CA2067197A1 - 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 R4 is independently H, C1-C6 alkyl, benzyl or an alternative biolabile ester-forming group; R1 is H or C1-C4 alkyl; R2 is C1-C6 alkyl substituted by C1-C4 alkoxy, aryl or aryloxy and R5 is defined to include a range of alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and substituted alkyl groups including in particular methoxyethyl, 2-methoxyethoxymethyl, N2-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

h lj ~ Y ( ~ 91/~7378 ~ pCT/~P90/01~87 ~ ~ .
m is invention relates to a series of cycloalkyl-substituted glutaramide derivatives which are diuretic agents having utilit~
in a variety of theraFeutic areas including the treatment of varicus cardiovascular disorders such as hypertension, heart failure and renal insufficiency.
According to the specification of our European patent applications E$-~-0274234 and EP-A-0343911 we describe and claim .
certain cycloalkyl-substitMted glutaramide derivatives~as diuretic agents. The present invention provides further related cc~poun~s having an extended substituent group in the amlnocycloalXanecarboxylate ring.
The cccpo~r~s are inh~bitors of the zinc~dependent, neutral endopeptidase E.C.3.4.24.11. This enzyme is involved in the bre~kdown of several peptide hormones, including atrial natriuretic factor (ANF), which is secreted by the heart and which has potent vascdilatory, diuretic and natriuretic activity. Thus, the ccn~x~D~ls of the invention, by inhibiting the neutral endopeptidase E.C.3.4.24.11, can pokentiate the biological effects of ANF, and in particular the compcunds are diuretic agents havin~
utility in the treatment of a number of disorders, including hyF~ nsi~n, heart failure, angina, renal insufficiency, premenstru~l s ~ e, cyclical oede~a, Meni~res disease, hypexaldosteronism (primary and secondary) pulmonary oedema, ascites, and hypercalciuria. In a~;tion, because of their ability to potentiate the effects of ANF the compourds have utility in the treatment of glauco~a. As a further result of their ability to inhibit the neutxal endopeptidase E.C.3.4.24.11 - :; ~

W O ~1/07378 ~2 ~ PC~/EP9010~8~/ ~

the compounds of the invention may have activity in cther ~i therapeutic areas including for example th~ treatment o~ asthma~
infla~mation, pain,~epilepsy, affective disorder~, dementia a~d geriatric confusion, obesity and gastrointe~ ~ disorders (especially diarrhoea and irritable kowel ~ e), the mLdNlation of gastric acid secretion and the treatment of hyi~n~ninaemia and leukemia.
The a mpounds of the present invention are of the formulao , A ~ R1 wherein A c~pletes a 4 to 7 m~mbered car~o~clic ring ~ihic~h m~y be saturated or mon~saturated and ~ich may optionally be fused to a further saturated or unsatura~d 5 or 6 membe~d car~o~clic ring;
B is (C~I2)m ~erein m is an integer of fro~n 1 to 3; :
each of R ar~l R4 is independerrtly H, C1~6 alkyl, be~l or an alt~rnative biolabile ester forming g~;

Rl is H or Cl-C4 alkyl;

. ~LC 513 R is C~C6 al}cyl substib~d by C1~4 aL'coxy, aryl or arylo~, or is phenyl;
and R is Cl-C6 a~l, C2~6 a~, 2 C6 al~l, :, C3~7 cycloalkyl, or C7 c~ycloalk~l, or R is Cl-C6 al~l subst ib.~ ~y halo, hy~ro~y, C~{6 al~coxy, Cl-C6 a~Sy(Cl-c6)alkos~ C3 C7 c~loal~l, ~:
C7 cycloal}~l, aryl, aryl~r, heter~clyl, ~6~7, ~R COR, ~R S02R10, {ONR6R or R6R N~ C6)a~r~
or R5 is Cl-C6 a~Syl s~titllt~ by a gr~p of t:he fo:~lao R12 :
_NRl~ R13 -NRllso ~_R13 or NRll~-Rl4 5TlTl)T~ r t).n WO 91/07378 . ~U ~ 719 7 ~ PCI/EP~0/0~18~7 wherein... R6 and R7 are each inde ~ ently H, ~ -C4 alkyl, C
cycloalkyl, aryl, aryl(Cl-C4)alkyl, C2-C6 alk~xyalkyl or heterocyclyl; or the two groups 6 7 together with the nitr~gen to which they are attached to form a pyrroli ~ yl, piperidino, mor~holino, piperazin~l or N~ C4)aIkyl-piperazinyl group;
R8 is H or ~ -C4 alkyl;
~ 4 alkyl, CF3, aryl, aryl(Cl-C4)alkyl~
arylt~ -C4)alkoxy, heteorcyclyl, Cl-C4 alkoxy or NR~7 wherein R6 and R7 are as previously defined;
R10 is Cl-C4 alkyl, C3 ~ cycloalkyl, aryl or heterocyclyl;
Rll is H, ~ -C6 alkyl, aryl or C3-C7 cycloalkyl;
R is RllC~NRll_, RllsO NRll_ R16R17N ( ~ ) . Rl10-, wherein each Rll is as previcusly defined aba~e, R13 and R14 are each indepenlently H or ~-~6 alkyll or R13 is H and R14 is Cl-C6 aIkyl which is sub6tituted ~y OH, Cl-C4 aIkoxy, SH, SCH3, NH2, aryl(Cl-C6)alkyl-OCONH-, ~ CO-, 002H, guanidino, aryl, or hst~ccyclyl;
or the two groups Rl and R1 ars joined together to form, with the car~on at~m to which they are attached, a 5 or 6 m~mbered car~ocyclic ring which may be saturated or mono-unsaturated and which may optionally be :: .~ ~ . . . . , .: . ,. .; .. ; : . . . . . .

~ 0 91/07378 2 ~ ~ 7i~9 ~ PCT/EP9~0~$~7 substituted by ~ -C4 alkyl or fused to a further 5 or 6 nx~ n3d saturated or unsaturated OE b~cyclic rin~;
or R13 is H and R12 and R14 are linked to form a ~- (N{!oRll-4~ ~lidiny~
R15 is R16R17NCo- RllOCO-, Rll ~ - or heterocyclyl7 wherain Rll is as pr~viously defined above;
R16 and R17 are each independently H or Cl-C6 alkylO ~;
and p is 0 or an integer of fm m 1 to 6;
and pharmaceutically a ~ le salts the~eof and bioprecursors therefor.
In the abcve definition, unless otherwise indicated, alkyl groups having three or more carbon atoms may be straight or branched-chain. The term aryl as used herein neans an arcmatic hydrocarbon group such as phenyl, naphthyl or biphenyl which may :
optionally be substituted with one or n~re OH, CN, CF3, Cl-C4 aIkyl, ~ -C4 aIkoxy grcups or halo atcms. Halo means fluoro~ ;
chloro, brom~ or io~o.

WO 91/07378 2 0 6 719 7 PCI/EP90/0~87 me term heterocyclyl 3neans a 5 or 6 n~e~ n~trogen, o~ygen or sulphur containing he~cyclic grwp ~ich, unless c~therwise stated, may b~3 saturated or unsaturatedl and ~ich may c~ptionally incl~}de a further oxygen or one to three nitrogen atans in the r mg an~l which nay aptionally be ~enzofused or s~bsti ~ t~d with for example, one or ~ore halo, ~ -C4 aIkyl, hydroxy, carbamoyl, benzyl, oxo, amino or mono or di~ C4 alkyl)amino ~
(~ -C4 alkanoyl)am mo groups. Particular examples of heterocy~l~s include pyridyl, pyraz myl, pyrimidinyl, pyrida~inyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, f ~ 1, tetrahydrofuranyl, tetrahydropyranyl, dioxanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, indolyl, isoindolinyl, quinolyl, quinoxalinyl, quinazolinyl and ~enzimidazolyl, each being optionally sukstituted as previously defined.
The conpounds of formula (I~ may contain several asymme~ric centres and thus they can exist as enantiomers and diast~recmers~
The inve~tion mcludes bcth mux~ures and the separated indi~.id~al i~omers. The substituents R2, an~ Co2R4 may have cis or trans geometry relative to the amide attachment.
The pharmace~tically aoceptable salts of the crmpiunu`s of formula (I) co ~ an acidic centre are those formed with bases which for~ non-toxic salts. Examples include the alkali metal salts such as the sodium, potassium or calcium salts or .
salts with amures such as diethylamune. ccmpcinds having a basic centre can also form acid addition salts with ~ceutically acceptable acids. Ex~les ~cl~de t~e hy~loride hy~r~r~Lide, sulphate or bisulEihate, E~06phate or hy~rogen phosp~ate, aoetate, citrate, f~rate, gluconat,e, lactake, : ' . .; ' ' . :' : ~.'::: - ' ': .. , .. ,, :.

9~ ~
0 91/07378 ~ PCT/E~90/018$7 maleate, succinate and tartrate salts.
Ihe teLm bioprecursor in the abcve definition means a pharma oe utically acceptable biologi~ally degradable derivative of the cc~pcund 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 prefexred grqup of crmpc~n~s of the for~la (I) are th~e wherein A iS ( ~ )4, Rl lS H and B is ( ~ )2' i.e. CG~pCAndS of the formula (II) below wherein R, R , R and R are as previously defined for formula (I):

CHc~2Q {~

(II) Also preferred are those ccmçcunds of formNlae (I) and (II) wherein R and R4 are both H (diacids) as well as biolabile mono and di-ester derivatives thereof where~n one or b~th of R and R4 .~ is a biolabile ester-forming group.
The term biolabile ester-forming gro~p is well understood Ln the art as n~aning a group which prcvides an ester which can be rea~;ly cleav~d in the body to liberate the corresponding diacid of formula (I) wherein R and R are both H. A number of such . :.: ~ : ....... :: -: . :, ~ ~ , . . , ;

i '; .'7 ~
W O 91tO7378 PCT/EPgO~0~$87 2a~7~97 ester grcups are well knswn, for example in the penic;llin c~rea or in the case of the AC$-inhibitor antihypertensive agentsi.
In the case of the ccmpounds of formulae (I) and (II) i~uch biolabile pro-drug esters are pc~rticularly advantageous m providing comçiunds of the formwla (I) ~uitable f.or oral adminiistration. m e isuiti~bilit~ of any particular ester-for group can be assessed by conventional a ~ or ln vitro enzyme hydrolysis studies. Thus, desirably for optimum effect, the ester i~hauld only be hydrolysed after cibsorption, ac ordingly, the es~er should be r~sistant to hydrolysis before absorption by digesti~e enzymes but should be readily hydrolysed by, for example, li~er enzymes. In this way the active diacid is released into the bloodstream following oral absorpkiQn.
In addition to lower alkyl esters ~partic~larly ethyl) and benzyl esters, suitable biolabile esters include aIkanoyloxyaIX~l esters, including alkyl, cycloalkyl and aryl su~stituted derivatives thereof, aryloxyalkyl esters, arcyloxyaIkyl este~sa arylaLkyloxyalkyl esters, arylesters, araIkylr~sters, and haloalkyl esters wherein said aIkanoyl or alkyl ~ s have frcm 1 to 8 carbon atoms and are branched or straight chain and said aryl groups are phenyl, naphthyl or indanyl optionally s~bstituted with one or more Cl-C4 alkyl or Cl-C4 aIkoxy groups or halo atoms.
Thus examples of R and R4 when ~hey are biolabile ester-forming groups other than ethyl and ben2yl include:
l-t2,2-diethylbutyryloxy)ethyl, 2-ethylpropion~loxymethyl, 1-(2-ethylprcpionyloxy)ethyl, 1-(2,~-dimethylbenzoyloxy)ethyl, 1-(benzoyloxy)benzyl, l-~benzoyloxy)ethyl, 2-methyl-1-propion~loxy-propyl, 2,4,6-trimethylbenzoyloxyme~hyl, 1-(2,4,6-trimethyl-.. . ., , ,: .,, .... , . .: , .

