PT95900A - PROCESS FOR THE PREPARATION OF UTILIZED CYCLOALKYL SUBSTITUTED GLUTARAMIDE DERIVATIVES AS DIURETIC AGENTS - Google Patents

Description

-4-

: The present invention relates to various cycloalkyl substituted glutaramide derivatives, and which are beneficial diuretic agents in a number of therapeutic domains, including the treatment of various disorders in the cardiovascular system, such as hypertension, cardiac collapse, and renal insufficiency.

In accordance with the specification of the European Patent Applications Nos. EP-A-0274234 and EP-A-0343911, we have provided and claimed certain cycloalkyl substituted glutaramide derivatives as diuretic agents. The present invention discloses additional related compounds which contain an extensive substituent group on the aminocycloalkane carboxylate ring.

These compounds are inhibitors of neutral endo-peptidase, dependent on the presence of zinc, E. C. 3.4.24.11. This enzyme intervenes in the decomposition of various peptide hormones, including the splitting of atrial natriuretic factor (ANP) that is secreted by the heart and exerts a powerful diuretic and natriuretic vasodilatory activity (sodium excretion). In this order, the compounds of the present invention - inhibiting endo-peptidase E.E. 3.4.24.11 neutral - are capable of potentiating the biological effects of ANF and in particular these compounds are diuretic agents useful in the treatment of a number of disorders, including hypertension, cardiac collapse, angina pectoris, renal insufficiency, premenstrual syndrome, cyclic edema, Menières's disease, hyperaldosteronism, pulmonary edema (primary and secondary), ascites as well as hyperealciuria. In addition, the compounds of the present invention are useful in the treatment of glaucoma by virtue of their ability to potentiate the effects of ANF. As a further corollary of its ability to inhibit neutral endopeptidase E.C. 3.4.24.11. The compounds of the present invention

The invention may be of benefit in other therapeutic domains, eg in the treatment of asthma, inflammation, pain, epilepsy, affective disorders, dementia and confusional states arising from advanced age (geriatrics), obesity and gastrointestinal disorders (especially diarrhea and intestinal hyperexcitation syndrome), regulation of gastric acid secretion, and treatment of hyperreninemia and leukemia.

The compounds according to the present invention have the following structure formula:

(I) wherein A completes a 4 to 7 ring carbocyclic ring, which may be saturated or mono-unsaturated, and which is optionally condensed with another saturated or unsaturated five-membered or hexagonal carbocyclic ring; B represents (VIII), wherein m represents an integer from 1 to 3; each of R 4 and R 4, independently of one another, represents hydrogen, C 1 -C 6 -alkyl, benzyl or an alternative biolabile ester-forming group; -6-

V

R 1 represents hydrogen or C 1 -C 4 -alkyl;

R1 represents C1 -C6 -alkyl substituted by C1 -C4 -alkoxy, aryl or aryloxy; 5 and R6 is C1 -C6 -alkyl, C2 -C6 -alkenyl, C2 -C6 -alkenyl, C1 -C4 -alkyl or C1 -C4 -cycloalkenyl; or R representa is C -C-C--alkyl substituted by halogen, hydroxy, C -C-C--alkoxy, C,-C--alkoxy- (C -C-C -) alkoxy, cycloalkyl, C -C-C--cycloalkenyl, aryl, aryloxy, heteroaryl, -NR R N, NR, COR, -NR ^SO₂, -CONR ouR or RRRNN- (C ,- -C) -alkoxy; R6 represents C1-6 alkyl substituted by a group of the formula:

-NR CO-C-R & _12

R 11 * 13

-NR 2 SO 3 -R 13 R 13 -aW 2 -c-R 14 -7-

Wherein R6 and R7 are independently hydrogen, C1 -C4 -alkyl, C1 -C4 -cycloalkyl, aryl, aryl- (C1 -C4) -alkyl, -C2 -C4 -alkyl, R 2 and R 2 together with the nitrogen atom to which they are attached form a pyrrolidinyl, piperidino, morpholino, piperazinyl or N- (C 1 -C 4) -alkoxy group, R 2 represents hydrogen or C 1 -C 4 -alkyl; R 4 is C 1 -C 4 -alkyl, CF 3, aryl, aryl- (C 1 -C 4) -alkyl, aryl- ( C -C-C--alkoxy, heterocyclyl, C -C-C--alkoxy, C -C-C -C-alkyl or NR¹R², wherein R¹ and R² are as defined above; R 11 represents hydrogen, C 1 -C 6 -alkyl, aryl or -cycloalkyl; R 11 represents hydrogen, C 1 -C 6 -alkyl, aryl or C 3 -C 7 -alkyl, aryl or heterocyclyl; cycloalkyl, R12 represents R3, R6, R7, R4, R7, R16, R17, N - (CH2) -, or R11 and R11 are each independently selected from the group consisting of: aminocarbonyl or C -C-C--alkyl; or R 2 represents hydrogen and R 1

R3 is C1 -C6 alkyl substituted by OH. C3 -C4 -alkoxy, SH3, NH3, aryl- (C1 -C6) -alkyl-OCONH-, NH2 CO-, CO2 H, guanidino, aryl or hetero-cyclic; Together with the carbon atom to which they are attached, a pentagonal or hexagonal carbocyclic ring which may be saturated or mono-unsaturated and which may optionally be substituted by C1-4 alkyl, Or may be condensed with another saturated or unsaturated five- or six-membered carbocyclic ring;

 € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒR1 or R3 is H, and R1 and R2 are joined to form a 2- (N-COOR4 -aminopyrrolidinyl) group; R 15 represents R 16 R 17 NCO-, R 1, R 2, R 1, CHOC- or heterocyclyl wherein R has the abovementioned meaning; R1 and R2, independently of one another, represent hydrogen or C1 -C6 -alkyl; and p is zero or an integer from 1 to 6; and refers to the pharmaceutically acceptable salts thereof, and to biological precursor compounds.

In the above definition and unless otherwise indicated, alkyl groups having 3 or more carbon atoms may be straight chain or branched. The term " aryl " used in the present invention means an aromatic hydrocarbon group, such as naphthyl naphthyl or biphenyl, optionally substituted by 1 or more OH, CN, CF 3, C 4 -C 4 -alkyl, C 1 -C 4 -alkoxy or atoms halogen atoms include halogen atoms. fluorine, chlorine, bromine or iodine. The term heterocyclyl means a 5- or 6-membered heterocyclic group containing a nitrogen, oxygen or sulfur atom which, unless otherwise indicated, may be saturated or unsaturated and which may further contain in its ring an additional oxygen atom or more 1 to 3 nitrogen atoms and may optionally be condensed with a benzene ring or may be substituted by eg 1 or more halogen atoms, or 1 or more C1 -C4 -alkyl, hydroxy, carbamoyl, benzyl oxo, amino or mono- or di (C1 -C4 -alkyl) -amino or -alkanoyl) -amino. • \.,

Specific examples of heterocyclic radicals include groups such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, furanyl, tetrahydrofuran. nyl, tetrahydropyranyl, dioxanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, indolyl, isoindolinyl, quinolyl, quinoxalinyl, quinazolinyl and benzimidazolyl, each of which is optionally substituted in the above manner.

The compounds of formula (I) may contain several centers of asymmetry and thus may be present in the form of enantiomers and diastereomers. The present invention relates to both the racemic mixtures and the individual, separated isomers. The R 1 and R 2 substituents may have the cis or trans configuration relative to their amide bond.

Pharmaceutically acceptable salts of the compounds of formula (I) containing an acyclic center are those which are formed with bases which are capable of producing non-toxic salts. Examples of such salts include alkali metal salts, such as sodium, potassium or calcium salts, or salts with amines, such as diethylamine.

Examples of such salts include the hydrochloride, bromate, sulphate or bisulfate, phosphate or hydrogen phosphate acetate, citrate, fumarate, gluconate, acetate, maleate salts, and the like. , succinate or tartrate. The term " biological precursor " -10-

- Λ ν ^ '. as defined in the definition above, means a pharmaceutically acceptable and biologically degradative derivative of the compound of formula (I) which, upon administration to a human animal or patient, undergoes transformation in the organism into a compound of the formula I).

A stated group of compounds of formula (I) comprises compounds wherein Δ represents (CH 2) 4, R 1 represents hydrogen and B stands for - (CH 2) 2, i.e. compounds of formula (II) below, and wherein R 4 and 5ΛR have the meanings mentioned in formula

R, R (I):

(II)

Also preferred are those compounds of formulas (I) and (II) wherein R and R4 are both H (diacids) as well as their biodegradable mono- and di-esterified derivatives, wherein either or both of R and R4 represents (m) a biodegradable esterifying group.

The term biodegradable esterifying group is well known in the art to which the present invention relates, meaning a group capable of giving an ester readily dissociable in the organism in order to release the corresponding di-acid of formula (I), wherein R1 and R2 are hydrogen.

Various ester groups of this type are well known eg in the penicillin area or in the case of ACE inhibitory antihypertensive agents.

In the case of the compounds of formulas (I) and (II) these biodegradable esters (pro-drugs) are especially advantageous in that they provide compounds of formula (I) suitable for administration by. oral. The suitability of a particular esterifying group may be assessed by standard in vitro or. in animals for enzymatic hydrolysis. Thus, in order to obtain an optimum effect, the corresponding ester should only undergo hydrolysis following its absorption, therefore. the ester should be resistant to hydrolysis before being absorbed by the digestive enzymes but should undergo rapid hydrolysis eg by liver enzymes. In this way, the active diacid will be released into the bloodstream following oral absorption.

In addition to the lower alkyl esters (especially the ethyl esters), and the benzyl esters, suitable biodegradable esters include alkanoyloxyalkyl esters, including alkyl, cycloalkyl and aryl substituted esters thereof, aryloxyalkyl esters, aryloxyalkyl esters, aralkyloxyalkyl esters, aryl esters, aralkyl esters and haloalkyl esters, wherein said alkanoyl or alkyl groups contain from 1 to 8 carbon atoms and have a straight or branched chain,

said aryl groups represent phenyl, naphthyl or indanyl, optionally substituted by 1 or more C1 -C4 -alkyl or C1 -C4 alkoxy groups or halogen atoms. 4

Thus, examples of R1 and R2 when they represent biodegradable esterifying groups other than ethyl and benzyl include: 1- (2,2-diethyl-butyryloxy) -ethyl-, 2-ethylpropionyloxymethyl, 1- (2-ethylpropionyloxy) ) -ethyl, 1- (2,4-dimethylbenzoyloxy) -ethyl, 1- (benzoyloxy) -benzyl, 1- (benzoyloxy) -ethyl, 2-methyl-1-propionyloxypropyl, 2,4apos; -trimethylbenzoyloxymethyl , 1- (2,4,6-trimethyl-benzyloxy) -ethyl, pivaloyloxymethyl, phenethyl, phenopropyl, 2,2,2-trifluoroethyl, 1- or 2-naphthyl, 2,4-dimethylphenyl, 4- t -butylphenyl, 5- (4-methyl-1,3-dioxolen-2-onyl) -methyl and 5-indanyl.

