IE911165A1 - Novel compounds - Google Patents

Abstract

Azalactam derivatives processes for their preparation and their use as collagenase inhibitors having structure (I).

Description

The present invention relates to novel phosphorus derivatives, processes for their preparation and their use in medicine. In particular, the present invention relates to their use as inhibitors of enzymes of the collagenase family of neutral metalloproteases, for treating arthritic and other diseases.

The mammalian collagenase family of enzymes comprises a number of proteases, exemplified by interstitial (type I) collagenase itself, the stromelysins (also known as proteoglycanases or transins), fibroblast and polymorphonuclear leucocyte gelatinases (also known as collagen-IV-ases), and 'pump-1' (putative metalloprotease 1, uterine metalloprotease). Membership of the mammalian collagenase family of proteases is evident by possession of a number of highly characteristic and experimentally verifiable properties. [Goldberg et al., J. Biol. Chem. 2610, 6600, 1986; Whitham et al., Biochem. J. 240, 913, 1986; Breathnach et al., Nucleic Acids Res., .15,, 1139, 1987; Muller et al., Biochem. J., 253, 187, 1988; Collier et al., J. Biol. Chem., 263, 6579, 1988; Murphy et al., Biochem. J., 258, 463, 1989; Quantin et al., Biochem.

(N.Y.), 28., 5327, 1989; Birkedal-Hansen, J. Oral Pathol., 17, 445, 1988].

The range of therapeutic applications of the invention described hereinafter reflects the fundamental role of collagen and other proteinaceous substrates of the collagenase family of enzymes in the connective tissue matrix throughout the body. Applications extend to clinical interventions in many diseases and phenomena involving the destruction of collagen and other connective tissue components, and also normal or disordered tissue remodelling.

IE 91 □ B2957 Inhibitors of the collagenase family of enzymes are considered to provide useful treatments for: arthritic diseases, such as rheumatoid and osteoarthritis, soft tissue rheumatism, polychondritis and tendonitis; bone resorption diseases, such as osteoporosis, Paget's disease, hyperparathyroidism and cholesteatoma; the enhanced collagen destruction that occurs in association with diabetes; the recessive classes of dystrophic epidermolysis bullosa; periodontal disease and related consequences of gingival production of collagenase, or of PMNL collagenase release following cellular infiltration to inflamed gingiva, including by combating the greater susceptibility of diabetes patients to periodontal disease; corneal ulceration, e.g. that induced by alkali or other burns, by radiation, by vitamin E or retinoid deficiency; ulceration of the skin and gastro-intestinal tract, and abnormal wound healing; postoperative conditions, including colonic anastomosis, in which collagenase levels are raised; cancer, where members of the collagenase family of enzymes have been implicated in the neovascularization required to support tumour growth and survival [P. Basset et al. Nature, 348, 699, 1990] in the tissue remodelling required to accommodate the growing primary and secondary tumours, and in the penetration of tumour cells through the basement membrane of the vascular walls during metastasis; and demyelinating diseases of the central and peripheral nervous systems, including syndromes in which myelin loss is the primary pathological event and those in which demyelination follows axonal atrophy. The degradation of myelin in these diseases, exemplified by multiple sclerosis, is mediated by members of the collagenase family of enzymes.

B2957 - 3 As a particular example of the therapeutic value of inhibitors of the collagenase family of enzymes such as are disclosed in the present invention, chronic arthritic diseases leading to extensive loss of the collagen, proteoglycan and elastin components of the cartilage, bone and tendons within the joints, should be amenable to treatment with inhibitors of the collagenases, proteoglycanases (stromelysins) and gelatinases currently thought to be the major enzymes involved.

These enzymes have been detected in extracts of synovial and cartilage tissue, and have also been extensively studied in tissue cultures of a wide range of connective tissues. Apart from control of the biosynthesis, secretion and activation of the enzymes, the most important natural regulation of these enzymes in normal and diseased states, is considered to be the endogenous production of inhibitors such as the family of Tissue Inhibitor of Metalloproteases (TIMPS), and alpha-2 macroglobulin. An imbalance between the local levels of the proteolytic enzymes and natural inhibitors will allow destruction of connective tissue components to occur.

The compounds described in the present invention, being synthetic and low molecular weight inhibitors of this family of enzymes, offer a therapeutically useful way in which a more normal or non-pathological balance between inhibition and enzymic activity can be restored: they thus act to complement and supplement the endogenous enzyme inhibitors. Indeed, because these enzymes usually act only within restricted pericellular environments, before being inactivated by inhibitors circulating in the blood and present in most inflammatory exudates, the low molecular weight inhibitors disclosed here may be more IE 9111S5 Β2957 - 4 effective than endogenous proteinaceous inhibitors that are excluded by their size from the localized regions of connective tissue destruction.

European Patent Application 88310492.9 (Beecham Group) discloses a class of phosphorus derivatives having activity as inhibitors of collagenase and utility in the treatment of rheumatoid arthritis and related diseases in which collagenolytic activity is a contributing factor.

Novel structurally related compounds have now been discovered, which are collagenase inhibitors and thus of potential utility in the treatment of diseases in which collagenolytic activity and tissue remodelling is implicated.

According to the present invention there is provided a compound of general formula (I), or a salt, solvate or hydrate thereof: R, (I) in which, R is hydrogen, alkyl or optionally substituted benzyl; R^ is hydrogen or C-j__g alkyl; - 5 Ε911165 Β2957 Rg is Cg_g alkyl; and Rg and R4 are joined together as - (CHg)p-X-(CHg)g- where p is an integer from 1 to 9, q is an integer from 2 to 10, and the moiety -(CHg)p- is adjacent to the carbon atom bearing Rg marked with an asterisk in formula (I), and X is -NRg- where Rg is selected from hydrogen, Cg_galkyl, Cg_galkanoyl, Cg_galkoxycarbonyl, aroyl, aralkyl or aralkyloxycarbonyl in each of which the aryl· moiety is optionally substituted.

Unless otherwise specified, each alkyl group is preferably a Cg_g group, more preferably Cg_g, and may be a straight chain or branched. An aryl moiety is preferably phenyl.

Optional substituents for an aryl moiety may be selected from OH, Cj_g alkyl, Cg_g alkoxy and halogen.

R is preferably hydrogen, methyl, ethyl or benzyl, especially hydrogen.

Values for Rg include hydrogen, methyl, ethyl, isopropyl and n-butyl. As an alkyl group, Rg is preferably methyl or ethyl.

Rg is preferably a Cg alkyl group, such as n-butyl, iso-butyl or sec-butyl, especially iso-butyl Values for Rg and Rg together include -(CHg)p-X-(CHg)gwhere p and q have values such that Rg and Rg form part of an 11- or 13 to 16-membered azalactam structure, and X is -NRg- where Rg is hydrogen, methyl, benzyl, t-butoxycarbonyl or benzyloxycarbonyl.

B2957 - β Most preferred are compounds where R3 and are joined together as - (CH2) -X-(CH2)g- where ρ is 4 and q is 5 or p is 3 and q is 6 or p is 4 and q is 6 or p is 4 and q is 3 and X is -NR^- where is hydrogen or methyl.

The compounds of formula (I) may form salts with bases e.g. sodium hydroxide. The compounds of formula (I) have a basic nitrogen atom and may form acid addition salts e.g. with hydrochloric acid. Such compounds form part of the present invention.

Where compounds of formula (I), or salts thereof, form solvates or hydrates, these also form an aspect of the invention .

The compounds of formula (I) have at least two, and may have three or more asymmetric centres and therefore exist in more than one stereoisomeric form. The invention extends to all such forms and to mixtures thereof, including racemates, and diastereoisomeric mixtures.

Preferred isomers are those having the S.-configuration at the chiral centre bearing R2 and the ^-configuration at the chiral centre bearing R^, marked with an asterisk in formula (I).

The compounds of formula (I) or their salts, solvates or hydrates are preferably in pharmaceutically acceptable form. By pharmaceutically acceptable form is meant, inter alia, of a pharmaceutically acceptable level of purity excluding normal pharmaceutical additives such as diluents and carriers, and including no material considered toxic at normal dosage levels.

IE 911' B2957 The compounds of formula (I) or their salts, solvates or hydrates are preferably in substantially pure form.

A substantially pure form will generally contain at least 50% by weight, preferably 75%, more preferably 90% and still more preferably 95% or 99% or more of the compound of formula I or its salt or solvate.

Compounds of formula (I) or their salts, solvates or hydrates may be isolated as crystalline solids or in the form of foams or gums.

A preferred pharmaceutically acceptable form is the crystalline form.

The present invention provides the compounds of formula (I) or pharmaceutically acceptable salts, solvates or hydrates thereof for use as active therapeutic agents, particularly as agents for treatment of conditions in which degradation of connective tissue and other proteinaceous components of the body occurs, such as musculo-skeletal disorders resulting from collagenoiytic activity, particularly rheumatism and/or arthritic conditions, and tissue remodelling.

Compounds of formula (I) also have potential utility in the treatment of cancer; for preventing myelin degradation in the central and peripheral nervous system; and in other conditions in which members of the collagenase family of neutral metalloproteases have pathological or other roles.

The present invention also provides a process for the preparation of a compound of formula (I) which comprises converting a group R^q to hydrogen by cleaving a group R^q from a compound of formula (II) : B2957 R R ·? O = P OR OR / R '4 R (II) wherein R2q is alkyl, optionally substituted phenyl, or optionally substituted benzyl and R21 is hydrogen, alkyl, optionally substituted phenyl, or optionally substituted benzyl and R^, R2, R3 and R^ are as defined in formula (I), and where necessary, converting R21 to hydrogen.

