a salt thereof. The reaction can be carried out using the reagents and conditions described above in the preparation of the compounds of formula (I) from the starting compounds of formulas (II) and (III). 22 -

In another interconversion process the compounds of formula (I) in which R 2 represents -CO 2 H and / or X contain a -OO H group may be prepared by hydrolysis of the corresponding esterified compound (for example, when R 2 is - and / or X contains an analogous group) using standard procedures, for example by treatment with a base, for example with an alkali metal hydroxide, such as lithium hydroxide, in a solvent such as an aqueous alcohol, for example aqueous methanol , or by treatment with an acid such as a mineral acid, for example hydrochloric acid, in the presence of a solvent, for example dioxane.

Similarly, the esters of formula (I), for example when R 2 represents a group -CH 2 R 3 and / or X contains a group -CO 2 R 5, can be prepared by reacting the corresponding acids in which R 2 represents a - H and / or X contains a -GGIII group or an active derivative thereof, with an alcohol, of formula OH, using standard conditions.

The compounds according to the invention are potent and selective inhibitors of gelatinase. The activity and the selectivity of the compounds may be determined by the use of appropriate enzymatic inhibition tests, for example as described in Example A below. In our assays using this working step the compounds according to the invention have been shown to inhibit gelatinase with IL values in the picomolar-nanomolar range, and to have a selectivity for gelatinase relative to stromelysin of about 40 fold, or greater, and have a selectivity for gelatinase relative to collagenase of about 20-fold or greater. The ability of the compounds of the invention to prevent invasion of tumor cells can be demonstrated in the standard mouse assay model.

Thus, briefly,

of a tumor cell line exhibiting gelatinase-dependent invasion, and the ability of the compounds of the invention to reduce subsequent collection of lung tumor can be assessed according to a standard procedure. In our assays the compounds according to the invention, when administered intravenously at the rate of 1 mg / kg to rats in the test model described above, reduced the colonization of lung tumor to negligible levels.

The compounds according to the invention can therefore be shown to be useful for the prevention of tumor cell metastases and invasion. The compounds may therefore be used in the treatment of cancer, particularly in combination with radiotherapy, chemotherapy or surgery, or in patients presenting with prior tumors, to control the development of tumor metastases. Thus, according to a further aspect of the invention, there is disclosed a compound of formula (I) for use in the treatment of cancer, to control the development of tumor metastases. Cancers can include, in particular, breast cancer, melanoma, lung cancer, cachexia, neck, or bladder.

For use according to this aspect of the invention the compounds of formula (I) may be conventionally formulated, optionally with one or more physiologically acceptable carrier substances, diluents or excipients.

Thus, according to a further aspect of the invention there is shown a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable diluent, carrier or excipient.

In yet another aspect of the invention there is disclosed a process for the preparation of a pharmaceutical composition comprising contacting a compound of formula (I)

pharmaceutically acceptable carrier, diluent or excipient.

The compounds for use according to the present invention may be formulated for oral, buccal, parenteral or parenteral 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 tablets prepared by conventional means with pharmaceutically acceptable excipients such as binders (for example pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); eargas (for example lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (for example magnesium stearate, talc or silica); disintegrants (for example potato starch or sodium glycol); or wetting agents (for example sodium lauryl sulphate). The tablets may be coated by methods known in the art. Liquid compositions for oral administration may take the form of, for example, solutions, syrups or suspensions, or may be presented in the form of a dry product for constitution with water or other suitable carriers prior to use. These liquid compositions may be prepared by conventional means with pharmaceutically acceptable additives, such as suspending agents, emulsifiers, nonaqueous carrier substances; and preservatives. The compositions may also contain buffers, flavoring, coloring and sweetening salts, as appropriate.

Compositions for oral administration may conveniently be formulated so as to provide a controlled release of the active compound. - 25 -

Compositions for oral administration may take the form of tablets or lozenges formulated conventionally.