~yVO 91/07378 PCT/EP90/018$7 . I- ;. t .`

kenzyloxy)ethyl, pivaloyloxymethyl, phenethyl, phenpropyl, 2,2,2-trifluoroethyl, 1- or 2-naphthyl, 2,4-dimethylphenyl, 4-t-butyl phenyl, [5-(4-methyl-1,3-dioxolen-2 onyl)]methyl and 5-indanyl.
Ccmpounds of the formulae (I) and (II) wherein R is benzyl or t-butyl and R4 is ethyl are valuable interm2diates for the prepara~ion of the diacids wherein R and R are ~oth H.
In a further preferred gxcup of compoonds R5 is methylen~
substituted by a gxoup of the formula -NHCocRl2Rl3Rl4~
parti~llarly where R12 is NH2, RllCONH- or RllSO NH- R13 is H and R14 is -(CH2)4NH2. Parti~11arly preferred are such groups deri~ed from S-lysine; thus especially preferred R5 sub6titutents of this type include N2-a oe tyl-S-lysylaminomethyl, * -~ethanesulphon~l-S-lysyl-amlnomethyl, N2-phenylsulphonyl-S-lysyl-aminomethyl an~
N2 ~ clcibut~1car~onyl-S-lysyl-aminanethyl.
In further groups of preferred oaTpc~ds R5 is Cl-C6 alkyl s~bstituted by Cl-C6 alkoxy, particularly methoxyethyl; or R5 is -C6 alkyl substituted by Cl-C6 alkoxy~ -C6)alk~xy, particula~ "7 2-methyoxyethoxymethyl; or wherein R5 is ~ -C6 alkyl substitu~ed by -NHaocRl2Rl3Rl4 wherein R12 is NH2, R13 is CH3 and R14 is H or CH3 including in particul æ R-alanyl-aminomethyl and (2-amino-2-methylpropanoyl)amuncmethyl.
R2 is preferably phenyl, kenzyl, phenethyl, methoxypropyl or ethoxypropyl.
Particularly preferred inlividL~. ccLpcunds of the inve~ion include:
2S~ methanesul p onyl-S-lysylamuncmethyl)-3-~1-[cis-4-carboxy-3-cis~3-ethoxypropy1)-cyclohexyl)carbamcyl]cyclopentyl~-propanoic acid, :

- . , , . , ~ , . ;

_. . .~1 W O 91/07378 PCT/EP90/018$7 ~
- 20~7~
2-(N2-methanesulphonyl-S-lysylaminomethyl)-3~ [(cis-4-carboxy-3-cis-ph~ethyl-cyclbhexyl)carbamoyl]cyclopentyl~pr3panoic acid, 2S-(2 methoxyethoxymethyl)-3-~1-[(cis-4-car~oxy-3-cis-~3-ethoxypropyl~cyclahexyl)carbamoyl]cyclqpentyl~propanoic acid and 2S-(2-~ethoxyethoxymethyl)-3-~ (cis-4-carboxy-3-cls-~3-methoxyprcpyl~-cyclchexyl~carbamoyl]cyclopentyl}propanoic acid, and biolabile ester derivatives thereof.
m e co~poun~s of formula (I) are p ~ by a number of dif~erent p ~ s. 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. me carboxylic acid group in the amine, if free, or any reactive groups in R5, may require protection during the ccupling step and such prokecting grcups are removed in the final stage of the prooess.
The synthetic route is illustrated in scheme 1 where m A~ B, Rl and R2 are as previausly defined, R5 is as definQd for R5 with r any r~eactive group therein prokected i~ n~cessary and RlB and R
are as defined for R and R4 excluding H, or they are conventional carboxylic acid protectLng groups:

~/001/~17378 . ~ CI/EP91)/01887 Scheme _ R5 l ( C~ R2 CIICR2 / \ CO H I El 11 {~

R O2C (III) (IV Co2Rlg R ~ 3~ 2 \ C~CR / \CON~ R

R18O C (V) C0 R~ 9 J, (I) :.

The reacticn of the compcunds of formula (III) and (IV) is achieved us m g oonventional amide ccupling techniques. Th~s in ~.
one process the reaction is achieved wnth the reactants dissol~ed in an organic solvent, e.g. dichlorsmethane, using a diimide oondensing agent, for example l-ethyl-3-(dimethyl ~ rcpyl)- ;
carbodiimide, or N,N'-dicyclohexylcarbrdiimide, advantageously in -the presence of l-hydroxykenzotriazole and an organic base suc~ as ~ .
N-methylmDrphol m e. The reaction is generally complete a~ter a period of frcm 12 to 24 hours at rocm temperature and the product is then isolated by conventional procedures, i.e. ~y washing with water or filtration to rem we the urea biprcduct and evaporation of ~he sol~ent. ~he prodNct may be furth2r purified by ~' .

W O 91/07378 PCT/EP90/OD887 ~ ~
20~7~97 ~ i-'12 1 crystallisation or chromatography, if neoessary. The compcunds of formLla (V) include cn}pcunl s of formula (I) wherein R and R4 are -C6 alkyl or benzyl.
In some cases the ocupled product, in prot~cted form, may ke subjectel to conventional chemical txansformation reactions to allow preparation of ~urther ccmpounds of formula ~V). Ihus for example co-pourds of formula (V~ wherein R5 conta ms an estRr ! ::
may be hydrolysed or hy*rogenated to generate the carboxylic acid which may be further reac~ed, for example with an amine; to give amide derivatives.
Simil æ ly oo pc~nds whereLn R5 oontains a ~ubstituted or ~ :
protected amino gxoup (for example a ben2ylam mo, dibenzylamino, benzyloxycarbonylamino or t-butyloxycarbonylamLno group) may be oonverted.to the free amLnes by hydrogenation or protonolysis as appropriate. Ihe amines produc~d may be further reacted, thus or example reaction with a sulphonyl halide yields the correspondi~
sulphonamides, acylation with an acid chloride or anhydride yield~s the corresponding amides, reaction with an isocyan~te yields urea derivatives and reaction with a chloroformate yields the aIkoxy-carbonylamino and products respectively. A11 these transformations are entirely conventional and appropriate conditions and reagents for their performanoe will be well known to those skilled in the art as will cther variations and possibilities.
m e diesters of formNla (V) may be further react~ to give .~the monoester of diacid deriva~ives of formula (I) wherein one or bokh of R and R are H. The conditions used will depend on ~he precise nature of the groups Rl and R present in the compound - , : . . ,. . :

YVO 91107378 2 0 6 7 ~ 9 7 PCT/EP9~ $8'7 of formula ~V) and a number of variations are possible. Thus for example when both of R18 and Rl9 are benzyl, hyc~nogenation of ~he produc~ will yield the ~;~cid of formula (I) wheJnein R and ~ ar~
both H. Alternatively if one of A 8 and Rl9 is benzyl and the other is aIkyl, hyc~nogenation will yield a LosKr:ster produc~O
qhis can be hydrolysed, i desired, again t~ yield ti~e diacid product. When c~e of R18 an~ R19 is t-b ~ l treatmen~ of th campcun~ of formula (V) with trifluoroa oetic acid yields the corresponding acid. The diester product wherein R18 and Rl9 are benzyl or lcx~r alkyl c~n also be treated witih trimethylsilyl iodide to prc~duce the dicarboxylic acid product. If some other carboxylic acid protecting grcup is used for R18 or Rl9 then clearly appropriate conlditions for its rem~val m~st be emplc~ed in the final step to give the ester or diacid product of form~la ~I). In the case where the ring A or the substituent R5 is unsaturated, the deprotection m~st be effected by non-reducti~e methods, thus for example lf ei~her of R and R4 is benzyl, ~he~
may be removed by treatment with trimethylsilyl iodide.
As well as remaving any protecting grcup which may be presen~
in R5 , a numker of chemical transformation reactions are po6sible 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 an~ isolated in ~A1t form.

W O 91/07378 2 0 6 7 i 9 r~ PCT/EP90/01887 ~ 1 In a varian~ of the above procedure, compounds of the formula (I) where m R5 is ~ ~C6 aIkyl substituted by -NRBCOR9, NR3So2RlO, NRllcoCR12Rl3Rl4 or _NRllso2cRl2Rl3R are prepared by a process which involves acylating or sulphonylating a co~pcund of the formula:

\ CHCH C \ CON~ ~ RC02R19 (VI) wh~rein R20 lS as defined for R8 or ~ll, Rl8 and R19 are as previcusly defined and Y is a Cl-C6 aLkyl grcup; by reaction with ~:
an acid of the formula R9Co2H, RlOSO3H, R12Rl3Rl4Coo2H, or Rl2Rl3Rl4Cso3H, or an activated derivative thexeof. ~he ~esultLng amide or sulphonamide product i5 then deprctected if required and the diester product cleaved to yield the carboxylic acids of formNla ~I) wherein R and R4 are each H as previously described.
The co~çcLnds of formula (VI) are prepared following the procedures shown in scheme l but using a compound of formula (III) ha~in~ R5 as a protected amine derivative. Thus, for example R5 can contain a bis-(lS)-phenylethylam mo su~stituent.
Hydrogenation of the ccupled product gives the corresponding free amine of formula ~VI) wherein * is H and Y is C~2. This route is of particular value for the preparation of compor nds having 2(S) stereochemistry in the glutarzmide bacXbone.

.
, . . . ~ , , .............................. , . . ,~
: ; . ,, ., , : , O 9l/07378 2 0 ~ ` PC~/EP90/6D~7 i, ,., " ~ '~ i -:

The starting cycloalkyl-substituted glutaric acid m~no esters of formula III may be prepared as described in cur ~ n paten~
applications EP-A-0274234, 89305180.5 and 89304698.7.
m e amunes of formula (IV~ are generally novel ccrpcunds (particularly whien the substituents have defined st~reochemistry) and they are preparel by a w rcpriate synthetic p~oc.dures in accordance with lit~rature preo;derts. Thus in one procedure ~he correspon~ing ketone is converted to the amine via re~uction of the oxime. In an alternative process ~he corresponding alcohol is reacted with methyl toluenesulphonate in the presenoe of diethyl diazodicarboxyla~e and triphenylphosphine; the res~ting 4-methylbenzenesulphonyloxy derivative is reacte~ wlth sodium azide and the pro~uct reduKed,to yield the amine. These .
prooedurrs are illustrated in the Preparations give hereafterO i:
Apprapriate coupling and protectln~ methods for all of the above steps and alternative variations and prooedur~s will be known to those skilled in the iart by reference to standard kooks and to the examples provided herea*ter.

.. . . .

W O 91/07378 ~ I PCT/EP90/0~887 As previcusly mentioned, the conpoun~s of the invention are potent inhibitors of the neutLal endopeptidase (E~C~3~4~24~11) This enzyme is involv~d in the breakdcwn of a rnnDber of pepkide hormones and, in particular it is involved in ~ breakdown of atrial natriuretic factor (ANF). This hormone a~nsists of a family of related natriuretic pepkides, secreted by the heartO of which the major ci ~ ating form in humans is kncwn to be the 28 amino,acid peptide re~erred to as~-hhNP. Thus, by preventing the de~raclation of ANF, by endc~epkidase E.C.3.4.24.11, the oompourds of the m vention can potentiate its biological effects and the oo pounds are thus diuretic and natriuret.ic agents of utility in a number of disorders as previously describ~d.
ALtivity against neutral endcpeptidase E.C.3.4.24.11 is assessed using a procedlre based on the assay described by JoTo Gafford, R~A. Skidgel, E.G. Erdos and L.B. Hersh, Biochemistry, 1983, 32, 3265-3271. The method involves determining the concentration of compcund ~ to reduce by 50~ the ra~e of relea æ of radiolabelled hippuric acid from hippuryl-L, phen~lalanyl-Lrarginine by a neutral endopeptidase preparation ~:
fr~m rat kidney.
m e activity of the corpcunds as diuretic agents is determined by measurLng their ability to increase urine output and sodium ion excretion Ln saline loaded conscicus mi oe. In this test, male mi oe (Charles River CDl, 22-28 g) are acclima~ised and .: : ,. . , , . ., ~ . :

. . - . - . . : .

starved overm~ght m metabcwls. The mioe are dosed intravencusly via tlle tail vein, with the test ~ d dissolve1 in a volume of saline solution equivalent to 2.5~ of body we:ight. Urine samples are coll~cted each hvur for two hcurs in pre-w~ighed tukes and analysed for electrolyte concentration. Urine volume and sodium ion concentration from the test animals ara c3mpared to a con~rol grcup which received only saline.
For admQnistration to man in t~e curative or prcph~lactic treatment of hypertension, congestive heart failure or renal insufficiency, oral dosages of the cc~pcun~s will genRrally b~ In the range of fr~m 4-800 mg ~;ly for an average adult patient ~70 kg). Thus for a typical adult patient, individual tablets or capsules contain frc~m 2 to 400 mg of active ccmpcund, in a suitable pharmaoeutically acceptable vehicle or carrier for administration singly, or in mwltiple doses, once or several tL~5 a day. Dosages for intravenous admDnistration would typicall~ ~æ
within the range 1 to 400 mg per sLngle dose as required. In practioe the physician will determine the actual dosage which will be m~st suitable for an individual patient and it will vary wi~h the age, weight and response of the particular patient. The above dcsages are exemplary of the average case kut there can, of ccurse, be indivîdual inst30ces where higher or lower dosage ranges are merited, and such are within the scope of this invention.
' 5U135TITl,3TE: SH ~:T

.