Cleavage of R2q, anci wilere necessary R2-j., may he carried out in aqueous acid or alkali or using a trimethylsilyl halide, preferably bromotrimethylsilane, in an inert solvent, for example dichloromethane or acetonitrile. Benzyl esters may alternatively be removed by hydrogenolysis or other standard debenzylation procedures. Phenyl residues may be removed by hydrogenation over platinum oxide.

When both R2q and R21 are al^yl' cleavage of R2q only, to give to a compound of formula (II) in which R2q is hydrogen and R2^ alkyl, which is a compound of formula (I) in which R is alkyl, may be carried out by treatment with excess alkali under mild conditions, for example with aqueous sodium hydroxide in an alcoholic solvent at room temperature.

Similarly, where R2q is optionally substituted benzyl and R22 is alkyl, the benzyl group only may be cleaved by B2957 - 9 hydrogenation to give a compound of formula (II) in which R.2Q is hydrogen and R21 is alkyl.

Cleavage of an R2j_ alkyl group may thereafter be carried out as described above to give a compound of formula (I) in which R is hydrogen.

When R in a compound of formula (I) is hydrogen and R2^ in a compound of formula (II) is not hydrogen, then cleavage of both R22 and R2q is conveniently effected in a single reaction. Preferably R2q and R2^ are both alkyl, such as methyl or ethyl, or benzyl.

It will be appreciated that compounds of formula (II) in 15 which R2i is hydrogen are themselves compounds of the invention of formula (I).

Compounds of formula (II) may be prepared by treating a compound of formula (III): in which R^, R2, R2q and R2^ are as defined in formula (II) (except that R2]_ is not Η) , with a compound of formula (IV): (IV) - Λ Λ ι ΙΕ 9Π5 Β2957 in which Rj and R4 together are as defined in formula (I) The reaction is preferably carried out in the presence of a coupling agent,, such as dicyclohexylcarbodiimide or l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride in the presence of 1-hydroxybenzotriazole, or using 1,1'-carbonyldiimidazole, in an inert solvent such as dichloromethane or acetonitrile.

Selective cleavage of the group R£^ may then be carried out using the procedures described above for the preparation of compounds of formula (I) to give compounds of formula (II) in which R21 is hydrogen.

The intermediate compounds of formula (III) may be prepared by treating a compound of formula (V) or a salt thereof: °*20 — P °*21 NH2 (V) in which Rp R£q and r21 are as defined in formula (III), with a compound of formula (VIA) or (VIB) or a salt thereof: R11 CC>2r12 (VIB) (VIA) Β2957 - 11 in which Rg is as defined in formula (I), Rgg is a leaving group such as halogen, methanesulphonyloxy or trifluoromethanesulphonyloxy and Rgg is hydrogen or a carboxyl protecting group, and thereafter removing an Rgg carboxyl protecting group. Preferred method is the reaction of (V) with (VIA).

When a compound of formula (VIB) is used, the reductive amination may be carried out by hydrogenation over a noble metal catalyst such as palladium on carbon or by reaction with sodium cyanoborohydride at pH 6 to 7. Lower alkyl alcohol solvents such as methanol and ethanol are suitable for both reactions. These reactions may be carried out in the presence of molecular sieves.

A hydrogenation reaction is preferred but this process precludes the use of compounds of formulae (V) and (VIB) in which any of Rgg, R21 or R12 benzyl. Preferably a carboxyl protecting group is a methyl or ethyl ester.

Ester protecting groups may be removed under standard basic hydrolysis conditions using dilute base such as 1 Normal aqueous sodium hydroxide in methanol or aqueous potassium hydroxide in 1,4-dioxane.

When the compound of formula (V) is in the form of the free base, the compound of formula (VIB) is suitably an α-keto ester (Rgg = slkyl).

When the compound of formula (V) is a salt, such as the hydrochloride salt, the compound of formula (VIB) is suitably a salt of an α-keto acid (Rg2 = H) > for example the sodium salt.

B2957 - 12 The preparation of compounds of formula (III) using a compound of formula (VIA) may be carried out under standard alkylation conditions. A halogen leaving group is preferably bromine and an oxygen-based leaving group is preferably trifluoromethanesulphonyloxy.

Compounds of formula (III) may alternatively be prepared by condensing a compound of formula (VII) or a salt thereof: *2 H2N CO2Ri2 (VII) in which R2 is as defined in formula (I) and R12 is a carboxyl protecting group with an aldehyde, R^-CHO in which is as defined in formula (I) and treating the condensation product with an appropriate dialkyl or trialkyl phosphite, for example dimethyl phosphite, and thereafter removing the carboxyl protecting group. The carboxyl group is conveniently protected as an alkyl or benzyl ester which may be removed using standard hydrolysis or hydrogenation conditions.

As described above in connection with reductive amination of compounds of formula (VIB), where a benzyl protecting group R12 is removed by hydrogenation then R2g and R21 are restricted to alkyl.

Alternatively, compounds of formula (II) in which R2g and R21 are alkyl or optionally substituted benzyl may be prepared by the reaction of a compound of formula (VIII): B2957 R, (VIII) in which R2, R3and R4 are as defined in formula (I), with a compound of formula (IX): in which R^ is as defined in formula (I), R2q and R21 are alkyl, optionally substituted phenyl, or optionally substituted benzyl and is a leaving group as defined for formula (VIA), in the presence of a base such as triethylamine or Proton Sponge (1,8-bis(dimethylamino)-naphthalene) , or using anhydrous potassium carbonate in an alcoholic solvent.

Where R^·^ is an oxygen-based leaving group, for example trifluoromethanesulphonyloxy, which is preferred, displacement of the leaving group is conveniently carried out in the presence of Proton Sponge in an inert solvent such as acetonitrile or dichloromethane, over a period of several days in the absence of light.

B2957 - 14 A further alternative preparation of compounds of formula (III) may be carried out by reacting a compound of formula (IX) as hereinbefore defined with a compound of formula (VII) in which R12 is a carboxyl protecting group, using conditions as described for the reaction of compounds of formula (VIII) with compounds of formula (IX), and thereafter removing the protecting group Rg2.

Suitable carboxyl protecting groups include alkyl, benzyl, trialkylsilyl and trialkylsilylethyl groups. A trialkylsilyl protecting group, for example trimethylsilyl, is especially useful in that it may be readily incorporated, in situ, for example by addition of hexamethyldisilazane to the reactants in acetonitrile in the presence of triethylamine, and selectively removed in aqueous methanol, without imposing any limitations on the value of R2q and R21· Other silylating agents include trimethylsilyl chloride and N, N-diethyltrimethylsilylamine.

An Rgg alkyl carboxyl protecting group may be removed by base hydrolysis, for example using sodium hydroxide in aqueous methanol or potassium hydroxide in aqueous 1,4dioxane .

It will be appreciated that where the carboxyl protecting group Rgg is alkyl, Rgg and Rgg may be alkyl, phenyl or benzyl derivatives, but where Rgg is a benzyl group, R2q and Rgg are limited to alkyl and phenyl.

When compounds of formula (III) are prepared by this route, it is preferred that R2q and R2l are benzyl and Rgg is trifluoromethanesulphonyloxy in the compound of formula (IX) and Rg2 is trimethylsilyl or methyl in the compound of formula (VII).

B2957 - 15 Compounds of formula (VIII) may be prepared by treating a compound of formula (VII): H2N CO2Ri2 (VII) in which R2 is as defined in formula (I), R12 is hydrogen and wherein the amino group is optionally protected, with a compound of formula (IV) as hereinbefore defined, in the presence of a coupling agent as hereinbefore described for the preparation of compounds of formula (II) from compounds of formulae (III) and (IV).

Compounds of formula (IX) may be prepared from hydroxyalkylphosphonate derivatives by conversion of the hydroxyl group to the leaving group R^ by conventional methods.

For example, where Rjj is trifluoromethanesulphonyloxy, trifluoromethanesulphonic anhydride may be added to a solution of the hydroxyalkylphosphonate in an inert solvent such as dichloromethane, the reaction being carried out at reduced temperature under an inert atmosphere, according to the general method of E. Vedejs et al. , Journal of Organic Chemistry 50, 2165, (1985) .

Hydroxyalkylphosphonate compounds may in turn be prepared by reaction of the corresponding phosphite, for example dibenzylphosphite, with an aldehyde R^-CHO in which R-j_ is as defined in formula (I) according to the general method of F. Texier-Boullet and A. Foucaud, Synthesis, 916 (1982). Benzyl and alkyl phosphites are either commercially available compounds or can be prepared from commercially available starting materials by standard methods.

B2957 Intermediate compounds of formula (V) are either known compounds or may be prepared from known aminoalkyl phosphonic acid derivatives using standard procedures to introduce R2q and R21 as required.

Protection of the amine function during these reactions may be necessary.

Introduction of an R2q or R2j methyl group may be effected 10 by reaction with diazomethane in a suitable inert solvent.

Compounds of formula (V) of fixed configuration may be prepared by the general method of R. Jacquier et al., Phosphorus and Sulfur 36, 73, (1988) .

Compounds of formula (IV) may be prepared by oxidising the primary alcohol function in a compound of formula (X): H I Y NH H I N-(CH2)q-OH (X) wherein p and q are as defined in formula (I), Y is a nitrogen protection group, and Z is R^ to give the corresponding aldehyde, followed by removal of Z where is an acyl group; cyclisation and reduction; and thereafter, as necessary, removing the nitrogen protection group Y and interconverting . ¢911165 B2957 - 17 Suitable nitrogen protection groups include t-butoxycarbonyl (BOC) and benzyloxycarbonyl groups.

