IE904122A1 - Cycloalkyl-substituted glutaramide diuretic agents - Google Patents

Abstract

Compounds of formula (I), wherein A completes a 4 to 7 membered carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be fused to a further saturated or unsaturated 5 or 6 membered carbocyclic ring; B is (CH2)m wherein m is an integer of from 1 to 3; each of R and R<4> is independently H, C1-C6 alkyl, benzyl or an alternative biolabile ester-forming group; R<1> is H or C1-C4 alkyl; R<2> is C1-C6 alkyl substituted by C1-C4 alkoxy, aryl or aryloxy and R<5> is defined to include a range of alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and substituted alkyl groups including in particular methoxyethyl, 2-methoxyethoxymethyl, N<2>-substituted-S-lysylaminomethyl, R-alanylaminomethyl and (2-amino-2-methyl-propanoyl) aminomethyl; are atriopeptidase inhibitors of utility in the treatment of hypertension, heart failure, renal insufficiency and other disorders.

Description

This invention relates to a series of cycloalkyl-substituted glutaramide derivatives which are diuretic agents having utility in a variety of therapeutic areas including the treatment of various cardiovascular disorders such as hypertension, heart failure and renal insufficiency.

According to the specification of our European patent applications EP-A-0274234 and EP-A-0343911 we describe and claim certain cycloalkyl-substituted glutaramide derivatives cis diuretic agents. The present invention provides further related compounds having an extended substituent group in the aminocycloalkanecarboxylate ring.

The compounds are inhibitors of the zinc-dependent, neutral endqpeptidase E.C.3.4.24.11. This enzyme is involved in the breakdown of several peptide hormones, including atrial natriuretic factor (ANF), which is secreted by the heart and which has potent vasodilatory, diuretic and natriuretic activity. Thus, the compounds of the invention, by inhibiting the neutral endopeptidase E.C.3.4.24.11, can potentiate the biological effects of ANF, and in particular the compounds are diuretic agents having utility in the treatment of a number of disorders, including hypertension, heart failure, angina, renal insufficiency, premenstrual syndrome, cyclical oedema, Meni^res disease, hyperaldosteronism (primary and secondary) pulmonary oedema, ascites, and hypercalciuria. In addition, because of their ability to potentiate the effects of ANF the compounds have utility in the treatment of glaucoma. As a further result of their ability to inhibit the neutral endopeptidase E.C.3.4.24.11 PLC 513 the carpounds of the invention may have activity in other therapeutic areas including for exanple the treatment of asthma, inflammation, pain, epilepsy, affective disorders, dementia and geriatric confusion, obesity and gastrointestinal disorders (especially diarrhoea and irritable bowel syndrome), the modulation of gastric acid secretion and the treatment of hyperreninaemia and leukemia.

The compounds of the present invention are of the formula: ro2c (i) wherein A completes a 4 to 7 membered carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be fused to a further saturated or unsaturated 5 or 6 membered carbocyclic ring ; B is (CI^)^ wherein m is an integer of from 1 to 3; A each of R and R is independently H, C^-Cg alkyl, benzyl or an alternative biolabile ester-forming group; R1 is H or Cj-C4 alkyl; PLC 513 and or or .

R is C^-Cg alkyl substituted by C^-C^ alkoxy, aryl or aryloxy; .

R is C^-Cg alkyl, C2~Cg alkenyl, C2-Cg alkynyl, C3~C7 cycloalkyl, or cycloalkenyl, R is Cj-Cg alkyl substituted by halo, hydroxy, C^-Cg alkoxy, C^-Cg alkoxy(C^-Cg)alkoxy, C^-C? cycloalkyl, C^-C? cycloalkerryl, aryl, aryloxy, heterocyclyl, -NR6R7, -NR8OOR9, -NR8SO2R10, -OONR6R7 or R^N-^-Cg)alkoxy; R is CL-C- alkyl substituted by a group of the formula: 1 o R12 -NR^CCHC-R13 R12 * 13 -NR SO -C-R or R13 -aONRU-C-R14 PLC 513 wherein R6 and R7 are each independently H, C-^-C^ alkyl, cycloalkyl, aryl, aryl(C^-C^)alkyl, C2~Cg alkoxyalkyl, 7 or heterocyclyl; or the two groups R and R are taken together with the nitrogen to which they are attached to form a pyrrolidinyl, piperidino, morpholino, piperazinyl or N-(C1-C4)alkyl-piperazinyl group; R8 is H or alkyl; R9 is (^-C^ alkyl, CF3, aryl, aryl (Gj-C^ alkyl, aryl(C^-cpalkoxy, heteorcyclyl, C|-C4 alkoxy or NR6R7 wherein R and R are as previously defined; R10 is c1_c4 alkyl, C^-C^ cycloalkyl, aryl or heterocyclyl; R11 is H, Cq-C6 alkyl, aryl or Clj-C^ cycloalkyl; R12 is R11C0NR11-, R11SO2NR11-, R16R17N-(0^)^, or R110~, wherein each R11 is as previously defined above; R13 and R14 are each independently H or C^-Cg alkyl; or R13 is H and R14 is C,-C^ alkyl which is substituted by A Ό CM, alkoxy, SH, SOt^, NH^, aryliC^-CgJalkylOCONH-, NH2OO-, O02H, 13 or the two groups R guanidino, aryl, or heterocyclyl; 14 . . and R are joined together to form, with the carbon a tern to which they are attached, a 5 or 6 membered carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be PLC 513 substituted by C^-C4 alkyl or fused to a further 5 or 6 membered saturated or unsaturated carbocyclic ring; or R^3 is H, and R12 and R14 are linked to form a 2-(N-1^-4-amincpyrrolidinyl) group; R15 is R16R17NOO-, R-^OCO-, R11OCH2- or heterocyclyl, wherein is as previously defined above; 17 R and R are each independently H or C^-Cg alkyl; and p is 0 or an integer of from 1 to 6; and pharmaceutically acceptable salts thereof and bicprecursors therefor.

In the above definition, unless otherwise indicated, alkyl groups having three or more carbon atoms may be straight or branched-chain. The term aryl as used herein means an aromatic hydrocarbon group such as phenyl, naphthyl or biphenyl which may optionally be substituted with one or more CH, CN, CF^, C^-C^ alkyl, C^-C^ alkoxy groups or halo atoms. Halo means fluoro, chloro, bromo or iodo.

PIC 513 The term heterocyclyl means a 5 or 6 membered nitrogen, oxygen or sulphur containing heterocyclic group which, unless otherwise stated, may be saturated or unsaturated and which may optionally include a further oxygen or one to three nitrogen atoms in the ring and which may optionally be benzofused or substituted with for exanple, one or more halo, C^-C4 alkyl, hydroxy, carbamoyl, benzyl, oxo, amino or mono or di-(Cj-C4 alkyl) amino or (C1-C4 alkanoyl)amino groups. Particular exanples of heterocycles include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, furanyl, tetrahydrofuraryl, tetrahydropyrany1, dioxanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, indolyl, isoindolinyl, quinolyl, quinoxaliryl, quinazolinyl and benzimidazolyl, each being optionally substituted as previously defined.

The compounds of formula (I) may contain several asymmetric centres and thus they can exist as enantiomers and diastereomers.

The invention includes both mixtures and the separated individual 2 4 isomers. The substituents R , and CO2R may have cis or trans geometry relative to the amide attachment.

The pharmaceutically acceptable salts of the compounds of formula (I) containing an acidic centre are those formed with bases which form non-toxic salts. Exanples include the alkali metal salts such as the sodium, potassium or calcium salts or salts with amines such as diethylamine. Compounds having a basic centre can also form acid addition salts with pharmaceutically acceptable acids. Exanples include the hydrochloride hydrobromide, sulphate or bisulphate, phosphate or hydrogen phosphate, acetate, citrate, fumarate, gluconate, lactate, PIC 513 maleate, succinate and tartrate salts.

The term bioprecursor in the above definition means a pharmaceutically acceptable biologically degradable derivative of the compound of formula (I) which, upon administration to an animal or human being, is converted in the body to produce a compound of the formula (I).

A preferred group of compounds of the formula (I) are those wherein A is (O^) 4 5 the formula (II) below wherein R, R , R and R are as previously defined for formula (I): ., R1 is H and B is (Qk)2» ΐ·β· compounds of (II) Also preferred are those compounds of formulae (I) and (II) wherein R and R are both H (diacids) as well as biolabile mono and di-ester derivatives thereof wherein one or both of R and R is a biolabile ester-forming group.

Ihe term biolabile ester-forming group is well understood in the art cis meaning a group which provides an ester which can be readily cleaved in the body to liberate the corresponding diacid of formula (I) wherein R and R are both H. A number of such PLC 513 ester groups are well known, for exanple in the penicillin area or in the case of the ACE-inhibitor antihypertensive agents.

In the case of the compounds of formulae (I) and (II) such biolabile pro-drug esters are particularly advantageous in providing compounds of the formula (I) suitable for oral administration. The suitability of any particular ester-forming group can be assessed by conventional animal or in vitro enzyme J hydrolysis studies. Thus, desirably for optimum effect, the ester should only be hydrolysed after absorption, accordingly, the ester should be resistant to hydrolysis before absorption by digestive enzymes but should be readily hydrolysed by, for exanple, liver enzymes. In this way the active diacid is released into the bloodstream following oral absorption.

In addition to lower alkyl esters (particularly ethyl) and benzyl esters, suitable biolabile esters include alkanoyloxyalkyl esters, including alkyl, cycloalkyl and aryl substituted derivatives thereof, aryloxyalkyl esters, aroyloxyalkyl esters, arylalkyloxyalkyl esters, arytesters, aralkylesters, and haloalkyl esters wherein said alkanoyl or alkyl groups have from 1 to 8 carbon atoms and are branched or straight chain and said aryl groups are phenyl, naphthyl or indanyl optionally substituted with one or more C^-C4 alkyl or alkoxy groups or halo atoms.