The compounds of the formulas (I) and (II) wherein R 2 represents benzyl or t-butyl and R 2 is ethyl are valuable intermediates for the preparation of the compounds. di-acids in which R e and R representam are hydrogen.

In another preferred group of compounds, R 1, 12-13 of the formula -NHCORR JR 4 represents methylene substituted by a group 12 in particular, wherein R 13

R 1 is H or NH 2, NH 2, and R 2 represents hydrogen, CH 2 N 4 NH 2. Especially those groups deriving from S-lysine are especially preferred; Thus, especially preferred substituents of this type include N-acetyl-2-S-lysylaminomethyl, N-methanesulfonyl-S-lysylaminomethyl 2 N -phenylsulfonyl-S-lysylaminomethyl and N-cyclobutylcarbonyl-S-lysyl- aminomethyl.

In other groups of compounds. Preferred R5 is C1 -C6 -alkyl substituted by C1 -C6 -alkoxy, in particular methoxyethyl; or represents -13-

C1 -C6 -alkyl substituted by C1 -C6 -alkoxy- (C1 -C6) -alkoxy, especially 2-methoxyethoxymethyl; or wherein R 2 represents C 1 -C 4 -alkyl substituted by -NHCOCR 2 R 2 R 3, wherein R A is R 1 represents CH 2 and R 2 represents hydrogen or methyl, including, in particular , R-alanyl-aminomethyl and (2-amino-2-methyl-propanoyl) -aminomethyl.

Preferably, R is phenyl, benzyl, phenethyl, methoxy, propyl or ethoxypropyl.

Especially preferred individual compounds of the present invention include: 2S- (N '-methanesulfonyl-S-lysylaminomethyl) -3- [1- -4-carboxy-3-ethoxypropyl) -cyclohexyl) -carbamoyl] -cyclopentyl-propanoic acid, 2- (N-methanesulfonyl-S-lysylaminomethyl) -3- [1-

-4-carboxy-3-cis-phenethyl-cyclohexyl) -carbamoyl] -cyclopentyl] propanoic acid, 2S- (2-methoxyethoxymethyl) -3- [cis -4-carboxy-3-cis-3-ethoxypropyl] ) cyclohexyl) carbamoyl] cyclopentyl] propanoic acid and 2S- (2-methoxyethoxy) ethyl] -3- [1- (4-carboxy-3-ethoxy) -3-methoxypropyl] cyclohexyl) carbamoyl] cyclopentyl] propanoic acid, and their biodegradable esterified derivatives. 1-

The compounds of formula (I) may be prepared by a number of different procedures. The basic procedure involves the preparation of a partially protected, cycloalkyl substituted glutaric acid derivative which is attached to an amine giving the desired glutaramide. The optionally free carboxylic acid group present in the amine or any existing reactive group in R may require protection during the coupling step, and the protecting groups used for this purpose may be removed during the final stage of the process. Scheme 1 shows the synthesis process, wherein A, B, R, and R have the meanings given above, R corresponds to the meaning assigned to R, any reactive group present being protected, this is necessary, and R1 and R2 are as defined in R1 and R2, except for the meaning of hydrogen, or they represent usual protecting groups of carboxylic acids. i-

Scheme 1

The reaction of the compounds of formula (III) and (IV) is achieved by standard coupling procedures for the preparation of amides. Thus, according to a suitable process, the reaction proceeds when the reactants are dissolved in an organic solvent, for example, with the use of a diimide as the condensing agent eg 1-ethyl-3- (diterethylaminopropyl) -carbodiimide, or N, N'-dicyclohexylcarbodiimide, and advantageously in the presence of 1-hydroxybenzo-triazole and an organic base, such as N-methylmorpholine. Generally, the reaction is terminated after a reaction period of 12 to 24 hours at ambient temperature, whereupon the reaction product is isolated by usual methods i.e. by washing with water or filtration for removal

of the secondary urea product, and by evaporation of the solvent used. The final product may be further purified by means of crystallization or chromatography if this is required. Compounds of formula (V) include, compounds of formula (I) wherein R 1 and R 2 represent C 1 -C 6 -alkyl or benzyl.

In some cases, the coupling product present in protected form may undergo chemical / customary reaction reactions in order to prepare other compounds of formula (V). Thus, for example, compounds of formula (V) in which R contains an ester group may be hydrolyzed or hydrogenated to give the carboxylic acid, which in turn may be subjected to other reactions, with an amine, to give amide derivatives of the carboxylic acid.

Similarly, compounds in which R contains a substituted or protected amino group (e.g., a benzylamino, dibenzylamino, benzyloxycarbonylamino or t-butyloxycarbonylamino group), may be converted to the free amines by hydrogenation or protonation, as is more appropriate. The amines obtained can be brought to react again; Thus, for example, the reaction with a sulfonyl halide gives rise to the corresponding sulfonamides, the acylation with an acid chloride or acid anhydride, provides the corresponding amides, the reaction with. an isocyanate gives rise to. urea derivatives and the reaction with a chloroformate results in the formation of the alkoxycarbonylamino end products. All such transformations are well known, and the conditions and reagents suitable for their accomplishment are well known to those skilled in the art, as are any further variations and hypotheses of synthesis. -17-

The di-esters of formula (V) may be subjected to subsequent reactions to give the monoesters of the dihydrate derivatives of formula (I) wherein one or both of R and R 4 are hydrogen. The reaction conditions adopted depend on the particular nature of the R and R groups present in the compound of formula (V), various variations of the synthesis process being possible. Thus, for example, if both R2 and R3 represent benzyl the hydrogenation of the product will provide the di-acid of formula (I) wherein R4 and R4 are both hydrogen. 18

In contrast, if one of R 19 and R is benzyl and the other is alkyl, hydrogenation will give a monoester as the final product.

Gaso if you wish, this one. The product may be hydrolyzed to give the di-acid again. If one of R1 and R2 is t-butyl, treating the compound of formula (V) with trifluoroacetic acid gives the corresponding acid. The final di-ester, wherein R e and R benz are benzyl or lower alkyl, may also be treated with trimethylsilyl iodide, to form the corresponding dicarboxylic acid. If any other carboxylic acid protecting group 18 is used for R or R, then the most suitable conditions for removal in the final step will be chosen to form the final ester or di-acid of formula (I ). In case 5 of ring A or substituent R is unsaturated, deprotection should be by non-reducing methods; Thus, for example, if any of R1 and R2 is benzyl, they may be removed by treatment with trimethylsilyl iodide.

As already mentioned above, in addition to the removal of any protecting groups optionally present in R, a number of chemical transformations can still be carried out on the final products

(mono-ester or di-acid). In either case the podule may be obtained in the form of the free carboxylic acid, or it may be neutralized with an appropriate base and isolated in the form of its salt.

According to a variant of the above process, compounds of formula 5 (I) are prepared, in which R represents C 1 -C 6 -alkyl substituted with -NR 8 COR 9, -NR 8 SO 2 R 10 11 -NR COCR RR Or -NR S02CR R RX41 through a process which involves acylation or sulfonylation of a compound of the formula:

(VI): wherein R 2 and R 2 are as defined above and Y represents a C 1 -C 6 -alkyl group, alkyl; by reaction with an acid of the formula or with one of its activated derivatives. If necessary, the protecting group is removed from the resulting product (amide or sulfoidamide), and the obtained di-ester is cleaved to form the carboxylic acids of formula (I), wherein R4 and R4 represent hydrogen, such as above is indicated. -19-

(VI) according to the procedures indicated in scheme 51, but using a compound of formula (III) containing R as protected amino derivative. Thus, for example, R may contain a bis- (1S) -phenylethylamino substituent. Hydrogenation of the coupled product gives rise to the corresponding free amine of formula (VI), wherein R represents hydrogen and Y means CH2

This synthetic pathway acquires particular importance in the preparation of compounds having the stereochemical configuration 2 (S) in its glutaramide base structure.

The starting compounds, the sky-alkyl substituted glutaric acid monoester esters of formula III may be prepared as described in our European patent applications Nos. EP-A-0274234, 89305180.5 and 89304698.7.

In general, the amines of formula (IV) are novel compounds (especially when the substituents have a defined stereochemical configuration) and may be prepared by suitable synthetic methods according to the teachings contained in the corresponding literature. Thus, according to one of the methods, the corresponding ketone is converted to the amine by reduction of the oxime. According to an alternative process, the corresponding alcohol is reacted with methyl toluene sulfonate in the presence of diethyl diazodicarboxylate and triphenylphosphine; The resulting 4-methylbenzenesulfonyloxy derivative is reacted with sodium azide, and the read-out product is reduced to the amine. These processes are explained in the Examples of Preparations set forth below. -20-

Referring to the manuals of the skill and on the basis of the examples below, those skilled in the art will readily be able to appreciate the protection and coupling processes for all alternative steps and variants as well as general synthetic processes.