The compounds of formula (I) may be formulated for parenteral administration by ingestion, for example by global injection or continuous infusion. Formulations for injection may be in unit dosage form. Injection compositions may take such forms as suspensions, solutions or emulsions in both oily and aqueous carrier substances, and may contain formulatory agents such as suspending agents, stabilizers and / or dispersing agents. As an alternative the active ingredient may take the form of powder for constitution with an appropriate carrier, for example sterile, pyrogen-free water, prior to use.

The compounds of formula (I) may also be formulated into rectal compositions, such as suppositories or retention enemas, for example containing conventional suppository bases, such as cocoa butter or other glycerides.

In addition to the formulations described above, the compounds of formula (I) may also be formulated into depot compositions. These long-acting formulations may be administered by implantation or by intramuscular injection.

For nasal administration or administration by inhalation the compounds for use in accordance with the present invention are conveniently provided in the form of an aerosol composition for pressurized packs or by nebulizer using an appropriate propellant, for example dichlorodifluoromethane, trichlorofluoromethane, dichloro-

rotortrafluoroethane, carbon dioxide or other suitable gases.

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

Doses of the compounds of formula (I) used to control the development of tumor metastases vary according to the condition of the patient to be treated, but in general may range from about 0 to 5 mg to 50 mg / kg of body weight body, in particular from about 1 mg to 40 mg / kg body weight. The dosage units may vary depending on the route of administration of the compound, according to standard practice.

DESCRIPTION OF SPECIFIC EMBODIMENTS The invention is illustrated below in the following non-limiting examples. In the examples the following abbreviations are used: BGGI-β, β-dicyclohexyl-carbodiimide DMF-dimethylformamide TH1- tetrahydrofuran trifluoroacetic acid RPHPI0- high performance liquid chromatography, reverse phase HOBT- N-hydroxybenzotriazo1. - 27 -

Reference Example C 4-N- (hydroxyamino) -2R-isobutyl-succinyl] -1-valine-L-alanine-amide R, S-isobutyl succinic acid

Sodium ethoxide was prepared by adding sodium metal (2.5 g, 108 mmol) to dry ethanol (150 mL) under nitrogen atmosphere. Triethyl 1,1,2-ethanetricarboxylate (26.6 g, 25 ml, 108 mmol) was added and the mixture was stirred at room temperature (1 A) for 20 minutes. Isobutyl bromide (19.12 g, 15 ml, 108 mmol) was added dropwise over 1 hour and the solution was boiled under reflux overnight. The precipitated sodium bromide was removed by filtration and the filtrate was concentrated in vacuo. The residue was treated with cold water (200 ml) and extracted with diethyl ether (3 x 100 ml). The organic phase was dried over sodium sulfate and concentrated to give a clear oil (32.2 g) which was heated to reflux with concentrated hydrochloric acid for 96 hours. After cooling a white crystalline solid precipitated which was isolated by filtration, washed with ice-water and dried in vacuo to give the title compound K (11.0 g). 1H NMR (CDCl3) Î'0.85 (3H, d, J = 6Hz), 0.90 (3H, d, J = 6Hz), 1.3-1.45 (1H, m), 1.55-1.75 (2H, m), 2.50 (1H , dd, J = 6 and 18Hz), 2.70 (1H, dd, J = 9 and 18Hz), 2.85-2.95 (1H, m).

28 -

3- (RtS) -isobutyl-succinic acid trihydrate L

The diacid K (10.21 g, 59 mmol) was treated with acetyl chloride (27 mL, 376 mmol) at reflux for 2.5 hours. The volatiles were removed under reduced pressure to give (37 g, 100%) as a tan oil. (3H, d, J = 6Hz), 1.05 (3H, d, J = 6Hz), 1.48-1.90 (3H, m), 2.65 (1H, dd, J = 7 and 18Hz ), 3.10 (1H, dd, J = 9 and 18Hz), 3.15-3.25 (1H, m). acid

4- (H-benzyloxyamino) -2-R, S-isobutyl] succinic acid M

O-Benzylhydroxylamine (7.8 g, 63.4 mmol) in anhydrous tetrahydrofuran (50 mL) was added dropwise (over 1 hour) to a solution of the anhydride I (9.37 g, 60.0 mmol mmol) in anhydrous tetrahydrofuran (100 ml) at -20 ° C. After stirring for an additional hour the volatiles were removed in vacuo and the residue was taken up in ethyl acetate. After washing with - 29