Thus exanples of R and R when they are biolabile ester-forming groups other than ethyl and benzyl include: 1-(2,2-diethylbutyryloxy) ethyl, 2-ethylpropionyloxymethyl, 1-(2-ethylpropionyloxy)ethyl, l-(2,4-dimethyIbenzoyloxy)ethyl, 1(benzoyloxy) benzyl, 1- (benzoyloxy) ethyl, 2 -methyl-1 -propionyloxypropyl, 2,4,6-trimethylbenzoy loxymethy 1, 1-(2,4,6-tr imethy 1IE 904122 PLC 513 benzyloxy)ethyl, pivaloyloxymethyl, phenethyl, phenpropyl, 2,2,2trif luoroethyl, 1- or 2-naphthyl, 2,4-dimethylphenyl, 4-t-butylphenyl, [5-(4-methyl-l,3-dioxolen-2-onyl) ] methyl and 5-indanyl. compounds of the formulae (I) and (II) wherein R is benzyl or 4 t-butyl and R is ethyl are valuable intermediates for the , 4 preparation of the diacids wherein R and R are both H.

. In a further preferred group of compounds R is methylene substituted by a group of the formula -NHC0CR12R^3R14' particularly where R^2 is NH^ R^^CCWH- or R^SO^NH-, R^3 is H and R14 is -(CH2)4NH2. Particularly preferred are such groups derived 5 from S-lysine; thus especially preferred R substitutents of this type include N2-acetyl-S-lysylaminomethyl, Lp-methanesulphonyl-Slysyl-aminomethyl, N^-phenylsulphonyl-S-lysyl-amirranethyl and N2 -cyclctoutylcarbonyl-S-lysyl-aminomethyl.

In further groups of preferred compounds R is c^~Cg alkyl 5 . substituted by C^-Cg alkoxy, particularly methoxyethyl? or R is C^-Cg alkyl substituted by C^-Cg 2-methyoxyethoxymethyl; or wherein R is C^-Cg alkyl substituted by -NHCOCR12R13R14 wherein R12 is NH2, R13 is CH3 and R14 is H or CH3 including in particular R-alanyl-amincmethyl and (2-amino-2methylpropanoy 1) amincmethy 1.

R is preferably phenyl, benzyl, phenethyl, methoxypropyl or ethoxypropyl.

Particularly preferred individual compounds of the invention include: 2S- (N2 -methanesulphonyl-S-lysylamincmethyl) -3-(l-[cis-4carboxy-3-cis {3-ethoxypropyl) -cyclohexyl) carbamoyl ] cyclopentyl} propanoic acid, PIG 513 2-(112 -methanesulphonyl-S-lysylaminomethyl)-3-{l-[ (cis-4carboxy-3-cis-phenethyl-cyclohexyl) carbamoyl] cyclopentyl} propanoic acid, 2S—(2-methoxyethoxymethyl)-3-{1-[(cis-4-carboxy-3-cis-(3ethoxypropyl}cyclohexyl)carbamoyl]cyclopentyl}propanoic acid and 2S—(2-methoxyethoxymethyl)-3-(1-[(cis-4-carboxy-3-cis-{3methoxypropyl} -cyclohexyl) carbamoyl ] cyclopentyl) propanoic acid, and biolabile ester derivatives thereof.

The compounds of formula (I) are prepared by a number of different processes. The basic procedure involves the synthesis of a partially protected cycloalkyl-substituted glutaric acid derivative which is coupled to an amine to give the desired glutaramide. The carboxylic acid group in the amine, if free, or any reactive groups in R , may require protection during the coupling step and such protecting groups are removed in the final stage of the process.

The synthetic route is illustrated in scheme 1 wherein A, B, ' . 5 .

R and R are as previously defined, R is as defined for R with 19 any reactive group therein protected if necessary and R and R 4 are as defined for R and R excluding H, or they are conventional carboxylic acid protecting groups: PIC 513 Scheme 1 I (I) The reaction of the compounds of formula (III) and (IV) is achieved using conventional amide coupling techniques. Thus in one process the reaction is achieved with the reactants dissolved in an organic solvent, e.g. diohloromethane, using a diimide condensing agent, for exanple 1-ethyl-3-(dimethylamincpropyl) carbodiimide, or N,Ν'-dicyclohexylcarbodiimide, advantageously in the presence of l-hydroxybenzotriazole and an organic base such as N-methylmorpholine. The reaction is generally complete after a period of from 12 to 24 hours at room temperature and the product is then isolated by conventional procedures, i.e. by washing with water or filtration to remove the urea biproduct and evaporation of the solvent. The product may be further purified by PLC 513 crystallisation or chromatography, if necessary. The compounds of 4 formula (V) include compounds of formula (I) wherein R and R are C-.-C, alkyl or benzyl.

JL D In some cases the coupled product, in protected form, may be subjected to conventional chemical transformation reactions to allow preparation of further compounds of formula (V). Thus for . 5' example carpounds of formula (V) wherein R contains an ester group may be hydrolysed or hydrogenated to generate the carboxylic acid which may be further reacted, for example with ah amine, to give amide derivatives.

’ Similarly ccnpounds wherein R contains a substituted or protected amino group (for exanple a benzyl amino, dibenzylamino, benzyloxycarbonylamino or t-butyloxycarbonylamino group) may be converted to the free amines by hydrogenation or protonolysis as appropriate. The amines produced may be further reacted, thus for example reaction with a sulphonyl halide yields the corresponding sulphonamides, acylation with an acid chloride or anhydride yields the corresponding amides, reaction with an isocyanate yields urea derivatives and reaction with a chloroformate yields the alkoxycarbonylamino and products respectively. All these transformations are entirely conventional and appropriate conditions and reagents for their performance will be well known to those skilled in the art as will other variations and possibilities.

The diesters of formula (V) may be further reacted to give the monoester of diacid derivatives of formula (I) wherein one or 4 both of R and R are H. The conditions used will depend on the T8 19 precise nature of the groups R and R present in the compound PIG 513 of formula (V) and a number of variations are possible. Thus for 18 19 example when both of R and R are benzyl, hydrogenation of the . . . . . 4 product will yield the diacid of formula (I) wherein R and R are 19 both H. Alternatively if one of R and R is benzyl and the other is alkyl, hydrogenation will yield a monoester product.

This can be hydrolysed, if desired, again to yield the diacid 18 19 product. When one of R and R is t-butyl, treatment of the compound of formula (V) with trifluoroacetic acid yields the 18 19 corresponding acid. The diester product wherein R and R are benzyl or lower alkyl can also be treated with trimethylsilyl iodide to produce the dicarboxylic acid product. If some other 18 19 carboxylic acid protecting group is used for R or R then clearly appropriate conditions for its removal must be employed in the final step to give the ester or diacid product of formula . . 5 .

(I). In the case where the ring A or the substituent R is unsaturated, the deprotection must be effected by non-reductive . . 4 . methods, thus for example if either of R and R is benzyl, they may be removed by treatment with trimethylsilyl iodide.

As well as removing any protecting group which may be present ' ... in R , a number of chemical transformation reactions are possible on the final mono-ester or diacid products as previously described. In each case the product may be obtained as the free carboxylic acid or it may be neutralised with an appropriate base and isolated in salt form.

PLC 513 In a variant of the above procedure, compounds of the formula C Q Q Q Ί n (I) wherein R is C^-Cg alkyl substituted by -NR COR , -NR SO2R , -NR11OOCR12R13R14 or -NR11SO2CR12R13R14 are prepared by a process which involves acylating or sulphonylating a compound of the formula: (VI) 8 11 18 19 wherein R is as defined for R or R , R and R are as previously defined and Y is a C^-Cg alkyl group; by reaction with 9 10 12 13 14 an acid of the formula R C02H, R SO3H, RRRCOC>2H, or R12R13R14CSO3H, or an activated derivative thereof. The resulting amide or sulphonamide product is then deprotected if required and the diester product cleaved to yield the carboxylic acids of 4 formula (I) wherein R and R are each H as previously described. Hie compounds of formula (VI) are prepared following the procedures shewn in scheme 1 but using a compound of formula (III) ' ... 5' having R as a protected amine derivative. Thus, for example R can contain a bis-(lS)-phenylethylamino substituent.

Hydrogenation of the coupled product gives the corresponding free . 20 . amine of formula (VI) wherein R is H and Y is Hus route is of particular value for the preparation of compounds having 2(S) stereochemistry in the glutaramide backbone.

PLC 513 The starting cycloalkyl-substituted glutaric acid mono esters of formula III may be prepared as described in cur European patent applications EP-A-0274234, 89305180.5 and 89304698.7.

The amines of formula (IV) are generally novel compounds (particularly when the substituents have defined stereochemistry) and they are prepared by appropriate synthetic procedures in accordance with literature precedents. Thus in one procedure the corresponding ketone is converted to the amine via reduction of the oxime. In an alternative process the corresponding alcchol is reacted with methyl toluenesulphonate in the presence of diethyl diazodicarboxylate and triphenylphosphine; the resulting 4-methylbenzenesulphonyloxy derivative is reacted with sodium azide and the product reduced to yield the amine. These procedures are illustrated in the Preparations give hereafter.

Appropriate coupling and protecting methods for all of the above steps and alternative variations and procedures will be well known to those skilled in the art by reference to standard text books and to the exanples provided hereafter.

PLC 513 As previously mentioned, the compounds of the invention are potent inhibitors of the neutral endopeptidase (E.C.3.4.24.11). This enzyme is involved in the breakdown of a number of peptide hormones and, in particular it is involved in the breakdown of atrial natriuretic factor (ANF). This hormone consists of a family of related natriuretic peptides, secreted by the heart, of which the major circulating form in humans is known to be the 28 amino-acid peptide referred to as £<-hANP. Thus, by preventing the degradation of ANF, by endopeptidase E.C.3.4.24.11, the compounds of the invention can potentiate its biological effects and the compounds are thus diuretic and natriuretic agents of utility in a number of disorders as previously described.

Activity against neutral endopeptidase E.C.3.4.24.11 is assessed using a procedure based on the assay described by J.T. Gafford, R.A. Skidgel, E.G. Erdos and L.B. Hersh, Biochemistry. 1983, 32, 3265-3271. The method involves determining the concentration of compound required to reduce by 50% the rate of release of radiolabelled hippuric acid from hippuryl-Lphenylalanyl-L-arginine by a neutral endopeptidase preparation frcan rat kidney.

Ihe activity of the compounds as diuretic agents is determined by measuring their ability to increase urine output and sodium ion excretion in saline loaded conscious mice, in this test, male mice (Charles River CDl, 22-28 g) are acclimatised and jxj starved overnight in metabowls. The mice are dosed intravenously via the tail vein, with the test compound dissolved in a volume of saline solution equivalent to 2.5% of body weight. Urine samples are collected each hour for two hours in pre-weighed tubes and analysed for electrolyte concentration. Urine volume and sodium ion concentration from the test animals are compared to a control group which received only saline.