The oxidation may be carried out using pyridinium 5 chlorochromate, or under Swern oxidising conditions, for example by treatment with dimethylsulphoxide and an acyl halide followed by triethylamine, as described by D. Swern et al., J. Org. Chem., 43, 2480 (1978). The cyclisation and reductive amination step may be effected by catalytic hydrogenation over a suitable noble metal catalyst, for example palladium on carbon, or by reaction with sodium cyanoborohydride or sodium borohydride. In some cases the yield of azalactams may be increased by carrying out the reductive amination step under acidic conditions.

Nitrogen protection groups may be removed by standard methods. A t-butoxycarbonyl group may be removed by treatment with trifluoroacetic acid at reduced temperature. Where Z is a nitrogen protection group, it may be selected to undergo concomitant cleavage during the cyclisation reaction to give a compound in which R^ is hydrogen. For example, when Z is a benzyloxycarbonyl group, it will be readily removed by catalytic hydrogenation .

An Rc hydrogen may be interconverted to an R^ C-j__galkyl, aralkyl or aryl group. The secondary amine group in the azalactam ring may be alkylated to form an alkyl group. For example, the amine group may be methylated to form an Rc> methyl group. The methylation step may be effected by catalytic hydrogenation over a suitable noble metal catalyst, for example palladium on carbon, in the presence of aqueous formaldehyde. Other suitable methylation procedures are described by E. Askitoglu et al., Helv.

B2957 - 18 Chim. Acta., .68., 750, (1985); E. Engler et al. , Helv. Chim. Acta., 68, 789, (1985); and M. Lennon et al., J. Chem. Soc. (Perkin I), 622, Compounds of formula (X) may compound of formula (XI): (1975) be prepared by reacting a H I Z-N-(CH2)P Y ι NH OH wherein ρ, Y and Z are as defined for formula (X), with a compound of formula (XII): H2N-(CH2)q-OH (XII) wherein q is as defined for formula (X).

The reaction may be carried out using standard procedures for forming an amide from a carboxylic acid and an amine, for example using a coupling agent such as 1,1'carbonyldiimidazole, 1,3-dicyclohexylcarbodiimide or 1-(3dimethylaminopropyl)-3-ethylcarbodiimide.

Compounds of formula (XI) are di-aminoalkanoic acid derivatives. These are known compounds or may be prepared from known starting materials by standard methods.

For example the compound of formula (IV) in which R^ and R4 together are -(CH2) -X-(CH2)q- where p is 3, q is 6 and B2957 - 19 X is -NH- is prepared from a compound of formula (XI) derived from ornithine which is commercially available.

The compounds of formula (IV) in which Rg and R4 together 5 are -(CH2)p-X-(CH2)q- where p is 4, q is 3, 5 or 6 and X is -NH- are prepared from a compound of formula (XI) derived from the amino acid lysine. The compound of formula (XI) , derived from S.-lysine, in which Y is t-butoxycarbonyl and Z is benzyloxycarbonyl, is commercially available.

Similarly, the compound of formula (IV) in which Ro and R^ together are - (CH2)p-X-(CH2)q- where p is 1, q is 8 and X is -NH- may be prepared from 2,3-diaminopropionic acid.

The compounds of formula (VII) are either known amino acid derivatives or can be made from these derivatives by known methods. Compounds of formula (VIA) and (VIB) are either known compounds or may be prepared from known compounds by known methods .

The intermediates of formulae (II), (III), and certain intermediates of formula (V) disclosed herein are novel compounds and form an aspect of the present invention as do the described processes for their preparation.

Where obtainable, pharmaceutically acceptable salts of the compounds of formula (I) may be formed conventionally by reaction with the appropriate acid or base. Solvates may be formed by crystallization from the appropriate solvent.

B2957 - 20 As mentioned previously, the compounds of formula (I) exist in more than one diastereoisomeric form. Where the processes of the invention produce mixtures thereof, the individual isomers may be separated one from another by chromatography e.g. HPLC.

Alternatively, separate diastereoisomeric compounds of formula (I) can be obtained by using stereoisomerically pure starting materials or by separating desired isomers of intermediates at any stage in the overall synthetic process, and converting these intermediates to compounds of formula (I).

It will be appreciated that where a single diastereoisomer of a compound of formula (I) is prepared by more than one process variant as hereinbefore described, each of which allows a different chiral centre to be defined, it may be possible to deduce the configuration at a chiral centre which is not pre-determined using a particular process variant.

Furthermore, it will be appreciated that although the absolute configuration at a particular chiral centre may not be known, it is possible to characterise a given diastereoisomer relative to its epimer by reference to the direction in which the plane of polarised light is rotated.

The present invention further provides a pharmaceutical composition, which comprises a compound of formula (I),or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

B2957 - 21 A composition of this invention is useful in the treatment of musculo-skeletal disorders, particularly arthritic diseases and for modulation of tissue remodelling.

A composition of the invention also has potential utility in the treatment of cancer; for preventing myelin degradation in the central and peripheral nervous system; and in other conditions in which members of the collagenase family of neutral metalloproteases have pathological or other roles.

A composition of the invention, which may be prepared by admixture, may contain a diluent, binder, filler, disintegrant, flavouring agent, colouring agent, lubricant or preservative in conventional manner. These conventional excipients may be employed in conventional manner, for example as in the preparation of compositions of related peptide enzyme inhibitors, such as the ACE inhibitor enalapril.

A composition of the invention may be adapted for oral, topical, rectal or parenteral administration but oral administration is preferred. Parenteral compositions may be administered intravenously, intramuscularly, intra25 articularly, intradermally, subcutaneously or into the cerebro-spinal fluid.

Preferably, a pharmaceutical composition of the invention is in unit dosage form and in a form adapted for use in the medical or veterinarial fields. For example, such preparations may be in a pack form accompanied by written or printed instructions for use as an agent in the treatment or prophylaxis of any of the disorders mentioned above .

B2957 - 22 The suitable dosage range for the compounds of the invention may vary from compound to compound and may depend on the condition to be treated. It will also depend, inter alia, upon the relation of potency to absorbability and the mode of administration chosen.

The compound or composition of the invention may be formulated for administration by any route, the preferred route depending upon the disorder for which treatment is required, and is preferably in unit dosage form or in a form that a human patient may administer to himself in a single dosage.

Compositions may, for example, be in the form of tablets, capsules, sachets, vials, powders, granules, lozenges, reconstitutable powders, or liquid preparations, for example solutions or suspensions, or suppositories.

The compositions, for example those suitable for oral administration, may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine; tableting lubricants, for example magnesium stearate; disintegrants, for example starch, polyvinylpyrrolidone, sodium starch glycollate or microcrystalline cellulose; or pharmaceutically acceptable wetting agents such as sodium lauryl sulphate.

Solid compositions may be obtained by conventional methods of blending, filling, tableting or the like. Repeated blending operations may be used to distribute the active agent throughout those compositions employing large quantities of fillers. When the composition is in the 911165 B2957 - 23 form of a tablet, powder, or lozenge, any carrier suitable for formulating solid pharmaceutical compositions may be used, examples being magnesium stearate, starch, glucose, lactose, sucrose, rice flour and chalk. Tablets may be coated according to methods well known in normal pharmaceutical practice, in particular with an enteric coating. The composition may also be in the form of an ingestible capsule, for example of gelatin containing the compound, if desired with a carrier or other excipients. For example, in a hard gelatin capsule containing the required amount of a compound of the invention in the form of a powder or granulate in intimate mixture with a lubricant, such as magnesium stearate, a filler, such as microcrystailine cellulose, and a disintegrant, such as sodium starch glycollate.

Compositions for oral administration as liquids may be in the form of, for example, emulsions, syrups, or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminium stearate gel, hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; aqueous or nonaqueous vehicles, which include edible oils, for example almond oil, fractionated coconut oil, oily esters, for example esters of glycerine, or propylene glycol, or ethyl alcohol, glycerine, water or normal saline; preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid; and if desired conventional flavouring or colouring agents .

B2957 - 24 The compounds of this invention may also be administered by a non-oral route. In accordance with routine pharmaceutical procedure, the compositions may be formulated, for example for rectal administration as a suppository or for parenteral administration in an injectable form. For injection, for example by intraarticular injection or by injection into the cerebrospinal fluid or via other routes which will gain access to sites of demyelination, such as by intramuscular, intradermal or subcutaneous injection, as freely soluble solutions or as poorly dispersed depot stores, the compounds of the invention may be presented in an aqueous or non-aqueous solution, suspension or emulsion in a pharmaceutically acceptable liquid, e.g. sterile pyrogen15 free water or a parenterally acceptable oil or a mixture of liquids, which may contain bacteriostatic agents, antioxidants or other preservatives, buffers or solutes to render the solution isotonic with the blood, thickening agents, suspending agents or other pharmaceutically acceptable additives. Such forms will be presented in sterile unit dose form such as ampoules or disposable injection devices or in multi-dose forms such as a bottle from which the appropriate dose may be withdrawn or a solid form or concentrate which can be used to prepare an injectable formulation.

For topical and percutaneous administration, the preparations may also be presented as an ointment, cream, lotion, gel, spray, aerosol, wash, skin paint or patch.