As discussed above, the compounds of the present invention are powerful inhibitory agents of the neutral endo-peptidase (E.C.3.4.24.11). This enzyme intervenes in the decomposition of a number of peptide hormones, and, in particular, conditions the degradation of atrial natriuretic factor (ANF). This hormone is a set of related natriuretic peptides - secreted by the heart - it being known that its most important form in the bloodstream of humans is a peptide containing 28 amino acids, designated alpha-hANP. Thus, by preventing the degradation of ANF by the endopeptidase E.C. 3.4.24.11, the compounds of the present invention may potentiate their biological effects whereby these compounds are diuretic and natriuretic entities with beneficial action on a number of disorders already mentioned above. The activity against the neutral endopeptide E.C. 3.4.24.11 can be evaluated by a method based on an assay described by J.T. Gafford, R.A. Skidgel, E.G. Erdos and L.B. Hersh, Biochemistry, 1983, 32, 3265-3271. This method determines the required concentration of a given compound to reduce the release rate of hippuric acid radiolabelled from hipuryl-L-phenylalanyl-L-arginine by a 50% neutralization mediated by a neutral endopeptidase preparation obtained from rat kidneys. -21-

The activity of the compounds as diuretic agents is determined by measuring their ability to increase urine and sodium ion excretion in conscious rats subjected to sodium chloride overload. In this assay, male rats (Charles River CD1, 22-28 g) are acclimatized and maintained in metabolism cages overnight without food. The mice receive the compounds intravenously through the caudal vein, the test compound being dissolved in a volume of saline equivalent to 2.5% of body weight. Urine samples are collected every hour for a period of 2 hours in previously weighed tubes and the samples are analyzed for their electrolyte content. The urinary volume and the sodium ion concentration of the test animals are compared with the data from a control group that was only treated with a sodium chloride solution.

For the curative and prophylactic treatment of hypertension, heart attack or renal insufficiency in humans, oral doses of the compounds of the invention may be administered between 4 and 800 mg per day in the case of a normal adult patient (70 kg). Thus, for a typical adult patient, the individual tablets or capsules contain from 2 to 400 mg of active compound, contained in a suitable pharmaceutically acceptable carrier or carrier, for single or multiple multiple dose administration administered once or several times daily . Generally, and according to need, the doses for an intravenous administration are between 1 and 400 mg per single dose. In clinical practice, the attending physician will decide on the most effective dosage schedule for the patient, which will vary, obviously depending on the age, body weight and response of the patient concerned. The above dosages are examples of average cases; however, individual cases may arise which require more effective dosage ranges.

or low rails. and these cases are also encompassed within the scope of the present invention.

For administration to human patients, the compounds of the present invention of formula I may be administered alone, although generally they are administered in admixture with a pharmaceutical carrier, chosen depending on the intended mode of administration and the usual pharmaceutical practice.

Thus, e.g. , the compounds may be administered orally in the form of tablets containing excipients, such as starches or lactose, or in capsules or ovules either alone or in admixture with excipients, or as elixirs or suspensions containing flavorings or dyes. The compounds may be injected parenterally, e.g., intravenously, intramuscularly or subcutaneously. For parenteral administration, the compounds may advantageously be used in the form of a sterile aqueous solution, which may contain further subtances, eg sufficient amounts of salts or glucose, to provide solutions isotonic to blood.

The compounds of the present invention may be administered alone but may also be administered together with other formulations or compounds which the attending physician specifies for the purpose of optimizing blood pressure control or treating heart attack due to hyperemia, renal failure or other disorders occurring in a patient, according to proven medical practice. Thus, the compounds may be administered together with a whole. of cardiovascular preparations, e.g. , with an ACE inhibitory agent, such as captopril or enalapril, in order to facilitate the regulation of blood pressure-

in the treatment of hypertension; or together with digitalis or other cardiac tonics or with an AGE inhibitor, in the treatment of hyperemia cardiac collapse. In other cases, the compounds may be administered concurrently with a calcium antagonist (eg, nifedipine, amlodipine or diltiazem), a beta-blocker (e.g., alpha-nolol) or an alpha-blocker (e.g., prazosin or doxazosin) as indicated by the physician as being appropriate for the treatment of the patient or the condition in question.

In addition, the compounds may also be administered together with exogenous ANF, or a derivative thereof, or a related peptide or a related peptide fragment, which have diuretic / natriuretic action, or other gene related peptides. of the ANF (e.g., as reported by D.L. Vesely et al., Biochem Biophys Res. Comm., 1987, 143, 186).

Thus, according to a further aspect of the present invention there is shown a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt or precursor compound thereof, together with a pharmaceutically acceptable diluent or excipient. The present invention also relates to compounds of formula (I) or to a pharmaceutically acceptable salt or prodrug derivatives thereof, for use in the field of medicine, in particular in the form of diuretic preparations for treatment of hypertension, cardiac collapse due to hyperemia or renal failure occurring in human patients. The present invention further provides the use of a compound of formula (I)

" X * tion of a medicament for the treatment of hypertension, cardiac collapse, angina pectoris, renal insufficiency, premenstrual syndrome, cyclic edema, Menières's disease, hyperaldosteronism, pulmonary edema, ascites, hipercalciuria, glaucoma, asthma , inflammation, painful conditions, epilepsy, affective disorders, dementia and confusional states in old age, obesity, gastrointestinal disorders (including diarrhea), hyper-reninemia, leukemia, as well as for the regulation of gastric acid secretion. The preparation of the compounds of the present invention will now be demonstrated in more detail by the following practical examples, in which the practical examples (preparations) 1 to 8 describe the preparation of the starting amines of formula (IV) and examples 1 to 45 refer synthetic methods of the compounds of formula (I). The purity of the compounds was usually assessed by thin layer erytheography using silica gel plates. The solvents system ss-1 is a mixture of dichloromethane, methanol and glacial acetic acid (90:10 : 1), the solvent system ss-2 is the upper phase of a mixture of methyl iso-butyl keto, glacial acetic acid and water (2: 1: 1), and the solvent system ss-3 is a mixture of ethyl acetate and hexane (1: 1). The nuclear magnetic resonance spectra of H were recorded by a Nicolet QE-300 spectrometer, revealing, in all cases, equivalence to the predicted structure.

PREPARATION 1

Ethyl 4-amino-c-2- (2-phenylethyl) cyclohexane-R-1-carboxylate hydrochloride i. Ethyl 4-QXO-2- (2-phenylethyl) cyclohex-2-ene carboxylate

In an atmosphere of nitrogen, a 80% dispersion of sodium hydride in oil (307 mg, 10 mmol) was added to a stirred, ice-cold mixture of ethyl 3-oxo-5-phenylpentanoate (28.4 g , 129 mmol) and absolute ethanol (1 mL). Then, methyl vinyl ketone (10.7 ml, 129 mmol) was added dropwise over 1/2 hour and, after ice-cooling, and after stirring for 1 hour at room temperature Acetic acid (1.4 ml) was added pyrrolidine (775 mg), ethanol (17 ml), " and 2 ml of water. After keeping the mixture boiling at reflux for a period of 2 hours the solvent was evaporated in vacuo and the residue was dissolved in diethyl ether. Washing successively with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and water, drying over magnesium sulfate and evaporation resulted in the formation of a yellow liquid (34.6 g). After chromatography on a silica gel plate (600 g), elution with a mixture of diethyl ether and hexane (1: 3, by volume), the desired enone was obtained as a clear liquid (18, 54 g), Rf 0.35 (1: 1 diethyl ether: hexane).

Found: C, 74.88; H, 7.40. C 17 H 9 N 4 requires the presence of C, 74.97; H, 7.40%.

ii. Ethyl 4-oxo-cis-2- (2-phenylethyl) cyclohexanecarboxylate At a pressure of 50 p.s.i. (3.45 bar) the enone (18.44 g, 6.76 mmol) obtained in (i) in ethanol (80 ml) containing 3 ml of 2N hydrochloric acid was hydrogenated using as palladium-on-charcoal catalyst to 5% (500 mg). After a further 3 hours, an additional 250 mg of the catalyst was added and the hydrogenation continued for an additional 2 hours. The resulting suspension was filtered through " Avicel " and the obtained filtrate was evaporated in vacuo. The residual oil was dissolved in diethyl ether, washed with a saturated aqueous solution of sodium bicarbonate and water was dried over magnesium sulfate and evaporated, yielding a clear (18, 52 g). After chromatography on silica gel (600 g) eluting with an increasing amount of diethyl ether in hexane (2: 8-4: 6) the desired ketone was obtained as a liquid. (Yield 16.78 g, 90%). Rf 0.4 (1: 1, diethyl ether: hexane). -X. ν: iii. Ethyl 4-methoxyimino-2- (2-phenylethyl) cyclohexanecarboxylate

The ketone obtained in (ii) (3.02 g, 11 mmol) methoxylamine hydrochloride (1.19 g, 14.3 mmol) was refluxed for a period of 5 hours in ethanol (50 ml) and acetate (1.17 g, 14.3 mmol). The solvent was evaporated in vacuo and the residue was dissolved in diethyl ether and washed with saturated aqueous sodium bicarbonate solution and water. After drying over magnesium sulfate and evaporation in vacuo the 0-methyloxime was obtained 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. C18 H25 NO3 requires: C, 71.26; H, 8.30; N, 4.62%. -28-

iv. c-4-Butoxycarbonylamino-c-2- (2-phenylethyl) cyclohexane-1-carboxylate 4.2 ml (54 mmol) of trifluoroacetic acid in anhydrous tetrahydrofuran (10 ml) were added dropwise , under ice-cooling, was added dropwise to a stirred suspension of sodium borohydride (2.04 g, 54 mmol) in anhydrous tetrahydrofuran. The temperature was maintained at 5 to 10 ° C and after stirring for 15 minutes the O-methyl oxy ma obtained in (iii) (3.28 g, 10.8 mmol) in tetrahydrofuran (10 mL) was added dropwise over a period of 10 minutes. The temperature rose to 20øC and, after stirring for 5 hours, a volume of water was carefully added under ice-cooling, and the obtained mixture was partitioned between ethyl acetate and water. The aqueous phase was extracted 2 times with ethyl acetate. The combined organic extracts were washed with brine, dried over magnesium sulfate and evaporated to give a gummy substance (3.25 g). The unprocessed amine was dissolved in methylene chloride (50 ml), 2.2 g (21.6 mmol) of N-methylmorpholine and 4.7 g (21.6 mmol) of di-iodomethyl bicarbonate and the obtained mixture was allowed to stand at room temperature for a period of 48 hours. The solvent was evaporated in vacuo, the residue was partitioned between diethyl ether and water, and the organic extract was washed successively with hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and water. Then, drying over magnesium sulphate and evaporation gave 3.9 g of a gummy substance. The required cis compound was separated from the smaller, more polar trans-isomer by chroma-

(400 g). Elution with a mixture of diethyl ether in hexane (3: 7 by volume) gave the formation of the protected amine as an oil (1.94 g g, 48%). 0.35 (3: 7, diethyl ether: hexane).