For administration to man in the curative of prophylactic treatment of hypertension, congestive heart failure or renal insufficiency, oral dosages of the ocnpounds will generally be in the range of from 4-800 mg daily for an average adult patient (70 kg). Thus for a typical adult patient, individual tablets or capsules contain from 2 to 400 mg of active compound, in a suitable pharmaceutically acceptable vehicle or carrier for administration singly, or in multiple doses, once or several times a day. Dosages for intravenous administration would typically be within the range 1 to 400 mg per single dose as required. In practice the physician will determine the actual dosage which will be most suitable for an individual patient and it will vary with the age, weight and response of the particular patient. The above dosages are exemplary of the average case but there can, of course, be individual instances where higher or lcwer dosage ranges are merited, and such are within the scope of this invention.

PLC 513 For human use, the compounds of the formula (I) can be administered alone, but will generally be administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For example, they may be administered orally in the form of tablets containing such excipients as starch or lactose, or in capsules or ovules either alone or in admixture with i excipients, or in the form of elixirs or suspensions containing flavouring or colouring agents. They may be injected parenterally, for example, intravenously, intramuscularly or subcutaneously. For parenteral administration, they are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood.

The compounds may be administered alone but may also be administered together with such other agents as the physician shall direct to optimise control of blood pressure or to treat congestive heart failure, renal insufficiency or other disorders in any particular patient in accordance with established medical practice. Thus the compounds can be co-administered with a variety of cardiovascular agents, for exanple with an ACE inhibitor such cis captopril or enalapril to facilitate the control of blood pressure in treatment of hypertension; or with digitalis, or another cardiac stimulant or with an ACE inhibitor, for the treatment of congestive heart failure. Other possibilities include co-administration with a calcium antagonist (e.g.

PLC 513 nifedipine, amlodopine or diltiazem) a beta-blocker (e.g. atenolol) or an alpha-blocker (e.g. prazosin or doxazosin) as shall be determined by the physician as expropriate for the treatment of the particular patient or condition involved.

In addition to the above, the compounds may also be administered in conjunction with exogenous ANF, or a derivative thereof or related peptide or peptide fragment having I diuretic/natriuretic activity or with other ANF-gene related peptides (e.g. as described by D. L. Vesely et al, Biochem. Biophys. Res. Camm., 1987, 143. 186).

Thus in a further aspect the invention provides a pharmaceutical composition comprising a compound of the formula (I), or a pharmaceutically acceptable salt thereof or bioprecursor therefor, together with a pharmaceutically acceptable diluent or carrier.

The invention also includes a compound of the formula (I), or a pharmaceutically acceptable salt thereof or bioprecursor therefor, for use in medicine, particularly for use as a diuretic agent for the treatment of hypertension, congestive heart failure or renal insufficiency in a human being.

The invention further includes the use of a compound of the formula (I) for the manufacture of a medicament for the treatment of hypertension, heart failure, angina, renal insufficiency, premenstrual syndrane, cyclical oedema, Menieres disease, hyperaldosteronism, pulmonary oedema, ascites, hypercaciuria, glaucoma, asthma, inflammation, pain, epilepsy, affective disorders, dementia and geriatric confusion, obesity, FUC 513 gastrointestinal disorders (including diarrhoea), hyperreninaemia, leukemia, and the modulation of gastric acid secretion.

The preparation of the compounds of the invention will new be more particularly illustrated by reference to the following experimental examples, in which Preparations 1-8 describe preparation of the starting amines of formula (IV) and Exanples 1-45 describe preparations of compounds of the formula (I). The purity of compounds was routinely monitored by thin, layer chromatography using Merck Kieselgel 60 F254 plates. Solvent system ss-1 was a mixture of didhlorcmethane, methanol and glacial acetic acid (90:10:1), ss-2 was the upper phase of a mixture consisting of methyl iso-butyl ketone, glacial acetic acid and water (2:1:1), and ss-3 was a mixture of ethyl acetate and hexane (1:1)· ^Ή-Nuclear magnetic reasonance spectra were recorded using a Nicolet QE-300 spectrometer and were in all cases consistent with the proposed structures.

PLC 513 PREPARATION 1 c-4-Amino-c-2- (2-phenvlethvl) cvclohexane-r-1-carboxylic acid ethyl ester hydrochloride i. 4-Oxo-2-f2-rhenvlethvl)cyclohex-2-ene carboxylic acid ethyl ester An 80% dispersion of sodium hydride in oil (307 mg, 10 mmole) was added under nitrogen to an ioe cold stirred mixture of 3-oxo-5-phenyl pentanoic acid ethyl ester (28.4 g , 129 mmole) and absolute ethanol (1 ml). Methyl vinyl ketone (10.7 ml, 129 mmole) was then added dropwise over half an hour with continued ice cooling and, after stirring for an hour at room temperature, acetic acid (1.4 ml), pyrrolidine (775 mg), ethanol(17 ml) and water (2 ml) were added. After refluxing for 2 hours, the solvent was evaporated under vacuum and the residue dissolved in diethyl ether. Washing sequentially with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate and water, drying over MgSO^ and evaporation gave a yellow liquid (34.6 g). Chromatography on silica gel (600 g) eluting with a mixture of diethyl ether and hexane (1:3 by volume) gave the required enone as a clear liquid (18.54 g,) Rf 0.35 (1:1 diethyl ether:hexane). Found: C,74.88; H,7.40. 0 requires C,74.97; H,7.40%.

PLC 513 ii. 4-Oxo-cis-2-(2-phenvlethvl)cyclohexanecarboxvlic acid ethyl ester The enone (18.44 g, 6.76 mmole) from part (i) in ethanol (80 ml) containing 2N hydrochloric acid (3 ml) was hydrogenated over 5% palladium on charcoal catalyst (500 mg) at 50 p.s.i. (3.45 bar) pressure. After three hours a further amount of catalyst (250 mg) was added and reduction was continued for a further two hours. The suspension was filtered through Avicel and the filtrate evaporated under vacuum. The residual oil was dissolved in diethyl ether, washed with saturated aqueous sodium bicarbonate and water, dried over MgSO^ and evaporated to give a clear liquid (18.52 g). Chromatography on silica gel (600 g), eluting with an increasing proportion of diethyl ether in hexane (2:8 - 4:6) gave the required ketone as a liquid. (16.78 g,90%). Rf 0.4 (1:1 diethyl ether:hexane). iii. 4-Methoxvimino-cis-2-(2-phenylethyl)cyclohexane carboxylic acid ethyl ester The above ketone (3.02 g, 11 mmole), methoxylamine hydrochloride (1.19 g, 14.3 mmole) and sodium acetate (1.17 g; 14.3 mmole) were refluxed for five hours in ethanol (50 ml), the solvent was evaporated under vacuum, the residue taken up in diethyl ether and washed with saturated aqueous sodium bicarbonate followed by water. Drying ever MgSO4 and evaporation under vacuum gave the 0-methyl oxime as an oil (3.32 g, 99%) Rf 0.55 and 0.66 (1:1 diethyl ether, hexane). Found: C,71.23; H,8.33; N,4.88. ClgH25NO3 requires C,71.26? H,8.30; N,4.62%.

PIC 513 iv. c-4-Butoxycarbonvlamino-c-2- (2-phenvlethvl) -cyclohexaner-l-carboxylic acid ethyl ester Trifluroacetic acid (4.2 ml, 54 mmole) in dry tetrahydrofuran (10 ml) was added dropwise with ice cooling to a stirred suspension of sodium bonohydride (2.04 g, 54 mmole) in dry tetrahydrofuran. Ihe temperature was kept between 5 and 10°C and after stirring for fifteen minutes the 0-methyloxime from part (iii) (3.28 g; 10.8 mmole) in tetrahydrofuran (10 ml) was added dropwise over ten minutes. The temperature rose to 20°C and, after stirring for five hours, water was very carefully added with ice cooling and the mixture was partitioned between ethyl acetate and water. The aqueous layer was extracted twice with ethyl acetate, the combined organic extracts washed with saturated salt solution, dried over MgSC>4 and evaporated to give a gum (3.25 g). The crude amine was dissolved in methylene chloride (50 ml) N-methylmorpholine (2.2 g, 21.6 mmole) and di-t-butyl dicarbonate (4.7 g, 21.6 mmole) were added and the mixture allowed to stand at room temperature for 48 hours. The solvent was evaporated under vacuum, the residue partitioned between diethyl ether and water and the organic extract washed sequentially with 0.5N hydrochloric acid, water, saturated aqueous sodium bicarbonate and water.

Drying over MgSO4 and evaporation gave a gum (3.9 g). The required cis- compound was separated from the minor more polar trans-iscmer by chromatography on silica gel (400 g). Elution with a mixture of diethyl ether in hexane (3:7 by volume) gave the protected amine as an oil (1.94 g, 48%). Rf 0.35 (3:7 diethyl ether:hexane). Found: C,70.43; H,8.75; N,3.45. C22^33^4 requires C,70.37; H,8.86; N,3.73%.

PLC 513 v. c-4-Amino-c-2-(2-phenvlethyl)cyclcthexane-r-1-carboxylic acid ethyl ester hydrochloride An ioe cold stirred solution of the above ester (1.94 g, .17 nmole) in diethyl ether was saturated with hydrogen chloride. After three hours the solvent was blown off with nitrogen and the residue triturated with diethyl ether and collected by filtration giving the title amine salt as a white solid (1.44 g, 89%). m.p. 213-214°C. Rf 0.5 (ss-1). Found; C,65.84; H,8.74; N,4.43. Cj^gNO^HCl requires C,65.48; H,8.40; N,4.49%.

PREPARATION 2 c-4-Amino-c-2-(3-methoxvpropyl)cvclohexane-r-1-carboxvlic acid ethyl ester hydrochloride Ihe procedure of Preparation 1 was followed but using 3-oxo-6-methoxy-hexanoic acid ethyl ester as the starting material in part (i) to give the title amine as white solid, m.p. 206-208°C. Rf 0.2 (ss-1). Found: C,55.15; H,9.11; N,5.34. C13H25NO3· HCl, 0.2 ^0 requires C,55.09; H,9.39; N,5.94%.