A unit dose for treating diseases in which enzymes of the collagenase family are involved will generally contain from 10 to 1000 mg and preferably will· contain from 10 to 500 mg, in particular 10, 50, 100, 150, 200, 250, 300, 350, 400, 450 or 500 mg. The composition may be B2957 - 25 administered one or more times a day, for example 2, 3 or 4 times daily, so that the total daily dose for a 70 kg adult will normally be in the range 10 to 3000 mg. Such a dosage corresponds to approximately 0.15 to 50 mg/kg per day. Alternatively, in particular for injection, the unit dose will contain from 2 to 200 mg of a compound of the invention and be administered in multiples, if desired, to give the desired daily dose.

The present invention additionally provides a method of treating conditions in which degradation of connective tissue and other proteinaceous components of the body occurs, such as rheumatism and/or arthritic conditions in mammals, such as humans, which comprises administering to the mammal in need of such treatment an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in the treatment of conditions in which degradation of connective tissue and other proteinaceous components of the body occurs such as rheumatism and/or arthritic conditions.

The following Descriptions and Examples illustrate the preparation of compounds of the invention. All temperatures are expressed in °C.

B2957 - 26 Description 1 Dibenzyl (1-hydroxypropyl)phosphonate (Dl) O (PhCH2O)2P OH C2H5 The general method of F. Texier-Boullet and A. Foucaud [Synthesis, 1982, 916] was employed. A mixture of dibenzyl phosphite (31.13 ml, 0.14 mole) and propionaldehyde (10.21 ml, 1 equiv.) was stirred at room temperature and basic alumina (70g) added in one portion. After standing overnight at room temperature chloroform was added and the alumina collected and washed with chloroform. The filtrate was evaporated to dryness and the resulting clear oil chromatographed on silica gel (600g) with gradient elution (ether - 5% methanol/ether). The title compound was obtained as a clear oil which solidified on standing (27.82g, 64%). A sample was recrystallized from ether/pentane to give a white crystalline solid, m.p. 81-82°C.

Found: C, 64.09; H,6.71. C17H21°4P1 reciuires C, 63.74; H,6.61%. δ (CDC13) 1.04(3H,t, J=7Hz) , 1.6-1.95 (2H,m), 2.27(1H, brs), 3.8(1H, 2 overlapping triplets,J=5 and 10Hz), 4.97-5.18 (4H,m) , 7.34(10H,s). •Ε 911165 Β2957 Description 2 Dibenzyl ((1-trifluoromethanesulphonyloxy)propyl)phosphonate (D2) O (PhCH2O)2P C2H5 O The title compound was prepared by the general method of E. Vedejs et al. [J. Org. Chem. 1985, 50(12), 2165]. A solution of dibenzyl (1-hydroxypropyl)phos-phonate (DI) (24.97g, 0.078 mole) in methylene chloride (180 ml) was cooled to -50°C under N2 . 2,6-Lutidine (11.12 ml, 0.095 mole) was added followed by trifluoro- methanesulphonic anhydride (15.1 ml, 0.0898 mole) keeping the temperature -50°C. The mixture was allowed to warm slowly to 0°C and then taken into cold ether. The solution was subjected to a rapid aqueous work-up by washing the organic layer with ice-cold water, dilute hydrochloric acid (x2) and finally brine. The organic layer was dried (anhydrous MgSO^) and evaporated to dryness to give the title compound as a pinkish orange oil (33.77g, 96%) which was used without further purification.

(CDC13) 1.08 (3H,t,J=7Hz) , 1.88(2H,m), 4.94(1H,2 overlapping triplets,J=5.5 and 7Hz), 4.88-5.22 (4H,m) and 7.35 (10H,m) . - 28 B2957 Description 3 N-(1-(R)-Dibenzyloxyphosphinylpropyl)-(S)-leucine (D3A) and N-(1-(S)-Dibenzyloxyphosphinylpropyl)-(S)-leucine (D3B) Method A Following the general method of US 4808741 for the preparation of leucine trimethylsilyl ester a mixture of (S)-leucine (1.15g, 0.0088 mole), hexamethyldisilazane (1.75 ml), and triethylamine (1.38 ml) in acetonitrile (13.5 ml) was heated at reflux for a total of 4h.

Dibenzyl ( (1-trifluoromethanesulphonyloxy) propyl)phosphonate (D2) (4.5g, 0.01 mole) was then added and the mixture maintained at 40-42°C for 48h. The reaction can also be carried out at ambient temperature. After cooling the mixture was filtered, washed with methanol and the filtrate evaporated to dryness. The residue was taken up in chloroform and washed with dilute HCl (x2) and finally water. The chloroform layer was dried (anhydrous Na2SO4), filtered and evaporated to dryness to give an orange gummy solid (3.67g). The crude product was triturated with the minimum volume of ether/pentane to give a white crystalline solid which after collection, washing with a little cold ether/pentane and drying gave the title compound, R,S isomer (D3A) (0.47g, 11%), m.p. 112-115°C.

B2957 - 29 Observed Desorption CI (NH3) MH+ 434. C23H32NO5P requires M 433. [a]D20 = -23.09° (c=0.97 MeOH).

Found: C,63.73; H,7.42; N,3.23. C23H32NO5P requires 5 C,63.73; H,7.44; N,3.23%. δ (CDCI3): 0.89 (6H,t), 1.03 (3H,t), 1.25-2.0 (5H,m), 2.74 (lH,m), 3.28 (2H, br s), 3.73 (IH, br t) , 4.9-5.15 (4H, m), 7.35 (10H, s).

The other isomer, N-(1-(S)-dibenzyloxyphosphinylpropyl) (S)-leucine (D3B), can be obtained by preparative HPLC using a Hamilton PRP-1 column, 300 x 7.0mm, 264R with a 40 : 60 acetonitrile:water eluent mixture and a flow rate of 4.0 ml/min. Under these conditions the R,S isomer (D3A) elutes first with a retention time of 34.6 min and the S,S isomer (D3B) is well separated at 42.7 min.

For the isomer (D3B): Observed FAB (M+H)+ 434. ^23^32^5^ requires M 433. δ (CDC13): 0.88 (6H,dd), 0.98 (3H,t), 1.4 (lH,m), 1.52-1.9 (4H,m), 2.72 (lH,m), 3.38 (lH,m), 4.9-5.15 (4H,m), 7.32 (10H,s) .

The S,S isomer (D3B) on coupling with (S)-amino acid derivatives leads to the S,S,S, series.

Method B A mixture of (S)-leucine methyl ester hydrochloride (0.543g; 0.003 mole), dibenzyl (1-trifluoromethanesulphonyloxy)propyl)phosphonate (D2) (1.35g; 0.003 mole) and anhydrous potassium carbonate (l.Og) in methanol (2 ml) was heated at 50°C, with stirring, for 4 hours and then left at room temperature overnight.

B2957 - 30 The reaction mixture was evaporated to dryness in vacuo, and dissolved in chloroform (5 ml) and filtered. The filtrate, and washings, were combined and chromatographed on silica gel 60 ,(50g) using ethyl acetate-pentane (1:1) as the eluent, to afford a mixture of N-(1-(R)-dibenzyloxyphosphinylpropyl)-(S)-leucine methyl ester and N-(1-(S)-dibenzyloxyphosphinylpropyl) -(S)-leucine methyl ester as an oil (0.55g). The esters can be separated into individual diastereomers by column chromatography on silica gel with initially 50% diethyl ether/pentane as eluent, rising to 100% diethyl ether.

The above mixture of esters (l.lg, 0.0025 mole) in methanol (4.0 ml) was treated with a solution of sodium hydroxide (O.llg; 0.00275 mole) in water (1.5 ml), and the solution was stirred at room temperature overnight. It was evaporated to one third volume, in vacuo, taken in water and extracted with ether. The aqueous fraction was acidified with citric acid to pH 3-4 and then extracted (5x) with chloroform. The chloroform fraction was dried (Na2SO4) and evaporated to dryness in vacuo to give a mixture of the title compounds (D3A) and (D3B) as an oil that slowly solidified.

Trituration of the product with ether gave N-(1-(R)-dibenzyl-oxyphosphinylpropyl) -(S)-leucine (D3A) (0.34g) as a white crystalline solid, identical to the product obtained by Method A.

Alternatively, the single isomer can be hydrolysed separately. For example N-(1-(S)-dibenzyloxyphosphinylpropyl) -(S)-leucine methyl ester on hydrolysis by the above method gave N-(1-(S)-dibenzyloxyphosphinylpropyl) - (S) -leucine (D3B), m.p. 71-73°C. - 31 B2957 Description 4 Na-tert-Butoxycarbonyl-Ne-benzyloxycarbonyl-(S)-lysine- (5hydroxy)pentylamide (D4) O To a solution of Na-tert-butoxycarbonyl-Ne-benzyloxycarbonyl-(S)-lysine (7.8g, 21 mmol) in anhydrous dichloromethane (150 ml) maintained at 0°C was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.3g, 22.5 mmol) and 1-hydroxybenzotriazole (3.6g, 26.5 mmole). The mixture was stirred for 0.5h at 0°C, 5-aminopentan-l-ol (2.3g, 22.5 mmol) added and stirring continued at room temperature. After 3h the mixture was washed with saturated aqueous NaHCOg (60 ml), dried over anhydrous magnesium sulphate and evaporated in vacuo to afford a viscous oil. Purification by flash chromatography [(CHClg:MeOH) (20:1) v/v] gave the title compound (D4) as a clear oil (8.01g).

Observed (M+H)+ 466. <~:24Η39θ6Ν3 rec5u+res M 465.

