CA2337261A1 - Utilisation d'inhibiteurs de prenyltransferases pour preparer un medicament destine a traiter les pathologies qui resultent de la fixation membranaire de la proteine g heterotrimerique - Google Patents

Abstract

L'invention concerne l'utilisation d'inhibiteurs de prényltransférases pour préparer un médicament destiné à traiter les pathologies qui résultent de la prénylation de la sous-unité .gamma. de la protéine G. Ces maladies comprennent en particulier des maladies liées aux fonctions biologiques ou désordres suivants: odorat, goût, perception de la lumière, neurotransmissio n, neurodégénérescence, fonctionnement des glandes endocrines et exocrines, régulation autocrine et paracrine, tension artérielle, embryogénèse, infecti on virale, fonctions immunologiques, diabète et obésité.

Description

Use of prenyltransferase inhibitors for ~re~aring a medicament intended to treat pathologies resulting from membrane fixing of the heterotrimeric G protein The present invention relates. to the use of prenyltransferase inhibitors for preparing a medicament intended to trea.t pathologies resulting from the membrane fixing of the heterotrime,ric G protein. These diseases include in particular diseases linked to the following biological functi.ons or disorders: smell, taste, perception of light, neurotransmission, neurode;generation, endocrine and exocrine gland fonctions, autocrine and paracrine regulation, arterial tension, embryogenesis, viral infection, immunological fonctions, diabetes and obesity.
The heterotrimeric G protein coupled with receptors with 7 transmembrane domains is a mediator of the extracellular information into the cell. Several intracellular effectors of the G protein are identified as adenylate cyclase, phospholipase C or also ionic channels (cf. Gilman, A.G. Biorci. Rein. 15, 65-97 (1995)).
The activity of the adenylate cyclase is modulated by the different G proteins (Gs, Gi, Gq, Go) which thus regulate; the biosynthesis of the cyclic AMP (cAMP). It is thus known that, in order to modu~late the adenylate cyclase, it is necessary for the G protein transitionally to be in a heterotrimeric form, in which form the monomer constituted by a sub-unit a is associated with the dimer constituted by the sub-units(3 and y. It is also known that for the G protein to be functional, it must be fixed to the membrane by its sub-unit y, this fixing being possible when the sub-unit Y is prenylated. Unly in this situation can the ligand activate the sub-unit a of the G protein, outside the cell, via the rece~ptors with seven transme:mbrane domains. Alter disassociation, the a sub-unit can modulate the adenylate cyclase and modulate the production of cAMP.
The harmful effects of an abnormal cAMP level are also known and occur in particular at the level of the following biological fonctions: smell, taste, perception of light, neurotransmission, neurodegeneration, endocrine and exocrine gland fonctions,

-2-autocrine and paracr~ine regulation, arterial tension, embryogenesis, benign cell proliferation, oncogenesis, viral infection, immunological fonctions, diabetes and obesity.
Prenyltransferase inhibitors are already used in the field of cancer treatment (cf. Sebti et al., Pharmacol. Ther. 74, 103-114 (1997); Sepp-Lorenzino et al., Cancer.Res.
55, 5302-5309 (1997)). The useful,ness of prenyltransferase inhibitors in this type of treatment would appear to derive from their action in preventing prenylation at the level of the Ras substrate. However, the prenylation of certain forms of Ras is not modified by prenylation inhibitors (Lerner et al., Oncogene, 15, 1283-1288, 1997).
The Applicant has now discovered that prenyltransferase inhibitors can also be used to modulate the action of the; heterotrimeric G protein, and thus treat all sorts of diseases linked with it.
The invention firstly relates to the use of prenyltransferase inhibitors to prepare a medicament intended to treat pathologies resulting from membrane fixing of the heterotrimeric G protein. In particular, it relates to the use of said inhibitors to prepare medicaments intended to treat diseases linked to the following biological fonctions or disorders: smell, taste, perception of light, neurotransmission, neurodegeneration, endocrine and exocrine ~;land fonctions, autocrine and paracrine regulation, arterial tension, embryogenesis, viral infection, immunological fonctions, diabetes and obesity.
More particularly, the invention relates to the use of certain prenyltransferase inhibitors for the preparation of a medicament intended to treat cholera, Acquired Immune Deficiency Syndrome (~.IDS), travel diarrhea and familial masculine precocious puberty.
Among the prenyltransferase inhibitors which can be used for the invention, there can in particular be mentioned those chosen from a group constituted by the following compounds: derivatives of general formula (GPI) defined below (in particular those descr~ibed in Patent Application WO 97/21701, such as the compound of formula (I~
defined hereafter, and those described in Patent Application WO 97/16443, such as the compound of formula (~~I) defined hereafter), thiazole derivatives such as those described in Patent Application WO 98/00409 (for example the compound of formula (I) defined below), peptïdomimetic derivatives of 4-aminobenzoic acid (for example the compounds of formulae (lI) and (II)a defined hereafter) or peptidomimetic analogues such as those described in Patent Applicatïon WO 96/21456 (for e:xample the compound of formula (III) defined hereafter), tricyclic amides such as those described in Patent Application WO 97/24378 (for example the compound of formula (~

-3-defined hereafter), polyacids such as those described in Patent Application WO 97/17321 (for exmr~ple the compound of formula (VII) defined hereafter), piperidine peptidomimetic derivatives such as those described in Patent Application WO 97/18813 (for example the compound of formula (VIII) defined hereafter), benzodiazepine peptidomimetic derivatives such as those described in US Patent 5,532,359 (for example th.e compounds of formulae (IX) and (IX)a defined hereafter), tripeptidic derivatives of phosphonic or phosphinic acids such as those described in US
Patents Nos. 5,523,430 and 5,510,510 (for example the compounds of formulae (X) and (X)a defined hereafter), pseudodipeptides based on a N-carbonylpyrazine structure such as those described in Patent Application WO 95/00497 (for example the compound of formula (XI) defined hereafter), and tetrapeptidic analogues of the type C:ys-wherein Cys is Cystein, A1 and A2 are aliphatic aminoacids and A3 represents any aminoacid (such analogues being described for example in US patent 5,627,202).
Preferably, the inhibitors of prenyltransferases will be chosen from the group constituted by the compounds corresponding to general formula (GPI) shown below R,i /
H3C~N
Ri (~,)\ ~ I ~ I