IE 904122 PLC 513 PREPARATION 3 c-4-Amino-c-2- (3-ethoxvpropyl) cvclohexane-r-l-carfeoxyl ic acid ethyl ester hydrochloride The procedure of Preparation 1 was followed but using 3-oxo-6-ethoxyhexanoic acid ethyl ester as the starting material in part (i) to give the title amine as a white solid, m.p. 201- 203°C. Rf 0.2 (ss-1). Found: C,55.50; H,9.52; N,4.33. Cl4H2?NO3.HC1.0.5H20 requires c,55.52; H,9.65; N,4.63%.

PREPARATION 4 c-4-Amino-c-2-(2-phenylethvl)cvclohexane-r-1-carboxylic acid methvl ester hydrochloride A solution of the ethyl ester from Preparation 1 (1.4 g; 4.48 mmole) in dry methanol (50 ml) was saturated with hydrogen chloride and heated at 55 to 60°C for four days and then refluxed for a further 3 days. Hydrogen chloride was periodically introduced to maintain saturation. The solution was evaporated to dryness under vacuum, the residue dried azeotropically with methylene chloride, and the residue triturated with diethyl ether to give the methyl ester as a white powder (1.29 g, 97%) m.p. 202— 4°C. Rf 0.15 (ss-1). Found: C,63.70; H,8.13; N,4.74.

HCl.0.25 requires 0,63.56; H,8.17; N,4.63%.

PLC 513 PREPARATION 5 c-4-Amino-t-2-( 2-phenylethyl) cyclohexane-r-1-carboxylic acid ethyl ester i. cis-7-(2-Phenylethyl) -1,4-dioxaspiror 4.51decane-8carboxylic acid ethyl ester Ihe ketone of Preparation 1 part ii (10.97 g; 40 mmole), ethylene glycol (2.73 g, 44 mmole) and 4-toluenesulphonic acid (100 mg) were refluxed in benzene (80 ml) using a Dean-Stark water trap. After eight hours the solution was cooled, diluted with diethyl ether and washed sequentially with saturated aqueous sodium bicarbonate and water. Drying over MgSO^ and evaporation under vacuum gave a clear liquid (12.22 g, 96%) Rf 0.32 (3:7, diethyl ether, hexane). Found: C,71.45; H,8.30. C^gH2604 requires C,71.67; H,8.23%.

PLC 513 ii. trans-7- (2-Phenylethyl) -1,4-diaxaspino Γ 4,51 decane-8 carboxylic acid ethyl ester Potassium t-butoxide (1.9 g, 17 mmole) was added to a solution of the above ester (12.17g, 38.2 nmole) in dry t-butanol (90 ml) and the mixture refluxed under nitrogen for forty eight hours. Neutralisation with 2N hydrochloric acid and evaporation under vacuum gave a light brown oil which was dissolved in diethyl ether and washed with water. Drying over and evaporation gave an oil (12.09 g) which was chromatographed cn silica gel (600 g). Elutions with a mixture of diethyl ether and hexane (3:7 by volume) gave the required trans-isomer as an oil (8.44 g, 69%). Rf 0.29 (3:7 diethyl ether, hexane). Found: C,71.60; H,8.14. ClgH2604 requires C,71.67; H,8.23%. iii. 4-Oxo-trans-2-(2-phenvlethvl) cvclohexanecarboxylic acid ethyl ester Ihe above ester (8.40 g, 26.4 nmole) was refluxed in IN sulphuric acid (50 ml) amd ethanol (70 ml). After three hours most of the ethanol was evaporated off under vacuum, water was added and the suspension extracted with diethyl ether. Ihe organic extract was washed sequentially with water, saturated aqueous sodium bicarbonate and water, dried over MgSO4 and evaporated under vacuum to give a clear oil (6.46 g, 89%) Rf 0.38 (1:1 diethyl ether, hexane). Found: C,74.25; H,8.07. C^7H22O3 requires C,74.42; H,8.08%.

PLC 513 iv. t-2- (2-Fhenylethyl) -c-4-phenylroethvlamino-cvclchexane-rl-cartooxylic acid ethyl ester 3A molecular sieve (3 g) was added to a stirred solution of the ketone from part iii (3.2 g, 11.66 mmole) and benzylamine (1.27 ml, 11.66 mmole) in ethanol (15 ml). After three and a half hours at room temperature, the solution was pipetted off into a dry hydrogenation vessel washing the sieves with a little ethanol (2x5 ml). Platinum oxide (500 mg) was added and the reduction completed in three hours at 50 p.s.i. (3.45 bar). Ihe mixture was filtered through artoaoel, evaporated under vacuum and the residual liquid chromatographed on silica gel (200 g). Elution with increasing proportions of ethyl acetate in hexane (7:3 - 1:1 by volume) gave the required cis-isomer as a clear oil (3.17 g, 74%) Rf 0.3 (ss-3) Found: C,78.85; H,8.53; N,4.86. C^H^NO requires C,78.86; H,8.55; N,3.83%. v. c-4-Amino-t-2-(2-phenvlethvl)cyclohexane-r-1-carboxylie acid ethyl ester A solution of the above amine (3.13 g, 8.56 mmole) in ethanol (80ml) containing IN hydrochloric acid (9.42 ml, 9.42 mmole) was hydrogenated over 20% palladium hydroxide and charcoal catalyst (600 mg) at 50 p.s.i. (3.45 bar) pressure. Further amounts of catalyst (500 mg) were added after six and twenty four hours.

After a further twenty two hours the suspension was filtered through arbacel and the filtrate evaporated to dryness under __ ~ ~ a Λ _ IE 9041« PLC 513 vacuum. The residue was recrystallised from a mixture of ethanol and diethyl ether to give the required amine as a white solid (2.03 g, 76%) m.p. 175-7°C. Rf 0.25 (ss-1). Found: C,64.42; H,8.50; N,4.52. Cp^gNO^HCl.0.251^0 requires C,64.54; H,8.44; N,4.43%.

PREPARATION 6 c-4-Amino-c-2-phenvlcvclchexane-r-l-carboxvlic acid methyl ester hydrochloride i. 4-Oxo-2-phenvlcyclohex-2-ene-carboxylic acid methvl ester 2-Hydrcxy-4-Qxo-2-phenylcyclohexane carboxylic acid methyl ester (17.5 g, 70.48 mmole) [J. Org. Chem., 39 2427 (1974)] was stirred for 15 minutes at 80°C with polyphosphoric acid (17.5 g). On cooling the mixture was partitioned between ethyl acetate and water. The organic extract was washed sequentially with water, saturated aqueous sodium bicarbonate and water, dried ever MgSO4 and on evaporation gave an oil. Chromatography on silica gel eluting with increasing proportions of ethyl acetate in hexane (2:8 - 3:7 by volume) gave the required compound as a golden oil (9.0 g, 56%) Rf 0.5 (ss-3). ii. 4-Oxo-2-phenylcyclohexanecarboxylic acid roether ester The enone of part 1 (9.0 g, 39.08 mmole) in methanol (40 ml) containing 2N hydrochloric acid (1.5 ml) was hydrogenated over 5% palladium on charcoal catalyst at 50 p.s.i. (3.45 bar). After three hours the suspension was filtered through Arbaoel and evaporated to dryness under vacuum. Ihe residual oil was chromatographed on silica gel (400 g). Elution with increasing proportions of ethyl acetate in hexane (2:8 - 4:6 by volume) gave PLC 513 the required ketene as a white solid (4.04 g, 49%) Rf 0.55 (ss-3) Found: C,72.39; H,6.95. C,,H,rO, requires C,72.39; H,6.94%. lo J iii. c-4-Hydroxy-c-2-t±ienvlcyclchexane-r-l-carboxvlic acid methyl ester Sodium borohydride (2.33 g, 61.51 mmole) was added in small portions to a stirred solution of the ketone of part ii (3.78 g, 16.27 mmole) in methanol (80 ml), keeping the temperature belcw °C. After fifteen minutes the solvent was evaporated under vacuum, the residue dissolved in methylene chloride and washed with saturated salt solution. Drying over MgSO4 and evaporation under vacuum gave a white solid. Chrcmatography on silica gel (250 g) eluting with diethyl ether in hexane (1:1 by volume) gave the required ester as a foam (2.93 g, 77%). Found: C,71.84; H,7.82. C. .Η1θΟ_ requires C,71.77; H,7.74%. ±4 lo J iv. t-4- (4-Methvlbenzenesulphonyloxy) -c-2-thenylcyclohexane-r-l carboxylic acid methyl ester Diethyl diazodicarboxylate (4.08 g, 23.47 mmole) in tetrahydrofuran (10 ml) was added under nitrogen to a stirred solution of the ester of part iii (4.4 g, 18.78 mmole), triphenylphosphine (6.16 g, 23.47 mmole) and methyl 4-toluene sulphonate (4.37 g, 23.47 mmole) in tetrahydrofuran (60 ml) keeping the temperature between 5 and 10°C. After stirring overnight at rocxn temperature the mixture was re-cooled to 0°C and further amounts of triphenylphosphine (1.23 g, 4.7 mmole) and methyl 4-toluene sulphonate (874 mg, 4.7 mmole) were added followed by diethyl diazodicarboxylate (816 mg, 4.7 mmole). After stirring at room temperature for a further four hours, the mixture IE 904122 “ PLC 513 was evaporated to a small volume, the residue redissolved in methylene chloride and applied directly to a silica gel (400 g) chromatography column. Elution with increasing proportions of ethyl acetate in hexane (1:9 - 3:7 by volume) gave the required 4-toluenesulphonate as an oil (3.45g, 47%). Rf 0.45 (4:1 ethyl acetate, hexane) Found: C,64.71; H,6.26. C^H^O^-S requires; C,64.93; H,6.23%. v. c-4-Azido-c-2-phenvlcvclchexane-r-l-carboxvlic acid methyl ester Sodium azide (1.16 g, 17.76 mmole) was added to a stirred solution of the 4-toluenesulphonate from part iv (3.45 g, 8.88 mmole) in dimethylformamide, and the mixture was stirred at 60°C for eighteen hours. On cooling water was added and the suspension extracted with diethyl ether. The organic extract was washed with water, dried over MgSO^ and evaporation gave a clear oil which was chromatographed on silica gel (300 g). Elution with increasing proportions of ethyl acetate in hexane (1:20 - 1:4 by volume) gave the azide as waxy solid (2.11 g, 92%). Rf 0.55 (4:1 hexane, ethyl acetate). Found: C,65.20; H, 6.60; N,15.32. C14H17N3O2 requires C, 64.85; H,6.61; N,16.20%. - —— Λ Λ Λ Λ.'! r _ _ ityu«n«PLC 513 vi. c-4-Amino-c-2-thenvlcyclchexane-r-l-carboxylic acid methyl ester hydrochloride Ihe azide from part v (2.1 g, 8.1 m mmole) in methanol (50 ml) was hydrogenated over 10% palladium on charcoal catalyst (210 mg) at 50 p.s.i. (3.45 bar). After two and a quarter hours the mixture was filtered through a short arbacel column and evaporated under vacuum. Ihe residue was dissolved in ethyl acetate and acidified with IN hydrogen chloride in diethyl ether. Ihe precipitated salt was filtered and washed with ethyl acetate to give the title amine salt as a white solid (1.88 g, 94%) m.p. 255-6°C (dec). Rf 0.2 (ss-1). Found: C,62.64; H,7.74; N,5.74.