IE 911165 B2957 Description 5 Na-tert-Butoxycarbonyl-Ne-benzyloxycarbonyl-(S)lysine-(4-formyl)butylamide (D5) H3C h3c V To a stirred solution of oxalyl chloride (1.47g, 12 mmol) in anhydrous dichloromethane (40 ml) maintained under an atmosphere of nitrogen at -60°C was added dimethylsulphoxide (1.21g, 15 mmol) dropwise, such that the temperature remained below -50°C. The mixture was left stirring at -60°C for 15 minutes, alcohol (D4) (3.6g, 7.7 mmol) diluted in anhydrous dichloromethane (10 ml) was added, and allowed to warm up to -25°C over lh. The mixture was then cooled down to -60°C, triethylamine (4.7g, 46 mmol) added slowly such that the internal temperature remained below -50°C. On completion of addition, the mixture was gradually warmed up to room temperature, washed with water (30 ml) and sat. aq. NaCl (30 ml). The aqueous washes were back extracted with dichloromethane (2x30 ml) and the combined organic fractions were dried over anhydrous magnesium sulphate and evaporated in vacuo to yield a viscous clear oil. Purification by flash chromatography [(EtOAc:MeOH)(20:l)v/v] afforded the title compound (D5) as an oil (2.8g).

Observed (M+H)+ 464. ^24^37^6^3 recIuires M 463 . •Ε 911165 - 33 Β2957 Description 6 (S)-3-(N-tert-Butoxycarbonyl)amino-8-(N-benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one (D6) Method A The aldehyde (D5) (1.8g, 3.88 mmol) was dissolved in ethanol (180 ml) and hydrogenated over 5% palladium on charcoal (200 mg) at atmospheric pressure and 35°C for 72h. The suspension was filtered through Kieselguhr and evaporated in vacuo to give crude 3-[N-tert-butoxycarbonyl]amino-(S)-1,8-diazacyclotridecan-2-one. The crude amine was dissolved in a mixed solvent system of tetrahydrofuran/water, (6:20 ml) v/v cooled to 0°C and treated with benzylchloroformate (0.66g, 3.88 mmol) and excess sodium carbonate to maintain a pH between 10 and 11. The mixture was left stirring at room temperature overnight, washed with ethyl acetate (3x25 ml), and the combined organic fractions dried over anhydrous magnesium sulphate and evaporated in vacuo to afford a clear oil. Purification by flash chromatography [(EtOAc:MeOH)(20:l)v/v] yielded the title compound (D6) as a white solid (0.2g).

Observed M+ 447. C24H3-7O5N3 requires M 447. - 34 B2957 Method B The aldehyde (D5) was hydrogenated at about 100 psi over 5% palladium on charcoal in methanol, and then in acidic methanol to afford crude (S)-3-[N-tertbutoxycarbonyl ] amino-1,8-diazacyclotridecan-2-one.

The amine was treated with benzylchloroformate and purified as described in Method A to yield the identical title compound (D6).

Description 7 (S)-3-Amino-8-[N-benzyloxycarbonyll-1,8-diazacyclotridecan-2-one, trifluoroacetate salt (D7) A cooled (0°C) solution of the lactam (D6) (0.19g, 0.42 mmol) in dichloromethane (5 ml) was treated with trifluoroacetic acid (2 ml). After 0.5h the solvent was evaporated under reduced pressure, the residue diluted with dichloromethane (15 ml) and washed with sat. aq. NaCl (10 ml). The organic fraction was dried over anhydrous magnesium sulphate and evaporated in vacuo to give crude title compound (D7) as an oil. This was used as such without further purification.

B2957 Description 8 (S)-3-[Ν-[N-(R)-(1-Phosphonopropyl)-(S)-leucyl]]amino8- (N-benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester (D8) CH3 A solution of N-(1-(R)-dibenzyloxyphosphinylpropyl)(S)-leucine (D3A) (0.194g, 0.45 mmol) in anhydrous dichloromethane (20 ml) maintained at 0°C was treated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.085g, 0.45 mmol) and 1-hydroxybenzotriazole (0.079g, 0.58 mmol). After stirring for 0.5h the reaction mixture was sequentially treated with lactam (D7) (0.15g) and N,N-diisopropyl-ethylamine (0.12g, 0.89 mmol). Stirring was continued for 18h at room temperature, then the mixture was washed with sat. aq. NaHCO3 (2x20 ml), and sat. aq. NaCl (2x20 ml) . The aqueous washes were back-extracted with dichloromethane and the combined organic fractions dried over anhydrous magnesium sulphate, and evaporated in vacuo to yield an oil. On purification by flash chromatography (2% methanol in chloroform), the title compound (D8) was obtained as a clear oil (O.llOg).

Observed (M+H)+ 763. C42H59N4°7P requires M 762.

,E 911165 B2957 δ (CDC13): 0.96(6H,t), 1.2(3H,t), 1.25-1.93(18H,m), 2.70(lH,m), 2.90(lH,m), 3.18(2H,m), 3.39 <2H, dt) , 3.62(2H,m), 4.23(lH,m), 4.92-5.12 (6H, m) , 6.0(lH,brs), 7.28-7.42 (15H,m) ., Description 9 Na-tert-Butoxycarbonvl-N£-benzyloxycarbonyl-(S)-ornithine (6-hydroxy)hexylamide (D9) A solution of Na-tert-butoxycarbonyl-Ne-benzyloxycarbonyl (S)-ornithine (12g, 0.033 mol) in anhydrous dichloromethane (200 ml) maintained at 0°C, was treated sequentially with 1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (7.5g, 0.039 mol), and 1hydroxybenzotriazole (5.3g, 0.039 mol). The solution was stirred at 0°C for 1 h, treated with 6-aminohexan-l-ol (4.6 g, 0.039 mol) and left stirring overnight at room temperature. The mixture was then washed with saturated aqueous NaHCO3, dried over anhydrous magnesium sulphate, and evaporated in vacuo to afford a viscous oil. Purification by flash chromatography [(CHC13:MeOH) (20:1)v/v] gave the title compound (D9) as a clear oil (9.lg) . + 466. c24H39°6N3 recIuires M 465.

Observed (M+H) IE 911165 B2957 - 37 δ (CDC13) : 1.25-1.4(5H,m) , 1.4(9H,S), 1.42-1.62 (7H,m) , 1.78 7.3 (5H,m) .

Description 10 Na-tert-Butoxycarbonyl-N£-benzyloxycarbonyl-(S)-ornithine(5-formyl)pentylamide (DIO) The alcohol (D9) (6.5g, 13.9 mmol) dissolved in dichloromethane (50 ml) was added to a vigorously stirred mixture of pyridinium chlorochromate (9g, 41.8 mmol) and 4A molecular sieves (20g) in dichloromethane (200 ml).

Additional portions of pyridinium chlorochromate (4g) were added after 30 minutes and 45 minutes. After lh total reaction time, the mixture was poured into ether (200 ml) and the reaction flask rinsed with ether (3x100 ml). The combined organic fractions were filtered through kieselguhr, and concentrated in vacuo to afford a yellow oil. Purification by flash chromatography [(EtOAc:MeOH) (49:l)v/v then (20:l)v/v] yielded the title compound (D10) as a viscous oil (3.5g).

Observed (M+H) + 464. ^24^37^3^6 re<4uires M 463.

,E 91U 65 B2957 Description 11 (S)-3-(N-tert-Butoxycarbonyl)amino-7-(Nbenzyloxycarbonyl)-1,7-diazacyclotridecan-2-one (Dll) N O OCH2Ph The aldehyde (DIO) (2.5g) in methanol (300 ml) was treated with 5% palladium on charcoal (2.5g) . The suspension was hydrogenated at 100 psi and ambient temperature for 48h, treated with 2.5M aqueous hydrochloric acid (2 ml) and hydrogenation continued at the said pressure for a further 24h. The suspension was filtered through kieselguhr and evaporated in vacuo to give crude 3- (N-tert-butoxycarbonyl)amino-(S)-1,7-diazacyclotridecan-2-one. The crude amine was dissolved in a mixed solvent system of tetrahydrofuran/water, (10:40 ml) v/v cooled to 0°C and treated with benzylchloroformate (0.92g) and excess sodium carbonate to maintain a pH between 10 and 11. The mixture was left stirring at room temperature for 4h, solvent partly evaporated in vacuo and the residue extracted with dichloromethane (3x50 ml). The combined organic fractions were dried over anhydrous magnesium sulphate and evaporated in vacuo to afford a clear oil. Purification by flash chromatography [(EtOAc:MeOH) (50:1) v/v] yielded the title compound (Dll) as a white solid (0.27g).

Observed M+ 447. (-24Η37θ5Ν3 reGuires M 447.

Description 12 (S)-3-Amino-7-[N-benzyloxycarbonyll-1,7B2957 diazacyclotridecan-2-one, trifluoroacetate salt (D12) Η O A cooled (0°C) solution of the lactam (Dll) (0.13g) in dichloromethane (5 ml) was treated with trifluoroacetic acid (3 ml). After lh the solvent was evaporated under reduced pressure, to afford crude title compound (D12) as an oil. This was used as such without further purification.

Description 13 (S) - 3-[Ν-[N-(R)- (1-Phosphonopropyl) -(S)-leucyl]) amino-7(N-benzyloxycarbonyl)-1,7-diazacyclotridecan-2-one, dibenzyl ester (D13) H O (PhCH2O)2P' I I o N A O OCH2Ph A solution of N-(1-(R)-dibenzyloxyphosphinylpropyl)-(S)leucine (D3A) (0.115g) in anhydrous dichloromethane (20 ml) maintained at 0°C was treated with 1- (3IE 911165 B2957 -40dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.056g) and 1-hydroxybenzotriazole (0.036g). After stirring for 0.5h the reaction mixture was sequentially treated with the crude lactam (D12), N,N-diisopropyl5 ethylamine (0.075g) and stirring continued at room temperature for 4h. The mixture was washed with water (20 ml) and sat. aq. NaHCOg (2x20 ml). The aqueous washes were back-extracted with dichloromethane and the combined organic fractions dried over anhydrous magnesium sulphate and evaporated in vacuo to yield an oil. On purification by flash chromatography (2% methanol in chloroform), the title compound (D13) was obtained as a white foam (0.125g).