I

(GPI) in which:
R~ represents OH, SH, NH2 or CH20H;
RZ represents a lower alkyl radical or CORS;
R3 and R4 represent independently a halogen atour or an OH radical;
RS represents a lower alkyl radical, a substituted or non-substituted carbocyclic or heterocyclic arylalkyl or aryl radical or a saturated heterocyclic radical comprising _5 to 6 members, laid members being chosen from CH2, O, NH
and S; and Y represents CO or CS or is absent;

-4-and compounds corresponding to one of the following formulae:
SH ~S S ~
N
NH~
HZN~ O NH O~
O
(I) HS
OR
H

(II) : R = CH 3 (II) a . R = H
HS
H2N ,,,~H
NH

H
(III)

-5-I

(IV) BI' r CI
N

N N' '' O
v v --N
HO /~/N
Cl C1~ v u ~N-I

(VI)

-6---° \
O ~ O
/
oH

OH
/ ~~ N O
O
N~ CH3 O
OH
(VII) NC
\.
N
H O
N N N
N O O
(VIII) N
H3C~ ,;
N
~ O O
H2N~~ HN
_ O
~"~CH2R
SH HOO ~C
(IX) . R = CH 2 SCH
(IX) a . R = iPr _ 'j _ H H
O~ N N~COOH
N
H O ~~SCH3 X
I
HO iP' \ \ \
O
(X) . X = CH 2 (X) a . X = O
SH
O
H2N.
O
(XI) or S
S~ O /
H.,N \
(XII) More particularly, the prenyltransferase inhibitors may be chosen frorr~ the group constituted by the followüng compounds : the compounds of general formula (GPI), thiazole derivatives such as those described in Patent Application WO 98/00409 (for example the compound of formula (I)), peptidomimetic derivatives of 4-aminobenzoic acid (for example the compounds of formulae (II) and (II)a) or peptidomimetic analogues such as those de;scribed in Patent Application WO 96/21456 (for example the compound of formula (III)), 1-[(2R)-amino-3-mercaptopropyl]-(2S)-(2-mercaptoethyl)-4-(1-naphtoyl)-piperazine-1,2-cyclodisulfide (XII), or pseudodipeptides such as those described in Patent Application WO 95/00497.

_g_ More preferably, the prenyltransferase inhibitors may be chosen from the group constituted by the compounds corresponding to general formula (GPI) shown below R~;

I

(GPI) in which:
R1 represents OH, SH, NH2 or CHZOH;
R2 represents a louver alkyl radical or CORS;
R3 and R4 represent independently a halogen atour or an OH radical;
RS represents a lover alkyl radical, a substituted or non-substituted carbocyclic or heterocyclic arylalkyl or aryl radical or a saturated heterocyclic radical comprisïng 5 to 6 members, said members being chosen from CH2, O, NH
and S; and Y represents CO or CS or is absent;
and compounds corresponding to one of the following formulae:
SH ~S S ~
N
NH..~
H2N~ O NH O~
O
(I) HS
OR

O
(II) . R = CH 3 (II) a . R = H
HS
HZN ,,~~H
NH

H
(III) SH
H2N. %~ O
O
(XI) or s S~
N N
H.,N
(XII) According to a preferred variant of the invention, the prenyltransferase inhibitors used for the invention correspond to general formula (GPI) and can in particular be chosen from the following compounds:
- 1-acetyl-3-(3-chlorophenyl)-5-[(4-chlorophenyl)hydroxy-(1-methyl-1H-imidazolyl)methyl]indole (GPI~);
- 1-(4-morpholinecarbamoyl)-3-(3-chlorophenyl)-5-[(4-chlorophenyl)hydroxy-(1-methyl-1H-imidazolyl)methyl]indole (GPI2);
- 4-(3-chlorophenyl)-6-[(4-chlorophenyl)hydroxy]-(1-methyl-1H-imidazol-5-yl)methyl-2(1H)-quinolinone (GPI3);
- 4-(3-chlorophenyl)-6-[(4-chlorophenyl)hydroxy]-(1-methyl-1H-imidazol-5-yl)methyl-2( 1 H)-quinolinone (GPI4).
According to another pref~rred variant of the invention, the prenyltransferase inhibitors will be one of the compounds of formula (I), (II), (II)a, (III) or (XII):
SH ~S S ~
N
NH.~
H2N~ O NH O~
O
(I) -Il-HS
H~I~f H OR
O
(II) . R = CH 3 (II) a : R = H
HS
H2N ,...H
NH

H
(III) S
O
~' ~i ~ I
H2N w (XII) Among the compounds previously mentioned, one of the following farnesyltransferase inhibitors of formula (I), (II) or (II)a, or (XII) will preferably be used as the prenyltransferase inhibitor:
SH ~ S ~
N
NH
H2N~ O NH O\
O
(I) H
HS N
N
OR

O
(II) . R = CH
(II) a . R = H
S
S~ O /
U
H.,N \
(XII) A particularly preferred compound for use according to the invention is the compound of formula (XII):
O /
H-, \
(XII) The compounds of general formula (GPI) can be prepared according to the methods described in Patent Application PCT WO 97/21701 when Y represents CO or CS, or according to the methods described hereafter when Y is absent. The 1-[(2R)-amino-3-mercaptopropyl)-(2S)-( 2-m ercaptoethyl)-4-( 1-naphthyl)-piperazine-1,2-cyclodisulphide (XII) can be prepared as described in the experimental part.
The compounds of gen.eral formula (GPI) are described in Patent Application US 09/098141 dated l6th June 1998 as well as in a very recently filed subsequent continuation-in-part of this Application, said Application having been the subject of a PCT Patent Application No. US 99/xxxxx not yet published at the date of this Application. Part of the content of this PCT Application is repeated in the experimental part.
The pharmaceutical compositions containing a compound according to the invention can be in solid form, for example powders, granules, tablets, gelatin capsules, liposomes or suppositories. The appropriate solid supports can be, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine and wax.
The pharmaceutical compositions containing a compound of the invention. can also be presented in liquid form, j~or example, solutions, emulsions, suspensions or syrups. The appropriate liquid supports can be, for example, water, organic solvents such as glycerol or glycols, as well as their mixtures, in varying proportions, in water.
The administration of a rnedicament according to the invention can be carried out by topical, oral or parenteral route, by injection (intramuscular, sub-cutaneous, intravenous, etc.), etc. The administration route will of course depend on the type of illness to be treated.
The administration dose envisaged for a medicament according to the invention is comprised between 0.1 rn;g and 10 g according to the type of pathology to be treated.
Unless defined differently, all the technical and scientific ternis used here have the sanie meaning as that normally understood by an ordinary specialist in the field to which this invention belongs. Similarly, all publications, patent applications, patents and all other references mentioned here: are incorporated by way of reference.
The following examples are presented in order to illustrate the above procedures and should in no event be considered to limit the scope of the invention.