1.0 M hydrochloric acid (3 times) the organic phase was dried over magnesium sulfate and evaporated to give a white solid. The crude solid was dissolved in hot diethyl ether and filtered. Colorless crystals of the acid M were deposited on standing (6.7 g, 41%). (3H, m), 2.1-2.4 (2H, m), 2.8-3.0 (1H, m), 4.85 (2H, s) , 7.3 (5H, bs), 8.6 (1H, bs). (502 mg, 1.8 mmol) was dissolved in anhydrous tetrahydrofuran (20 ml) and cooled to -78øC. The combined organic extracts were dried over sodium sulfate and evaporated to dryness. -20Â ° C. Ethyl chloroformate (245 mg, 233 Âμl, 1.8 mmol) and N-methylmorpholine were added and the suspension was allowed to stand for one hour at -20Â ° C. A solution in dimethylformamide (10 ml) of 1-valine-1-alanine amide (500 mg) was added dropwise. After this addition was completed, the cooling bath was removed and the reaction mixture was allowed to warm to room temperature overnight. The organic solution was poured into 10% hydrochloric acid and extracted with ethyl acetate (3 times). The organic phase was dried over magnesium sulfate and concentrated in vacuo to give a solid.

This solid was dissolved in degassed methanol (20 ml) and subjected to hydrogenolysis using 5% palladium-carbon and hydrogen gas. After one hour at room temperature the catalyst was isolated by filtration and the product was purified by RPHPLO using 0.1% -> 0.1% IFA / OH] OI (43:57) under isocrylic conditions to give the title compound. 1 H NMR (CD3 OD) Î'0.95 (12H, m), 1.35 (3H, d, J = 6Hz), 1.95 (3H, m), 2.20 (1H, m), 4.35 (2H, m). - 30 -

The following compounds of examples 1 to 4 were prepared in a manner analogous to that of the reference compound, using the appropriate starting compounds.

Example 1 4-N- (hydroxyamino) -2R-phenylethyl-succinyl] -leucine-1- (2-phenylethyl) -amide 1H NMR (Q3 &gt; 3OD) 7.15-7.30 (10H, mult) ; 4-40 (IR, mult, ECHCO); (2H, mult, OCH2), 2.20-2.85 (7H, mult, CHCO + CH2ir), 1.50-1.95 (5H, mult, CSC) 0.95 (6H, dd,

Example 2 n- (N-hydroxyamino) -2R-phenylpropylsuccinyl] -L-leucine-E- (2-phenylethyl) -amide 4-NMR (CDCl3): 7.1-7.3 (10H, mult, br); 4.30 (1H, dd, EGHOG), 3.35 (2H, mult, 2H2D, 2.15-2.80 (7H, mult, GHCQ + CH2), 1.5-1.75 (7H, mult, CHC), 0.95 (6H, dd, CH3)

Example 3 [4- (1-Hydroxyamino) -2 (R) -isobutylsuccinyl] -1H-pyrazole-1 H-NMR (GD 3 OD) δ 7.65 (1H, d), 7.35 (1H, d), 6.95-7.15 (3H, m), 4.65 (1H, dd), 3.05-3.2 (1H, m), 2.7-2.85 (2H, m), 2.0-2.2 (2H, m) 1.5 (2H, m), 1.05-1.15 (1H, m), 0.7 (3H, d), 0.65 (3H, d)

Example 4 (R) - (E-hydroxyaryain) -2 (R) -isobutylsuccinyl] -β-valine-amide (CD3 OD) <4.15 (3R, d), 2.8-2.9 (1H, m) 2.2-2.35 ( 1H), 1.95-2.15 (2H, m), 1.45-1.55 (2H, m), 1.1-1.2 (1H, m), 0.8-1.05 (12H, m)

Example A The activity and selectivity of the compounds of the present invention may be determined in the manner described below.