Observed (M+H)+ 763. C42H59N4°7P rec(uires M 762.

Description 14 Na-tert-Butoxycarbonyl-N£-benzyloxycarbonyl-(S)-lysine- (620 hydroxy)hexylamide (D14) A solution of Na-tert-butoxycarbonyl-Ne-benzyloxycarbony1(S)-lysine (15.8g, 0.042 mol) in anhydrous dichloromethane (200 ml) maintained at 0°C, was treated sequentially with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (9.96g, 0.051 mol) and 1-hydroxybenzotriazole (7.0g, 0.051 mol). The solution was stirred ¢911165 B2957 - 41 at 0°C for lh, treated with 6-aminohexan-l-ol (4.7g, 0.046 mol), and left stirring overnight at room temperature.

The mixture was then washed with saturated aqueous NaHCOg, dried over anhydrous magnesium sulphate, and evaporated in vacuo to afford a viscous oil. Purification by flash chromatography [(CHCI3:MeOH) (20:1) v/v] gave the title compound (D14) as a clear oil (16g).

Observed (M+H) + 480. ^25^41^3^6 rec5uires M 479.

Description 15 Na-tert-Butoxycarbonyl-Ne-benzyloxycarbonyl-(S)-lysine- (5formyl)pentylamide (D15) Η O O A stirred solution of dimethyl sulphoxide (3.63g, 0.046 mol) in anhydrous dichloromethane (100 ml) maintained at -60°C, was treated with oxalylchloride (2.58g, 0.0198 mol) diluted in dichloromethane (10 ml) at such a rate, so as to ensure temperature remained below -50°C. After stirring for 20 minutes, the alcohol· (D14) (6.35g, 0.013 mol) dissolved in dichloromethane (50 ml) was added dropwise over 5 mins. The mixture was stirred at -60°C for 15 mins, warmed up to -35°C, stirred for a further 10 mins then cooled down to -60°C. The solution was treated with triethylamine (8g, 0.08 mol), warmed up to room B2957 - 42 temperature, washed with water (2x100 ml), dried over anhydrous magnesium sulphate and solvent evaporated under reduced pressure to afford a viscous oil. Purification by flash chromatography [(EtOAc:MeOH) (30:1) v/v] gave the title compound (D15) as an oil (5g).

Observed (M+H) + 478. ^25Η39Ν3θ6 requires M 477.

Description 16 (S)-3-(N-tert-Butoxycarbonyl) amino-7-(Nbenzyloxycarbonyl)-1,8-diazacyclotetradecan-2-one (D16) H’CX°v H3C CH3o The aldehyde (D15) (5.0g) in methanol (450 ml) was treated with 5% palladium on charcoal (5.5g). The suspension was hydrogenated at 140 psi and ambient temperature for 48h, treated with 2.5M aqueous hydrochloric acid (3 ml) and hydrogenation continued at the said pressure for a further 24h. The suspension was filtered through Kieselguhr and evaporated in vacuo to give crude (S)-3-(N-tert-butoxycarbonyl)amino-1,8-diazacyclotetradecan-2-one. The crude amine was dissolved in a mixed solvent system of tetrahydrofuran/water, (10:40 ml) v/v cooled to 0°C and treated with benzylchloroformate (2.85g) and excess sodium carbonate to maintain a pH between 10 and 11. The mixture was left stirring at room temperature for 4h, solvent B2957 - 43 partially evaporated in vacuo and the residue extracted with dichloromethane (3x100 ml). The organic fraction was dried over anhydrous magnesium sulphate and evaporated in vacuo to afford a clear oil. Purification by flash chromatography [(EtOAc:MeOH) (50:1) v/v] yielded the title compound (D16) as a white solid (2.0g) m.p. 131.5-134.0°C.

Observed M+ 461. ^^Η^θΝ^Ο^ requires M 461.

Description 17 (S)-3-Amino-8-[N-benzyloxycarbonyl]-1,8diazacyclotetradecan-2-one, trifluoroacetate salt (D17) A cooled (0°C) solution of the lactam (D16) (0.175g) in dichloromethane (5ml) was treated with trifluoroacetic acid (3 ml). After lh the solvent was evaporated under reduced pressure, to afford crude title compound (D17) as an oil. This was used as such without further purification. - 44 B2957 Description 18 (S) -3-[N-[N-(R)-(1-Phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacyclotetradecan-2-one, dibenzyl ester (D18) CH3 A solution of N-(1-(R)-dibenzyloxyphosphinylpropyl) -(S)leucine (D3A) (0.133g) in anhydrous dichloromethane (10 ml) maintained at 0°C was treated with 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.065g) and 1-hydroxybenzotriazole (0.046g). After stirring for 0.5h the reaction mixture was sequentially treated with the crude lactam (D17) and N,N20 diisopropylethylamine (O.lg) and stirring continued at room temperature for 4h. The mixture was washed with water (10 ml) and sat.aq. NaHCO^ (2x10 ml). The aqueous washes were back-extracted with dichloromethane and the combined organic fractions dried over anhydrous magnesium sulphate and evaporated in vacuo to yield an oil. On purification by flash chromatography (2% methanol in chloroform), the title compound (D18) was obtained as a white foam (170 mg).

Observed (M+H) + 777. C^^Hg^N^O-yP requires M 776.

IE 911165 - 45 B2957 Description 19 (S)-3-(N-tert-Butoxycarbonyl)amino-8-(N-methyl)-1,8diazacyclotridecan-2-one (D19) A solution of diazalactam (D6) (0.35g) in methanol (20 ml) was treated with 5% palladium on charcoal (0.2g) and hydrogenated for 24h at atmospheric pressure and room temperature. The suspension was filtered through kieselguhr, and the filtrate diluted with methanol to a total volume of 50 ml. The solution was treated with 5% palladium on charcoal (0.3g) followed by 40% aqueous formaldehyde (1 ml), and the resulting suspension hydrogenated for 48h at a pressure of 100 psi. The suspension was filtered through Kieselguhr, solvent evaporated in vacuo, to yield an oil which on exposure to diethyl ether solidified. Purification by flash chromatography [(CHCI3:MeOH:NH3) (90:9:0.5) v/v] afforded the title compound (D19) as a pale yellow solid (0.14g). m.p. 135-137°C.

Observed M+ 327. Ci7H33N3°3 requires M 327. - 46 B2957 Description 20 (S)-3-Amino-8-[N-methyl]-1,8-diazacyclotridecan-2-one, trifluoroacetate salt (D20) A cooled solution of the diazalactam (D19) (0.175g) in dichloromethane (5 ml) was treated dropwise with trifluoroacetic acid (3 ml). After lh the solvent was evaporated under reduced pressure to afford crude title compound (D20) as an oil. This was used as such without further purification.

Description 21 (S)-3-[Ν-[N-(R)- (1-Phosphonopropyl) -(S)-leucyl]]amino-8(N-methyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester (D21) A solution of the N-(1-(R)-dibenzyloxyphosphinylpropyl)(S)-leucine (D3A) (0.133g) in anhydrous dichloromethane (10 ml) maintained at 0°C was treated with 1-(3IE 911165 B2957 - 47 dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.065g) and 1-hydroxybenzotriazole (0.046g). The mixture was allowed to stir for 0.5h at room temperature, then treated with amine (D20) dissolved in dichloromethane (10 ml), followed by N,N-diisopropylethylamine (O.lg).

After stirring for 18h, the mixture was washed with sat. aqueous NaCl (2x10 ml), dried over anhydrous magnesium sulphate and solvent evaporated in vacuo to afford an oil. Purification by flash chromatography (2% methanol in chloroform) gave the title compound (D21) as a white foam (0.17g).

Observed (M+H) + 643. C35H55N4O5P requires M 642.

Description 22 Na-tert-Butoxycarbonyl-N£-benzyloxycarbonyl-(S)-lysine-(3hydroxy)propylamide (D22) A stirred solution of Na-tert-butoxycarbonyl-Nebenzyloxycarbonyl-(S)-lysine (17.2g) maintained at 0°C in anhydrous dichloromethane (250 ml) was treated with 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.5g) and 1-hydroxybenzotriazole (5.3g). The solution was stirred at 0°C for lh, treated with 3-aminopropan-l-ol (4.08g) dissolved in dichloromethane (50 ml) and left B2957 stirring for 18h at room temperature. The solution was then washed with sat. aqueous NaHCO^, dried over anhydrous magnesium sulphate, and evaporated in vacuo to afford a viscous oil. Purification by flash chromatography [(EtOAc:MeOH) (49:1) v/v] gave the title compound (D22) as a clear oil (15.2g).

Observed (M+H) + 438. C22H35N3°6 requires M 437.

Description 23 Na-tert-Butoxycarbonyl-N£-benzyloxycarbonyl-(S)-lysine- (2formyl)ethylamide (D23) H O The title compound (D23) was prepared following the procedure as described for the synthesis of Na-tertbutoxycarbonyl-Ne-benzyloxycarbonyl-(S)-lysine-(4formyl)butylamide (D5) (yield: 59%).