W O 91/07378 ~ ~ ~ 719 ~ PCTtEP90/01~7 For h moan use, the ccmpcunds of the formlla (I) can be adminis~ered alone, but will generally be administered in -~
admixture with a pharma oe utical carrier selected with regard t~
the intended rcu~e of admdnistration and standard p~armaceutical practi oe. For example, they may be administ~red orally in the form of tablets containing such excipients as starch or lactc~e~
or in capsules or oNules either alone or in admixture with excipients, or in the form of elixirs or suspensions cor~air~rg flavauring or colouring agents. m ey may ~e injected parenterally, for e ~ le, intravencusly, in¢ramuscwlarly or subcutaneously. For parenelral administration, they are best used in the form of a sterile aque~us so~ution which may contain okher substances, for example, enough salts or glucose to make the solution isctonic with blood.
The c~cFa nds may be administered alone but may also be ad=unistmred together with such other agen~s as the physician . shall direct ~o opkimuse control of blood pressure or to trea~ ~
congestive heart failure, renal insufficiency or other disorde~s in an~ particular patient in aocordance with established n~dical practice. Thus the ccmpcunds can be co-admlnistered with a variety of cardiovascular agents, for example with an A OE
inhibitor such as captcpril or enalapril to fac;litate the oontrol of blood pressure in treatme~t of hypertension; or with digitalis, or ancther cardiac stimulant or with an ACE inhibitor, for the treatment of congestive heart failure. Other possibilities include co-administ~ation with a calcium antagsnist (e.g.

~ O 9t/07378 2 ~ 6 7 19 ~ PCI/EP9~/~l887 nif~;pine, amlodcpLne or diltiazem) a bet~-blocker (e.g.
atenolol) or an alpha-blocker (e.g, prazosin or doxazosin) as shall be determined by the physician as appropriate for the treatment of the particular patient or conditiaII involved.
Xn addi~ion to the abcve, the crmpcon~s may al50 be admilisdlar~t Ln conjunction with exogencus ANF, or a deri~ati~e there~f or related peptide or pep~ide fragment having diuretic/natriuretic activity or with okher ANF-gene related peptides (e.g, as described by D. L. Vesely et al, BiochemO
Biophys. Res. Comm., 1987, 143, 186).
~ hu~s in a further aspect the inven~ion pr~vides a pbarmaceutical composition co~prising a oompcund of the formula (I), or a ph2rma oeutically acceEtable salt thereof or bioprecl~or , ' therefor, tcgether w,ith a pharmaceutically acceptable ~;luent or carrier.
The invention also includes a compcunl of ~he formula (I~ 9 or a pharmQ oe utically ac oeptable salt thereof or bicprecursor ' therefor, for use in nYxlicine, particularly for use as a diure~ic agent for the treatment of hypertension, congestive heart failure or renal insufficienc.y in a human being.
The invention further includes the u~se of a ocmpound of the formula (I) for the manufacture of a m~dicament for the treatment of hypertension, heart failure, ang ma, renal insufficiency, prrm~qstrual syndrome, cyclical oedema, Menieres disease, hyperaldosteronism, pulmonary oedema, a~cites, hypercaciuria, glaucoma, asthma, infl a tion, pain, epilepsy, affective disorders, dementia and geriatric oonfusion, obesity, W O 91/0737R 2 ~ 6 719 7 PCT/EP90/01887 .
gastrointestinal disorders (including diarrhoea), hyperrenunaemia, leukemia, and the mcdulation of ~astric acid secretion.
Ihe preparation of the cn poonds of the ir~ention will ncw be more particularly ill~skrated by xeference to the followm ~ I
experimental examples, in which Preparations l--8 describe , .
preparation of the starting amQnes of formula (IV) and Examples 1-45 desGribe pr ~ tions of ccmpourl s of the formula (I). The purity of ccmpcunls was rcut mely monitoxed by thin layer chromatography using Merck Kieselgel 60 F254 plates. Solvent syste~ ss-l was a mixtlre of dichloro~ethane, methanol an~ glacial acetic acid (90:10:1), ss-2 was the upper phase of a mixture consisting of methyl iso-butyl ketone, glacial a oetic acid and water (2:1:1), and ss-3 was a nix*ure of ethyl a oetate and hexane (1:1). ~ -NUC1 OE magnetic reasccanDs spectra were recorded using a Nicolet QE-300 spect~c=eter and were in all cases oons~sten~
with the proposed structures.

O 91/07378 2 0 6 7~ 9 ~ PCr/EP90/01887 PRE~AR~ON 1 '' .

c-4-Aminorc-2-(2-phenvlethy~is~_ohexane-r-1-carboxYlic acid ethYl ester h~dLcchlor e i. 4-Oxo-2-f2-phenvle~hvl)cvclchex-2-ene carboxylic acid ethvl An 80% dispersion of sodium hydride in oil (307 mg, 10 le) was added under nitrogen to an ice cold stirred mixture of 3-oxo-5-phenyl pentanoic acid ethyl ester (28.4 g , 129 mmole) and absolu~e ethanol (1 ml). Methyl vinyl ketol~e (10.7 ml, 129 le) was then added drcpwise over half an hcur with continued ioe cooling and, after stirring for an hour at room temperature, acetic acid tl.4 ml), pyrrDlidine (775 mg), ethanol(l7 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 hy ~ oric acid, water, saturated aqueous sodium bicarbonate and water, drying over MgS0 and evaporation gave a yellow liquid (34.6 g). ~ atography on silica gel (600 g) elutLng with a mixture of diethyl ether and hexane (1:~ 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- C17 ~ 003 requires C,74.97; H,7.40%.

.

WO 91/07378 2 0 6 7 1 9 ~ PCI/EP90/01887 22 ' ii. 4 oxO-cis-2_(2-Phe~ylethvl)cyclcalexan~ar~xvlic acid ethvl ester The enone (18.44 g, 6.76 Dle) fr~n part l~i) in ethanol (80 ml) corrtain~ng 2N hydro~loric acid (3 ml) was hy~mgena~ed ov~ 5% palladi~n on ~arco~ catalyst ~500 m~) at 50 p.s.i. (3045 bar) pressure. ~ ~ree ha~r~; a furthex ama~t of catalyst (250 m~ was added and reduction was conti~ Eor a further two ho~ s. I~e s ~ ion was filtered thra~gh Avicel and the filtrate evaPorated under vacuum. The residual oil was dissol~ed in diethyl ether, washed with saturated aqueous sodium bicartDrate and water, dried over MgS04 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 ais a liquid. (16.78 g,90~). Rf 0.4 (1:1 diethyl ether:hexane~. ;
iii . 4-Methoxyi~nir~is-2- (2-phersYlethvll cYclollexane ca~oxvlic acid ethvl ester The above ketone (3.02 g, 11 mmole), methoxylamine h~vdrochloride (1.19 g, 14.3 mmole) and solium a oe tate (1.17 g;
14.3 mmole) were refluxed for five hours in ethanol (50 ml). the solvent was evaporated under vacuNm, the resi~ue taken up m diethyl ether and washed with saturated aqueous sodium bicarbonate follow3d by water. ~rying over MgS04 and evaporation under vacuNm gave the O-mlethyl oxime as an oil (3.32 g, 99%) Rf 0.55 and 0066 (1:1 diethyl ether, hexane). Found: C,71.23; H,8033; N,4.88.
C18~ 5N03 reguirrs C,71.26; H,8.30; N,4.62%.

. , . -: . . .. .

. : .... . . :

`: ;
~09~/07378 2~719~ 1 Pcr/lEP90/~

iv. ~4-Butox~car~or~la~2-t2~herr$~1ethYl)~lQhe~
r~ oxvlic acid ethyl ester Irifluroaoetic acid (4.2 ml, 54 le) in clry tetrahyd:rofuran tlO ~) was added dr~i~se with ioe cooling to a st~red su~sion of sodimn bo~ide (2.04 g, 54 ~le) in dry te:tr ~y~rofl~ran. Ihe te ~ ture was l ~ t ketween 5 and 10C a~l after stirring for fifteen minLtes the 0-methyloxIme from Fart (iii) (3.28 g; 10.8 mmole) m tetrahydr~furan (10 ml) wc~s added dropwise over ten ninLtes. The temperature rose to 20C c~nd, after stirring for five hours, water was very c ~ efully added with ice cooling and the mixture was partitioned between e~hyl a oe tate :
and water. m e aqueous layer was extracted twi oe with ethyl a oetate, the ccmbined organic extrac~s washed with saturated ~lt solution, dried oYer MgS04 and evaporated to give a gum (3~25 g)~ ~:
Ihe crude c~mine ~as 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) w~re added and the mixbure allowed to stand a~
room temperat~re for 48 hcurs. ~he solve~t was evaporated under vacuum, the residue partitioned between diethyl ether and water a~d the organic extract washed sequentially with 0.5N hydrochloric acid, water, saturated aqueous sodium bicarbonate and water.
Drying over M~S04 and evaporation gave a gum (3.9 g). The required cis- oompcund was separated frc~ the munor more polar trans-isomer by chrcmatography on silica gel (400 g). Elution with a n~ rre of diPthyl ether in hexane (3:7 by volume) gave the protected amune as an oil (1.94 g, 48%). Rf 0.35 (3:7 diethyl ether:hexane). Fcund: C,70.43; Hr8.75; N,3.45. C22H33N34 requires C,70.37; H,8.86; N,3.73%.

, : :,: : . :. ; .

W O 91/07378 ~ PCTiEP90/01887 v. c-4-Aminorc-2-(2-PhenYlethvl)cvclQhexane-r-l-carboxylic acid ethYl ester hYdrochloride An i oe cold stirre1 solution of the ~bove ester ~1.94 g, 5.17 le) in diethyl ether was saturated with :hydrogen chloride.
After three hcurs the solvent was blo~ off with nitrogen and the residue triturated with diethyl ether and oollectel by filtration giving the ~itle amune ~lt as a white solid (1.44 g, 89%). m.p.
213-214C. Rf 0.5 (ss-l). FQUnd; C,65.84; H,8.74; N,4.43.
C17H25NO2.HCl requires C,65.48; H,8.40; N,4.49%.

PREPAR~IqON 2 c-4- ~ 2-~ hoxvprooYl)c~clohexane-r-l-carboxvlic acid ethYl ester hYdrochloride Ihe procedure of Preparation 1 was follcwed but using 3-oxo-6-methoxy-hexanoic acid ethyl ester as the starting material in part ~i) to give the title amune as white solid, m.p.
206-208 &. Rf 0.2 (ss-l). Fw nd: C,55.15; H,9.11; N,5.34.
C13H25N03. HCl, 0.2 ~ 0 requires C,55.09; H,9.39; N,5.94%.

~ 0 91/07378 2 ~ ~ 7 1 ~ 7 i ij, !;' PCT/EP90/01887 " ~
.. . ., ., ~, .

~K~ARAIION 3 c-4-Aminotc-2-~3-ethoxyproPyl)cvclchex~n~-r-l-carboxylic acid ethyl ester h~kcchloride Ihe procedure of ~ ation 1 was followed but us mg 3-oxo-6-ethoxyhexanoic acid ethyl ester as the i ~ material in part (i) to give the title amine as a whi~e solid, m.p.
201-203 &. Rf 0.2 (ss-l). Found: C,55.50; H,9.52; N,4.33.
.HCl.O.5H20 requires C,55.52; H,~.65; N,4.63~.