EXPERIMENTAL PART
Preparation of certain compounds ased in the inventïon Preparation A:
Compounds of general formula (GPI) in which Y does not exist:
Said compounds are prepared according to the methods described in Patent Application PCT US 99/xxxxx, under priority of a Patent Application US 09/098141 dated l6th June 1998 as well as a subsequent continuation-in-part of this Application.
Part of the preparation protocols for the products of this PCT Application is repeated hereafter.
"" The invention (the subject of PCT Application US 99/xxxxx) relates to the compound of general fornmla (A) R' R R1 R Ra N
R3 1 R6 (' )p R10 (T ~m ( I )n (-r4 ~ R 11 ,, Z____ Ra (A) or a pharmaceutically acceptable sait of said compound, said general formula (A) being such that:

------- represents an optional bond, it being understood that only one of the optional bonds is presented in a compound of general formula (A);
m, n, p, and q are all independently 0 or 1;
each tune T1, T2, T3 and 'T4 are involved, they are chosen from the group composed of CR26R27, S, O, C(O), S(O)2 and NR2g;
X is N-Y, O or S, Y being chosen from the group composed of H, CR14R1sR16, s(~)R17~ s(~)2R18, C(C~W19~ C(~)~20R21, C(s)~22R23~ C(~)~R24~ C(S)~R2s~
S(O)NR29R3o and S(O)2NR31 R~2;
Z is chosen from the group composed of H, hydroxy, alkoxy, aryloxy, cyano, halo, CR14R1sR16, s(O)R17~ s(O)2R18~ C(~)R19~ C(~)~20R21~ C(~)~R24~ C(s)~22R23~
S(O)NR29R3o, C(S)OR2s and S(O)2NR31R32~ it being understood that when the optional bond to Z is present then ~'. is an oxygen or sulphur atour;
each time Rl, R2, R3, 1Z4, R5, Rb, R11, R12, R13~ R14~ Rts~ R16~ R26 Wd R27, are involved, they are each independently chosen from the group composed of H, halo, hydroxy, thio and cyano, or an optionally substituted radical chosen from the group composed of the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, aryl, arylalkyl, alkyloxy, aryloxy, alkylthio, arylthio, alkylamino, arylamino and alkyl caibonyl amino radicals;
or R1 and R2 when they aire in adjacent positions, or R4 and Rs, or R11 and R12 when they are in adjacent positions, form together a divalent radical chosen from the group composed of -O-CH2-O-, -O-CH2-CH2-O-, -O-CH=CH-, -O-CH2-CH2-, -O-CH2-CH2-CH2- and -CK33=CR34-CR3s=CR36-;
R7, R8 and R9 are each ündependently chosen from the group composed of H, halo, amino, cyano, hydroxycarbonyl, or an optionally substituted radical chosen from the group composed of the aryl, alkyl, alkyloxy, alkylthio, aryloxy, arylthio, mono- or di-alkylamino, alkoxycarbonyl, alkyl-S(O)-alkyl, alkyl-S(O)2-alkyl, cyanoarylalkyl and arylalkyl radicals;
Rlo is chosen from the group composed of H, amino, azido, hydroxy, halo, alkyl, substituted alkyl, cyano, hydroxycarbonyl, mono- or di-alkylaminoalkyl, rnono-or di alkylamino, alkyloxy, alkylcarbonylalky~, alkyloxy-carbonylalkyl, carboxyalkyl, cycloalkyl, cycloalkylamino, cycloalkyloxy, imidazolyl, substituted imidazolyl, aminocarbonylalkyl, aryloxy, thio, alkylthio, arylthio, OS(O)2R1H, OC(O)R19, OC(O)NR2oR21, OC(S)NR22R23, OS(O)NR29R30 and OS(O)2NR31R32~
each tune R17, Ri g, R19, R2o, R21~ R22, R23, R24, R2s, R28, R29, R3o~ R31~
R32 and R37 are involved, they are eaclz independently chosen from the group composed of H
and of an optionally substituted radical chosen from the group composed of the alkyl, alkenyl, cycloalkyl, aryl and arylalkyl radicals;

or R2p and RZ1, or Rz2 and R23, or R2ç and R3p, or R31 and R32 form together a divalent radical chosen from the group composed of the -(CHZ)r NR3~-(CHZ)S-, -(CHZ)r O-(CH2)S- and -(CR38R39)t- radicals in which r and s represent independently integers from 1 to 3 and t represents an integer from 2 to 6;
and R33, 1234, R3s~ R3f» R3s and R39 are each independently chosen from the group composed of H, amine, halo, cyano, alkyl, substituted alkyl, aryl, substituted aryl, alkyloxy, aryloxy, alkylth.io, arylthio, mono- or di-alkylamino, arylamine, hydroxy and thio.
The following definitions are used for formula (A) above:
- In the part of the compound of formula (A) in which two optional bonds are indicated, only one of these optional bonds can be present in a compound at a tune. When the optional bond directly attached to the variable Z is present, then Z is an oxygen or sulphur atour.
- The terni "alkyl" refers to non-substituted linear or branched hydrocarbon drains contairllng 1 to 20 carbon atours, and preferably 1 to 7 carbon atours.
The terni "substituted alkyl" refers to an alkyl radical group substituted, for example, by one to four substituents, such as halo, hydroxy, alkoxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amine, alkylamino, arylamine, aralkylamino, disubstituted amines in which the 2 substituents of the amine fonction are chosen from alkyl, aryl or aralkyl;
alkanoylamino, aroylamino, aralkanoylamino, substituted alkanoylamino, substituted arylamine, substituted aralkanoylamino, thiol, alkylthio, arylthio, aralkylthio, alkylthiono, arylthiono, aralkylthiono, alkylsulphonyl, arylsulphonyl, aralkylsulphonyl, sulphonamido, e.g. SOZNH2, substituted sulphonamido, nitre, cyano, carboxy, carbamyl, for example CONH2, substituted carbamyl for example CONf-I alkyl, CONH
aryl, CONH aralkyl or in the case in which there are two substituents on the nitrogen they are chosen from al:kyl, aryl and aralkyl; alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocycles, such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and analogues. When it is indicated that the substituent is itself substituted, it is substituted by an alkyl, alkoxy, aryl or aralkyl radical.
- The terni "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.
- The terni "aryl" refers to monocyclic or bicyclic aromatic groups having 3 to 12 carbon atours and 0 to 2 nitrogen atours and 0 to 1 sulphur atour in their cyclic part such as phenyl, naphthyl, biphenyl, imidazoyl, pyridyl and diphenyl, each being optionally substituted.
- The terni "arylalkyl" refers to an aryl group directly linked by an alkyl group, such as benzyl.