C14HigNO2.HC1 requires C,62.33; H,7.47; N,5.19%.

FLL OXJ PREPARATION 7 c-3-Antino-c-5-phenvImethvlcvclohexane-r-l-carboxylic acid methyl ester hydrochloride i. cis-5-tert-Butvldimethvlsilvloxv cvclchexane-cis-1,3dicarboxylic acid dimethyl ester t-Butyldimethylsilyl chloride (16.58 g, 110 mmole) in dry methylene chloride (65 ml) was added dropwise over half an hour to a stirred solution of cis-3-hvdroxycvclohexane-cis-l. 3dicarboxylic acid dimethyl ester (21.5 g, 99 mmole) [J. Org.

Chem., 38, 1726 [1973], triethylamine (11.13 g, 110 mmole and 4-dimethylaminopyridine (488 mg, 4 mmole) in methylene chloride (80 ml), with ice cooling keeping the temperature between 5 and 10°C. After standing overnight at room temperature further amounts of t-butyldimethylsilyl chloride (1.6 g), triethylamine (1.1 g) and dimethylaminopyridine (450 mg) were added and stirring continued for sixty-eight hours. Ihe mixture was then washed sequentially with water, saturated aqueous ammonium chloride and water, dried over MgSO4 and evaporated under vacuum to give a liguid (34.4 g) which was chromatographed on silica gel (600 g). Elution with a mixture of diethyl ether in hexane (1:3 by volume) gave the required product as a liquid (32.6 g, 99%) Rf 0.3 (1:3 diethyl ether, hexane). Found: C,57.89; H,9.20. Ό^^Η^θΟ^Ξΐ requires C,58.15; H,9.15%.

PLC 513 ii cis-5-tert-ButyldimethylsilyloxvcyclcAiexane-cis-l, 3dicarboxylic acid mono-methyl ester IN Sodium hydroxide (94 ml) was added to an ice cooled solution of the above diester (30.87 g, 93.4 mmole) in methanol (500 ml). The mixture was stirred at 5°C for 3 hours and then allowed to stand at room temperature for eighteen hours. Most of the solvent was evaporated off under vacuum, water was added and the suspension extracted with diethyl ether to recover unreacted diester (3.86 g). The aqueous phase was acidified with 2N hydrochloric acid, extracted with diethyl ether, and the organic extract washed with water, dried over MgSO4 and evaporated under vacuum to give a clear gum (25.5 g). Chromatography on silica gel (500 g) eluting with a mixture of diethyl ether in hexane (7:3 by volume) gave the required mono-ester as thick oil (20.0 g, 68%).

Rf 0.75 (diethyl ether). Found: C,56.75; H,8.71. C15H28C>5Si requires C,56.93; H,8.92%. iii c-3-tert-Butyldimethylsilyloxy-c-5-hvdro>xymethyl-cyclohexaner-l-carboxvlic acid methyl ester A IM solution of diborane in tetrahydrofuran (59 ml) was added dropwise under nitrogen to a stirred solution of the mono ester from part ii (17.0 g, 53.72 mmole) in tetrahydrofuran keeping the temperature at -5°C. The mixture was allowed to warm up to room temperature and after four hours a further amount of IM diborane solution (10.7 ml) was added and the mixture allowed to stand for eighteen hours. The solvent was evaporated under vacuum, diethyl ether was added and the solution washed sequentially with saturated salt solution, saturated aqueous PIC 513 sodium bicarbonate solution and again with saturated salt solution. Drying ever MgSO4 and evaporation under vacuum gave a clear oil which was chrwatographed on silica gel (500 g).

Elution with a mixture of ethyl acetate and hexane (1:1 by volume) gave an oil (15.18 g, 93%). Rf 0.42 (ss-3). Found: C,59.24; H,9.67. C15H3QO4Si requires C,59.56; H,10.0%. iv. c-3-tert-Butyldimethvlsilvloxy-c-5- (4-methvlbenzenesulphonvloxyroethyl)cyclohexane-r-l-carboxylic acid methvl ester 4-Methylbenzenesulphonyl chloride (14.18 g, 74.39 mmole) was added to an ice cold stirred solution of the ester from part iii (15.0 g, 49.59 mmole) in dry pyridine (130 ml). After stirring at room temperature for twenty four hours the mixture was poured onto ice, extracted with diethyl ether and the extract washed sequentially with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and water. Drying ever MgSO4 and evaporation under vacuum gave an oil (22.0 g) which was chromatographed on silica gel (600 g). Elution with increasing proportions of ethyl acetate in hexane (1:9 - 1:4 by volume) gave the required product as an oil (21.78 g, 96%). Rf 0.3 (4:1 hexane, ethyl acetate. Found: C,57.66; H,7.96. C22H36OgSSi requires C,57.86; H,7.95%.

PIT 513 v. c-3-tert-Butyldimethylsilyloxv-c-5-phenvlmethyl cyclchexane-r-1-carboxylie acid methyl ester 2M Ehenyl lithium in cyclohexane-ether (7:3) (24.5 ml, mmole) was added under nitrogen to a stirred suspension of cuprous iodide (4.67 g, 24.5 mmole) in dry diethyl ether (50 ml) keeping the temperature below 5°C. The resulting dark green solution was stirred at roam temperature for one and three quarters of an hour, cooled to -70°C and a solution of the ester from part iv (3.7 g, 8.1 mmole) in ether (10 ml) and cyclohexane (40 ml) added dropwise. Ihe temperature was maintained belcw -60°C during the addition, and then reduced to -70°C for fifteen minutes. Ihe mixture was then allowed to warm up to room temperature, stirred for an hour and the reaction quenched with saturated aqueous ammonium chloride with ioe cooling. Ihe organic phase was washed with saturated salt solution, dried over MgSO4 and on evaporation under vacuum gave an oil. Chrcmatography on silica gel, eluting with increasing proportions of diethyl ether in hexane (1:20 - 1:10 by volume) gave the required title product (1.4 g, 48%) Rf 0.2 (1:9 diethyl ether, hexane).

PLC 513 vi. t-3-(4^fethylbenzenesulphonvloxv) -c-3-phenvlmethvl cvclchexane-r-1-carboxvlic acid methyl ester 40% Aqueous hydrofluoric acid (0.5 ml, 10 mmole) was added to a stirred solution of the above ester (3.6 g, 9.93 mmole in acetonitrile (40 ml). After an hour the solution was diluted with diethyl ether, washed with saturated aqueous sodium bicarbonate solution and water, dried over MgSO4 and evaporated under vacuum to give the required alcohol as a clear oil (2.36 g,. 96%). Rf 0.2 (1:1 diethyl ether hexane). Ihe product (2.39 g, 9.62 mmole) was treated as described in Preparation 6, part iv to give the trans-4-methyIbenzene sulphonate title ocmpcund as a pale yellow gum (2.44 g, 63%). Rf 0.4 (1:1 diethyl ether hexane). Found: C,65.59; H,6.54; C^H^OgS requires C,65.65; H,6.51%. vii c-3-Amino-c-5-rhenvlmethylcyclchexane-r-l-carboxylic acid methyl ester hydrochloride The trans-4-methylbenzenesulphonate from part iv (2.41 g, .99 mmole) was treated with sodium azide as described in Preparation 6, part v to give the corresponding azide as an oil (1.44 g, 88%), Rf 0.72 (1:1 diethyl ether, hexane). Ihe azide (1.41 g, 5.16 mmole) was then reduced as described in Preparation 6, part vi to give the required amine salt as a foam (1.44 g, 99%), Rf 0.2 (ss-1). Found: C,63.3l; H,7.76; N,4.56.

C15H21N°2’HC1 C,63.48; H,7.81; N,4.94% PIC 513 PREPARATION 8 cis and trans-3-Amino-c-4-rhenvlmethvlcvclohexane-r-lcarboxylic acid methvl ester hydrochloride i 3-Oxo-4-chenvlmethylenecyclohexanecarboxylic acid methvl ester A mixture of ethyl (trimethylsilyl) acetate (16.01 g, 99.9 mmole) and 3-oxocyclohexanecarboxylic acid methyl ester (13.0 g, 83.24 mmole) was slcwly added dropwise under nitrogen to a stirred solution of 1.0M tetrabutylammonium fluoride in tetrahydrofuran (2.78ml) keeping the temperature between 0 and 10°C. Ihe resulting pale brown solution was stirred at room temperature for seventeen hours, further amounts of tetrabutylammonium fluoride (0.3 ml) and ethyl (trimethylsilyl) acetate (5 ml) were added and after three hours the reaction was shown to be completed by n.m.r. The mixture was diluted with n-hexane (50 ml), stood over 3A molecular sieves for an hour, filtered and the solvent evaporated under vacuum to give the silylenol ether as a golden oil (19.97 g, 100%). The silylenol ether product in dry methylene chloride (50 ml), was added drcpwise under nitrogen to a stirred suspension of titanium tetrachloride (9.15 ml, 83.24 mmole) in benzaldehyde (9.31 ml, 91.56 mmole) and dry methylene chloride (250 ml) at -70°C.