~KL~AR~IION 4 c-4-Aminotc-2-~2-phenylethvl)cyclohexan~-r-1-carboxylic acid methYl ester hYdrochloride A solution of the ethyl ester from Preparation 1 (1.4 g;

4.48 mm~le) in dry methanol (50 ml) was isat~rated with hy~rogen chloride and heated at 55 to 60 & for four days and ~hen refluxed for a further 3 days. Hydrogen chloride was periodically in~roduced to maintain saturation. The solution was evaporated to dryness under vac~um, the residue dried azectr3pically with methylene chloride, and the residue triturated with diethyl ether to give the methyl ~ster as a white powder (1.29 g, 97%) m.p.
202-4C. ~f 0.15 (ss-l). Found: C,63.70; H,8.13; N,4.74.
C16~ 3N02. HClØ25 H20 requixes C,63.56; H,8.17; N,4.63%.

.: .. ,.:, - :.

:, . , . , : .. :,. ., : : : .:

W O 91/07378 !,, ' ', `~ PCT/EPgO/3~87 ~
2~7~9~ - .

c-4-Amino-t-2-(2-Phenyl~thYl)cyclohexane-r-l-car~ox~lic acid ethyl ester i. cis-7-(2- ~ enylethyl~-1,4-dioxasl?~ror4,51de 8-carboxvlic acid ethvl ester m e 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 wat~r trap. After eight hours the solution was oooled, diluted with diethyl ether and washed sequentially with saturated aquecus sodium bicarbonate and water. DryLng over M4S04 an~ evaporation un~er vacuum gave a clear liquid (12.22 g, 96%) Rf 0.32 (3:7, diethyl ether, hexane). Found: C,71.45; H,8.30. ~ 9~ 64 requires C,71.67; H,8.23%.

. , ~ , . . . :

~ 91/07378 2 0 6 ? 1j~7 7~ ~ PCT/EP90/01887 ii. trans-7-(2-Phen~lethvl)-1,4_dioxas~Lror4,51decane-8 carboxylic acid ethYl ester Pokassium t-butoxide (1.9 g, 17 mmole) was added to a solution of the above ester (12.17g, 38.2 mmole) in dry t-butanol (90 ml~ and the ~ixture refluxed under nitrogen for forty eight hours. Neu~ralisation with 2N ~ydrochloric acid and evaporation un~er vacuum r~ave a light brcwn o;l which was dissDlved in diethyl ether and washed wqth water. Drying over MgS04 and evaporation gave an oil (12.09 g) which was chromatograFhed on s;lica gel (600 g)~ Elutions with a nixture 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 die~hyl ether, hexane). Found: C,71.60; H,8.14.

9H264 requires C,71.67; H,8.23%.

iii. 4-Oxo-trans-2-~ -Phenvlethyl)cvclohex necarbox~ic acid ethYl ,~er ~' ' The above ester (8.40 g, 26.4 D le) was refluxed in IN
sulphuric acid (S0 ml) amd èthanol (70 ml). After three hours most of the ethanol was evaporated off under vacuum, water was added and the suspension extracted with diethyl ether. The organic extract was washed sequentially with water, satNrated aqueous sodium bicarbonate and water, dried over MgS04 and evaporated under vacuum to give a clear oil (6~46 gt 89%) Rf 0.38 (1:1 diethyl ether, hexane). Found: C,74.25; H,8.07. C1 ~ 203 requires C,74.42; H,8.08%.

: :., . , ,.. , , : : .- : . .

W O 9l/07378 2~6~i~7 PCr/EP90/0l887 iv. t-Z-f2-PhenvlethVll-c-4-Phenylmethvlamino-cvclohexane-r=
l-carboxvlic acid ethvl 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 benzylamune ~1.27 ml, 11.66 mmole) in ethanol (15 ml). After three and a half hcurs at room ~ ature, the solutiQn was pipetted of~ into a dry hydrogenation ves ~ washing the si~ves with a little ethanol (2xS ~1). Platinum oxide (500 mg) was added and the reduction completed in three hours at 50 p.s.i. (3.45 bar). The mixbure was filtered t ~ arbacel, evaporated under vacuum and the residual liquid chromatographed on silica gel (200 g). Elution with increasing proportions of ethyl aoetate in hexane (7:3 - 1:1 by volume) gave the requlred CiS-LSomer as a cl OE oil (3.17 g, 74%) Rf 0.3 (ss-3) Found: C,78.85; H,8.53; N,4.86. C24H31N02 requi~es .
C,78.86; H,8.55; N,3.83%.
v. c-4-Aminc-t-2-L2-~henY--thvl~cvclohexane-r-l-carboxy-ic acid ethvl ester A solution of the above amune ~3.13 g, 8.56 mmole) in ethanol (80ml) containing IN hydrcchloric acid (9.4~ ml, 9.42 mmole) was hydrogenated over 20% palladium hydroxide and char~cal catalyst (600 mg) at 50 p.s.i. (3.45 ~ar) pressure. Further amcunts of catalyst (500 mg) were added after six and twenty fcur hcurs.
After a further twenty two hcurs the suspension was filtered through arbacel and the filtrate evaporated to dryness urder ~ yVO 91/07378 2 0 ~ 7 ~ 9 7 PCT/EP90/0~$87 vacuum. The residue was re~rystdllised frcm a ILLXture of ethanol and diethyl ether to give the required amlne as a white solid (2.03 y, 76%) m.p. 175-7 &. Rf 0.25 (ss-l). ~Dnd: C,64.42;
H,8.50; N,4.52. ~ 7 ~ ~ 2.HC1Ø2 ~ 0 requires C,64.54; H,8.44;
N,4.43%.

c-4-~ninY-c-2-phenylcyclohexane-r-1-carboxylic acid methyl ester hydr~chloride i. 4-Oxo-2-phenylcy~lQ~ex-2-ene-cartDxylic acid methyl ~
2-H ~ y-4-oxo-2- ~ lcyclohexane carboxylic acid methyl ester (17.5 g, 70.48 mmole) [J. Org. Chem., 39 2427 (1974)~ was stirred for 15 minutes at 80 & with polyphosphoric acid (17.5 g).
on oooling the mixture was partitioned between ethyl a oe tate and water. The organic ex~ract was washed sequentially with water, saturated aqueous sodium bicarbonate and water, dried over ~SO4 and on evaporation gave an oil. Chromatogra~hy on silica gel eluting wi~h increasing proportions of ethyl a oe tate in hexane t2:8 - 3:7 by volume) ga~e the reguir0d compcund as a golden oil (9.0 g, 56%) Rf 0.5 (55-3).

ii. 4-Q~r~ a~y~cyclohexanvcarboxylic acid ~ether ester The enone o~ part 1 (9.0 g, 39.08 mmole) in methanol (40 ml) contai m ng 2N hydrochloric acid (1.5 ml) was hydrogenated over 5%
palladium on ch2rcoal ca~alyst at 50 p.s.i. (3.45 bar). After three ho~rs the suspension was filtered through Arbacel and evapora~ed to dryness under vacuum. The residual oll was chromatographed on silica gel (400 g). Elution wlth increasing proportions of ethyl acetate in hexane !2:8 - 4:6 by volum~) gave W O 9l/07378 ` ` )~' PCT/EP90/0l$8/
2~71~7 the required ketone as a white solid (4,04 g, 49~) Rf 0.55 (ss-3) Found: C,72.39; H,6.95. C14H1603 requires C,72.39; H,6.94%.
lii. c-4-HydroxY-c-2-PhemlcYclchexane-r-l-car~Dxylic acid methy ester ~ odium borohydride (2.33 g, 61.51 mmDle) was added in small portions ~D a stirred solution of the kebDne of part ii (3.78 g, 16.27 ~m~le) in methanol (80 ml), keeping the temperature below 5 &. After fifteen minNtes the sDlven~ was eYaporated under vacuum, the residue dissolved in methylene chloride and washed with saturated ~lt sDlution. Drying over MgS04 and evaporation under vac~um gave a white solid. Chromatsgraphy on silica gel (250 g) eluting with diethyl ether in hexane ~ y volume) gave the required ester as a foam (2.93 g, 77%). Fcund: C,71.84;

14 ~ ~3 reqYire5 C,71.77; H,7.74%.

iv. t-4-t4-MethYlbenzenesul~honYloxv~-c-2-phenylcyclbhexane-r~l carboxvlic acid methvl ester : Diethyl diazcdicarboxylate (4.08 g, 23.47 mmole) in tetrahydrofuran (10 ml) was added under ni ~ en to a sti 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 ~ ature ketween 5 and 10C. After StirrLllg ~vernight at r~m temperature ~he mixture was re-oooled to 0C and further amGunts of triphenylphosphine (1.23 g, 4.7 le) and methyl 4-toluene sulphonate (874 mg, 4.7 mm~le~ were added followed by diethyl diazodicaLboxylate (816 mg, 4.7 mmole). After stirrLng at room temçerature for a further four hours, the mixture ~71 ~ ~
~VO 91/07378 ~ ~ , PCT/EP90/0~8$'7 ~ ' '':. :

was evaporated to a small volume, the residue redissolved Ln methylene chloride and applied directly to a silica gel (400 g) chr~mato~raphy column. Elution with incre~sing 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 eth~l acetate, hexane) Fcund: ~,64.71, H,6.260 C21~ 405S
C,64.93; H,6.23%.

v. c-4-Azido-c-2-~henYlcvclQhexane-r-l-carboxylic acid ~eth~
ester Sodium azide (1.16 g, 17.76 mmole) was added to a stirred solution o~ the 4-toluenesulphonate fro~ part iv (3.45 g, 8.88 mmole) in dimethylformamide, and the mixture was stirr3d at 60C
for eighteen hcurs. On cooling water was added an~ the suspension extracted with diethyl ether. Ihe organic extract was washed with water, dri~d aYer MgSO4 and evaporation gave a clear oil which was chromatographed on silica gel (300 g). Elution with increas m g prcportions of ethyl a oetate in hexane (1:20 - 1:4 by volume) gave the azide as wzxy solid (2.11 g, 92%). Rf 0.55 (4:1 hexane, ethyl aoetate). Fcund: C,65.20; H, 6.60; N,15.32. C14H17N302 requires C, 64.85; H,6.61; N,16.20%.

W O 91/07378 2 0 6 719 7; PC~/EP9~/01887 vi. c-4-AminKrc-2~phenv1cvclchexane-r-lecarbo~ylic acid methy ester hvdrcchloride qhe azide frcm part v (2.1 g, 8.1 m ~mnle) Ln ~ethanol (50 ~1) was hydrcgenated over 10% palladium on char~al catalyst (210 mg) at 50 p.s.i. (3.45 bar). After two and a quaxter hours the mixture was filtered through a shDrt arbacel column and evaporated under vacuum. The residue was dissolved in ethyl aoetate and acidified with IN hydrogen chloride in diethyl ether.
Ihe precipitated salt was filter~d an~ washRd with ethyl a oetate tD give the title amine salt as a white solid (1.88 g, 94%) m.p.
255-6C (dec). Rf 0.2 (ss-l). Found: C,62.64; H,7.74; N,5.74.
C14~ 9N02.HCl requires C,62.33. H17.47; N,5.19%.

. ., "; , -. , " . ., , ,:, , :: : ., ., :
.::: .: ~, : ~ : :, .. , -:~, :

91/07378 ~ ; PCT/EP90/01887 c-3-AminK-c-5-phenvlmethYlcvclRhexane-r-l-carboxylic acid methYl ester hvdLcchloride i. cis-5-tert-ButvldLmethylsilyloxv cyclohexane-cis-1,3-dicarboxvlic acid dimethyl ester t-Butyldimethylsilyl chloride (16.58 g, lIO ~mole) in dry methylene chloride (65 ml~ was added drqpwise over half an hcur to a stirred solution of cls-3-hydroxycyclohexane-cis-1,3-dicarboxylic acid dimethyl ester (21.5 g, 99 mmole) [J~ Org.
Chem., 38, 1726 [1973], triethylamlne (11.13 g, 110 le and 4-dimethylaminopyridine (488 mg, 4 mmole) in me~hylene chloride (80 ml), with i oe cooling keepLng ~he temperature between 5 and lo&. A~H~r standing overnight at rocm tençerature further amcints of t-butyldimethylsilyl chloride (1.6 g~, trieth~lamine ~1.1 g) and dim2thyl ~ idine (450 mg) w~re added and stirring continued ~or sixty-eight haurs. The mixture was then washed sequentially with water, saturated aqueous ammonium chloride an~
water, dried over MgS04 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 ~equired product as a liquid (32.6 g, 9~%) Rf 0.3 (1:3 diethyl ether, hexane~. Fcund: C,57.89; H,9.20. C1 ~3005Si requires C,58.15; H,9.15%.