_ 17_ - The tenu "substituted ar~l" refers to an aryl group substituted by, for example, one to five substituenis such as alkyl; substituted alkyl, halo, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, arylalkylamino, arylalkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, ureido, nitro, cyano, carboxy, carboxyalkyl, carbamyl, alkoxycarbonyl, alkylthiono, arylthiono, alkylsulphonyl, sulphonamido, aryloxy and analogues. The substituent cran itself be substituted by hydroxy, alkyl, alkoxy, aryl, substituted aryl, substituted alkyl, or arylalkyl.
- The terni " alkenyl" refc;rs to non substituted linear or branched hydracarbon drains containing 2 to 20 carbon atours, preferably 2 to 15 caubon atours, and more preferentially 2 to 8 carbon atours, having one to four double bonds.
The terni "substituted allcenyl" refers to an alkenyl group substituted by, for example, one to four substituents, such as halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, alkylthiono, alkylsulphonyl, sulphonamido, nitro, cyamo, carboxy, carbamyl, substituted carbamyl, guanidino, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyr-imidyl and analogues.
- The terni "alkynyl" refers to non substituted linear or branched hydrocarbon drains containing 2 to 20 carb~on atours, preferably 2 to 15 carbon atours, and more preferentially 2 to 8 carbon atours, having one to four triple bonds.
The terni "substituted alkynyl" refers to an alkenyl group substituted by one to four substituents, for examplc: a substituent such as halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, alkylthiono, alkylsulphonyl, sulphonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl, guanidïno and heterocyclo, for example imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and analogues.
- The terni "cycloalkyl" refers to optionally substituted saturated hydrocarbon rings or systems of saturated hydrocarbon rings, preferably comprising 1 to 3 rings and 3 to 7 carbon atours per ring, which can themselves be joined with an unsaturated C3-carbocyclic ring. Sach groups include for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl and adamantyl. The substituents include for example one or more alkyl groups as described above, or one or more groups described above as a substituent of an alkyl group.
- The ternis "heterocycle", "heterocyclic" and "heterocyclo" refer to an optionally substituted, saturated or unsaturated, aromatic or non-aromatic cyclic group, which is, for example, a monocyclic system containing 4 to 7 atours, a bicyclic system containing

7 to 11 atours, or a tricyclic system containing 10 to 15 atours, which contains at least one heteroatom in a ring comprising at least one carbon atour. Each ring of the heterocyclic group comprising a heteroatom can contain l, 2 or 3 heteroatoms chosen from nitrogen, oxygen and sulphur, the oxygen and sulphur atours being optionally oxidized and the nitrogen atours being optionally in the form of quaternary cations. The heterocyclic group can be attached to any carbon atour or heteroatom.
- Monocyclic heterocyclic groups include for example pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyra~zolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulphoxide, thiamorpholinyl sulphone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl, thüranyl, triazinyl, and triazolyl, and analogues.
- Bicyclic heterocyclic groups include for example benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, quinolinyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,1-b]pyridinyl) or furo[2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl, benzisoxaxolyl, benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl, benzopyrazolyl, dihydrolbenzofuryl, dihydrobenzothienyl, dihydrobenzathiopyranyl, dihydrobenzothiopyranyl sulphone, dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl, naphthyriclinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl, thienothienyl, and analogues.
- The substituents for the heterocyclic systems include for example one or more alkyl groups as described abovc:, or one or more groups described above as substituent of an alkyl group. Smaller heterocycles such as epoxides and aziridines are also i ncluded.
- The terni "heteroatom" includes oxygen, sulphur and nitrogen.
The compounds of formula (A) can be prepared according to the methods described hereafter or by analogy to these methods:
EXAMPLE 1: ~ -3- 3~-chloroPhen~ll-5 ~j~,4-chlorophenyl)h dy roxy(1-methyl-1H
imidazol-S~llmethyl] indole A solution of butyllithium in hexane (1.6 M, 430 pl) at approximately -78 °C is added dropwise to a solution of' 1-methylimidazole (53 mg) in anhydrous THF (3 ml).
The mixture is agitated at approximately -78 °C for approximately 15 minutes. A solution of chlorotriethylsilane in THF (1.0 M; 660 pl) is added dropwise to the solution. The mixture is brought progressively back to ambient temperature then agitated at ambient temperature for one hour. The mixture is cooled down to approximately -78 °C and a solution of' butyllithium in hexane (1.6 M; 430 ~tl) is added dropwise. The solution is agitated at approximately -78 °C for approxirnately I hour and, over the next 15 minutes, brought back to a temperature of approximately -15 °C. The solution is cooled down to approximately -78 °C. A solution of I-methylsulphonyl-3-(3-chlorophenyl)-5-(4-chlorobenzoyl)indole (95 mg, see preparation 7) in THF (2 ml) is added dropwise.
The mixture is brought progressively back to ambient temperature then agitated for approximately 2 hours. The solution is cooled down to approximately 0 °C and methanol and water are ad~ded. The mixture is agitated for approximately 2 hours. The solution is concentrated by evaporation under reduced pressure. The residue is dissolved in dichlorometh<me (DCM) and washed once with water. The organic phase is dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The crude product is purified by column chromatography, on silica, eluting with a DCMlMeOH mixture 95:5 in order to produce the expected compound (60 mg;
yield 63 %).
R~ 0.20 (silica, DCM/MeOH 9:1). MS (ES) 447.2 (Calc. MM = 447.4).
Alternatively, the compound of Example 1, (t)-3-(3-chlorophenyl)-5-[(4-chlorophenyl)hydroxy(1-m:~ethyl-1H-imidazol-5-yl)methyl]indole can be synthesized according to the procedure: hereafter. A solution of butyllithium in hexane (1.6 M, 430 pl) is added dropwise to a solution of 1-methylimidazole (53 mg) in anhydrous THF (3 ml) at approximately -78 °C. The mixture is agitated at approximately -78 °C for approximately 15 minutes. A solution of chlorotriethylsilane in THF (1.0 M;
660 p.l) is added dropwise to the solution. The mixture is brought progressively to ambient temperature then agitated pat ambient temperature for one hour. The mixture is cooled down to approximately -78 °C and a solution of butyllithium in hexane (1.6 M; 430 pl) is added dropwise. The solution is agitated at approximately -78 °C for approximately 1 hour and, over the next 15 minutes, brought to a temperature of approximately -15 °C.
The solutïon is cooled down to approximately -78 °C. A solution of methylsulphonyl-3-(3-chlorophenyl)-5-(4-chlorobenzoyl)indoline (95 mg, see prEparation 6) in THF (2 nnl) is added dropwise. The mixture is brought progressively to ambient temperature the:n agitated for approximately 2 hours. The solutian is cooled down to approximately 0 °'C and methanol and water are added. The mixture is agitated for approximately 2 hours. The solution is concentrated by evaporation under reduced pressure. The residue is dissolved in dichloromethane (DCM) and washed once with water. The organic phase is dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The crude product is purified by column chromatography, on silica, eluting with a DCM/MeOH mixture 95:5 in order to produce the expected compound.