Observed (M+H) + 436. ^22^33^3^6 reciuires M 435. - 49 B2957 Description 24 (S)-3-(N-tert-Butoxycarbonyl)amino-8-(Nbenzyloxycarbonyl)-1,8-diazacycloundecan-2-one (D24) The title compound (D24) was prepared following the procedure described for the synthesis of (S)-3-(N-tertbutoxycarbonyl) amino-7-(N-benzyloxy-carbonyl)-1,715 diazacyclotridecan-2-one (Dll) (Yield 11%), m.p. 172174°C.

Description 25 (S)-3-Amino-8-[N-benzyloxycarbonyl]-1,8-diazacycloundecan2-one, trifluoroacetate salt (D25) Η O tfa-h V- OCH2Ph O The title compound (D25) was prepared following the 30 procedure described for the synthesis of (S)-3-amino-7-[Nbenzyloxycarbonyl]-1,7-diazacyclo-tridecan-2-one, trifluoroacetate salt (D12), and used without purification . - 50 B2957 Description 26 (S)-3-[N-[N-(R)-(1-Phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacycloundecan-2-one, dibenzyl ester (D26) >-OCH2Ph O The title compound (D26) was prepared following the procedure described for the synthesis of (S)-3-[N-[N-(R)15 (1-phosphonopropyl) -(S)-leucyl]]amino-7-(Nbenzyloxycarbonyl)-1,7-diazacyclotridecan-2-one, dibenzyl ester (D13). (Yield: 44%).

Observed FAB [M-(PhCH2O)2PO]+ 473. c40H55N4°7p requires M 734.

Description 27 (S)-3-fN-[N-(S)- (1-Phosphonopropyl) -(S)-leucyl]1amino-825 (N-benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester (D27) B2957 - 51 A solution of N-(1-(S)-dibenzyloxyphosphinylpropyl)- (S)leucine (D3B) (0.27g) in anhydrous dichloromethane (10 ml) maintained at 0°C was treated with 1-(3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.125g) and 15 hydroxybenzotriazole (0.088g). After stirring for 0.5h the reaction mixture was sequentially treated with the lactam trifluoroacetate salt (D7) (0.75 mmol) and N,Ndiisopropylethylamine (0.16g). Stirring was continued for 18h at room temperature, then the mixture was washed with sat.aq. NaHCO^ (2x15 ml), and sat. aq. NaCl (2x15 ml).

The organic fraction was dried over anhydrous magnesium sulphate, and evaporated in vacuo to yield an oil. On purification by flash chromatography (2% methanol in chloroform), the title compound (D27) was obtained as a clear oil (0.31g).

Observed (M+H)+ 763. Ο42Η59θ7Ν4ρ requires M 762.

Description 28 (S)-3-[Ν-[N-(S)-(1-Phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacyclotetradecan-2-one, dibenzyl ester (D28) A solution of N- (1-(S)-dibenzyloxyphosphinylpropyl)-(S)leucine (D3B) (0.091g) in anhydrous dichloromethane (10 ml) maintained at 0°C was treated with 1-(3IE 911165 B2957 - 52 dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.04g) and 1-hydroxybenzotriazole (0.028g). After stirring for 0.5h the reaction mixture was sequentially treated with lactam trifluoroacetate salt (D17) (0.23 mmol) and N,N-diisopropylethylamine (0.054g). Stirring was continued for 18h at room temperature, then the mixture was washed with sat. aq. NaCl (2x10 ml). The organic fraction was dried over anhydrous magnesium sulphate, and evaporated in vacuo to yield an oil.

Purification by flash chromatography (2% methanol in chloroform) afforded the title compound as an oil (0.105g).

Observed (M+H)+ 777. ^43^^61^4^7^ requires M 776.

Description 29 (S)-3-[N-[N- (R)-(1-Phosphonoethyl)-(S)-leucyl]]amino-8-(Nbenzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester (D29A) and (S)-3-[N-[N-(S)-(1-phosphonoethyl)-(S)leucyl]]amino-8-(N-benzyloxycarbonyl)-1, 8diazacyclotridecan-2-one, dibenzyl ester (D29B) A solution of the isomeric mixture of acids (D29A) and (D29B) (0.28g) in anhydrous dichloromethane (20 ml) maintained at 0°C was treated with 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.137g) and 1-hydroxybenzotriazole (0.097g). After B2957 - 53 stirring for 0.5h the reaction mixture was sequentially treated with lactam (D7) (0.072 mmol) and Ν,Νdiisopropylethylamine (0.185g). Stirring was continued for 18h at room temperature, then the mixture was washed with sat. aq. NaCl (20 ml). The organic fraction was dried over anhydrous magnesium sulphate, and evaporated in vacuo to yield an oil. On purification by flash chromatography (2% methanol in chloroform), the title compound (D29) was obtained as oil (0.32g).

Observed FAB (M+H) + 749. C41H57N4O7P requires M 748.

Description 30 N-((R)-Dibenzyloxyphosphinylethyl)-(S)-leucine (D30A) and N-((S)-1-dibenzyloxyphosphinylethyl)-(S)-leucine (D30B) The title mixture of diastereoisomers (D30) was prepared analogously to the method in Description 3, Method B, as a white solid.

Observed FAB (M+H) + 420. ^22^30^^5^ re<4uires M 419. - 54 IE 911160 B2957 Example 1 (S)-3-[N-[N-((R)-1-Phosphonopropyl)-(S)-leucyl]]amino1, 8-diazacyclotridecan-2-one (El) A solution of phosphonic diester (D8) (O.llg) in ethanol (30 ml) was hydrogenated over 10% palladium on charcoal at atmospheric pressure for 24h. The solution was filtered through Kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound as a white solid, 0.057g, m.p. 178.5-180.5°C.

Observed FAB (M+H)+ 449. c20H41°5N4P requires M 448. δ (CD3OD): 0.96(6H,t), l.l(3H,t), 1.25-2.0(17H,m), 2.66(lH,m), 3.05(5H,m), 3.3 (lH,m overlapping with MeOD), 3.58(IH, m) , 4.38 (IH,m) . - 55 B2957 Example 2 (S)-3-[N-[N-((R)-1-Phosphonopropyl) -(S)-leucyl]]amino-1,7diazacyclotridecan-2-one (E2) A solution of phosphonic diester (D13) (0.09g) in methanol (25 ml) was hydrogenated over 5% palladium on charcoal at atmospheric pressure for 24h. The solution was filtered through kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound (E2) as a white solid (0.038g).

Observed (M+H) + 449. C2qH^^O^N^P requires M 448. δ (CDC13): 0.96(6H,d), 1.2(3H,t), 1.35-1.9(17H,m), 2.49(lH,m), 2.9(lH,m), 2.98 (3H, m) , 3.12 (1H, m) , 3.42(lH,m), 3.7(lH,t), 4.33(lH,t).

IE 911165 - 56 B2957 Example 3 (S)-3-[N-[N-((R)-1-Phosphonopropyl)-(S)-leucyl]]amino-1,8diazacyclotetradecan-2-one (E3) A solution of phosphonic diester (D18) (0.09g) in methanol (20 ml) was hydrogenated over 5% palladium on charcoal (0.08g) at atmospheric pressure'for 24h. The solution was filtered through kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound (E3) as a white solid (0.05g).

Observed FAB (M+H)+ 463. C21H43N4°5P reciuires M 462. δ (CD3OD): 0.98(6H,t), 1.12(3H,t), 1.3-2.1(19H,m), 2.7-3.25(6H,m) , 3.72(lH,m), 4.45(lH,m), 4.55(lH,m). - 57 B2957 Example 4 (S)-3-[N-[N-((R)-1-Phosphonopropyl)-(S)-leucyl]]amino-8(N-methyl)-1,8-diazacyclotridecan-2-one (E4) A solution of phosphonic diester (D21) (0.09g) in methanol (15 ml) was hydrogenated over 5% palladium on charcoal at atmospheric pressure for 24h. The solution was filtered through kieselguhr and the solvent evaporated in vacuo.

The residue was triturated with diethyl ether (5 ml) to give the title compound (E4) as a white solid (0.051g).

Observed (M+H) + 463. ^21Η43Ν4θ5Ρ requires M 462.

(CD3OD) : 0.98(6H,t), l.l(3H,t), 1.4-2.1 (17H,m) , 2.7(lH,m), 2.88(3H,s), 3.05-3.4 ( 6H, m) , 3.65(lH,m), 4.4 (lH,m) . - 58 B2957 Example 5 (S)-3-[N-[N-((R)-1-Phosphonopropyl) -(S)-leucyl]]amino-1,8diazacycloundecan-2-one, hydrochloride salt (E5) A solution of phosphonic diester (D26) (0.07g) in ethanol (10 ml) was treated with 5% palladium on charcoal (0.06g) and hydrogenated for 18h at atmospheric pressure. The suspension was filtered through kieselguhr, and the solvent evaporated in vacuo to give a pale brown foam.

The foam was dissolved in ethanol, and treated with 1M ethereal HCI to afford the title compound (E5) as a white solid. δ (CD3OD) : 1.0(6H,t), 1.18(3H,t), 1.58(2H,m), 1.7-2.1(llH,m), 2.9-3.2(6H,m), 3.6(lH,m), 4.45(lH,m), 4.55(lH,m).

Example 6 (S)-3- [N- [N- ( (S)-1-Phosphonopropyl)-(S) -leucyl] ]amino-1,8diazacyclotridecan-2-one (E6) B2957 - 59 A solution of phosphonic diester (D27) (0.29g) in methanol (40 ml) was hydrogenated over 5% palladium on charcoal (0.25g) at atmospheric pressure for 24h. The solution was filtered through kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound (E6) as a white solid (0.16g).