W O 91/07378 ~ , , PC~/EP90iO~$$7 ii cis-5-tert-ButyldimethYlsilvloxvcxclohf~ane-cis-l,3-dicarboxvlic acid mono-methvl ester lN 5Odium hydroxide (94 ml) was added to an i oe cooled solution of the above diester (30.87 g, 93.4 nn~le) ln meth~ ol (500 ml). m e mixture was stirred at 5C for 3 hcurs and thf~n allcwed to stand at roon temperature for eighteen hours. ~u6t of the solvent was f~vapora~ed ~ff under vacuum, water was added a~d ~e suspension extracted with diethyl ether to ~er unreacted diester (3.86 g). IhR aqueous phase was addified with ',~
hy~ loric acid, extracted with diethyl ether, an~l the organic extract washed with water, dried over ~S04 an~ evaporat~d ~mder vacu~n to give a clear g~n ~25.5 g). ~r~[at~3raF~y on silica gel (500 g) eluting with a mixb~re of diethyl ether in he~ne (7:3 by volume) gave the required mrnc-ester as thick oil (20.0 g, 68%)o Rf 0.75, (diethyl ether) . Fou~d: C,56.75; H,8.71. Cl5H~8o5si requires C,56.93; H,8.92%.

iii c-3-tert~ silYloxy ~ 5-h~ rox ~thyl ~ clG~exane r-l car~oxylic acid methYl ester A lM soluti.on of diborane in tetrahydrofuran (59 Il~) was added dropwise under nitrogen t~ a stirred solution of the mono ester Erc~ p~rt ii (17.0 g, 53.72 mmole) in tetrahydrofuran keeping the temperature at -5C. The mixture was allowed to warm up to room temperature and after fcur hours a further amount of IM
di~orane solution (10.7 ml) was ad~ed and the mixture allowed to stand for eighteen hours. The solvent was evap~rated under vacuum, diethyl ether was added and the solution washed se~uentially with sat~rated salt solution, saturated aqueous ~ O 9l/07378 2 0 6 7 1 9 `7 PCT/EP~0/~$~7 sodium bicarbDnate solution and agaLn with saturated :~alt solution. Drying over M~SO4 an~ evaporation under ~acuum g~ve a cl OE oil which was chromatrgraFhed on silica gel (SoO g).
Elution with a nixtore of ethyl a oetate and hexane (1:1 by v~l~me~
gave an oil (15.18 g, 93%). Rf 0.42 (ss-3). Found: C,59.24~o H,967- cl5H30o4si requires C,59.56; H,lQ.0%

iv. c-3-tert-ButYldimethvlsilYloxY-c-5-f~-methyIben2enesNl~hon~l-oxymethvl)cYcl ~ r-l-carboxylic acid methYl ester :~
4-MethyIbenzenRsulphonyl chloride (14.18 g, 74.39 mmole) was added to an i oe cold stirred solution of the ester from ~art iii :
(15.0 g, 49.59 mmole) Ln dry pyridine (130 ml). After gLrrillg at room t ~ ture for twenty f~ur hours the mL~bure was iourcd onto ice, extracted with diethyl ether and the extract washed sequentially wlth 2N hy*rochlorlc acid, wa~er, saturated aquecus ~:
sodium bicarbonate solution an~ water. Drying o~er MgSO4 and ~ oration under vacuum gave an oil (22.0 g) which was chromatographed on silica gel (600 g). Elution with i~creasing proportions of ethyl aoetate in hexane (1:9 - 1:4 by volume) gave the re~ir~ pr~duct as an o;l (21.78 g, 96%). Rf 0.3 (4:1 hexane, ethyl aoetate. Found: C,57.66; H,7-96- C22H36o6SSi requlres C,57.86; H,7.95~.

W O 91/07378 2 0 6 719 7 PCT/EP90/01~87 ~

v. c-3-tert-Butvldimet~ylsilyl~ ~5~eny~meth cvclohe ~ r-l-carboxvlic acid methvl ester 2M Fhenyl lithium in cyclohh xan:-ether (7:3) (24.5 ml, 49 mmole) was added under nitrogen to a stirred suspension of cuprcus iodide (4.67 g, 24.~ mmole) in dry diethyl e~her ~50 ml) keeping the ~ ature below 5C. The resulting dark green solution was stirre1 at room temperatNre for one and three quarters of an hour, cooled to -70C and a solution of the ester from part iv (3.7 g, 8.1 mm~le) in ether (10 ml) and cyclohe~ane (40 ml? added dropwise. The temperature was maintained below -60 & during the addition, and then reduced to -70C for fifteen minutes. ~he mixture was then allowed to warm up to room temperature, stirred for an hour and the reaction quenched with saturated aquecus ammDnium chloride with i oe cooling. The organic phase was wa~hed with saturated sal~ solution, dried over MgS04 and on evaporation under vacuum ga~e an oil. Chromatography on silica gel, eluting ~ith increasing proportions of diethyl ether in h~xane (1:20 - 1:10 by volume) gave the required title pro*uct (1.4 g, 48%) Rf 0.2 (1:9 diethyl ether, hexane).

.: .: , ~ ; . . i.

~ ~ - , .:. - ~ . : : .

0 91/07378 .s..r ~ ~;. PCT/~90/~8~7 vi. t-3-(4-Methvlbenzenesulphonyloxv)-c-3-phenvlmethvl cyclohexane-r-l-carboxYlic acid methYl ester 40~ AL~Ieous hydrofluoric acid (0.5 n~, 10 n~ole) was added t~
a st ~ d solution o~ the above ester (3.6 g, 9.93 mmole in a oe tonitrile (40 ml). After an hour the solution was dilu~ed with diethyl ether, washed with satuxated aqueous sc~lium bicarbonate solution and water, dried over ~gS04 and evaporated under vacu~m to give the required alcohol as a clear oil (2.36 g, ~6%). Rf OD2 (1:1 diethYl ether hexane). The product (2.39 g, 9.62 ~mole) was treated as described in Preparation 6, part iv to give the trans-4-methyIbenzene sulphonate title cc~pound as a pale yellow gum (2.44 g, 63%~. Rf 0.4 (1:1 diethyl ether hexane). Found:
C,65-59; H,6-54; C22H2605S requlres C,65.65;

vii c-3-AminK-c-5-~henvlmethylcyclohexane-r-1-carboxvlic acid methYl ester hydrochloride The trans-4-methylbenzenesulphonate from part iv (2041 g, 5.99 mmole) was treated with sodium azide as descri~ed in Preparation 6, part v to give the c~rresponding azide as an oil (1.44 g, 88%), Rf 0.72 (1:1 diethyl ether, hexane). The azide (1.41 g, 5.16 mmole) was then reduced as described in Preparation 6, part vi to give the required am me salt as a foam (1.44 g, 99~), Rf 0.2 (ss-l). Found: C,63.31; H,7.76; N,4.56.
Cl ~ lN02.HCl requires C,63.48; H,7.81; N,4.94%

W O 91/07378 2 0 6 71~ 7 PCT/EP90/~1$~7 PREPARA~ION 8 cis and trans-3-~minc-c-4-phenylmethYlcvcl~lexane-r-l~
carboxylic acid methvl~a3E_hylrDchloride i 3-O~o-4-Phenylmethvlenecycloh ~ xvlic_acid methYl A mixture of ethyl(trim~thylsilyl) a oe tate (16.01 g, 99.9 ~mole) and 3-oxocycloheKanecarboXylic acid methyl es~er (13.0 g, 83.24 mmole) was slowly added dropwise under nitrogen ~o a stirred sDlution of l.OM tetrabutylammonium fluoride in tetrahydrofuran (2.78ml) keeping the ~emperature between O c~n1 10C. m e resNlting pale brown solution was stirred at room temperature for seventeen hours, further amounts of tetrabutylammonium fluoride (0.3 ml) and ethyl(trimethylsilyl) a oetate (5 ml) were added and after three hours the reaction was shown to ~e completed ~y n.m.r. m e mixture was diluted with n-hexane ~SO ml), stcod over 3A mDlecular sieves for an hour~
filtered and the ~Dlvent evaporatsd under vacuum to giv~ the silylenol ether as a golden oil ~19.97 g, 100%). The silylenol ether pr~duct in dry methylene chloride (50 ml), was add0d dropwise under nitrogen to a stirred suspension of titanium tetrachloride (9.15 ml, 83.24 le) in benzaldehyde (9.31 ml, 91.56 mmole) and dry methylene chloride (250 ml) at -70 &.
The mixture was allowed to warm up to roam ten~erature over ei~hteen hours and quenched with water (100 ml) with ioe coolLngo The organic phase was washed with water, dried over MgS04 and evaporated under vacuum. The residue was chroma ~ hed on silica gel, eluting with increasing prcportions of ethyl aoe tate in hexane (1:4 - 3:7 by volume) to give the required product as a , . :,, - , , , , ,, : , . .
. , . , :" ., .:........... :,: : , . . , , , . . .~. -: .
. ::: - ,.. , , , : ~ ;. .; .;. : ... :, O 91/07378 2 0 6 7 ~ PCT/EP~ 87 pale solid (6.89 g, 34%) m.p. 73-5 &. Rf 0.25 (4:1 hexan2, ethyl acetate). Fcund: C,73.20; H,6.54. C15~ 63 ~

requires C,73.21; H,6.63%.
'.

ii. 3-Oxo-cis-4-~henylmethylcvcl~ o~eK,~ L~ th~
,ester The enone from part i (6.55 g, 26.8 mm~le) in methan~l ~152 ml) and water (8 ml) was hydrogenated cver 5% palladium on charcoal catalyst (750 mg) at 20 p.s.i. ~1.4 bar). After three hours the s ~ sion was filtered and on evaporation under vacuum the residue was chramatographed on silica gel (600 g). Elution with increasing prcportions of ethyl a oetate in hexane (1:4 - 3~7 , by volume) gave the trans-isomer as a white solid m.p. 54-5C. Rf 0.35 (1:4 ethyl a oetate, hexane). Found: C,73.11; H,7.46.
183 reguires C,73.15; H,7.37%. Further elution gave the more polar cis-isomer as a white solid m.p. 71-2C. ~f 0.25 tl:4 e~
a oetate, hexane). Found: C,72.81; H,7.41. C1 ~ 8G3 requireso C,73.15; H,7.37%.

;.
iii. 3iMethoxYiminio-cis-4-phenvlmethvlc,vclohexane carboxvlic acid methyl ester m e cis-iso~er from part ii above (1.9 g, 7.71 mmole) on treatment with methoxylam m e hydroihloride as described in Preparation 1 part iii gave the title requir3d 0-me~hylox~ne as a "^
clear oil (2.0 g, 94%). R~ 0.4 and 0.45 (4:1 hexane, ethyl a oe tate). Fo~nd: C,69.53; H,7.69; N,4.94. Cl ~ lH~

C,69.79; H,7.F9; N~5-09%-.' ' :
~ .