The enantiomers of the compound of Example 1 can be separated using techniques known to a person skilled i,n the art, such as preparative HPLC on a chiral column.
EXAMPLE 2: ~ -1-methylsulphonyl-3-(3-chlorophenyll-5-(j4-chlorophen~l)hydroxy(1-rr~ethyl-1H imidazol-5-yl)methyl]indole A solution of butyllithium in hexane (1.6 M; 694 ~1) is added dropwise tu a solution of 1-methylimidazole (88 ml;) in anhydrous THF (3 ml) at approximately -78 °C. The mixture is agitated at approximately -78 °C for approximately 15 minutes. A solution of chlorotriethylsilane in THF (1.0 M; 1.08 ml) is added dropwise to the solution. The mixture is brought pro,gressively to ambient temperature then agitated at ambient temperature for one hour. 'The mixture is cooled down to approximately -78 °C and a solution of butyllithium in hexane (1.6 M, 694 pl) is added dropwise. The solution is agitated at approximatel:y -78 °C for approximately 1 hour then brought to a temperature of approximately -15 °C. It is agitated at approximately -15 °C for approximately 15 minutes. The solution is cooled down to approximately -78 °C. A
solution of 1-methylsulphonyl-3-(3-chlorophenyl)-5-(4-chlorobenzoyl)ïndoline (150 mg, see preparation 6) in THF (2 ml) is added dropwise. The mixture is brought progressively to ambient temperature then agitated for approximately 2 hours.
The solution is cooled down to approximately -78 °C. Methanesulphonyl chlaride ( 116 mg) is added dropwise. The solution is brought slowly to ambient temperature and agitated overnight. The solution is cooled down to approximately 0 °C. Water is added and the mixture is agitated for approximately 2 hours. The solution is diluted using DCM and the organic phase is separated and concentrated by evaporation under reduced pressure.
The crude product is purified by column chromatography, on silica, eluting with a CHC13/MeOH mixture 95:5 in order to produce the expected compound in the form of a solid (30 mg; yield 17 %). MS (ES): 525.1 (Calc. MM= 525.5).
Alternatively, the compound of Example 2, can be prepared as follows. A
solution of butyllithium in hexane (1.6 M; 694 pl) is added dropwise to a solution of 1-methylimidozole (88 mg) in anhydrous THF (1.5 ml) at approximately -78 °C . The mixture is agitated at approximately -78 °C for approximately 30 minutes. A solution of chlorotriethylsilane in '1CHF (1.0 M; I .11 ml) is added dropwise to the solution. The mixture is brought progressively to ambient temperature then agitated at ambient temperature for one hour. The mixture is cooled down to approximately -78 °C and a solution of butyllithium in hexane (1.6 M, 694 Vil) is added dropwise. The mixture is agitated at approximately -78 °C for approximately 1 hour then brought to approximately -15 °C for the next 30 minutes. The solution is cooled down to approximately -78 °C. A solution of 1-methylsulphonyl-3-(3-chlorophenyl)-5-(4-chlorobenzoyl)indole (150 mg, see preparation 7) in THF (I ml) is added dropwise.