All enzyme assays to determine IL values were performed using the peptide substrate Bnp-Pr o-Leu-Gly-leu-Trp-Ala-B-Ar-G-NE. Γ- !. Sharon Stock and Robert D. Gray JBC 264, 4277-81, 1989). Enzymes perform the peptide binding at the G-ly-leu bond, which can be followed fluorimetrically by measuring the increase in the emission of Trp fluorescence associated with the removal of the dinitrophenol blocking group (Dnp).

Essentially the enzyme (for example gelatinase, stromelysin or collagenase) at 0.08-2 nM; a range of inhibitor concentrations (0.1-50 Âμl) and the substrate (approximately 20 Âμm) are incubated overnight in 0.1 M Tris / HCl buffer, pH 7.5, containing 0.1 M NaO1, 10 mM GaO2 and 0.05% Brij 55 or at room temperature, or at 57 ° C, depending on the enzyme. The reaction is stopped by adjusting the pH to the value 4 using a 0.1 M sodium acetate buffer and the S fluorescence is read at an excitation wavelength of 280 nm and at an emission wavelength of 346 nm.

Values can be established using the equation for ultimate inhibition:

Vi = Vo 2ZX7

1 (app (¾ (app) Γ Γ Γ Γ Γ Γ í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í na na na na na -

inhibitor, T. is the initial rate in the presence of inhibitor, C & J ® is the total concentration of the enzyme, and n is the total concentration of the inhibitor in the reaction mixture.

For stromelysin and collagenase it was considered that it approached the actual value JL as it has been found for the hydrolysis substrate. For gelatin the value was determined by performing the analysis at several substrate concentrations. A graph of E ^ (app) in relation to jTS_7 allowed to obtain the actual IC value as the value of the intrasection of the I axis.

The results of the following table were obtained with the compounds according to the invention.

Ki (nM)

Colagenase Stromelysin-1 G-e Latins e-7 2ZB Reference Compound 303 129 33

Compound of example 2 79 16 0.17

- 33 -

Claims (5)

A compound of formula (I): â € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒR2 R5 X (I) in which R represents a -CONHOH, carboxyl (-C0o H) or esterified carboxyl group; R represents a hydrogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, heteroaralkyl or heteroarylthioalkyl atom; R 2 represents a phenylethyl, phenylpropyl or phenylbutyl group, optionally substituted; represents a hydrogen atom or an alkyl group; represents a hydrogen atom or an alkyl group; represents an optionally substituted alkyl or alkenyl group optionally interrupted by one or more -O- or -S- or -N (R ') - groups, wherein R' represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms carbon; X represents an amino (-NH 3) group or substituted amino, hydroxy or substituted hydroxyl; and their salts, solvates and hydrates. A compound according to claim 1 wherein R is -CONHOH or carboxyl (-q2h). - 34 - ........... A compound according to claims 1 or 2 characterized in that R 1, R 2 and R 3 are each a hydrogen atom. A compound according to any one of claims 1 to 3, characterized in that R represents an optionally substituted phenylpro pyl group. A compound according to any of the preceding claims characterized in that X represents an amino or substituted amino group. Compound of formula (la); R2 Η O in which R represents a group -COOH, carboxyl (-CO₂H) or esterified carboxyl; R 2 represents an optionally substituted phenylpropyl group, R 5 represents an optionally substituted alkyl or alkenyl group optionally interrupted by one or more -O- or -S- or -N (R 2) a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, carbon_7; X represents an amino (-NH 3) or substituted amino, M-hydroxyl or substituted hydroxyl; and their salts, solvates and hydrates. 7. A pharmaceutical composition comprising a compound of any one of claims 1 to 6 in combination with a pharmaceutically acceptable diluent, carrier or excipient. A process for the preparation of a compound of formula (I) as defined in claim 1, which process comprises: a) coupling of an acid of formula (II) or an active derivative thereof and / or a blocked derivative thereof, with an amine of formula (III) R (III) or a blocked derivative thereof, followed by removal of any blocking group; or (b) the decarboxylation of a compound of formula (IX) 5 (IX) in order to provide a compound of formula (I) in which R presenta has a group -00 HH; and / or (c) the interconversion of a compound of formula (1) 37