W O 91/07378 ~ ;; PCT/EP90/0l887 iv. c and t-3-(tert-Butox~carbonY1amino~-c-4-Ehenylmethyl cyclohexan2-r-l-carboxYlic acid methyl ester The 0-methyl oxlme ~ m part iii tl.97 g, 7.15 mmole) on treatment as descriked in Preparation 1, part iv gave the title product as a mixture of cis an1 trans-isomers which were separated by chromatography on silica gel (300 g) eluting with increasing proportions of ethyl a oetate in ~ e (1:3 - 3:7 by volume). q~le trans-iscmer was obtained as a gum (680 mg, 27%). Rf 0.5 (1:4, hexane, ethyl a oetate). Found: C,68.84; H,8.17; N,3.91.
C20H29N04 requires C,69.14; H?8.41; N,4.03%. The cis-is~mer was also obta med as a gum (345 mg, 14%). Rf 0.45 (1:4 hexane, ethyl a oetate). Fcun~: C,67.55; H,8.08; H,3.80, C2o~ 9N04Ø5 H20 requires C,67.39; H,8.48; N,3.93%.

v. t-3-Amino-c-4-bhenvlmethYlcvclohexane-r-l-carboxvlic acid methyl ester hvdrochloride ~he t~ans-iscmer fro~ Fart iv (650 mg, 1.87 ~mole) on treat~ent with HCl as described in Preparation 1, part v gave the title salt as a gum 1533 mg, 99%). Rf 0.25 (ss-l). Found:
C,62-41; H,7-78; N,4-77- C15H21N02. HClØ25 H20 requires C,62.49; H,7.87; N,4.86%.

vi. c-3-Aminotc-4-phen~lmethylcyclohexane-r-1-carboxvlic acid methvl ester hydrochloride The cis-isomer fr~m part iv ~325 ng, 0.94 mm~le) on treatment with HCl as descr ~ in Preparation 1, part v, gave the title r salt as a gum (263 mg, 99%). Rf 0.25 (ss-l). Found: C,59.45;
, ~ 5 21N02~ ~Cl. H20 requires C,59,69; H,8.01;
N,4.64%

:; ., . . :

~ O 91/07378 ~ 7 ~ PCT/E~ 7 ,, 3~ .[(cis-4 EthoxYcarbonyl-cis-3-~2~phenvlethyl~cyclohexvll carbamoyllcyclc~entyl~-2-(2-methoxyethyl)prcpanoic acid benzyl ester l-Rthyl-3-(3-dimethylaminDprcpyl)carbodii~ide hydrochloride (460 mg, 2.4 mmole) was added to an ioe oold stirred solution 3f 3-(l~carboxycyclGpentyl)-2-t2 methyoxyethyl)prcpanoic acid ben~yl ester (EP-A-0274234) t401 mg, 1.2 mmole), c 4 ~ 2 ~2-phenylethyl)cycl~hexane-r-l-carbQxylic acid ethyl ester hydrcchloride (343 mg, 1.1 mnole), l-hydroxybenzckriazole (162 ~g, 1.2 D le) and N-methyl-morpholine (354 ~g, 3.5 mmole) in dry methyl~ne chloride (15 ml). After half an hc~r the mixture was allowed to attain room temperature and stirred for twenty hours.
m e solvent was evaporated under vacuum a~d the residue partitioned between diethyl ether and water. The organic extract was washed s~qu~ntially with water, 2N hydrochloric acid, waker~ ~;
saturated aqueous sodium bicarbonate and water. Drying (MgS0 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 volumR) 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. C36H49N06 re ~ C,73.07; H,8.35; N,2.37~.

, , ,- ,; , : .

wn9l/07378 2~ii71!~7~ pcr/~p9o/o78a7 ~ 1 . EX~M~LES 2-5 1 :
m e following ccmFounds were prepared foll.owing the p ~ e of Example 1 using the appropriate amine of formula (IV).

C H3 O ~ :

~ ~ H R2 C6H5 ~o2c~ Co2R4 he following ccmpZ unds were prepared following the procedure of Example 1 ~ut using 3~ carboxycycl ~ 1~-2(S)-(2-methoxy-~ ethoxymethyl~prcpanoic acid t-butyl ester (EP application no.
: 8930469~.7) as the starting ~aterial and coupling with the appr~priate amlne of formula (IV~.
, . .

~3U02c~ Q~
", O

, ' .

,~WO 91/07378 2 ~ ~ 7 ~ ~j;7,. .. i` ;? PCI/El~ l$$7 a~r r~ u~r c~
I
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u~ c~ o~ C~ Co C~C~ tO
~.~ :~
~ ~ 1~1~ O~r ~
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u ~ ~ r- I`t~ f`t~ l`
.
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': ~
Y 5~ ~
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. ~ In _ .

. , . . . , . `; . .. ~,` . ~ .
.. , , ~ . . ~ . .. . . . ...... . . . .. . .

W O 91/07378 2 0 6 719 7 PCTtEP90/Q1887 _ , .
Example R2 Form Analysis ~/~
No isolated (theoretical in brackets) C H N
.

6-~cH2)3ocH3 oil 64.47 9.35 2.53 CiSRf 0.45 (ether) (64.84 9.61 2.52) _ 7-(CH2)30C2~5oil 65.17 9.44 2.51 i -c sRf 0.5 (etherl (65.35 9.73 2.46) i EXP~ELES 8-12 Ihe followLng ccspcunds were prepared follawln~ the procedure of Example 1 but using 2S-(~ -berzyloxycarbonyl-N6-t-kutyloxy-~!, carbonyl-S-lysylam m c;ethyl)-3-[1-(1-carboxycyclopentyl)]propanoic acid t-butyl ester (EP application 89305180) as the start mg material and coupl mg with the aFprcpriate amine o~ formula (lV).

!
C~Z NH

BOC-NH (CH2)4~0 ~, ~dU2c~ H 2 '. ~
;` ' .~ .

O 91/07378 ~ i 7 ~ PCI`/E~0/~ $~7 ~T~` 1~
z; co ul ~ ~ ~ ~ I~ ~ ~
,~ ~ W ~D ~D ~D ~ In ~D
~ '... , In O d' CO ~ I~ ~ l~ OD 1` : ::
,~ ~ ~ ~ ~,~ o ~ ~ -~ a) o~ cs~ co ca o~ cs) Cl~ 0~ ~
~, ~r ~D~ In o a~c~ o o o O ~ ~ O O ~~ 0~ ~` ~1 r~ t~ r-~ ~o ~ l`
~_ ~D ~ ~ ~ ~ D~ ~D ~D ~D
_ ..
l l l l l o~q E~ u~ ~n . ~ q~,~ ~0 ~In. ~ ~. .
. ~ o ~ o o o . ~ o~ ~ _ ~ ~ :~

~ ' ~D U,~ ~iD 3iD
~ ~ ~ l ~ ~1~
~1 u~
I -~1 ~1 ,Ull -~nl . _ o ~ ~ ::
~ ~ ~ 0 ~

..

W O 91/07378 2 0 6 719 ~ PCT/EP~0/0~$$7 ~X~MFIE 13 2(S)-~N6-t-Butoxycarbony1-S-ly~rl-am momethyl)-3-~1-rcis-4 ethoxycarhon~l-cis-3-l3-ethoxypropvl~cyclohexyl)carbamcvll c~cl~
pentvllprcp~y~e acid t-~utvl ester m e diester of Example 8 (734 ~g, 0.85 mmole) m methanol (18 ml) and water (2 ml) was h~drogenated at room temperature for ~hree hours over 5% palladium on charcoal catalyst (75 mg) at 50 p.s.i. (3.45 bar) pressure. m e s y iQn was filtered thr~
arbacel, and evaporation of the solvent under reduced pressure gave the title product as a white foam ~630 mq, 100~). Rf 0.66 (ss-l). Found: C,63.22; H,9.73; H,7-17- C38~ oN409 reqlLres C,62.78; H,9.70; 7.71%.

: -: ,: ,. .:: :, . :: . . ;,.. :: :: : .
: . ~ . : : : .. , : ;

, ,, , .. :: .. . : -:
. : . . - , , ~!O 91/07378 2 a ~ 71 9 7 `

- E~ 14-17 ~ o follawing a~s were prepar~ fmlr Examples 9-:L2 b~
hydn~genation follc~ng the pr~cedu~ of 13x~mple 13.

~ , H2N I .

BOC-NH (CH2)4~0 HN~ ~ H R2 2C~ Co2R4 ,', ' , '~ .

,......................................................................... .

.

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,:

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8 ~7`1~ ~7~ PCI/EP9B/018$'7 ~ l ~ ~,~ ~
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.
u~ co a~ ~ ~1 o a~ o r- o t` ~ t` OD
~ oD a~ o~ ~ oo a~ c~ c~ ' ': , ~.~
In o ~n ~ O cO o r~ ~o CO
~n In ~ ~ ~ ~ ~r ' ~ ~ D ~O ~O ~D ~O ~D ~ ~D :~
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::
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~ O 9l/07378 2 0 6 7 ~ ~ 7 PCT/EP~D~ 7 `~, . ...................... .

EX~MFIE 18 2S~ ithan~sulE~onyl N6-t-butoxyc æ bonyl--S-lysyl-amlno-methyl)-3-(1- Ucis-4-ethoxycar~onyl-cls-3-f3-,e,thoxypr~pyl~cyclo-hexyl)carbamo~cYclopentyllpr3panoic acid t-~a~ r Methanesulphonyl chloride (0.14 ml, 1.8 mmole) was added dropwise to an ioe cold stirred solution of tbe amine from Exa~ple 13 (601 mg; 0.83 mmole) and N-methylmorpholine (0.2 ml, 1.8 mmole~
in dry methylene chloride (8 ml). me solution was stirred at lo& for 3 hours and the reaction mixture then quenched with saturated aqueous sodium hydrogencarbonate. m e organic phase was separated, the aqueous phase extracted with methelene chloride and the combined extracts dried over M~S04. Evaporation under vacuum gave a gum which was chr~matographed on silica gel (100 g) eluting with increasing p ~ ons of ethyl aoetate in hexane starting with a 1:1 mixture and finally with neat ethyl acetate.
Evaporation of relevant fractions gave the required product as a foalm (576.mg, 87%). Rf 0.56 (ss-l). FcunA: C,58.66; H,8091;
N,6-65- C39H72N4011S requires C,58.18; H,9.01; N,6.96%.

W O 91/07378 . - . PCT/EP90~ $7 EX~MPIES 19-23 The followin~ ccrFcL~I~t w~re prepared in a ~imilar manner to Example 18, using the apprcpriate amine of Examples 14 to 17 and reacting with the appropriate sulphonyl chloride or acyl chloride to yield the N2-substitNted-lysyl derivative.

B OC-NH(CH2)4~ ~ ~I
HN / \ `

t~UO2C~ N ~-- C02 O

'~

:~ ~

0 91/073782 0 6 7 ~ ~ 7 PC~D~ D~)11$$7 51 . `

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...,`
~ r- ~ o ô ~ ~ In r~-- r~ ~
d~ ~n In 1` ~ ~D 0~ 1` C~ ~ d' a~
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.~ o o o o` o ~ ~ ~ ~ ' r~ ~ o O O ~ ~
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_ _ ~ _ q_~ ~ _ ~ ~ 4~

~r~ ~ ~ ~ ~ ~
.' I ul ' I ul ~1 . _ In ~ I
_~' ~ ~ ~ 1, ~
~ 1 l ul tnl -ul 51 -~1 _ ~ N
--k; 5r ~ N ~ ~ > N 5 _ _ N j N ~ N ~ N

, . i .
.~ . . .

W 0 91/07378 `;; ~ ` ' PCT/EP90/0~887 2S-~bis-tlSl-PllenYlethyllamin ~ethY11-3-fl-~cis-4-e~hoxy-carbonvl-cis-3-(3-methoxypropyl)cvclohexyl)c ~ llcyclopentyl~-pro~anoic acid t-butYl ester 2S-{bis-[lS)-Phenylethyl]aminomethyl}-3-(1-carboxy-cyclcpentyl)prcpanoic acid and c-4-amino-c-2-(3-methoxyprcpy1~-cyclohexane-r-l-carboxylic acid ethyl ester (from Preparation 2) were coupled as described in Exa~ple 1 to give the title diester as a gum. Rf 0.1 (4:1 hexane, ethyl a oetate) Found: C,73.19;
H,9-13; N,4-01- C43H64N206 requires C,73.26; H,9.15; N,3.97%.

EX~MPLE 2S

2S-~bisrllS)-Pher~lethYllam mcmethY11-3-llrtcis-4-ethox~- -il carbonyl-cis-3-(2-Pheny~_thvl)cvclohexvl)carbamovllcYclopentvl~-propanoic acid t-~utvl ester ~ -The title co~pcund was p ~ as descriked in EXample 24 abave l~ciny c-4-am m o,c-2-(2-phe~ylethyl)cyclohexane-r-1-carboxylic acid ethyl ester hydr~chloride (from Preparation 1) as the amine c~mponent to yield the pro~uct as a white foam. Rf 0.55 (ss-3). Foundi: C,76.11; H,8.74; N,4.25. C47H64N205, 0.25~ 0 requires C,76.12; H,8.77; N,3.77%.

~ '' ;
~ . .

.~ , . ... ... .: , . , .. ~.