The mixture is brought progressively to ambient temperature then agitated at ambient temperature for 19 hours. The solution is cooled down to approximately -78 °C.
Methanesulphonyl chloride (163 mg) is added dropwise, followed by düsopropylethylamine (8'7 mg). The solution is brought to ambient temperature over one hour and agitated at ambient temperature for 3 hours. 4 ml of a 1N aqueous solution of HCI and 4 ml of THF are added to the solution. The solution is agitated at 0 °C for 1 hour 30 minul:es. The organic solvent is eliminated by evaporation under reduced pressure. The adueous solution is adjusted to pH 8 by adding a 6N
aqueous solution of KOH at 0 °C. The aqueous solution is extracted twice with DCM. The combined organic phases are washed once with sait water, dried over MgS04, filtered and reduced by evaporation under reduced pressure. The residue is purified by column chromatography, on silïca, eluting with a CH3Cl/MeOH/Et3N mixture, 98:2:0.1.
The expected compound is obtained in the form of a solid (44 mg; yield 25%).
MS(ES) 525.2 (Calc. MM= 525.3).
The enantiomers of the compound of Example 2 car be separated using techniques known to a person skilled in the art, such as preparative HPLC on a chiral column.
Example 3: f -1- rd-dimethylcarbamoyl)-3-(3-chlorophenyll-5-[(4-chlorophenyll )~droxy,~l -methvl-1 H-irnidazol-5-yl)methyl] indole This compound is synthesized in a marner analogous to the first method described for preparing the compound of Example 2, using dimethylcarbamyl chloride instead of the methanesulphonyl chloride. MS (electrospray): 518.2 (Calc. MM: 518.5).
Example 4: ~ -1-methylsulphonyl-3-(3-chlorophenyl)-5-[amino 4-chloro-phenyll(1-methyl-1H imidazol-5-vl)methyll indole Methanesulphonyl chloridle (66 mg) at -78 °C is added dropwise to a solution of (~)-1-methylsulphonyl-3-(3-chlorophenyl)-5-[(4-chloro-phenyl)hydroxy(1-methyl-1H
imidazol-5-yl)methyl] indole (100 mg) and triethylamine (58 mg) in anhydrous DCM (2 ml). The solution is brought slowly to ambient temperature then agitated at ambient temperature for 19 hours. The solution is diluted by adding DCM and washed twice with salt water, dried over MgS04, filtered and reduced by evaporation under reduced pressure. The residue is dissolved in DMF. Sodium azide (124 mg) is then added. The mixture is heated to approximately 60 °C overnight. The solvent is eliminated by evaporation under reduced pressure. The residue is dissolved in water and DCM.
The organic phase is separated, washed once with salt water, dried over MgS04, filtered and reduced by evaporation under reduced pressure. The residue is dissolved in an EtOAc/HOAc mixture 9:1. 200 mg of palladium on carbon (Pd/C 10%) is added. The mixture is agitated under HZ (40 psi) for approximately 3 hours. Thc; solvent is eliminated by evaporation under reduced pressure. The residue is purified by column chromatography, on silica~, eluting with a CH3C1/MeOH/Et3N mixture, 98:2:0.1, in order to produce the expected compound.
Example 21: ~ -1-acet~l-3-(3-chlorophenyll-5-((4-chlorophenyl)hydroxy(1-methyl-1H-imidazol-5-yl)methyllindole This compound is synthesüzed in a marner analogous to the first method described for preparing the compound of Example 2, using acetic anhydride instead of methanesulphonyl chloride;. MS (electrospray): 489.2 (Cale. MM: 489.4).
The following compounds car be prepared by procedures analogous to the procedures detailed in Examples 1 to 4 using the appropriate starting reagents and modifications which are well known to a person skilled in the art. The compounds of Examples 5, 6,

8, 10, 12, 16, 18, 20 and 2:Z car be synthesized in a marner analogous to the compound of Example 4. Examples 7, 9, 11, 15, 17 and 19 car be synthesized in a marner analogous to the compound of Example 2.
PRÉPARATION 1: 2-(3-chlorophenyl)-N-methoxy-N-methyl-acetamide A solution of 3-chlorophenylacetic acid (5.00 g; 29.3 mmol), l -(3-dimethyl aminopropyl)-3-ethylcarbodümide hydrochloride (6.18 g; 32.2 mmol), and 1-hydroxybenzotriazole (HOBt; 4.00 g; 29.3 mmol) in dichloromethane (DCM; 40 ml) is agitated at ambient temp~erature for approximately 10 minutes. The solution is cooled down to approximately 0 "C. N,O-dimethylhydroxylamine hydrochloride (2.86 g;
29.0 mmol) and düsopropylethylamine (DIEA; 3.80 g, 29.3 mmol) are added. The reaction mixture is brought to ambi.ent temperature and agitated for approximately 5 hours. The solution is diluted with 100 ml of DCM and washed with a saturated aqueous solution of NaHC03 (twice), a 1N aqueous solution of HCl (twice) and salt water (twice), dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The liquid obtained is purified by silica column chromatography eluting with an EtOAc/hexane mixture 1:1. The expccted compound is obtained in the form of a colourless liquid (5.60 g, 89%). Rf = 0.44 (silica, EtOAc/hexane 1:1 ).