Ihe mixture was allowed to warm up to room temperature ever eighteen hours and quenched with water (100 ml) with ice cooling. Ihe organic phase was washed with water, dried ever MgSO^ and evaporated under vacuum. The residue was chrcroatograpahed on silica gel, eluting with increasing proportions of ethyl acetate in hexane (1:4 - 3:7 by volume) to give the required product as a PUC 513 pale solid (6.89 g, 34%) m.p. 73-5°C. Rf 0.25 (4:1 hexane, ethyl acetate). Found: C,73.20; H,6.54. 0.11^0 requires C,73.21; H,6.63%. ii. 3-Oxo-cis-4-phenylirethvlcyclohexanecarfaoxylic acid methyl ester The enone from part i (6.55 g, 26.8 mmole) in methanol (152 ml) and water (8 ml) was hydrogenated over 5% palladium on charcoal catalyst (750 mg) at 20 p.s.i. (1.4 bar). After three hours the suspension was filtered and on evaporation under vacuum the residue was chromatographed on silica gel (600 g). Elution with increasing proportions of ethyl acetate in hexane (1:4 - 3:7 by volume) gave the trans-isomer as a white solid m.p. 54-5°C. Rf 0.35 (1:4 ethyl acetate, hexane). Found: C,73.11; H,7.46. requires C,73.15; H,7.37%. Further elution gave the more polar cis-isomer as a white solid m.p. 7l-2°C. Rf 0.25 (1:4 ethyl acetate, hexane). Found: C,72.81; H,7.41. requires: C,73.15; H,7.37%. iii. 3-Methoxviminio-cis-4-phenvlmethvlcyclchexane carboxylic acid methyl ester The cis-isomer from part ii above (1.9 g, 7.71 mmole) on treatment with methoxylamine hydrochloride as described in Preparation 1 part iii gave the title required 0-methyloxime as a clear oil (2.0 g, 94%). Rf 0.4 and 0.45 (4:1 hexane, ethyl acetate). Found: C,69.53; H,7.69; N,4.94. C^gH^HO^ requires C,69.79; H,7.69; N,5.09%.

PLC 513 iv. c and t-3-(tert-Butoxvcarbonylamino)-o-4-phenvlmethvl cyclohexane-r-l-carboxvlic acid methyl ester Ihe 0-methyl oxime from part iii (1.97 g, 7.15 mmole) on treatment as described in Preparation 1, part iv gave the title product as a mixture of cis and trans-isomers which were separated by chrcmatography on silica gel (300 g) eluting with increasing proportions of ethyl acetate in hexane (1:3 - 3:7 by volume). Ihe trans-isomer was obtained as a gum (680 mg, 27%). Rf 0.5 (1:4, hexane, ethyl acetate). Found: C,68.84; H,8.17; N,3.91. ^0^91¾ recT-1^res C,69.14; H,8.41; N,4.03%. Ihe cis-isomer was also obtained as a gum (345 mg, 14%). Rf 0.45 (1:4 hexane, ethyl acetate). Found: C,67.55; H,8.08; H,3.80, (^θΗ^ΝΟ^Ο.δ ^0 requires C,67.39; H,8.48; N,3.93%. v. t-3-Amino-c-4-phenvlinethylcvclcbexane-r-l-carboxvlic acid methyl ester hydrochloride Ihe trans-iscmer from part iv (650 mg, 1.87 mmole) on treatment with HCl as described in Preparation 1, part v gave the title salt as a gum (533 mg, 99%). Rf 0.25 (ss-1). Found: C,62.41; H,7.78; N,4.77. C-j^H^NC^. HCl.0.25 ^0 requires C,62.49; H,7.87; N,4.86%. vi. c-3-Amino-c-4-phenvlmethvlcyclchexane-r-l-carboxvlic acid methyl ester hydrochloride Ihe cis-isomer from part iv (325 mg, 0.94 mmole) on treatment with HCl as described in Preparation 1, part v, gave the title salt as a gum (263 mg, 99%). Rf 0.25 (ss-1). Found: C,59.45; H,7.44; N,4.77. Cj^H^NC^. HCl. ^0 requires C,59.69; H,8.01; N,4.64% PIC 513 EXAMP1E 1 3-( 1—Γ (cis-4-Ethoxvcarbonvl-cis-3-( 2-phenvlethvl )cyclohexvl) carbamoyl! cvclcpentvl )-2-(2-methoxyethvl) propanoic acid benzyl ester -Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (460 mg, 2.4 mmole) was added to an ice cold stirred solution of 3- (1-carbaxycyclopentyl) -2-(2-methyaxyethyl)propanoic acid benzyl ester (EP-A-0274234) (401 mg, 1.2 mmole), c-4-amino-c-2-(2phenylethyl)cyclohexane-r-l-carboxylic acid ethyl ester hydrochloride (343 mg, 1.1 mmole), 1-hydroxybenzotriazole (162 mg 1.2 mmole) and N-methyl-morpholine (354 mg, 3.5 mmole) in dry methylene chloride (15 ml). After half an hour the mixture was allowed to attain room temperature and stirred for twenty hours. The solvent was evaporated under vacuum and the residue partitioned between diethyl ether and water. Ihe organic extract was washed sequentially with water, 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate and water. Drying (MgSO^) and evaporation under vacuum gave an oil (640 mg), which was chromatographed on silica gel (50 g) eluting with a mixture of diethyl ether and hexane (7:3 by volume) to give the required diester as a colourless oil (550 mg, 85%) . Rf 0.32 (7:3 diethyl ether, hexane). Found: C,73.23? H,8.37; N,2.12. C36H4gN06 requires C,73.07; H,8.35; N,2.37%.

PLC 513 EXAMPLES 2-5 The following ccarpounds were prepared following the procedure of Example 1 using the appropriate amine of formula (IV).

CO2R4 EXAMPLES 6-7 The following cxanpounds were prepared following the procedure of Exanple 1 but using 3-(l-carboxycyclopentyl)-2(S)-(2-methoxyethoxyroethyl) propanoic acid t-butyl ester (EP application no. 89304698.7) as the starting material and coupling with the appropriate amine of formula (IV).

CH3 O , / lBuO2C s CO2 Cjj H 5 Ul to Ρω ι σι CO °5 cn O HItn ι ca A in Ω HIcn ι ca I s to LO to to Ul s Ol σ tn ι ca Ω HIcn ι ca έ tO J2 LO cn tn cn o HItn ι 4» έ to J2 ?s σ οι I LO ** ό to to c? Sc ca oo 0? I ca έ to 51 'J Ό to to • · 4* σ 4. ca 'J Ό to to • · 4»· O 4^ CO ~~1 M I—1 l-> • · 'J M O O σ σ co co o to oo to oo oo Ο H 4* O 00 Ο H· 4- tO ~J 00 • · CO H> O O oo oo • · co co tn to to to 4* cn Ό to to 4» cn 00 o to to tn o ca -i to to 4^ ca 4» co εν PLC 513 Exanple No R2 Form isolated Analysis % (theoretical in brackets) C H N 6 -(0^)3°¾ oil 64.47 9.35 2.53 cis Rf 0.45 (ether) (64.84 9.61 2.52) 7 -(0^0¾ oil 65.17 9.44 2.51 cis Rf 0.5 (ether) (65.35 9.73 2.46) EXAMPLES 8-12 Ihe following compounds were prepared following the procedure of Exanple 1 but using 2S- (N2-benzyloxycarix)nyl-N6-t-butyloxycartxanyl-S-lysylainincmethyl) -3-[l-(l-carboxycyclcpentyl) ] propanoic acid t-butyl ester (EP application 89305180) as the starting material and coupling with the appropriate amine of formula (IV). 4— CO2R^ 12 11 10 CO 00 Exanple No Ω Ω 1° |Ω Η· Η· Η· Η- Ιω Ιω 3 Ica Ica co σι 5 co CO 1 A ι ,Λ S to νΡ Q G1 1 S to cP So to Ο Sc σι to Π co Q Sc to Sc O to σι η σι κ Sc σι σι |Ω |Ω |Ω ΙΩ Η- Η' ο Η- Ιω ι Ιω Η- Ιω η· ι ω ι Ιω A CO A ι 4»· 1 4* A Λ 1 w έ NJ έ to Π to A cP cP to Ο co Π to w ο to w Μ σι κ σι σι S s S W ο S Η) · H- ο o ο tO Η) ο lx • Η> • t-h • ο σι ο • to ο Η· O co Η) Η· I—· o σι g σι g σι tojcg 4. Η) In Ίη 3 ο {η ω 3 s ω ul 1 ω ω ι ω ι CO co ω CO CO σ σ σ σ σ σ σ σ σ σ X—X ό σ σ ό •ο Ό ό σ 4» 4» π Et • · • · • • • · • · Ό oo to co ο Ο 00 1—1 ο Φ - ο ο Ο 00 co ο σι ω σ 4> II Ω ω g (-.. 00 00 00 00 00 00 οο ω 00 00 „ h tn • · • · • • • · • · to ο t—1 ω co co ω ι—1 CO Ό H· <*> Ό 00 Ό tO Ό to 00 4» ο σι 3 ί σ σ σ σι σ σ σ σ σ σ S • · • · • • • · • · 2 ίί CO CO to to to Η-1 σι οο tn co CO co σι co σ 4* t— σι ςν PLC 513 EXAMPLE 13 (S) - (N6-t-ButoxvcaiA)orivl-S-lvsvl-amincroethvl) -3-( 1- Γ cis-4 ethoxycarbonvl-cis-3-( 3-ethoxypropyl )cvclohexyl) carbamoyl 1 cyclopentyl) propanoic acid t-butvl ester The diester of Example 8 (734 mg, 0.85 mmole) in methanol (18 ml) and water (2 ml) was hydrogenated at room temperature for three hours over 5% palladium on charcoal catalyst (75 mg) at 50 I p.s.i. (3.45 bar) pressure. The suspension was filtered through arbacel, and evaporation of the solvent under reduced pressure gave the title product cis a white foam (630 mg, 100%). Rf 0.66 (ss-1). Found: C,63.22; H,9.73; H,7.17. requires C,62.78; H,9.70; 7.71%.

PLC 513 EXAMPLES 14-17 Ihe following compounds were prepared from Examples 9-12 by hydrogenation following the procedure of Exanple 13.