Found: C, 70.43; H, 8.75; N, 3.45. C22H33N04 and <3re the presence of C, 70.37; H, 8.86; N, 3.73%. v. C-4-amino-and-2- (2-phenylethyl) cyclohexane-1-carboxylic acid ethyl ester hydrochloride

A stirred, ice-cooled solution of the above ester (1.94 g, 5.17 mmol) in diethyl ether was saturated with hydrogen chloride. After 3 hours the solvent was evacuated with a stream of nitrogen and the residue was triturated with diethyl ether and collected by filtration to give the title amine as a white solid. as a white solid (1.44 g, 89%).

Melting point: 213-214 ° C. Rf 0.5 (ss-1).

Found: C, 65.84; H, 8.74; N, 4.43. C 17 H 26 NO 2 .HCl requires the presence of C, 65.48; H, 8.40 and N, 4.49%. -30-

PREPARATION 2

Ethyl c-4-amino-c-2- (3-methoxypropyl) -hexyhexane-1-carboxylate hydrochloride

The procedure adopted in Preparation 1 was followed, but ethyl 3-oxo-6-methoxyhexanoate was used as starting material in step (i) to give the title amine as a solid white solid having a melting point of 206-208 ° C.

Rf 0.2 (ss-1).

Found: C, 55.15; H, 9.11; N, 5.34. C13 H25 NO3 .HCl, 0.2 H2 O, assumes the presence of C, 55.09; H, 9.39; N, 5.94%. PREPARATION 3 ---

C-4'-Amino-c-2- (3-ethoxypropyl) cyclohexane-1-carboxylic acid ethyl ester hydrochloride

The procedure of Preparation 1 was followed, however, using ethyl 3-oxo-6-ethoxyhexanoate as the starting compound in step =. (i); the title amine was obtained as a white solid. Melting point: 201-203 ° C. Rf 0.2 (ss-1). Found: C, 55.50; H, 9.52; N, 4.33. C ^ fH ^ ^NO.. HC1.V2 assumes the presence of C, H and N in a percentage of 55.52; 9.65 and 4.63%, respectively. -6-

C4-Amino-2- (2-phenylethyl) cyclohexane-1-carboxylic acid methyl ester hydrochloride

A solution of the ethyl ester obtained in Preparation 1 (1.4 g, 4.48 mmol) in anhydrous methanol (50 ml) was saturated with hydrogen chloride and heated to 55-60øC for 4 days and then , maintained at reflux for an additional 3 days. To ensure a sufficient degree of saturation, hydrogen chloride was introduced at periodic intervals. The solution was evaporated to dryness in vacuo, the residue was dried under azeo-tropic conditions with methylene chloride, and the residue was triturated with diethyl ether to give the methyl ester as a white powder ( 1.29 g, 97%), mp 202-204 ° C. Rf 01.5 (ss-1).

Found: C, 63.70; H, 8.13; N, 4.74%.

HCl 0.25 assumes the presence of C, H and N in percentages of respectively 63.56; 8.17 and 4.63%. i-

PREPARATION 5 Ethyl e-4-amino-t-2- (2-phenylethyl) cyclohexane-1-carboxylate i. The ketone of Preparation 1, step (ii) (10.97 g, 40 mmol), ethylene-2-carboxylic acid (100 mg) were refluxed in benzene (80 ml) using a Dean-Stark water-collection flask. After 8 hours, the solution was cooled, diluted with diethyl ether and washed, in a saturated aqueous solution of sodium bicarbonate and water. After drying over magnesium sulfate and evaporation in vacuo, a clear liquid (12.22 g, 96%), Rf 0.32 (3: 7, diethyl ether: hexane) was obtained.

Found: C, 71.45; H, 8.30. The presence of C and H in a percentage of respectively 71.67 and 8.23% was observed. -33-

ii. Ethyl trans-7- (2-phenylethyl) -1,4-dioxaspiro [4.5] decan-8-carboxylate

Potassium t-butoxide (1.9 g, 17 mmol) was added to a solution of the above-referenced asber (12.17 g, 38.2 mmol) in anhydrous t -butanol (90 ml) and the obtained mixture was refluxed under nitrogen for a period of 48 hours. After neutralization with 2N hydrochloric acid and evaporation in vacuo a light brown oil was obtained, which was dissolved in diethyl ether and washed with water. After drying over magnesium sulfate and evaporation there was obtained an oil (12.09 g) which was chromatographed on silica gel (600 g). Elution operations 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. And the presence of C and H in a percentage of respectively 71.67 and 8.23%. -6-

iii. Ethyl 4-oxo-trans-2- (2-phenylethyl) cyclohexanecarboxylate The ester mentioned above (8.40 g, 26.4 mmol) was refluxed in 1N sulfuric acid (50 ml) and ethanol (70 ml) ml). After 3 hours, most of the ethanol was evaporated in vacuo; water was added and the suspension was extracted with diethyl ether. The organic extract was washed successively with water, saturated aqueous solution of sodium bicarbonate and water, dried over magnesium sulfate and evaporated in vacuo to give a clear oil (6.46 g, 89%), 0.38 (1: 1, diethyl ether: hexane).

Found: C, 74.25; H, 8.07. And the presence of C and H in a percentage of 74.42 and 8.08%, respectively. iv. ethyl 3- (2-phenylethyl) -c-4-phenylmethylamino-cyclohexane-1-carboxylate 3 g of a molecular sieve of 38 was added to a stirred solution of the ketone obtained in step iii (3, 2 g, 11.66 mmol) and benzylamine (1.27 mL, 11.66 mmol) in ethanol (15 mL).

After 3 hours at room temperature, the solution was pipetted into a dry hydrogenation vessel with washing the molecular sieves with a small volume of ethanol (2 x 5 mL). Platinum oxide (500 mg) was added, and the reduction was terminated within 3 hours at a pressure of 50 p.s.i. -35-

(3.45 bar). The mixture was filtered through &quot; Arbacel &quot;, was evaporated in vacuo, and the residual liquid was chromatographed on a silica gel plate (200 g). Elution with increasing amounts of ethyl acetate in hexane (7: 3-1: 1 by volume) resulted in the formation of 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%. C24H31N ° 2 Pressure and the Presence of C, H and N in percentages of respectively 78.86%, 8.55 and 3.83%. v. Ethyl c-4-amino-2- (2-phenylethyl) cyclohexane-1-carboxylate

A solution of the above amine (3.13 g, 8.56 mmol) in ethanol (80 mL) containing 1N hydrochloric acid (9.42 mL, 9.42 mmol) was hydrogenated over palladium hydroxide (20 mL). %) and charcoal as the catalyst (600 mg) at a pressure of 50 psi (3.45 bar). After 6 to 24 hours, new amounts of catalyst (500 mg) were added. After another 22 hours, the suspension was filtered through &quot; Arbacel &quot; and the filtrate was evaporated to dryness in vacuo. The residue was recrystallized from a mixture of ethanol and diethyl ether to give the required amine as a white solid (2.03 g, 76%). M.P. = 175-177 ° C. R 0.25 (ss-1).

Found: C, 64.42; H, 8.50; N, 4.52. C 17 H 25 NO 2 .HCl. 0.25 H2O assumes the presence of C, H and N in percentages of respectively 64.54, 8.44 and 4.43%. -36-

PREPARATION 6

C-4-amino-c-2-phenylcyclohexane-R-1-carboxylate hydrochloride i. 4-Oxo-2-phenylcyclohex-2-ene-2-ene-2-ene-2-carboxylate 17.5 g (70.48 mmol) of 2-hydroxy-4-oxo-2-phenylcyclohexane, J. Org. Chem. 3, 2427 (1974)] were stirred for 15 minutes at 80 ° C with 17.5 g of polyphosphoric acid.

After cooling, the mixture was partitioned between ethyl acetate and water. Then, the organic extract was washed successively with water, saturated aqueous solution of sodium bicarbonate and water, dried over MgSO 4 and, after evaporation, formed an oil. After chromatography on silica gel plates and elution with increasing amounts of ethyl acetate in hexane (2: 8-3: 7 by volume) the desired compound was obtained as a gold oil (9: 0 g, 56% yield). Rf 0.5 (ss-3).

ii. The enone obtained in step 1 (9.0 g, 39.08 mmol) in methanol (40 ml) containing 2N hydrochloric acid (1.5 ml) was hydrogenated over palladium- (5%) as catalyst, at a pressure of 50 psi (3.45 bar) After 3 hours the residue was filtered through Arbacel and evaporated to dryness in vacuo The remaining oil was chromatographed on silica gel plates (400 g) After elution with a gradient of ethyl acetate in hexane (2: 8-4: 6 by volume) gave the desired ketone as a white solid (4.04 g, 49%): 0.55 (ss-3 ).

Found: C, 72.39; H, 6.95. W

O- assumes the presence of C and H in a percentage of 72.39 and 6.94% respectively. -38-

iii. methyl bromo-2-phenylcyclohexane-1-carboxylate 2.33 g (61.51 mmol) of sodium borohydride were added in small portions to a stirred solution of the ketone obtained in step ii ( 3.78 g, 16.27 mmol) in methanol (80 mL); in this operation, the temperature was maintained below 5 ° C. After 15 minutes, the solvent was evaporated in vacuo, the residue was dissolved in methylene chloride and washed with a saturated sodium chloride solution. After drying over magnesium sulfate and evaporation in vacuo a white solid was obtained. The desired ester (2.93 g, 77%) was obtained as a foam after chromatography on silica gel plates (250 g) and elution with diethyl ether in hexane (1: 1, by volume).

Found: C, 71.84; H, 7.82. (2) assumes the presence of C and H in a percentage of 71.77 and 7.74% respectively. iv. t-4- (4-methylbenzenesulfonyloxy) -β-2-phenylcyclohexane-1-carboxylate 4.08 g (23.47 mmol) of diethyl diazodicarboxylate in tetrahydrofuran (10 ml) were added, To a stirred solution of the ester obtained in step iii (4.4 g, 18.78 mmol) triphenylphosphine (6.16 g, 23.47 mmol) and methyl 4-toluenesulfonate (4.37, 47 mmol) in tetrahydrofuran (60 ml); the temperature was maintained between 5 and 10 ° C. After stirring overnight at room temperature, the

The reaction mixture was cooled to 0øC and additional triphenylphosphine (1.23 g, 4.7 mmol) and methyl 4-toluenesulfonate (874 mg, 4.7 mmol) were added, after which diethyl diazodicarboxylate (816 mg, mg, 4.7 mmol). After stirring at room temperature for an additional 4 hours the mixture was evaporated to a flash volume, the residue was redistributed in methylene chloride and applied directly to a plug. silica gel plate (400 g) for column chromatography. After elution with increasing volumes of ethyl acetate in hexane (1: 9-3: 7 by volume) the required 4-toluenesulfonate was obtained as an oil (3.45 g, 47%), 0.45 (4: 1, ethyl acetate, hexane).