NMR 1H (300 MHz, CDC13): 7.18-7.34 (m, 4H); 3.76 (S, 2H); 3.66 (S, 3H);
3.22 (S, 3H).
PRÉPARATION 2: 2~3-chloro~phenylLacetaldehyde A suspension of LiAIH~ (1.90 g, 51 mmol) in anhydrous ether (250 ml) is agitated at ambient temperature under a nitrogen atmosphere for approximately 1 hour. The suspension is cooled down to approximately -45 °C. A solution of 2-(3-chlorophenyl) N-methoxy-N-methyl-acetarnide (8.19 g; 38.3 mmol; see Preparation 1) in 10 ml of anhydrous tetrahydrofuran (THF) is added dropwise. The mixture is brought to approximately 0 °C and ag;itated for approximately 3 hours. The solution is then cooled down to approximately -4-'i °C. A solution of KHS04 (13 g) in water (approximately 30 ml) is slowly added to this solution. The resulting mixture is filtered on Cl?LITE. The filtrate is concentrated by c:vaporation under reduced pressure, and the resulting solution is diluted with DCM and washed with a 1 N aqueous solution of HCl (twice), and salt water (twice), dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. 'The expected compound is obtained in the form of a liquid (5.80 g), which is used. immediately in the additional stage without additional purification.
Rf= 0.71 (silica, EtOAc/h~exane 1:3).
PRÉPARATION 3: 3 ~3-~chlorophenyl)indole A solution of 2-(3-chlorophenyl)-acetaldehyde (5.80 g; 37.5 mmol) and phenylhydrazine (6.22 g; 57.5 mmol) in glacial acetic acid (150 ml) is saturated with nitrogen by passing N2 th~rough the solution. The solution is then taken to reflux for approximately 2.5 hours. The solvent is eliminated by evaporation under reduced pressure and the residue o~btained is dissolved in DCM and washed with a 1N
aqueous solution of HCl (twice), a saturated aqueous solution of NaHC03 (twice) and sait water (twice), dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The c;rude product is purified by silica column chromatography eluting with an EtOAc/he:xane mixture 1:6. The expected compound is obt.ained in the form of a reddish oil (5.30 g; 62%). Rf= 0.26 (silica, EtOAc/hexane 1:4) PRÉPARATION 4: 3-~3_--chlorophen~l-indoline 3-(3-chlorophenyl)indole (5.30 g; 23.3 mmol) is dissolved in 50 ml of a 1M
solution of BH3 in THF. The mixture is cooled down to approximately 0 °C.
Trifluoroacetic acid (TFA, 50 ml) is then added slowly. Alter this addition, the solution is agitated for approximately 10 minute:.. A 1M solution of BH3 in THF (40 ml) is added slowly to the solution. The mixture is agitated for approximately 5 minutes then concentrated by evaporation under reduced pressure. The residue is purified by silica column chromatography eluting with an EtOAc/hexane mixture 1:6. The expected compound is obtained in the form of an oil (3.93 g; 74%).
Rf= 0.20 (Silica, EtOAc/hexane 1:4). MS (ES): 229.1 (Calc. MM = 229.7).
PRÉPARATION 5: 1-me hYlsu~honyl-3-(3-chlorophenyl indoline Methanesulphonyl chloride (2.13 g; 18.6 mmol) is added dropwise to a solution of 3-(3-chlorophenyl)indoline (3.8.8 g; 16.9 mmol) and DIEA (2.40 g; 18.6 mmol) i.n DCM (40 ml) at approximately 0 °C. The mixture is agitated for approximately one hour 30 minutes. The solution is; diluted with DCM and washed with a saturated aqueous solution of NaHC03 (twice), a 1N aqueous solution of HCl (twice) and sait water (twice) then dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The crude product is purified by silica column chromatography eluting with an EtOAc/hexane mixture 1:4. The expected compound is obtained in the form of an oil (4.40 g; 85°rb). Rf= 0.41 (silica, EtOAc/hexane 1:2).
NMR ~H (300 MHz, CDC',13): 7.52 (d, 1H); 7,24-7.34 (m, 3H); 7.20 (s, 1H); 7,02-7.14 (m, 3H); 4.59 (t, 1H); 4.38 (t, 1H); 3.87 (dd, 1H); 2.92 (s, 3H).
PRÉPARATION 6: 1-meth~phonyl-3-(3-chlorophenyll-5-(4-chlorobenzovll indoline AlCl3 (7.62 g; 57.2 mmol) is added by portions to a solution of 1-methylsulphonyl-3-(3-chlorophenyl)indoline (4.40 g; 14.3 mmol, see Preparation 5) and 4-chlorobenzoyl chloride (3.25 g; 18.6 nunol) at approximately 0 °C in CS2 (25 ml). A
brown precipitate immediately fonns. The mixture is agitated for approximately 2 hours. 100 ml of cold water containin.g 3 ml of concentrated HCl are added slowly to this mixture.
The solution is diluted with DCM and the organic phase is separated and washed with a 1N aqueous solution of HC',1 (twice), a saturated aqueous solution of NaHC03 (twice) and sait water (twice), then dried over anhydrous MgS04, filtered and concentrated by evaporation under reduced pressure. The crude product is purified by silica column chromatography eluting with an EtOAc/hexane mixture 1:2. The expected compound is obtained in the form of a solid (3.90 g; 61 %). R f = 0.24 (silica, EtOAc/hexane 1:2).
MS (ES): 445.2 (Calc. MM=445.4). NMR ~H (300 MHz, CDC13): 7,67-7.76 (m, 3H);
7,53-7.59 (m, 2H); 7.48 (s, 1H); 7.44 (s, 1H); 7,30-7.32 (m, 2H); 7.20 (m,, 1H); 7,09-7.14 (m, 1H); 4.65 (t, 1H); 4.50 (t, 1H); 3.98 (dd, 1H); 3.02 (s, 3H).
PRÉPARATION 7: 1-methylsul~honyl-3-(3-chlorophenyl)-5-(4-chlorobenzo~)indole A solution of 1-methylsulphonyl-3-(3-chlorophenyl)-5-(4-chlorobenzoyl)indoline (350 mg) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (356 mg) in dioxane (6 ml) is taken to reflux under a nitrogen atmosphere for approximately 6 hours then heated to approximately 95 °C overnight. The solvent is eliminated and the residue purified by silica column chromatography eluting with an EtOAc/hexane mixture, 1:4. The expected compound is obtained in the form of a solid (195 mg; 56%). MS(ES):
443.2 (talc. MM=443.4). Rf=0.38 (silica, EtOAc/hexane, 1:2).
,> >, Preparation B:
1-~(2R)-amino-3-mercaptopropylJ-(2S)-(2-mercaptoethyl)-4-(I-naphthoyl) ~iperazine-1,2-cyclodisulphide (XII):
a) Synthesis of 1-benzyl-3(S)-benzyloxycarbonylmethyl piperazine-2,5-dione:
A cold solution of dicyclohexylcarbodümide (DCC, 7.1 g) in 20 ml of CH2C12 is added to a solution cooled down by an ice bath of the ~3-benzylic ester of BO(:-aspartic acid (10 g), hydroxybenzotriaz,ole (HOBT, 4.2 g) and the ethyl ester of N-benzylglycine (6.4 g) in 80 ml of CH2C12.. The reaction medium is agitated for approximately 1 hour at a temperature of 0 to 5 °C, then overnight at ambient temperature.
The precipitate is eliminated by filtration and the filtrate is evaporated to dryness under reduced pressure.
The residue is dïvided beoween ethyl acetate and water. The organic phase is washed with 100 ml of aqueous NaHC03, then with water, and dried (MgS04). The solvent is eliminated by evaporation to dryness under reduced pressure in order to produce 16 g of a solid. Thin layer chromatography (TLC) (silica gel: CHC13/acetone = 9:1, Rf=
0.55).
The product obtained is th.en treated with 50% trifluoroacetic acid in CHCI_;
(40 ml) for approximately 1 hour and the volatile substances are eliminated by evaporation to dryness under reduced pressure. The residue is divided between ethyl acetate and a saturated aqueous solution of NaHC03. The organic phase is dried (MgS04) and the solvent is eliminated by evaporation to dryness under reduced pressure in order to produce 10 g. TLC (silica gel: CHCI3/acetone = 9:1, Rf= 0.14).
b) Synthesis of 4-benzyl- L-tert-butoxycarbonyl-2(S)-(2-hydroxyethyl) piperazine:
A 50% mineral dispersion of lithium aluminium hydride (LAH) (12.5 g) under a nitrogen atmosphere is ad<ied by portions to a solution of the product of Stage a) (9.73g) in 200 ml of tetrahydrofuran (THF) cooled down by an iced water bath. The reaction medium is maintained a1: reflux overnight. Alter cooling down in an ice bath, a saturated aqueous solution of Na2S04 is added dropwise in order to decompose the excess LAH and the white molasse obtained is eliminated by filtration. The filtrate is evaporated to dryness under reduced pressure and dissolved in dichlorornethane (55 ml), treated with tert-butyll dicarbonate (5.9 g), and agitated for approximately 1 hour.
A saturated aqueous solution of NaHC03 (25 ml) is added and the mixture is agitated for approximately 2 hours. The organic phase is washed [with aJ saturated aqueous solution of sodium chlorid~e and dried (MgS04). Alter evaporation of the solvent, the residue is chromatographed on silica gel ( 160 g) using a CHC13/MeOH rnixaure 19:1 as eluant. The appropriate fractions are collected, and the solvents are elïminated by evaporation to dryness undler reduced pressure in order to produce 10 g of a glass. TLC
(silica gel: CHCl3lMe01-I == 9:1, Rf= 0.56).
c) Synthesis of 1-tert-butoxycarbonyl-2-(S)-(2-hydroxyethyl) piperazine:
The product of Stage b (8.7g) is dissolved in ethanol (35 ml) and treated with Pd(OH)Z
on carbon (0.8 g) and acetic acid (3 ml). Hydrogenation is then carried out under a pressure of 30 p.s.i. ove:rnight. The reaction mixture is filtered and the solvent eliminated by evaporation to dryness under reduced pressure in order to produce the expected product.
d) Synthesis of 1-tert-butoxycarbonyl-2(S)-(2-hydroxyethyl)-4-(1.-naphthoyl) piperazme:
110 ml of a 1N aqueous solution of NaOH then a solution of 1-naphthoyl chloride (5.14 g) in acetonitrile (20 ml) are added to a solution of the product of Stage c) (8.4 g) in acetonitrile (40 ml). Aj~er approximately 3 hours under agitation, a large part of the acetonitrile is eliminated by evaporation under reduced pressure and the residual mixture is extracted with chloroform. The extract is dried (MgS04) and the solvent eliminated by evaporation to dryness under reduced pressure in order to produce 8.12 g of the expected product. TLC (silica gel: CHC13/MeOH = 9:1, Rf = 0.64).
e) Synthesis of 1-tert-butoxycarbonyl-2(S)-(2-triphenylmethylthioethyl)-4-(1-naphthoyl)-piperazine A solution of diethylazodicarboxylate (DEAD, 0.25 g) in 2 ml of THF is added dropwise to a solution of t.riphenylphosphine (0.53 g) in 5 ml of anhydrous THF cooled down by an ice bath. At~er agitation for approximately 30 minutes at a temperature of 0 to 5 °C, a solution of the product of Stage d) (0.4 g) and triphenylmercaptan (0.55 g) in 10 ml of THF is added dropwise. The mixture ïs agitated for approximately 1 hour at a temperature of 0 to 5 °C and for approximately 1 hour at ambient temperature. The solvent is eliminated by evaporation to dryness under reduced pressure and the residue is chromatographed on ~,ilica gel (40 g) using CHC13 as eluant. The appropriate fractions are collected and the solvent is eliminated by evaporation to dryness under reduced pressure in order to produce 420 mg of a pale yellow foam. Mass spectrometry (MS) (Electrospray) 665.2 (643 + 23(sodium)). TLC (silica gel: CHCl3/acetone =