Observed FAB (M+H)+ 449. (-'20H41N4(^5P requires M 448. δ (CD3OD): 1.0(6H,dd), l.l(3H,t), 1.4-2.0(17H,m), 2.45(lH,m), 2.85-3.2 (5H,m) , 3.38(lH,m), 3.58(lH,m), 4.49(lH,t).

Example 7 (S)-3-[N-fN-((5)-1-Phosphonopropyl)-(S)-leucyl]]amino-1,8diazacyclotetradecan-2-one (E7) A solution of phosphonic diester (D28) (0.085g) in methanol (20 ml) was hydrogenated over 5% palladium on charcoal (0.08g) at atmospheric pressure for 24h. The solution was filtered through kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound (E7) as a white solid (0.045g).

Observed FAB (M+H)+ 463. C21H43N4°5P requires M 462.

B2957 - 60 δ (CD3OD): 0.98(6H,m), 1.12(3H,t), 1.3-2.0(19H, m) , 2.85-3.25(6H,m), 3.65(lH,m), 4.12(lH,m), 4.49(lH,dd).

Example 8 (S) - 3-fN-[N-((R)-1-Phosphonoethyl)-(S)-leucyl]1 amino-1,8diazacyclotridecan-2-one (E8A) and (S)-3-[N-[N-( (S)-1phosphonoethyl)-(S)-leucyl]]amino-1,8-diazacyclotridecan2-one (E8B) A solution of phosphonic diester (D29) (0.26g) in methanol (30 ml) was hydrogenated over 5% palladium on charcoal (0.2g) at atmospheric pressure for 24h. The solution was filtered through Kieselguhr and solvent evaporated in vacuo. The residue was triturated with diethyl ether (5 ml) to give the title compound (E8) as a white solid as a mixture of diastereoisomers.

Observed FAB (M+H) + 435. ^!9Η39Ν4θ5^ requires M 434.

B2957 - 61 COLLAGENASE INHIBITOR ASSAY The test is performed essentially as in Cawston and Barrett, Anal. Biochem. 99, 340-345 (1979) . Compounds for testing are dissolved in methanol by sonication and added to collagenase (purified from culture supernatants from the human lung fibroblast cell line, WI-38) in buffer. After a 5 min pre-incubation at 37°C, the assay tubes are cooled to 4°C and ^H-acetylated rat skin type I collagen is added. The assay tubes are incubated at 37°C overnight. The H-collagen forms insoluble fibrils, which are the substrate for the enzyme.

To terminate the assay, the assay tubes are spun at 12000 rpm for 15 minutes. Undigested "Ή-collagen is pelleted, o while digested H-collagen is found as soluble peptides in the supernatant. A sample of the supernatant is taken for liquid scintillation counting.

The activity of collagenase inhibitors (IC^q: 50% inhibitory concentration) is expressed as that concentration of compound that inhibits a known (standard) concentration of enzyme by 50%.

The compounds of Examples E1-E7 had IC^q values in the range 6 χ 10~θ - 3 x 10~®M.

Claims (21)

Claims :

1. A compound of the formula (I) or a salt, solvate or 5 hydrate thereof 15 in which, R is hydrogen, C-[__g alkyl or optionally substituted benzyl; R^ is hydrogen or C-^.g alkyl; 20 R 2 is C 2 _g alkyl; and R^ and R^ are joined together as -(CH 2 ) -X-(CH 2 ) - where p is an integer from 1 to 9, q is an integer from

2. To 10, and the moiety -(CH 2 ) - is adjacent to the carbon atom bearing Rg marked with an asterisk in formula (I), and X 25 is -NR5- where Rg is selected from hydrogen, C]__galkyl, C 2 _galkanoyl, C^.galkoxycarbonyl, aroyl, aralkyl or aralkyloxycarbonyl in each of which the aryl moiety is optionally substituted. 30 2. A compound according to claim 1 in which R is hydrogen, methyl, ethyl or benzyl. B2957 - 63

3. A compound according to claim 1 or 2 in which is hydrogen, methyl, ethyl, isopropyl or n-butyl.

4. A compound according to any one of claims 1 to 3 in 5. Which R 2 is n-butyl, iso-butyl or sec-butyl.

5. A compound according to any one of claims 1 to 4 in which R 3 and R^ are joined together as -(CH 2 ) p - X- (CH 2 ) where p and q have values such that Rg and R 4 form part of 10 an 11- or 13 to 16-membered azalactam structure, and X is -NRg- where Rg is hydrogen, methyl, benzyl, tbutoxycarbonyl or benzyloxycarbonyl.

6. A compound according to any one of claims 1 to 5 in 15 which R is hydrogen, Rj is methyl or ethyl, R 2 is isobutyl, and Rg and R^ are joined together as -(CH 2 )p-X-(CH 2 )where p is 4 and q is 5, ρ is 3 and q is 6 or p is 4 and q is 6, or p is 4 and q is 3 and X is -NRg- where Rg is hydrogen or methyl.

7. A compound according to any one of claims 1 to 6, in which the chiral centres marked with an asterisk in formula (I) have the S-configuration when Rg is other than hydrogen .

8. A compound according to claim 1 which is: (S)-3-[N-[N-((R)-1-phosphonopropyl) -(S)-leucyl]]amino1, 8-diazacyclotridecan-2-one; (S)-3-[N-[N-((R)-1-phosphonopropyl)-(S)-leucyl]]amino30 1,7-diazacyclotridecan-2-one; (S)-3-[N-[N-((R)-1-phosphonopropyl)-(S)-leucyl]]amino1,8-diazacyclotetradecan-2-one; B2957 - 64 (S) - 3-[Ν-[Ν-((R)-1-phosphonopropyl)-(S)-leucyl]]amino8-(N-methyl)-1,8-diazacyclotridecan-2-one; (S) -3-[N-[N-((R)-1-phosphonopropyl)-(S)-leucyl]]amino1, 8-diazacycloundecan-2-one, hydrochloride salt; 5 (S)-3-[N-[N-((S)-1-phosphonopropyl)-(S)-leucyl]]amino1, 8-diazacyclotridecan-2-one; (S)-3-[N-[N- ( (S)-1-phosphonopropyl)-(S)-leucyl]]amino1.8- diazacyclotetradecan-2-one; (S) -3-[N-[N-((R)-1-phosphonoethyl)-(S)-leucyl]]amino10 1,8-diazacyclotridecan-2-one and (S)-3-[N-[N- ( (S)-1-phosphonoethyl)-(S)-leucyl]]amino1.8- diazacyclotridecan-2-one.

9. A process for the preparation of a compound according 15 to claim 1 which process comprises converting a group R£q to hydrogen by cleaving a group R20 from a compound of formula (II): (II) wherein R 20 is alkyl or optionally substituted benzyl and R2i_ is hydrogen or alkyl or optionally substituted benzyl and R lz R2, R3 and R 4 are as defined in formula (I), and where necessary, converting R21 to hydrogen.

10. A compound of the formula (II) as defined in claim 9 subject to the proviso that R21 is not hydrogen. ΙΕ 9111θ 5 B2957 - 65

11. A compound according to claim 10 which is (S) -3-[N-[N-(R)- (1-phosphonopropyl)-(S)-leucyl]]amino8-(N-benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester; 5 (S)-3-[N-[N-(R)-(1-phosphonopropyl)-(S)-leucyl]]amino-7(N-benzyloxycarbonyl)-1,7-diazacyclotridecan-2-one, dibenzyl ester; (S)-3-[N-[N-(R)-(1-phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacyclotetradecan-2-one, 10 dibenzyl ester; (S)-3-[N-[N-(R)-(1-phosphonopropyl)-(S)-leucyl]]amino-8(N-methyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester; (S)-3-[N-[N-(R)-(1-phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacycloundecan-2-one, 15 dibenzyl ester; (S)-3-[N-[N-(S)-(1-phosphonopropyl)-(S)-leucyl]]amino-8(N-benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester; (S)-3-[N-[N-(S)-(1-phosphonopropyl)-(S)-leucyl]]amino-820 (N-benzyloxycarbonyl)-1,8-diazacyclotetradecan-2-one, dibenzyl ester; (S)-3-[N-[N-(R)- (1-phosphonoethyl) - (S)-leucyl]]amino-8-(N benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, dibenzyl ester and 25 (S)-3-[N-[N- (S)-(1-phosphonoethyl)-(S)-leucyl]]amino-8-(N benzyloxycarbonyl)-1,8-diazacyclotridecan-2-one, .dibenzyl ester .

12. A pharmaceutical composition comprising a compound 30 according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof, and a pharmaceutically acceptable carrier. B2957 - 66

13. A compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof, for use as an active therapeutic substance. 5

14. A compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof, for use in the treatment of conditions in which degradation of connective tissue and other proteinaceous components of the body occurs.

15. The use of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof, in the manufacture of a medicament for the treatment of conditions in which degradation of 15 connective tissue and other proteinaceous components of the body occurs .

16. A compound of the formula (I) given and defined in claim 1 or a salt, solvate or hydrate thereof, substantially as hereinbefore described and exemplified.

17. A process for the preparation of a compound of the formula (I) given and defined in claim 1 or a salt, solvate or hydrate thereof, substantially as hereinbefore described and exemplified.

18. A compound of the formula (I) given and defined in claim 1 or a salt, solvate or hydrate thereof, whenever prepared by a process claimed in claim 9 or 17.

19. A compound of the formula (II) as claimed in claim 10, substantially as hereinbefore described.

20. A pharmaceutical composition according to claim 12, substantially as hereinbefore described.

21. Use according to claim 15, substantially as hereinbefore described.