Found: C, 64.71; H, 6.26. C21H24O5S assumes the presence of C and H in percentages of 64.93 and 6.23% respectively. v. Cis-4-Azido-cis-2-phenylcyclohexane-1-carboxylate 1.16 g (17.76 mmol) of sodium azide were added to a stirred solution of the 4-toluenesulfonate obtained from step iv (3.45 g , 8.88 mmol) in dimethylformamide, and the resulting mixture was stirred at 60 ° C for 18 hours. After cooling, water was added and the resulting suspension was extracted with diethyl ether. The organic extract was washed with water, dried over magnesium sulfate and, after evaporation, a clear oil was obtained which was chromatographed on silica gel (300 g). After elution with increasing volumes of ethyl acetate in hexane (1:20 - 1: 4 by volume) the azide was obtained as a waxy solid (2.11, 92%). 0.55 (4: 1, hexane, ethyl acetate).

Found: C, 65.20; H, 6.60; N, 15.32. C 14 H 17 N 3 O 2 of 64.85, assumes the presence of C, H and N in percentages 6.61 and 16.20%, respectively. saw. The azide obtained in step (v) (2.1 g, 8.1 mmol) in methanol (50 ml) was refluxed for 2 hours at room temperature. was hydrogenated over 10% palladium-on-charcoal (catalyst) (210 mg) at a pressure of 50 psi (3.45 bar). After 2 x 4 hours, the mixture was filtered through a short column of &quot; Arbacel &quot; and evaporated in vacuo. The residue was dissolved in ethyl acetate and acidified by the addition of 1N hydrogen chloride in diethyl ether. The 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. 255-256 ° C (decomposition) . Rf 0.2 (ss-1).

Found: C, 62.64; H, 7.74; N, 5.74. C14H19NC> 2. HCl assumes the presence of C, H and N in percentages of 62.33; 7.47 and 5.19%, respectively. PREPARATION 7

Cis -3-Amino-cis-5-phenylmethyl-cyclohexane-R-1-carboxylic acid methyl ester hydrochloride i. cis-5-tert-butyldimethylsilyloxy-cyclohexane-cis-1,3-dicarboxylate

Under ice-cooling, 16.58 g (110 mmol) of t-butyldimethylsilyl chloride in anhydrous methylene chloride (65 ml) were added dropwise over a period of 5 to 10 ° C. A solution of cis-3-hydroxycyclohexane-cis-1,3-dicarboxylic acid dimethyl ester (21.5 g, 99 mmol) (11.13 g, 110 mmol) and 4-dimethylaminopyridine (488 mg, 4 mmol) in methylene chloride (80 mL). After being allowed to stand overnight at ambient temperature, additional amounts of t-butyldimethylsilyl chloride (1.6 g), triethylamine (1.1 g) and dimethylaminopyridine (450 mg) were added to the mixture, and stirring was further maintained for a period of 68 hours.

The mixture was then washed successively with water, saturated aqueous ammonium chloride solution and water, dried over magnesium sulfate and evaporated in vacuo to give a liquid (34.4 g) which was chromatographed on silica gel plates (600 g). After elution with a mixture of diethyl ether in hexane (1: 3 by volume) the desired product was obtained as a liquid (32.6 g, 99%). Rf 0.3 (1: 3, diethyl ether, hexane).

Found: C, 57.89; H, 9.20. C 5 H 8 O 4 O requires the presence of C and H in percentages of 58.15 and 9.15% respectively. ii. cis-5-tert-butyldimethylsilyloxycyclohexane-cis -1,3-dicarboxylate 94 ml of 1N sodium hydroxide were added to an ice solution of the above-referenced diester (30.87 g, 93.4 mmol), in methanol (500 ml). The mixture was stirred at 5 ° C for a period of 3 hours, and then stood at room temperature for a period of 18 hours. Most of the solvent was evaporated off in a vacuum, water was added and the resulting suspension was extracted with diethyl ether to recover the unreacted di-ester (3.86 g). The aqueous phase was acidified by the addition of 2N hydrochloric acid, extracted with diethyl ether and the organic extract was washed with water, dried over magnesium sulfate and evaporated in vacuo to give a clear, gummy substance (25.5 g ). Chromatography on silica gel (500 g) and elution with a mixture of diethyl ether in hexane (7: 3 by volume) resulted in the formation of the required mono ester in the form of a solid oil (20.0 g, 68% ). 0.75 (diethyl ether). Found: C, 56.75; H, 8.71. C15 H28 O5 S The presence of C and H is found in a percentage of 56.93 and 8.92% respectively. -43-

iii) methyl cis-3-tert-butyldimethylsilyloxy-cis-5-hydroxymethyl-cyclohexane-R-1-carboxylate

A 1M solution of diborane in tetrahydrofuran (59 ml) was dropwise, under a nitrogen atmosphere, to a stirred solution of the monoester obtained in step ii (17.0 g, 53.72 mmol) in tetrahydrofuran while maintaining the temperature at &lt; 5 ° C. The mixture was allowed to warm to room temperature, and after 4 hours, a further amount (10.7 ml) of the 1M solution of diborane was added, and the mixture was allowed to stand for 18 hours. The solvent was evaporated in vacuo and diethyl ether was added and the solution obtained was washed successively with saturated sodium chloride solution, saturated aqueous sodium bicarbonate solution and again with saturated sodium chloride solution. sodium.

After drying (MgSO4) and evaporation in vacuo, a clear oil was obtained, which was chromatographed on silica gel (500g). After elution with a mixture of ethyl acetate and hexane (1: 1 by volume) an oil (15.18 g, 93%) was obtained.

R f 0.42 (ss-3). Found: C, 59.24; H, 9.67. C 15 H 30 O 4 S requires the presence of C and H in a percentage of 59.56 and 10.0% respectively. -44-

iv. cis-3-tert-butyldimethylsilyloxy-cis -5- (4-methylbenzenesulfonyloxymethyl) -cyclohexane-1-carboxylate 14.18 g (74.39 mmoles) of 4-methylbenzenesulfonyl chloride were added to an ice-cold solution , stirred solution of the ester obtained in step (iii) (15.0 g, 49.59 mmol) in anhydrous pyridine (130 ml). After stirring for 24 hours at room temperature the mixture was poured onto ice, extracted with diethyl ether and the extract washed successively with 2N hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and water. After drying over magnesium sulfate and evaporation in vacuo, an oil (22.0 g) was obtained which was subjected to silica gel chromatography (600 g). Elution with increasing volumes of ethyl acetate in hexane (1: 9-1: 4 by volume) resulted in the formation of 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. ^ 2 ^ 6 ^ 5 ^ assumes the presence of C and H in a proportion of 57.86 and 7.95% respectively. -4-

v. cis-3-tert-butyldimethylsilyloxy-cis-5-phenylmethyl-cyclohexane-1-carboxylate 24.5 ml (49 mmoles) of 2M phenyl lithium in cyclohexane ether (7: 3) in a nitrogen atmosphere, was added dropwise to a stirred suspension of cuprous iodide (4.67, 24.5 mmol) in anhydrous diethyl ether (50 ml), keeping the temperature below 5 ° C. The resulting dark green solution was stirred at room temperature for 1 3/4 hours, cooled to -70 ° C, a solution of the ester of step (iv) (3.7 g, 8.1 mmol) in 10 ml of ether and cyclohexane (40 ml) was added dropwise. The temperature was maintained below -60øC during this addition and then reduced to -70øC for 15 minutes. The mixture was then allowed to warm to room temperature, stirred for 1 hour and the reaction was quenched by addition of saturated aqueous ammonium chloride under ice-cooling, and the mixture was dried over sodium sulfate. magnesium and, upon evaporation, in a vacuum, resulted in the formation of an oil. After chromatography on silica gel, elution with increasing volumes 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). -4-

saw . 0.5 ml (10 mmol) of hydrofluoric acid at 40 ° C were added to a stirred solution of the above ester (3-methyl-3-methylbenzenesulfonyloxy) -cis-3-phenylmethylcyclohexane-1-carboxylate , 6 g, 9.93 mmol) in 40 ml of acetonitrile. After 1 hour the solution was diluted with diethyl ether, washed with a saturated aqueous solution of sodium bicarbonate and water, dried over magnesium sulfate and evaporated in vacuo to give the desired alcohol as an oil clear (2.36 g, 96%). Rf value: 0.2 (1: 1, diethyl ether, hexane). The resulting product (2.39 g, 9.62 mmoles) was treated as described in Preparation 6, step iv, whereby the title compound was obtained as a gently yellow gum (2 g). , 44 g, 63%). Rf value: 0.4 (1: 1, diethyl ether, hexane).

Found: C, 65.59; H, 6.54; C 22 H 26 O 5 S requires the presence of C and H in a proportion of 65.65 and 6.51% respectively. -4-

vii. 2.41 g (5.99 mmol) of the trans -4-methylbenzenesulfonate obtained in step iv were treated with sodium azide as a white solid. as described in Preparation 6, step v, to give the corresponding azide as an oil (1.44 g, 88%). 0.72 (1: 1, diethyl ether, hexane). The azide was then reduced (1.41 g, 5.16 mmol) as described in Preparation 6, step vi to form the required amine salt as a foam (1.44 g, 99%). Rf: 0.2 (ss-1).