~ O 91/07378 2 ~ ~ 719 ~ ~ PCT/EP90~1887 ~ 26 2S-fAminomethyl)-3-~1-[fcis 4-ethoxYcarbonvl-cis-3-~3-methox~r Prcpyl~cyclohexyl)carbamoyllc~clopentyll~rQeanoic acid t-butvl ester :
The diester from Example 24 above (992 mg, 1.4 mmole) in ethanol (20 ~1) was hy~rogenated over 20% palladium hydroxi~e on charcoal catalyst at 60 p.s.i. (4.1 bar) press~re. After twenty four hcurs the suspension was fil ~ thrcugh a short arbacel column and the .solvent was evaporated under vacuum. me residue was dried azeotropica~ly with methy~ene chloride to give the title amine as a ~hick oil. (690 mg, 99%) Rf 0.35 (ss-l) Found:
C 65.69 H,9.88; N,5.19. C2 ~4~8N2O6 require5 N,5.64~.
EX~MPIE 27 2S-(A~L~x~methvl)-3-~1-rfcis-4-ethoxyca~bonv1-cis-3-(2-~heny~
ethyl~cyclohexyl~carbamoyllcyclopentvl~prcpanoic acid t-butYl ester The product of Example 25 was hydrogenated as descriked in Example 26 above to yield the title ocwpaund as a gum. Rf 0.35 (ss-l) Found: C,69.00; H,8.80; N,4.88. C31~48N205. 0.6 H20 requlres C,69.00; H,9.19; N,5.19.

W O 91/07378 2 ~ g ~ ` !` PCT/EPg~ $'~ ~
,.............................. ~' EX~MPIE 28 2S-(N iMet ~ phonvl-N -t~butox~carbonYl~-S-ly~yl-aminc-methYl)3-~1-[(cis-4-ethoxYcarbonvl-cis-3-~3-methoxYpropvl~yclo-hexYl)carbamoyllcyclcpentvl~propanoic acid t-t~¢~
N-me~lylmorpholine (0.34 ml, 3.06 mmole) was added to an ic~
cooled stirred mixture of the product from Example 26 (690 mg, 1.39 mmole) ~ -methanesulphonamido-N6-t-buboiyca~bonyl-S-lysine! ;
(406 mg, 1.39 mmole), l-hydroxybenzotriazole (188 mg, 1.39mmole) l-ethyl-3-(3 ~ ethylaminopropyl)car30diimide hydrcchloride (586 m~, 3.06 mmole) in dry methylene chloride ~10 ml). After six hcurs the mix*ure was diluted with methylene c~loride (20 ml) ~:;
washed with water and clried over Mg504. Evaporation of the solve~ under vacuum and c~romatography on silio~ gel, eluting with increasing proportions of e~hyl aoetate in hexane (50 90%) and finally with 5% ethanol in ethyl a oetate gave the required product as a c~am foam (880 mg, 81%). Rf 0.2 (i~s-3) Fcund:
c,sa.o7; H,9.06; N,6.74. C39H70N4011S requires C,58.33; H~807 N,6.98%.

s~VO 91/07378 ;~ PCT~EP90/0~$~7 ¦ ~
~67~7 EX~MPIE 29 2S-(N-t-Buty__xy~arb ny____ala~vlaminomethyl)-3-~1-[(cis-4-ethoxvcarbonyl-cis-3-(2-phenylethvl)c~clohexyl~carbamcyllcyclc pentYl~propanoic acid t-butyl ester Ihe procedure of Example 28 was ~ollowed startiny with the amine of Exampl~ 27 and coupling with N-t-butyloxycarbonyl-R-alanine to yield the title pr3duct as a foam. Rf 0.5 (ss-l)O i Found: C,67-22; H,8.86; N,5.66. C39H61N308 requires C,66.92;
H,8.78; N,6.00%.

EX~MPIE 30 2-rN-~2-t-Buty__ ycarbony1 ~ 2-meth~lprcpanovl)am mo-methvl~-3-~1- r (cis-4-ethoxvcarbonyl-cis-3-(2~E~envlethYl) hexYl)caIt~nK~ cycl~entv~ L~acid t-butvl ester m e procedure of Example 28 was followed starting with th~
amlne of Exalmple 27 and ccupling with 2-t-butyloxycarbonylamm o-2-m~ithyl-propanoic acid to give the title pro*uct as a foamO ~f 0.48 (ss-l). Found: C,67.06; H,8.94; N15.81. C40H63N308 requires C,67.29; H,8.89; N,5.89%

.

. . ; ;, .

W O 91/07378 ~ PCT/EI so/~l~a7 EX~MPIE 31 2S-(~`-MethanesulPhonyl-S-lysyl-a~nomethvl~-3~ r (cis-4 carboxv-cis-3-~2-PhenYlethyl~cyclohexYllcarbamoyllcYclopent prcpanoic acid Trifluoroa oetic acid (5 ml) was added to an ioe cold soluti~
of 2S-(N2-methanesulphonyl-N -t-butoxycarbonyl-S-lysyl-amino- ' methyl)-3-~1-[(cis-4-ethoxycarbonyl-cis-3-(2-phenylethyl)cyd ~ ~ -hexyl)carbamoyl]cycl~pentyl)propanoic acid-t-butyl ester (frcm ~ le 19) (445 mg, 0.53 mmole) in dry methylene chloride ~5 ml)O
l~e solution wzs allowed to warm to roo~ temperature and after three hcurs evaporated to dryness under vacuum. The residue was dried azeotrcpically twioe with toluene and dis~olved in lN sodium hydroxide. the sc,lution was stirred at 40-55C for sixty eight hcurs, cooled and passed down a sulphonic acid ion-exchange ~;
column. Elution with 10% aqueous pyridine and evaporation of the relevant fLdctions gave a foam which was crushed under a oe tonitrile and filtered to give a whi~e pcwder (279 mg, 30%) Rf 0.25 (ss-2) Found: C,58.66; H,7.92 N,8-49- C32H50N408S requlres C,59.06; H,7.74; N,8.61%. ;

~ YO 91/07378 2 0 6 7 1~9 7;` ~ PCr/~P9OiO1887 EX~MPIES 32-37 Ihe followLng compoun~s were prepared fram the appropriate N6-t-butyloxycarbonyl prctected diester of EXamples 18, Z0-23 or 28 by treatment w.ith trifluoroa oe tic acid followed by alX21ine hydr~lysis followin3 the proaedure of Example 31.

~,~12 ! -H2N(CH2)4~

HOIC ~ ~ C 2 H

.

WO 91/07378 2~ Q ~ 7 3~ ' ; `PCr/EP9~1l/01l887 ~ .

58 `
.___ __.___.
Z r~ I` ~ ~^ ,~ ~rr I` C a:~ Co ~ t~` ' ~ .
d~ coco coo~ coco t~o t;oCo 0~0~ ' ~
u~ ,~ ~ ~ a~ ~ ~ u~ ~D ~ U~ ~Oo ~r r` ' ' ,~.~$ ) o~ cooo oo t~ r~ 1~ 1~ c ~.~ ~ ~1 0~ n~'' InW ,r~,o~
1~ ~, u~ u~ ~ m u~ ~u~ u~ ~ ~ ~
. . . .
_ o~ , ~ ~ ,., '~o~ ~1 l c)u~ ~ ' ~:n .

1~ ~} ~ ~ ~L ~ o ~~ o ~ ~ lL o ~ ~ N
. . i . _ _ 1 ul ~1 1 1 1 -~,1 ,U~I , . __ ~

~ d,~ ~ ~ Cln ~

-ul ' I ~,1 ~1 ,U~I ,U~I
~" ~ . ~ '' <>

_~ r~ r~ .r ~-- i r :;:
~i~ r~ r~ ~ ~ r~ ~ .~
.

~'O91/07378 20 6 7~ 97 , i :~ . PCI/EP90/10~8$7 EX~ 38-39 Ihe follc~ng ~unds were preparsd fr~n the diesters of ExalTples 29 ar~ 30 ~y treatment with trifluoroacetic acid fol.lowed alXaline hy~rolysis follawing the prooe~ of E~{ample 31 l3 NH2.
R l o HO2C~ H ~ C6~15 o C2 H

. :...... . ... ..

- . . . ...... . . .

WO 91/07378 2 ~ ~ 7 ~L 9 7; ~ ' pCI`/FP9~ 1$~7 ; :~
_ _ o ~ ,~
a~a~ cs~cO . ' :~
~ . i':.
~ ~ 1~ . I
.~ coco a: 0 I :, 1~ u~ ~In, I , ~v . . . _ ',~' _ ~ ~ :
~ ~ 5~
o ~ o ~ o ., .~ ~ 'Q) ~
,4 1:4 0 o ~ R Q. O o~K
. ~.
'~' ~--, ~
~ .
:~ V~ , ~`
. '.' ~ ~ ' ~ . ' ' ,~
,~ . ~, _. _ : '.

~ 0 a~ :' ', ~ ~
_ ~ O 91/07378 2 ~ ~7;1 g 7 . . .

EX~MPIE 40 3-tl-rcis-4-CarboxY-cis-3-~2-phenvlethyl~cyclQhexyl) carbamoyllcyclopentyl~-2-t2-methoxyethyl)prc~anoic acid The diester of Example 1 (510 mg, 0.86 mmole) in ethanol ~25 ml) an~ water (10 ml) was hydrogenated a~ room temperature for three hcurs cver 10% palladium on charcoal catalyst (100 mg~ at 50 p.s.i. (3.45 bar) pressure. The ~ e was filtered thrcugh a short ~vicel column and the solvent evaporated under ~ ced pressure. Ihe monoester thus obtained was dissolved in lN sodium hydr~xide and the solution heated to 65C under a nitrcgen atmosphere for two days. On oooling the solution was ex~racted ~;
with diethyl ether, the aqueous phase acidified with 2N
hydrochloric acid and extracted with diethyl ether. m e organic phase was washed with wzter, dried cver M~SO4 and evaporated under reduced pressure to give the required title cliacid as a white foam, (300 mg, 75%). Rf 0.62 tss-l) Found: C,68.17; H,8.45;
, C27~ gNO6 requireS C,68.47, H,8.30; N,2.96%.

. . . ;:. ' ! , . . `

W09t/07378 ~ ; Pcr/~p90/on$s7: ~ ' 2 0 ~ ~ ~ 9 7 EX~ 41~44 Ihe follawing ~s were prepared fmm t~e a~r~priate diester of ~amples 2 to 5 }:~y hy~rogenation follaw~d by aL~l:lne hy~lysis foll~ing the pr~cedure of Example 40.
, O ~ ~ .

,.,. WO 91tO7378 ~ 7 ~ 9 7 PCT/EPg(D/~)1887 ,_ ~ ~ ~ ~ ~ a~
u~ r~ o ~ ~ o ~ a~ o ~ ~i ~l7 ~ ~ ~ ~ ~ ~
~n~
'~n ~ ~ a~ ~ ~D ~ r- In ~r ~ rl ~ O ~ OD ~ ~ ~ ~
~ a~ a~ t~l~ co oo c~ OD
'~, ~7 ~ ~a~
~t~ r ~ I~D ~r~
:~ C) ~ ~ ~ ~ ~ ~D t` ~`
_ ~-D ~0~ ~0 ~D

. ~.
Ul Uu)~ ou~
o ~ o In r~ o u~ ~
o o rro U~ O
~0~ ~ ~Oæ ~0 . .
.
. ~

~ -tl tl ~1 ~1 u~
o, ~ C~

c~l I l ~1 ~ .
~ ~ ~ ~ ~r ~ .

.. . . . ... . i , . .. - . ~, , . . , ;

... . ., . . .. ~. . ..... ~ . ~ . . .
.. ... . . .~, . . . .