O Η σ> t-* <_π θ ΗΙ(Ω I ΟΙ έ to ο σι ιο ΗΙω GJ ι CO CO Ω σι to Ω °9 σι CO θ Η· Ιω I ω έ to J2 Ο Η· 1(0 I ό CO Ω to W σι έ to θ Η· 1(0 to ΙΟ «? έ to W ο ο · i-b • 'JO σι οι g 3 Λ 8 θ % ω Η) 2^1 ο Sh ο σι Μ) . ο I So ο σι Μι &?=1 ο Ιο (0 £ £“ θ’ σι σι • · ό σι Ο 03 00 < ΙΟ to ό σι σι σι σι to σι σι to to • · to σι ο Ό ιο οο CO Ό Ο Η· σι σι • · ΙΟ 00 I—1 ο 8V σι σι w ω ιο οο σι ο ιο οο ο Μ W ΙΟ 'J Ό • · to Ό σι σι σι σι • · ω ό σι μ ΙΟ 00 • · ο ιο οο σι Ό Ό • · W Η1 Η PLC 513 EXAMPLE 18 2S-(N2-Methanesulphonyl-N6-t-butoxvcarbonvl-S-lvsvl-aminomethyl) -3-( 1- Γ fcis-4-ethoxycarbonyl-cis-3-( 3-ethoxypropyl) cyclohexyl) carbamoyl Icvclopentvl) propanoic acid t-butvl ester Methanesulphonyl chloride (0.14 ml, 1.8 mmole) was added dropwise to an ice cold stirred solution of the amine from Example 13 (601 mg; 0.83 mmole) and N-methylmorpholine (0.2 ml, 1.8 mmole) ί in dry methylene chloride (8 ml). Ihe solution was stirred at 10°C for 3 hours and the reaction mixture then quenched with saturated aqueous sodium hydrogencarbonate. The organic phase was separated, the aqueous phase extracted with methelene chloride and the combined extracts dried over MgSO^. Evaporation under vacuum gave a gum which was chromatographed on silica gel (100 g) eluting with increasing proportions of ethyl acetate in hexane starting with a 1:1 mixture and finally with neat ethyl acetate.

Evaporation of relevant fractions gave the required product as a foam (576 mg, 87%). Rf 0.56 (ss-1). Found: C,58.66; H,8.91; N,6.65. C H? N O s requires C,58.18; H,9.0l; N,6.96%.

PLC 513 EXAMPTES 19-23 Ihe following compounds were prepared in a similar manner to Exanple 18, using the appropriate amine of Exanples 14 to 17 and reacting with the appropriate sulphonyl chloride or acyl chloride to yield the N2-substituted-lysyl derivative. co2r τς PLC 513 EXAMPLE 24 2S— (bis- (IS) -Phenylethyl 1 aminoroethvl) -3- (1- Γ cis-4-ethoxycarbonyl-cis-3- (3-methoxypropvl) cyclohexyl) carbamoyl 1 cyclopentyl) propanoic acid t-butyl ester 2S— (bis- [ IS) -Phenylethyl ] amincmethy1 )-3-( 1-carboxycyclopentyl) propanoic acid and c-4-amino-c-2-(3-methoxyprcpyl)cyclohexane-r-1-carboxylie acid ethyl ester (from Preparation 2) were coupled as described in Exanple 1 to give the title diester as a gum. Rf 0.1 (4:1 hexane, ethyl acetate) Fcund: C,73.19; H,9.13; N,4.01. C.,H,.N_O- requires C,73.26; H,9.15; N,3.97%. 4J o4 2 o EXAMPLE 25 2S- (bis Γ (IS) -Phenylethyl 1 amincmethy 1) -3- (1Γ (cis-4-ethoxycarbonyl-cis-3- (2-phenylethyl) cyclohexyl) carbamoyl 1 cvclopentyl) propanoic acid t-butyl ester Ihe title compound was prepared as described in Exanple 24 above using c-4-amino-c-2-(2-phenylethyl) cyclohexane-r-lcarboxylic acid ethyl ester hydrochloride (from Preparation 1) as the amine component to yield the product as a white foam. Rf 0.55 (ss-3). Found: C,76.11; H,8.74; N,4.25. 0.25H20 requires C,76.12; H,8.77; N,3.77%.

PLC 513 EXAMPLE 26 2S-(Aminamethvl) -3-( l-Γ fcis-4-ethoxvcarbonvl-cis-3-( 3-methoxvpropyl)cyclohexyl)carbamoylIcvclopentvlIpropanoic acid t-butvl ester Ihe diester from Exanple 24 above (992 mg, 1.4 mmole) in ethanol (20 ml) was hydrogenated over 20% palladium hydroxide on charcoal catalyst at 60 p.s.i. (4.1 bar) pressure. After twenty four hours the suspension was filtered through a short arbacel column and the solvent was evaporated under vacuum. Ihe residue was dried azeotrcpically with methylene chloride to give the title amine as a thick oil. (690 mg, 99%) Rf 0.35 (ss-1) Found: C,65.69 H,9.88; N,5.19. C27H48N2C>6 requires C,65.29? H,9.74; N,5.64%.

EXAMPLE 27 2S-(Aminomethyl)-3-(l-Γ fcis-4-ethoxycarbonyl-cis-3-f2-phenylethvl) cyclohexyl) carbamoyl 1 cyclopentyl) propanoic acid t-butvl ester Ihe product of Exanple 25 was hydrogenated as described in Example 26 above to yield the title compound as a gum. Rf 0.35 (ss-1) Found: C,69.00? H,8.80; N,4.88. C31H48N2O5. 0.6 H20 requires C,69.00; H,9.19; N,5.19.

PLC 513 EXAMPLE 28 2S- (N2-Methanesulchonvl^6-t-butoxvcarbonyl-S-lvsyl -aminomethyl) 3-(l-Γ (cis-4-ethoxycarbonyl-cis-3-( 3-methoxvpropyl(cyclohexyl) carbamoyl lcvclcpentyl (propanoic acid t-butyl ester N-methylmorpholine (0.34 ml, 3.06 mmole) was added to an ice cooled stirred mixture of the product from Example 26 (690 mg, 1.39 mmole) N2-methanesulphonamido-N6-t-butoxycarbonyl-S-lysine (406 mg, 1.39 mmole), 1-hydrpxybenzotriazole (188 mg, 1.39mmole) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (586 mg, 3.06 mmole) in dry methylene chloride (10 ml). After six hours the mixture was diluted with methylene chloride (20 ml) washed with water and dried over MgSOj. Evaporation of the solvent under vacuum and chromatography on silica gel, eluting with increasing proportions of ethyl acetate in hexane (50 - 90%) and finally with 5% ethanol in ethyl acetate gave the required product as a cream foam (880 mg, 81%). Rf 0.2 (ss-3) Found: C,58.07; H,9.06; N,6.74. OUS requires C,58.33; H,8.79; N,6.98%.

IE 904122 “ PLC 513 EXAMPLE 29 2S-(N-t-Butvloxvcarbonvl-R-alanvlamincmethvl) -3-( l-Γ (cis-4ethoxycarbonyl-cis-3- (2-phenylethyl) cyclohexyl) carbamoyl 1 cyclopentyl) propanoic acid t-butyl ester Ihe procedure of Exanple 28 was followed starting with the amine of Exanple 27 and coupling with N-t-butyloxycarbonyl-Ralanine to yield the title product as a foam. Rf 0.5 (ss-1). Found: C,67.22; H,8.86; N,5.66. C^Hg^Og requires C,66.92; H,8.78; N,6.00%.

EXAMPLE 30 2-ΓΝ- (2-t-Butyloxycarfaonvlamino-2-methvlpropanoyl) aminomethyl 1-3-( 1-Γ (cis-4-ethoxycarbonyl-cis-3-(2-rheny lethyl) cyclohexyl) carbamoyl 1 cvclopentyl) propanoic acid t-butyl ester Ihe procedure of Exanple 28 was followed starting with the amine of Exanple 27 and coupling with 2-t-butyloxycarbonylamino2-methyl-propanoic acid to give the title product as a foam. Rf 0.48 (ss-1). Found: C,67.06; H,8.94; N,5.81. requires C,67.29; H,8.89; N,5.89 PLC 513 EXAMPLE 31 -fN2-Methanesulphonyl-S-lvsyl-arniincroethyl)-3-(1-Γ (cis-4carboxy-cis-3- (2-phenylethvl} cvclohexyl) carbamoyl 1 cyclopentyl} propanoic acid Trif luoroacetic acid (5 ml) was added to an ice cold solution of 23-(112-methanesulphonyl-N6-t-butoxycarbonyl-S-lysyl-amiinomethyl)-3-{l-[(cis-4-ethoxycarbonyl-cis-3-(2-phenylethyl)cyclohexyl) carbamoyl]cyclopentyl}propanoic acid-t-butyl ester (from Example 19) (445 mg, 0.53 mmole) in dry methylene chloride (5 ml). Ihe solution was allowed to warm to room temperature and after three hours evaporated to dryness under vacuum. Ihe residue was dried azeotrcpically twice with toluene and dissolved in IN sodium hydroxide, the solution was stirred at 40-55°C for sixty eight hours, cooled and passed down a sulphonic acid ion-exchange column. Elution with 10% aqueous pyridine and evaporation of the relevant fractions gave a foam which was crushed under acetonitrile and filtered to give a white pcwder (279 mg, 80%) Rf 0.25 (ss-2) Found: C,58.66; H,7.92; N,8.49. C^Hg^O S requires: C,59.06; H,7.74; N,8.61%.

PLC 513 EXAMPLES 32-37 Ihe following compounds were prepared from the appropriate N6-t-butyloxycarbonyl protected diester of Examples 18, 20-23 or 28 by treatment with trifluoroacetic acid followed by alkaline hydrolysis following the procedure of Example 31.

PUC 513 EXAMPLES 38-39 Ihe following compounds were prepared from the diesters of Examples 29 and 30 by treatment with trifluoroacetic acid followed by alkaline hydrolysis following the procedure of Example 31. w KD GJ Η» GJ GJ « o oo σ R> · · ο ro GJ -J 2 Ho win CLuQ to « £8 σ σ ω gj 4- Η Ul 03 00 GJ U> σ ό 00 ui σ σ σ to NJ ύι ίη 4* 4* 00 NJ GJ 4> 4» kO kD H> NJ NJ O PLC 513 EXAMPLE 40 3-( 1-f cis-4-Carboxv-cis-3-( 2-phenylethyl )cyclohexyl) carbamoyl 1 cyclopentyl )-2-( 2-methoxyethvl) propanoic acid Ihe diester of Example 1 (510 mg, 0.86 mmole) in ethanol (25 ml) and water (10 ml) was hydrogenated at room temperature for three hours ever 10% palladium on charcoal catalyst (100 mg) at 50 p.s.i. (3.45 bar) pressure. Ihe mixture was filtered through a short Avicel column and the solvent evaporated under reduced pressure. The monoester thus obtained was dissolved in IN sodium hydroxide and the solution heated to 65°C under a nitrogen atmosphere for two days. On cooling the solution was extracted with diethyl ether, the aqueous phase acidified with 2N hydrochloric acid and extracted with diethyl ether. The organic phase was washed with water, dried ever MgSO4 and evaporated under reduced pressure to give the required title diacid as a white foam, (300 mg, 75%). Rf 0.62 (ss-1) Found: C,68.17; H,8.45; N,2.95. C2?H3gNO6 requires C,68.47; H,8.30; N,2.96%.