Found: C, 63.31; H, 7.76; N, 4.56. The presence of C, H and N in a ratio of 63.48, 7.81 and 4.94%. PREPARATION 8

3-Amino-cis-4-phenyl-methylcyclohexane-R-1-carboxylic acid methyl ester hydrochloride (cis and trans). i. Methyl 3-oxo-4-phenylmethylcyclohexanecarboxylate

A mixture of ethyl acetate (trimethylsilyl) (16.01 g, 99.9 mmol) and methyl 3-oxocyclohexane carboxylate (13.0 g, 83.24 mmol) was slowly added dropwise to a solution of nitrogen atmosphere was added to a stirred solution of 1.0M tetrabutylammonium fluoride in 2.78 ml of tetrahydrofuran, maintaining the temperature between 0 and 10 ° C. The resulting light brown solution was stirred at room temperature for 17 hours; additional tetrabutylammonium fluoride (0.3 ml) and ethyl acetate (trimethylsilyl) (5 ml) were added and after 3 hours monitoring by NMR showed the reaction to be complete. The mixture was diluted with n-hexane (50 ml) distributed over molecular sieves: 48 for 1 hour and filtered; the solvent was evaporated in vacuo to give the silylenole ether as a gold oil (19.97 g, 100%). The product (silylenole ether) in anhydrous methylene chloride (50 ml) was dropwise, in a nitrogen atmosphere, to a stirred suspension of titanium tetrachloride (9.15 ml, 83.24 mmol) in benzaldehyde (9.31 ml, 91.56 mmol) and anhydrous methylene chloride (250 mL) at -70 ° C. The mixture was allowed to warm to room temperature over a period of 18 hours, and the mixture was poured into 100 ml of water under ice-cooling. The organic phase was washed with water, dried over magnesium sulfate and evaporated in vacuo. The residue was chromatographed on silica gel, eluting with increasing volumes of ethyl acetate in hexane (1: 4: 3: 7 by volume) to give the desired product as a solid (6.89 g, 34%). M.P. = 73-75 ° C. : 0.25 (4: 1, hexane, ethyl acetate).

Found: C, 73.20; H, 6.54. C15 H16 O3 Â · H2 O The presence of C and H at a ratio of 73.21 and 6.63% was observed. ii. Methyl 3-oxo-cis-4-phenylmethylcyclohexanecarboxylate The enone obtained in step 1 (6.55 g, 26.8 mmol) in methanol (152 ml) and water (8 ml) was hydrogenated over palladium-on-charcoal (5%) (750 mg) at a pressure of 20 psi (1.4 bar). After 3 hours the suspension was filtered and, after evaporation in vacuo, the residue was chromatographed on silica gel (600 g). Elution with increasing volumes of ethyl acetate in hexane (1: 4: 3: 7 by volume) gave the formation of the trans-isomer as a white solid, m.p. 54-55 ° C. 0.35 (1: 4 ethyl acetate, hexane).

Found: C, 73.11; H, 7.46. and the presence of C and H in a ratio of 73.15 and 7.37% respectively. Continuing the elution gave the more polar cis-isomer as a white solid with M.P. of 71-72Â ° C. 0.25 (1: 4, ethyl acetate, hexane).

Found: C, 72.81; H, 7.41; 15.48%; and the presence of C and H in a ratio of 73.15 and 7.37% respectively. iii. Methyl 3-methoxyimino-cis-4-phenylmethylcyclohexanecarboxylate The cis-isomer obtained in step ii. (1.9 g, 7.71 mmol) when treated with methoxylamine hydrochloride as outlined in Preparation 1, step iii, was converted to the desired compound. O-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. C16H21H03 PressuP ° and the presence of C, H and N in proportions of 69.79, 7.69 and 5.09% respectively. iv. The methyl O-oxime obtained in step iii. The tert-butoxycarbonylamino- (1.97 g, 7.15 mmol) when treated as in Preparation 1, step iv. the title compound was transformed as a mixture of cis- and trans-isomers, which were separated by silica gel chromatography (300 g) and eluting with increasing volumes of ethyl acetate in hexane (1 : 3 - 3: 7 by volume). The trans-isomer was obtained as a resin (680 mg, 27%).

Rf 0.5 (1: 4 hexane, ethyl acetate). Found: C, 68.84; H, 8.17; N, 3.91 and 20H29NO4 PressuP ° and the presence of C, He N in proportions of 69.14, 8.41 and 4.03% respectively. The cis-isomer was also obtained as a resin (345 mg, 14%). 0.45 (1: 4, hexane, ethyl acetate). Found: C, 67.55; H, 8.08; N, 3.80. C 20 H 29 N 4 O 5. 5 H 2 O requires the presence of C, H and N in proportions of 67.39, 8.48 and 3.93% respectively. -51-

ν.

Methyl t-3-amino-c-4-phenylmethylcyclohexane-R-1-carboxylate hydrochloride The trans isomer obtained in step iv. (650 mg, 1.87 mmol) when treated with HCl as outlined in Preparation 1, step v, resulted in the formation of the title salt as a resin (533 mg, 99%).

Rf (ss-1). Found: C, 62.41; H, 7.78; N, 4.77. C15H21NO2 · HCl.0.25 H2O assumes the presence of C, H and N in proportions of 62.49, 7.87 and 4.86%. saw. cis-3-Amino-cis-4-phenylmethylcyclohexane-R-1-carboxylate (Hydrochloride) The cis-isomer from step iv. (325 mg, 0.94 mmol) when treated with HCl as outlined in Preparation 1, step v, was converted into the title salt as a resin (263 mg, 99%). Rf 0.25 (ss-1) Found: C, 59.45; H, 7.44; N, 4.77. C15H21NO2.HCl assumes the presence of C, H and N in ratios of 59.69, 8.01 and 4.64 respectively. Benzyl 4-ethoxycarbonyl cis-3- (2-phenylethyl) cyclohexyl) carbamoyl] cyclopentyl} -2- (2-methoxyethyl) propanoate 460 mg (2.4 mmol) of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride were added to a stirred, chilled solution of benzyl 3- (1-carboxy-cyclopentyl) -2- (2-methoxyethyl) -propanoate ( EP-A-0274234) (401 mg, 1.2 mmol) ethyl c-4-amino-c-2- (2-phenylethyl) cyclohexane-1-carboxylate hydrochloride (343 mg, 1.1 mmol ) 1-hydroxybenzotriazole (162 mg, 1.2 mmol) and N-methylmorpholine (354 mg, 3.5 mmol) in anhydrous methylene chloride (15 ml). After V2 hour the mixture was allowed to warm to room temperature and was stirred for 20 hours. The solvent was evaporated in vacuo, and the residue was partitioned between diethyl ether and water. The organic extract was washed successively with water 2N hydrochloric acid, water, saturated aqueous solution of sodium bicarbonate and water. After drying (MgSO4) and evaporation in vacuo, an oil (640 mg) was obtained, which was chromatographed on silica gel (50 g) eluting with a mixture of diethyl ether and hexane (7: 3 by volume) to give the desired compound (di-ester) as a colorless oil (550 mg, 85%). Rf 0.32 (7: 3, diethyl ether, hexane).

Found: C, 73.23; H, 8.37; N, 2.12. assumes the presence of C, H and N in proportions of 73.07, 8.35 and 2.37% respectively. -53-

EXAMPLES 2-5

The following compounds were prepared according to the procedure of example 1, using the appropriate amine of formula (IV)

C02Ff

EXAMPLES 6-7

The following compounds were prepared according to the procedure adopted in example 1 but using t-butyl 3- (1-carboxycyclopentyl) -2- (S) - (2-methoxyethoxy-methyl) -propanoate (European patent application N2 89304698.7) as the starting compound, and upon connection to the appropriate amine of formula (IV). CH3o

or co2 CjH5

Analysis% (theoretical values between parentheses) C Η N <T> ^ 3 * r ~ &gt; * &quot; K CM CM cm in σ »σ» k K co CO CM CO CM O * K 0 \ U) 10 Sw · 71,70 8,10 2,07 (71,70 7,90 2,53) Ο C0 ιη οΓ οΓ γμ ri Ο ». * ω ω σι • «ίο ·> 3 · *. Ο Ο--Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο Ο »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» »» ». κ ΓΜ ΓΜ Γ * Isolated Form 1 *. n 1 t / 1 u 'w /. co g m 1 ° a r &gt; 1 ω ω Φά &amp; ο ιη ιη «*. ο rd jm * &quot; &gt; &lt; 1 &gt; 1 &lt; / RTI &gt; gum η 1 01 01 Ν- 'ιο Ο · S η in 2 X η r &gt; υ CM X κ CM u υ υ Ω CM Ο) CM 0 PS or Ο ο τ rsj r \ 9 9? η? Ο g 1/51 011 ΰιι -H • Η • Η J-I 01 οι ο | V 10 ~ ΓΜ η ιη δ X Τ ιη νο η Τ Τ Τ V V V CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM CM ι ι ι ι ι ι ι ι ι ι ι ι ι ι ι ι ι -ι • Η -Η οι ο | ο | -Ρ 0 rH Qj S 0 ΓΜ r &gt; ιη (U 2 X w -56-

Example No R2 Form, isolated Analysis% f (Theoretical values in parentheses) CHN 6 - (O) 30% oil, 64.47 9.35 2.53 cis Rf 0.45 (64.84, 9.61, 2.52) 7 -> 30 ° oil 65.17 9.44 2.51 cis Rf 0.5 (65.35 9.73 2.46)

EXAMPLES 8-12

The following compounds were prepared according to the procedure of example 1 using 2S- (N-benzyloxycarbonyl-N-t-butyloxycarbonyl-S-lysylaminomethyl) -3- [1- (1-carboxycyclopentyl) ) T-butyl 7-propanoate (European Patent Application No. 893051 -80) as the starting compound, and by attachment to the appropriate amine of formula (IV).

C &amp; Z NH

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CM Ο μ? Ι011 Η | (I.e.

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* -N Μ1 10 Η cn in cn m cn Η CM c \ CM t ». Tet cn cn - - - - - - - - - - - - - - * The RVR is 10 vo in vo ve ** 00 &lt; Ν t-cn r- CO t &quot; «Η η η n cn h or CM 4. * ν • S · \ • v *. • v C0 00 C0 CO CO 00 »CO CO η ιη Ο cn CO o o C0 ο ο n CM C0 r *** rH * Ν» &gt; φ · r r Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ Τ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ &lt; S &lt; RTI ID = 0.0 &gt; 3 &lt; / RTI &gt; 3 &lt; / RTI &gt; ol <M cn tu W W s w S H δ ιη ιη S a CM cn cn CM Ο a K U CM u 0 CM 8 o ™ CM Υ τ? YI CO 1 0) 1 1 οι • Η 011 01 • Η ο • η ul η η ΙΤ ΙΤ ΙΤ ΙΤ in in in in in in in in in in in in in in in in in in in in in in in in in in Ύ 1 V η ã in 1 r Ò 01 Ò Ò Ò Ò Ò Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η Η.