W O 91/07378 20~ `i PCT/EP~70/01887 EX~MPLE 45 2S-t2-MethoxvethoxYmethyl~-3~1-r(cis-4-carboxy-cis-3-~3-methoxy-prcpvllcvclQhexyLcarkamovllcYclopentyllpropanoic acid Trifluoroa oe tic acid t5 ml) was added to an i oe cold solution ' :
of the diest~r of Example 6 (480 mg; 0.77 mmole) in dry methylene -~
chloride (3 ml). After standing at room temperature for three hcurs 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 wat~r (xg) and the resulting monc-ester extracted into lN sodium hYdroxide solution (2 x 10 ml). The aqueo~s extract was heated at 65-70C under ;
nitrogen for 24 hours, cooled saturated with salt, acidified with concentrated hydrochloric acid and extracted with ethyl acetate.
Ihe organic extract was washed with water, dried over MgS04 and on evaparation gave the required diacid as a gum (320 mg 88%).
Found: C,60.55; H,8.54; N,2.80, C24H41NO~. 0.4H20 requlres C,60.20; H,8.80; N,2.93%.
EX~MPLE 46 2S-(2-Meff~oxyethoxvmethvl)-3-f1-rlcis-4 car~oxy-cis-3-~3-ethox pro~Yl)cvlohexYl)carkamoYllcYclo~entYl)prcpanoic acid The title product was prepared fr~m the diester of EXample 7 as described in ~ le 45 above and was obtained as a gum.
Found: C,61.26; H,8.66; N,2.83. C25H43N08. 0.3H20 C,61.15; H,8.95; N,2.85%~

Claims (22)

1. A compound having the formula:- (I) wherein A completes a 4 to 7 membered carbolic 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 R4 is independently H, C1-C6 alkyl, benzyl or an alternative biolabile ester-forming group;
R1 is H or C1-4 alkyl;

R2 is C1-C6 alkyl substituted by C1-C4 alkoxy, aryl or aryloxy, or is phenyl;
and R is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, or C7 cycloalkenyl, or R5 is C1-C6 alkyl substituted by halo, hydroxy, C1-C6 alkoxy, C1-6 alkoxy(C1-C6)alkoxy, C3-C7 cycloalkyl, C7 cycloalkenyl, aryl, aryloxy, heterocyclyl, -NR6R7, -NR8COR9, -NR8SO2R10, CONR6R7 or R6R7N-(C1-C6)alkoxy;
or R5 is C1-C6 alkyl substituted by a group of the formula:

or wherein R6 and R7 are each independently H, C1-C4 alkyl, C3-C7 cycloalkyl, aryl, aryl(C1-C4)alkyl, C2-C6 alkoxyalkyl, or heterocyclyl; or the two groups R6 and R7 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 C1-C4 alkyl;
R9 is C1-C4 alkyl, CF3, aryl, aryl(C1-C4)alkyl, aryl(C1-C4)alkoxy, heterocyclyl, C1-C4 alkoxy or NR6R7 wherein R6 and R7 are as previously defined;
R10 is C1-C4 alkyl, C3-C7 cycloalkyl, aryl or heterocyclyl;
R11 is H, C1-C6 alkyl, aryl or C3-C7 cycloalkyl;
R12 is R11CONR11-, R11SO2NR11-, R16R17N-(CH2)p-, or R11O-, wherein each R11 is as previously defined above;
R13 and R14 are each independently H or C1-C6 alkyl; or R13 is H and R14 is C1-C6 alkyl which is substituted by OH, C1-C4 alkoxy, SH, SCH3, NH2, aryl(C1-C6)alkyl-OCONH-, NH2CO-, CO2H, guanidino, aryl, or heterocyclyl;
or the two groups R13 and R14 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 substituted by C1-C4 alkyl or fused to a further 5 or 6 membered saturated or unsaturated carbocyclic ring;
or R13 is H, and R12 and R14 are linked to form a 2-(N-COR11-4-aminopyrrolidinyl) group;
R is R16R17NCO-, R11OCO, R11OCH2- heterocyclyl, wherein R11 is as previously defined above;
R16 and R17 are each independently H or C1-C6 alkyl;
and p is o or an integer of from 1 to 6;
and pharmaceutically acceptable salts thereof and bioprecursors therefor.

2. A compound according to claim 1 wherein A is (CH2)4, R1 is H
and B is (CH2)2 having the formula-:

(II) wherein R, R2, R4 and R5 are as previously defined.

3. A compound as claimed in claim 1 or claim 2 wherein R and R4 are both H.

4. A compound as claimed in claim 1 or claim 2 wherein one or both of R and R4 is a biolabile ester-forming group and said group is ethyl, benzyl, 1-(2,2-diethylbutyryloxy)ethyl, 2-ethyl-propionyloxymethyl, 1-(2-ethylpropionyloxy)ethyl, 1-(2,4-dimethyl-benzoyloxy)ethyl, 1- (benzoyloxy)benzyl, 1-(benzoyloxy)ethyl, 2-methyl-1-propionyloxypropyl, 2,4,6-trimethylbenzoyloxymethyl, 1-(2,4,6-trimethylbenzyloxy)ethyl, pivaloyloxymethyl, phenethyl, phenpropyl, 2,2,2-trifluoroethyl, 1- or 2-naphthyl, 2,4-dimethylphenyl, 4-t-butylphenyl, [5-(4-methyl-1,3-dioxolen-2-onyl)]methyl or 5-indanyl.

5. A compound according to any one of claims 1 to 4 wherein R5 is methylene substituted by a group of the formula -NHCOCR12R13R14, where R12 is NH2, R11CONH- or R11SO2NH-, R13 is H
and R14 is -(CH2)4NH2.

6. A compound according to claim 5 wherein R5 is N2-acetyl-S-lysylaminomethyl, N2-methanesulphonyl-S-lysyl-aminomethyl, N2-phenylsulphonyl-S-lysyl-amiomethyl or N2-cyclobutylcarbonyl-S-lysyl-aminomethyl.

7. A compound according to any one of claims 1 to 4 wherein R5 is methoxyethyl or 2-methoxyethoxymethyl.

8. A compound according to any one of claims 1 to 4 wherein R5 is C1-C6 alkyl substituted by -NHCOCR12R13R14 wherein R12 is NH2, R13 is CH3 and R14 is H or CH3.

9. A compound according to any one of claims 1 to 8 wherein R2 is phenyl, benzyl, phenethyl, methoxypropyl or ethoxypropyl.

10. A compound according to claim l wherein said compound is 2S-(N2-methanesulphonyl-S-lysylaminomethyl)-3-{1-[cis-4-carboxy-3-cis(3-ethoxypropyl}-cyclohexyl)carbamoyl]cyclopentyl}-propanoic acid;
2-(N2-methanesulphonyl-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 or 2S-(2-methoxyethoxymethyl)-3-(1-[(cis-4-carboxy-3-cis-{3-methoxypropyl)-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 formula-:

(V) wherein R18 and R19 are as defined for R and R4 excluding H, or they are conventional carboxylic acid protecting groups and R5' is as defined for R5 with any reactive groups therein optionally protected;
to a hydrolysis and/or hydrogenation and/or other deprotection reaction to remove any protective group present in R5' and to remove one or both of R18 and R19 to yield the corresponding dicarboxylic acid of formula (I) wherein R and R4 are both H, or to yield the corresponding mono-ester product wherein one of R and R4 is H and the other is a biolabile ester-forming group; and optionally forming a pharmaceutically acceptable salt of the product.

12. A process as claimed in claim 11 wherein R18 and R19 are selected from t-butyl, ethyl and benzyl and said groups are removed by treatment with trifluoroacetic acid, aqueous alkali or catalytic hydrogenation respectively, to yield the compound of formula (I) wherein R and R4 are both H.

13. A pharmaceutical composition comprising a compound of the formula (I) or (II) as claimed in any one claims 1 to 10 or a pharmaceutically acceptable salt thereof or bioprecursor therefor, together with a pharmaceutically acceptable diluent or carrier.

14. A compound of the formula (I) or (II) as claimed in any of claims 1 to 10 or a pharmaceutically acceptable salt thereof or bioprecursor therefor, for use in medicine, particularly for the treatment of hypertension, heart failure or renal insufficiency.

15. A process for preparing a compound having the 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 a 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 R4 is independently H, C1-C6 alkyl, benzyl, or an alternative biolabile ester-forming group;
R1 is H or C1-C4 alkyl;

R2 is C1-C6 alkyl substituted by C1-C4 alkoxy, aryl or aryloxy, or is phenyl;
and R5 is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, or C7 cycloalkenyl, or R5 is C1-6 alkyl substituted by halo, hydroxy, C1-6 alkoxy, C1-C6 alkoxy(C1-C6)alkoxy, C3-C7 cycloalkyl, C7 cycloalkenyl, aryl, aryloxy, heterocyclyl, -NR6R7, -NR8COR9, -NR8SO2R10, -CONR6R7 or R6R7N-(C1-C6)alkoxy;
or R5 is C1-C6 alkyl substituted by a group of the formula:

or wherein R6 and R7 are each independently H, C1-C4 alkyl, C3-C7 cycloalkyl, aryl, aryl(C1-C4)alkyl, C2-C6 alkoxyalkyl, or heterocyclyl; or the two groups R6 and R7 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 C1-C4 alkyl;
R9 is C1-C4 alkyl, CF3, aryl, aryl(C1-C4)alkyl, aryl(C1-C4)alkoxy, heterocyclyl, C1-C4 alkoxy or NR6R7 wherein R6 and R7 are as previously defined;
R10 is C1-C4 alkyl, C3-C7 cycloalkyl, aryl or heterocyclyl;
R11 is H, C1-C6 alkyl, aryl or C3-C7 cycloalkyl;
R12 is R11CONR11-, R11SO2NR11-, R16R17N-(CH2)p-, or R11O-; wherein each R11 is as previously defined above, R13 and R14 are each independently H or C1-C6 alkyl; or R13 is H and R14 is C1-C6 alkyl which is substituted by OH, C1-C4 alkoxy, SH, SCH3, NH2, aryl(C1-C6)alkyl-OCONH-, NH2CO-, CO2H, guanidino, aryl, or heterocyclyl;
or the two groups R13 and R14 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 substituted by C1-C4 alkyl or fused to a further 5 or 6 membered saturated or unsaturated carbocyclic ring;
or R13 is H, and R12 and R14 are linked to form a 2-(N-COR11-4-aminopyrrolidinyl) group;
R15 is R16R17NCO- R11OCO-, R11OCH2- or heterocyclyl, wherein R11 is as previously defined above;
R16 and R17 are each independently H or C1-C6 alkyl;
and p is 0 or an integer of from 1 to 6;
which comprises subjecting a compound of the formula (V) wherein R18 and R19 are as defined for R and R4 excluding H, or they are conventional carboxylic acid protecting groups and R5' is as defined for R5 with any reactive groups therein optionally protected;
to a hydrolysis and/or hydrogenation and/or other deprotection reaction to remove any protective group present in R5' and to remove one or both of R18 and R19 to yield the corresponding dicarboxylic acid of formula (I) wherein R and R4 are both H, or to yield the corresponding mono-ester product wherein one of R and R4 is H and the other is a biolabile ester-forming group; and optionally forming a pharmaceutically acceptable salt of the product.

16. A process as claimed in claim 15 wherein R18 and R19 are selected from t-butyl, ethyl and benzyl and said groups are removed by treatment with trifluoroacetic acid, aqueous alkali or catalytic hydrogenation respectively, to yield the compound of formula (I ) wherein R and R4 are both H.

17. A process according to claim 15 wherein A is (CH2)4, R1 is H
and B is (CH2)2 to give a compound having the formula-:

(II) wherein R, R2, R4 and R5 are as previously defined.

18. A process according to claim 15 wherein R5 is N2-acetyl-S-lysylaminomethyl, N2-methanesulphonyl-S-lysylaminomethyl, N2-phenylsulphonyl-S-lysylaminomethyl or N2-cyclobutylcarbonyl-S-lysylaminomethyl.

19. A process according to claim 15 wherein R5 is methoxyethyl or 2-methoxyethoxymethyl.

20. A process according to claim 15 wherein R5 is C1-C6 alkyl substituted by -NHCOCR12R13R14 wherein R12 is NH2, R13 is CH3 and R14 is H or CH3.

21. A process according to claim 15 wherein R2 is phenyl, benzyl, phenethyl, methoxypropyl or ethoxypropyl.

22. A process according to claim 15 wherein said compound of formula (I) produced is-:
2S-(N2-methanesulphonyl-S-lysylaminomethyl)-3-{1-[cis-4-carboxy-3-cis{3-ethoxypropyl}-cyclohexyl)carbamoyl]cyclopentyl}-propanoic acid;
2-(N2-methanesulphonyl-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 or 2S-(2-methoxyethoxymethyl)-3-{1-[(cis-4-carboxy-3-cis-{3-methoxypropyl}-cyclohexyl)carbamoyl]cyclopentyl}propanoic acid, or a biolabile ester derivative thereof.