PUC 513 EXAMPLES 41-44 Ihe following cxxnpounds were prepared from the appropriate diester of Examples 2 to 5 by hydrogenation followed by alkaline hydrolysis following the procedure of Exanple 40. 44 43 42 41 Example No O Ω Ω H H· Η- Η- A Ιω Ιω Ιω σ ω CJ σι A έ to A St S a a η Ul to to to Ω Sc CJ Sc Ul 8 NJ Ul a Ul |O rf Η- ΙΩ |Ω K Ιω Η- Η- 1 3 I Ιω Ιω Q g ω ι 4» ι 4* 0 to 1 1 | a CJ s θ H) t-b SP8 So1 BPS’ rb · NJ fi σι 1 ' μ 8 O Ul 5 ο Ο 3 LkF σι ο .47 U1 ο 8 S’ o K χ—χ χ—X Χ-Χ» Ρ» 3 ω {» ω ω (+ ω ω ω ω Φ I I ι ι 1—1 1—1 1-· I-> Χ_Χ· X—x σ» σ σ σ σ σ σ σ X—X Ό Ό σ σ Ό Ό to to o ti • • • · • * • · Ex nj 4» σ ό NJ CJ Ό 00 δ kO NJ 4* kO ω ko Ο 4» I H- 5? £ 00 00 00 00 Ό < kO kO £3 • • • · • · • · Η» 15 Η* 1—1 Ό kO 00 κο σ nj Ο Η* KO NJ 5'ui H· σω tf Λ> U> NJ NJ CJ NJ CJ CJ C0 {§ • • • · • · • · z π· o 00 kO ο ko l·-· Ο Ό tn NJ kO KO CJ kO 4» U1 4* €9 PIG 513 EXAMPLE 45 2S-(2-Methoxyethoxymethyl)-3(l-f (cis-4-carboxy-cis-3-(3-methoxypropylIcyclohexy)carbamoyl1cyclopentyl)propanoic acid Trifluoroacetic acid (5 ml) was added to an ice cold solution of the diester of Example 6 (480 mg; 0.77 mmole) in dry methylene chloride (3 ml). After standing at room temperature for three hours the solution was evaporated to dryness under vacuum and the residue was dried azeotropically with toluene. The residual oil was dissolved in diethyl ether, washed with water (x9) and the resulting mono-ester extracted into IN sodium hydroxide solution (2 x 10 ml). Ihe aqueous extract was heated at 65-70°C under nitrogen for 24 hours, cooled saturated with salt, acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic extract was washed with water, dried over MgSO4 and on evaporation gave the required diacid as a gum (320 mg 88%).

Found: C,60.55; H,8.54; N,2.80, Ο^Η^ΝΟθ. 0.41^0 requires C,60.20; H,8.80; N,2.93%.

EXAMPLE 46 2S-(2-Methoxyethoxvmethvl)-3-(l-Γ (cis-4-carboxy-cis-3-(3-ethoxypropyl ) cylohexy1) carbamoyl 1 cyclopentyl) propanoic acid Ihe title product was prepared from the diester of Example 7 as described in Exanple 45 above and was obtained as a gum.

Found: C,61.26; H,8.66; N,2.83. C25H ΝΟθ. 0.3H20 requires C,61.15; H,8.95; N,2.85% PLC 513

Claims (11)

1. A compound having the formula:- R0 2 C (I) Therein A completes a 4 to 7 membered carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be fused to a further saturated or unsaturated 5 or 6 membered carbocyclic ring; B is m is an integer of from 1 to 3; each of R and R is independently H, C^-Cg alkyl, benzyl or an alternative biolabile ester-forming group; R 1 is H or C^-C alkyl; PLC 513 and or or R is C^-Cg alkyl substituted by C^-C* alkoxy, aryl or aryloxy; R 5 is Cj-Cg alkyl, C 2 ~Cg alkenyl, C 2 ~Cg alkynyl, cycloalkyl, or cycloalkenyl, R is Cj-Cg alkyl substituted by halo, hydroxy, C^-Cg alkoxy, C^-Cg alkoxy(C^-Cg)alkoxy, C^-C? cycloalkyl, 6 7 C 3 ~Cj cycloalkenyl, aryl, aryloxy, heterocyclyl, -NR R Q Q Q In 6 7 ' 7 -NR COR , -NR SO 2 R , -CONR R or R R N-^-Cg) alkoxy; R is C,-C alkyl substituted by a group of the formula x D R 12 -NR 11 CD-C-R 13 R 12 -NR 11 ^ -C-R 13 il4 or R 13 -CONR^-C-R 14 PLC 513 wherein 6 7 R and R are each independently H, C^-C 4 alkyl, (^-G? cycloalkyl, aryl, aryl(C-^-C 4 )alkyl, C 2 ~Cg alkoxyalkyl, 6 7 or heterocyclyl; or the two groups R and R are taken together with the nitrogen to which they are attached to form a pyrrol idinyl, piperidino, morpholino, piperazinyl or N-(C 1 ~C 4 ) alkyl -piperazinyl group; Q R is H or C^-C 4 alkyl; R 9 is C^-C^ alkyl, CF 3 , aryl, aryl(C^-C^)alkyl, 6 7 aryl (C^-C^) alkoxy, heterocyclyl, C^-C 4 alkoxy or NR R 6 7 wherein R and R are as previously defined; R 10 is C 1 ~C 4 alkyl, cycloalkyl, aryl or heterocyclyl; R 11 is H, C^-Cg alkyl, aryl or c 3~ C j cycloalkyl; R 12 is R 11 OONR 11 -, R 11 SO2NR 11 -, R 16 R 1? N-(CH2) p -, or rH0-, wherein each is as previously defined above; R^ 3 and R 3-4 are each independently H or C^-Cg alkyl; or 13 14 R is H and R is C,- OH, C^-C 4 alkoxy, SH, SCH 3 , N^, aryl (C^-Cg) alky 1OCONH-, NHgCO-, 00 2 H, guanidino, aryl, or heterocyclyl; or the two groups R and R are joined together to form, with the carbon atom to which they are attached, a 5 or 6 membered carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be PIG 513 substituted by C^-C 4 alkyl or fused to a further 5 or 6 membered saturated or unsaturated carbocyclic ring; or R^ 3 is H, and R 12 and R 14 are linked to form a 2. - (N-COR^^-4-aminopyrrolidinyl) group; R 15 is R 16 R 17 NOO-, R 13 OOO-, R 11 OCH 2 - or heterocyclyl, wherein R^ is as previously defined above; Σ6 17 R and R are each independently H or C^-Cg alkyl; and p is 0 or an integer of from 1 to 6; and pharmaceutically acceptable salts thereof and bicprecursors therefor.

2. A compound according to claim 1 wherein A is (CIL,)^ R 1 is H and B is (CH^^ having the formula-: RO 2 C (II) wherein R, R z 4 5 R and R are as previously defined. PLC 513 . . . 4

3. A compound as claimed in claim 1 or claim 2 wherein R and R are both H. 4. A compound as claimed in claim 1 or claim 2 wherein one or

4 . . . both of R and R is a biolabile ester-forming group and said group is ethyl, benzyl, l-(2,2-diethylbutyryloxy)ethyl, 2-ethylprop ionyloxymethyl , l-(2-ethylpropionyloxy)ethyl, l-(2,4-dimethylbenzoyloxy)ethyl, 1- (benzoyloxy)benzyl, 1-(benzoyloxy)ethyl, 2-methyl-l-propionyloxypropyl, 2,4,6-trimethylbenzoyloxymethyl, 1- (2,4,6-trimethylbenzyloxy)ethyl, pivalcyloxymethyl, phenethyl, phenprcpyl, 2,2,2-trifluoroethyl, 1- or 2-naphthyl, 2,4-dimethylphenyl, 4-t-butylphenyl, [5-(4-methyl-1,3-dioxolen2- onyl) ]methyl or 5-indanyl. . 5

5. A conpound according to any one of claims 1 to 4 wherein R is methylene substituted by a group of the formula -NHC0CR 12 R 13 R 14 , where R 12 is Nl·^, R n C0NH- or R^SC^NH-, R 13 is H and R 14 is -(CH 2 ) 4 NH 2 .

6. A compound according to claim 5 wherein R 5 is i^-acetyl-Slysylaminomethyl, f^-methanesulphonyl-S-lysyl-amincmethyl, N 2 -phenylsulphonyl-S-lysyl-aminomethyl or N 2 -cyclobutylcarbonyl-Slysyl-aminoroethyl.

7. A compound according to any one of claims 1 to 4 wherein R is methoxyethyl or 2-methoxyethoxymethyl.

8. A compound according to any one of claims 1 to 4 wherein R is C, —C, alkyl substituted by -NHCOCR 12 R 13 R 14 wherein R 12 is NH , R 13 is CH3 and R 14 is H or CH3· _ Pic 513

9. A compound according to any one of claims 1 to 8 wherein R is phenyl, benzyl, phenethyl, methoxypropyl or ethoxypropy1.

10. A compound according to claim 1 wherein said compound is 2S— (N^methanesulphonyl-S-lysylaminomethyl) -3-{ 1- [cis-4carboxy-3 -cis {3-ethoxypropyl) -cyclohexyl) carbamoyl ] cyclopentyl} propanoic acid; 2-(11 2 -methanesulphonyl-S-lysylamironethyl) —3—{1—[ (cis-4carboxy-3-cis-phenethyl-cyclohexyl) carbamoyl ] cyclopentyl) propanoic acid; 2S- (2-methoxyethoxymethyl) -3-{l-[ (cis-4-carfooxy-3-cis-{ 3ethoxypropyl} cyclohexyl) carbamoyl ] cyclopentyl} propanoic acid or 2S-(2-methoxyethoxymethyl)-3-{l-[(cis-4-carboxy-3-cis-{3methoxypropyl) -cyclohexyl) carbamoyl ] cyclopentyl} propanoic acid, or a biolabile ester derivative thereof.

11. A process for preparing a compound of the formula (I) as claimed in claim 1 which comprises subjecting a compound of the