EXAMPLE 13 2 (S) - (N '- t -Butoxycarbonyl-S-lysylaminomethyl) -3- [cis -4-ethoxycarbonyl-cis -3- (3-ethoxypropyl) cyclohexyl) The di-ester of Example 8 (734 mg, 0.85 mmol) in 18 mL of methanol and water (2 mL) was hydrogenated at ambient temperature for 3 hours using t-butyl dicarboxylate as a palladium-on-charcoal catalyst (75 mg) at a pressure of 50 psi (3.45 bar). The suspension was filtered through &quot; Arbacel &quot; and after evaporation of the solvent under reduced pressure, the title product was obtained as a white foam (630 mg, 100%).

Rf: 0.66 (ss-1).

Found: C, 63.22; H, 9.73; N, 7.17 C 38 H 13 N 4 O 9 PressuP ° and the presence of G, H and N in percentages of 62.78, 9.70 and 7.71% respectively.

EXAMPLES 14-17

The following compounds are prepared from the compounds of examples 9-12, by hydrogenation, and following the procedure used in example 13.

Analysis% (theoretical values in parentheses) C Η N X &quot; * ». ** H Η n #v ts t &quot; r- ιη ω σι ο * &gt; Ο co to to a a a a a a a a a a a C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C »Ο VO CM r &gt; Μ Μ Μ Μ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ ΙΟ M M M M M M M M U U U U U U U U U (Ss-1) is S ddm W * -> gt; H 1 cn U ] ns iH o 3 ta g -d * 3 m vo w% jn ΙΩ nxo ffi, CM CM uou U CM CM CM CM Q oooooo QTYTY n QM · M · 1 0 1 1 1 tfll T or UI • H ol ol ol 0 in jo in KX 10 vo vo or U in <N CM • OL K VO CM CN D u cm s δ Nm »1 1 sf wn 0 1 1 t Y n r> g in All w 5 ΰιι • Hj • H jj -H υ | 0 ol * * LO LO LO LO LO LO LO C C C C 62 62 62 62 62 62 62.

EXAMPLE 18 2S- (N -Methanesulfonyl-N-t-butoxycarbonyl-S-lysyl-aminomethyl) -3- [1 - [(4-ethoxycarbonyl-cis-3- [3-ethoxypropyl] cyclohexylcarbamoyl] cyclopentyl I t-butyl propanoate 0.14 ml (1.8 mmol) of methanesulfonyl chloride were added dropwise to a stirred, ice-cooled solution of the amine of example 13 (601 mg, 0.83 mmol) and The solution obtained was stirred at 10Â ° C for 3 hours and to the reaction mixture was added a saturated aqueous solution of methylene chloride The organic phase was separated, the aqueous layer was extracted with methylene chloride and the combined extracts were dried over magnesium sulfate. After evaporation in vacuo, a resin (gum) was obtained which was subjected to chromatography on silica gel (100 g), eluting with increasing volumes of ethyl acetate in hexane, starting with a 1: 1 ratio mixture and finally using only ethyl acetate. Evaporation of the relevant fractions provided the required product as a foam (576 mg, 87%) 0.56 (ss-1).

Found: C, 58.66; H, 8.91; N, 6.65. The presence of C, H and N in percentages of 58.18, 9.01 and 6.96% respectively. -63-

EXAMPLES 19-23

The following compounds were prepared in a similar manner to that adopted in Example 18 using the appropriate amine of Examples 14 to 17, and by reaction with the appropriate sulfonyl chloride or acyl chloride. 2, to give the N-substituted phenyl derivative.

-64-

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EXAMPLE 24 2S- [1S- (1S) - [Phenylethyl] aminomethyl] -3- [cis -4-ethoxycarbonyl-cis-3- (3-methoxypropyl) cyclohexyl) carbamoyl] cyclopentyl t-butylpropanoate

For the preparation of the title di-ester as a resin (gum), 2S-bis - [(1S) -phenylethyl] aminomethyl-3- (1-carboxycyclopentyl) - propanoic acid and ethyl c-4-amino-2- (3-methoxypropyl) cyclohexane-1-carboxylate (obtained in Preparation 2) as described in example 1.

Rf value: 0.1 (4: 1, hexane, ethyl acetate).

Found: C, 73.19; H, 9.13; N, 4.01. C43H64N2O6 Pressu &quot; puts the presence of C, H and N in percentages of 73.26, 9.15 and 3.97% respectively. EXAMPLE 25 2S- [bis - [(1S) -phenylethyl] aminomethyl] (Cis-4-ethoxycarbonyl-cis-3- (2-phenylethyl) -cylohexyl) -carbamoyl] -cyclopentyl-1-propanoate The title compound was prepared as described in Example 24, using the c-4-amino-c2- (2-phenylethyl) cyclohexane-1-carboxylic acid ethyl ester hydrochloride (from Preparation 1) as the amine compound, yielding product required as a white foam.

Rf value: 0.55 (ss-3).

Found: C, 76.11; H, 8.74; N, 4.25. And the presence of C, H and N in percentages of 76.12, 8.77 and 3.77% respectively. EXAMPLE 26 t-Butyl 2S- (Aminomethyl) -3- [1-tet-4-ethoxycarbonyl-cis -3- (3-propyl) cyclohexyl) carbamoyl] cyclopentyl] propanoate The di-ester of example 24 (992 mg, 1.4 mmol) in 20 mL of ethanol was hydrogenated over palladium hydroxide (20%) with charcoal as catalyst at a pressure of 60 psi (4.1 bar). After 24 hours, the suspension was filtered through a short column of &quot; Arbacel &quot; and the solvent was evaporated in vacuo. The residue was dried under azeotropic conditions 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. N in percentages C 27 H 48 N 2 O 6 assumes the presence of C, He of 65.29, 9.74 and 5.64%, respectively. -68-

EXAMPLE 27 t-Butyl 2S- (aminomethyl) -3- [cis - 4-ethoxycarbonyl-cis -3- (2-phenylethyl) cyclohexyl) carbamoyl] cyclopentyl] -4-propanoate Example 25 was hydrogenated as in Example 26 to give the title compound as a gum.

Rf value: 0.35. (ss-1).

Found: C, 69.00; H, 8.80; N, 4.88. C31 H48 N2 O5 H2 requires the presence of C, H and N in percentages of 69.00, 9.19 and 5.19 respectively. Cis-4-ethoxycarbonyl-cis-3- (3-methoxypropyl) cyclohexyl) carbamoyl] cyclopentyl] -N-t-butoxycarbonyl-S-lysyl aminomethyl- t-butyl propanoate 0.34 ml (3.06 mmol) of N-methylmorpholine were added to a stirred, ice-cooled mixture of the product obtained in Example 26 (690 mg, 1.39 mmol-266), N 1-hydroxybenzotriazole hydrochloride (188 mg, 1.39 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (40 mg, 1.39 mmol) 586 mg, 3.06 mmol) in anhydrous methylene chloride (10 mL). After 6 hours, the mixture was treated with methylene chloride (20 ml) and washed with water and dried over magnesium sulfate. After evaporation of the solvent in vacuo and chromatography on silica gel, elution with increasing volumes of ethyl acetate in hexane (50-90%) and finally with 5% ethanol in ethyl acetate gave the product required as a creamy foam (880 mg, 81%).

Value R: 0.2 (ss-3).

Found: C, 58.07; H, 9.06; N, 6.74%. C39 H70 N4 O11 S PressuP ° and the presence of C, H and N in percentages of 58.33; 8.79 and 6.98% respectively. EXAMPLE 29 2S- (Nt -Butyloxycarbonyl-R-alanylaminomethyl) -3- [1-tis-4-ethoxycarbonyl-cis -3- (2-phenylethyl) -cyclohexyl) -catba-moiety] -cyclopentyl] -propanoic acid t- butyl

The procedure of Example 28 was followed, starting from the amine of Example 27 and coupling with N-t-butyloxycarbonyl-R-alanine, to give the title product as a foam. Rf: 0.5 (ss-1).

Found: C, 67.22; H, 8.86; N, 5.66. The presence of C, H and N in percentages of 66.92, 8.78 and 6.00% respectively, were monitored. EXAMPLE 30 2 - [- N - (2-t-Butyloxycarbonylamino-2-methylpropanoyl) aminomethyl] -3 - [(cis-4-ethoxycarbonyl-cis -3- (2-phenylethyl) cyclohexyl) carbamoyl] cyclopentyl] propanoate

Following the process of

Example 28, the amine of Example 27 was used as the starting compound, coupling with 2-t-butyloxycarbonylamino-2-methylpropanoic acid to give the title product as a foam.

Rf value: 0.48 (ss-1).

Found: C, 67.06; H, 8.94; N, 5.81. The presence of C, H and N in percentages of 67.29, 8.89 and 5.89 respectively were observed. EXAMPLE 31 2S- (N '-methanesulfonyl-S-lysylaminomethyl) -3- [1 - [(cis-4-carboxyis-3- (2-phenylethyl) cyclohexyl) carbamoyl] cyclopentyl -propanoic acid 5 ml of trifluoroacetic acid was added to an ice-cold solution of 2S- (N-methanesulfonyl-N-t-butoxycarbonyl-S-lysylaminomethyl) -3- [ (4-ethoxycarbonyl-cis-3- (2-phenylethyl) cyclohexyl) carbamoyl] cyclopentyl] propanoate (obtained in Example 19) (445 mg, 0.53 mmol) in anhydrous methylene chloride ml). The solution was allowed to warm to room temperature and, after 3 hours, the solution was evaporated to dryness in vacuo. The residue was dried under azeotropic conditions (twice with toluene) and dissolved in 1N sodium hydroxide. The solution was stirred at 40-55 ° C over a period of 68 hours, cooled and induced to pass through an ion exchange column of sulfonic acid. After elution with 10% aqueous pyridine, and evaporation of the relevant fractions, a foam was obtained which was decomposed by the addition of acetonitrile and filtered to give a white powder (279 mg, 80%). R f 0.25 (ss-2).

Found: C, 58.66; H, 7.92; N, 8.49. C 32 H 50 N 2 O 2 S requires the presence of C, H and N in percentages of 59.06, 7.74 and 8.61 respectively.

EXAMPLES 32-37

The following compounds were prepared from the corresponding NÎ ± -t-butyloxycarbonyl protected di-ester of examples 18, 20-23 or 28 by treatment with trifluoroacetic acid followed by alkaline hydrolysis according to the procedure adopted in example 31. R12

H 2 N (CH 2) 4 ·

HN H 2 C

H,

C02 H

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-74-

EXAMPLES 38-39

The following compounds were prepared from the di-esters obtained in Examples 29 and 30 by treatment with trifluoroacetic acid, followed by alkaline hydrolysis according to the procedure of example 31.

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η rH «ο Μ λ ε <υχ η η η δ ω η σ η η -76-

EXAMPLE 40 3- (cis-4-Carboxy-cis-3- (2-phenylethyl) cyclohexyl) carbamylcyclopentyl] -2- (2-methoxyethyl) propanoic acid The di-ester of Example 1 (510 mg, 0.86 mmol) in 25 mL of ethanol and 10 mL of water was hydrogenated at room temperature for 3 hours over 10% palladium-on-charcoal (catalyst, 100 mg) at a pressure of 50 psi (3.45 bar). The mixture was filtered through a short column of &quot; Avicel &quot; and the solvent was evaporated under reduced pressure. The mono ester thus formed was dissolved in 1N sodium hydroxide, and the solution was heated to 65øC, in a nitrogen atmosphere, for 2 days. After cooling, the solution was extracted with diethyl ether, the aqueous phase was acidified by the addition of 2N hydrochloric acid and extracted with diethyl ether. The organic phase was washed with water, dried over magnesium sulfate and evaporated under reduced pressure to give the required title compound (di-acid) as a white foam (300 mg, 15%).

Rf: 0.62 C 27 H 39 NO 6 68.47; ss-1). Found: C, 68.17; H, 8.45; N, 2.95. assumes the presence of C, H and N in percentages 8.30 and 2.96 respectively. -77

EXAMPLES 41-44

The following compounds were prepared from the appropriate di-ester of Examples 2 to 5 by hydrogenation, followed by alkaline hydrolysis according to the procedure of Example 40.

R 2

C02H-78-Γ

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EXAMPLE 45

2S- (2-methoxyethoxymethyl) -3 - [(cis-4-carboxy-cis-3- (3-methoxypropyl) cyclohexyl) carbamoyl] cyclopentyl] propenoic acid 5 ml of trifluoroacetic acid were added to an ice-cooled solution of the di-ester of Example 6 (480 mg, 0.77 mmol) in methylene chloride (3 ml). Left standing for a period of 3 hours, the solution was evaporated to dryness in vacuo, and the residue was dried under azeotropic conditions with toluene. The remaining oil was dissolved in diethyl ether, washed with water (9x) and the resulting monoester was extracted into 1N sodium hydroxide solution (2 x 10 ml). The aqueous extract was warmed to 65-70øC in a nitrogen atmosphere for 24 hours, cooled and saturated with sodium chloride solution, acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic extract was washed with water, dried over magnesium sulfate and, after evaporation gave the desired di-acid as a gum (320 mg, 88%).

Found: C, 60.55; H, 8.54; N, 2.80. ^ 24 ^ 41 ^ 8 '^ 2® Assumes the presence of C, H, and N in percentages of 60.20, 8.80 and 2.93 respectively.

EXAMPLE 46 2S- (2-Methoxyethoxymethyl) -3 - [(cis-4-carboxy-cis-3- [3-ethoxypropyl] cyclohexyl) carbamoyl] cyclopentyl] propenoic acid The title product was gum. prepared from the di-ester of example 7, as in Example 45, is obtained as a

Found: C, 61.26; H, 8.66; N, 2.83 C25 H43 NO8.0.3H2 O requires the presence of C, H and N in percentages of 61.15, 8.95 and 2.85, respectively.

Claims (6)

-8- R REIV INDICATIONS: l. A process for the preparation of a compound of the formula: (I) wherein A completes a 4 to 7 ring carbocyclic ring which may be saturated or mono-unsaturated, and which is optionally condensed with another saturated or unsaturated pentene or hexagonal carbocyclic ring; B represents (CH2) m 'wherein m is an integer from 1 to 3, each of R1 and R2, independently of one another, is hydrogen, C1 -C6 -alkyl, benzyl or a biolabile alternative group R 2 represents hydrogen or C 1 -C 4 -alkyl; R 2 is C 1 -C 6 -alkyl substituted by C 1 -C 4 -alkoxy, aryl or aryloxy; C 2 -C 6 -alkenyl, C 2 -C 6 -alkynyl, C 1 -C 6 -cycloalkyl or C 1 -C 4 -cycloalkenyl, or represents C 1 -C 6 -alkyl substituted by halogen, hydroxy, C1 -C6 alkoxy, C1 -C6 alkoxy- (C1 -C6) -alkoxy, C1 -C6 -cycloalkyl, C1 -C4 -cycloalkenyl, aryl, aryloxy, heterocyclyl, R8 Q R 1, R 2, NR 3, COR 2, -NR 3 R 2, -CONR 3, or NR 3 NR 6 -C 11 -C 11 -alkyl; R 2 represents C 1 -C 6 -alkyl substituted by a group of the formula: in which R 1 is hydrogen or C 1 -C 4 -alkyl; that 15? -83- R1 and R2, independently of one another, represent hydrogen, C1 -C4 -alkyl, C1 -C4 -cycloalkyl, aryl, aryl- (C1 -C4) -alkyl, C2 -C4 -alkoxyalkyl or heterocyclyl, or the two groups and R together with the nitrogen atom to which they are attached form a pyrrolidinyl, piperidino, morpholino, piperazinyl or N- (C1 -C4) -alkyl-piperazinyl group; R3 is C1 -C4 -alkyl, CF3, aryl, aryl-) alkyl, aryl- (C1 -C4) -alkoxy, heterocyclyl, C1 -C4 -alkyl. -alkoxy or NR 2 R 3, wherein R 1 and R 2 are as defined above; R10 represents C1 -C4 -alkyl, C1 -C6 -cycloalkyl, aryl or heterocylyl; R3 is hydrogen, C1 -C6 -alkyl, aryl or C1 -C4 -cycloalkyl; R 12 represents R 11 CONR 11, R 11 SO 11 NR 11, R 16 R 17 N - (CH 2) - or 11, R 11 -, wherein each of R has the abovementioned meaning; R1 and R2, independently of one another, represent hydrogen or C1 -C4 -alkyl; or R 2 represents hydrogen and R 2 represents C 1 -C 6 -alkyl, substituted by OH, C 1 -C 4 -alkoxy, SH, SCH 3, NH 2, aryl- (C 1 -C 10 -alkyl-OCONH-, NH 2 CO-, C02H, guanidino, aryl or heterocyclyl, or both groups and together, 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 be substituted by C1-6 alkyl or may be fused to another saturated or unsaturated, 5- or 6-membered carboxylic or cyclic ringyl ring; or R¹ and R² are joined to form a 2- (N-CO-R-4-aminopyrrolidinyl) group; R 2 represents R 2, R 3, R 4, NCO-, R 2 -COO-, R 2 OCH 2 -, or heterocyclyl, wherein R has the abovementioned meaning; R1 and R2 independently of one another represent hydrogen or C1 -C6 -alkyl; and p is zero or an integer from 1 to 6; characterized in that a compound of formula in which R and R are as defined for R and R, with the exception of the meaning of H, or they represent usual protecting groups of carboxylic acids and R is as defined for R being the reactive groups, if desired present optionally protected; X to a hydrolysis and / or hydrogenation and / or other deprotecting reaction to remove any existing protecting group in R 1, and to remove one or both R 2 and R 3, the corresponding dicarboxylic acid of formula (I) wherein R 1 and R 2 represent hydrogen, or to form the corresponding final monoester, wherein one of R 4 and R 4 signifies hydrogen and the other represents a biolabile ester forming group; with optional formation of a pharmaceutically acceptable salt of the final product obtained. 2§. A process according to claim 1, wherein R 1 and R 2 are selected from the group consisting of t-butyl, ethyl and benzyl, and said groups are removed by treatment with trifluoroacetic acid, catalytic hydrogenation or aqueous alkali hydrolysis, respectively , yielding the compound of formula (II), wherein R1 and R2 are hydrogen. 3ã. A process according to claim 1, wherein A represents R &quot; &quot; represents hydrogen and B represents characterized in that if it is a compound of formula (II) -86- wherein R2, R3, R4, R4 and R4 are as defined above. 41. A compound according to claim 1 for preparing a compound of formula I, wherein R represents N-acetyl-S-lysylaminomethyl-, N-methanesulfonyl-S-lysylaminomethyl, N- phenylsulfonyl-2-S-lysylaminomethyl or N-cyclobutylcarbonyl-S-lysylammethyl. 51. A process according to claim 1, wherein a compound 5 in which R is methoxyethyl or 2-methoxyethoxymethyl. 61. A compound according to claim 1, wherein a compound of formula I wherein C represents substituted C 1-6 alkyl is replaced by -NHCOCR RR, where R represents ΝΗ-, R represents 14, Δ methyl and R is hydrogen or methyl. 71. A process according to claim 1, wherein a compound of formula I, wherein R is phenyl, benzyl, phenethyl, methoxypropyl or ethoxypropyl. A process according to claim 1, wherein said compound of formula (I) is: 2S- (N -methanesulfonyl-S-lysylaminomethyl) -3- [cis-4-carboxy-3- cis-3-ethoxypropyl] cyclohexyl) carbamoyl] cyclopentyl} propanoic acid; -87- 2- (N -methanesulfonyl-S-lysylaminomethyl) -3- [1- (cis-4-carboxy-3-cis-phenethyl-cyclohexyl) -carbamoyl] -cyclopentyl] -propanoic acid; 2S- (2-methoxyethoxymethyl) -3- [1-cis-4-carboxy-3-cis-3-ethoxypropyl] cyclohexyl) carbamoyl] cyclopentyl] propanoic acid, methoxypropyl] cyclohexyl) carbamoyl] cyclopentyl] propanoic acid, or a biolabile ester derivative thereof. Lisbon, November 15, 1990 J. PEREIRA DA CRUZ Official Agent for Industrial Property RUA VICTOR CÕRDON, 10-A 3 «1200 LISBOA