9:1, Rf= 0.53) fJ Synthesis of 2(S)-(2-triphenylmethylthioethyl)-4-(1-naphthoyl) piperazine

10 ml of trifluoroacetic acid (TFA) is added to an agitated solution of the product of Stage e) (2.2 g) in 30 ml of CHZC12. The mixture is agitated for approximately minutes. The volatile substances are eliminated by evaporation to dryness under reduced pressure. The residue is dissolved in CHCl3 (50 ml) and treated with triethylamine in excess (4 ml). The mixture is washed with water, then dried (MgS04) and the volatile substances are eliminated by evaporation to dryness under reduced pressure in order to produce 2.1 g of a pale yellow glass. TLC (silica gel:
CHC13/MeOH = 9:1, Rf = 0.63) g) Synthesis of 1-[2(F;)-N-tert-butoxycarbonylamino-3-triphenylmethylthiopropyl]-2(S)-(2-triphenylmethyl-thioethyl)-4-( 1-naphthoyl)-piperazine 4 g of 4~ molecular sieve then by portions Na(OAc)3BH (1 g) are added over a period of 30 minutes to a solution of the product of Stage f) (0.9g) and 2(R)-N-tert-butoxycarbonylamino-3-triphenylmethylthiopropan-al (1.2 g), prepared according to the procedure of O.P. Goel, et al., (Org. Syn. 1988, 67, 69-75), in CH2C12 (20 ml) containing 1 % of acetic acid. Alter agitation for approximately 2 hours, the mixture is filtered and the filtrate is washed with water, with a 5% aqueous solution of NaHC03, with water, then dried (M:gS04). The solvent is eliminated by evaporation to dryness under reduced pressure, and the residue is chromatographed on silica gel (60 g) using CHCl3 as eluant. The ap~propriate fractions are collected and the solvent is eliminated by evaporation to dryness, under reduced pressure in order to produce 0.6 g of a white foam. TLC (silica gel: CHCI~/acetone = 9:1; Rf= 0.55); MS (Electro Spray) 974.3.
h) Synthesis of 1.-[2(R)-amino-3-mercaptopropyl]-2(S)-2-mercaptoethyl)-4-(1-naphthoyl )-piperazine The product of Stage g) (450 mg) is treated for approximately 30 minutes with 50%
TFA in CHZCIz (10 ml) c;ontaining 1 ml of triethylsilane. The volatile substances are eliminated to dryness by evaporation under reduced pressure. The residue is triturated with ether, filtered, then dried in order to produce 280 mg of 1-[2(R)-amino-3-mercaptopropyl]-2(S)-(2-mercaptoethyl)-4-( 1-naphthoyl)-piperazine. MS
(electrospray) 390.3 i) Cyclizatïon of 1-[2(R)-amino-3-mercaptopropylJ-2(S)-2-mercaptoethyl)-4-(1-naphthoyl)-piperazine in order to form 1-[2(R)-amino-3-mercaptopropyl]-2(S)-2-mercaptoethyl)-4-(1-naphthoyl)-piperazine-1,2-cyclodisulphide (XII) 100 mg of the product of Stage a) is dissolved in 10 ml of an aqueous solution of CH3CN (H20/CH3CN - 7:3), and treated with 3 g of EKATHIOXC~ resin (0.34 mmoles/g). The reaction medium is agitated for approximately 6 hours at ambient temperature. The mixture is then filtered, the resin is washed with an aqueous solution of methanol (water/methanol 1:3), and most of the organic solvent is eliminated by evaporation under reduced pressure, retaining only a small volume. The concentrate is subjected to preparative HPLC chromatography casing 0.1 % aqueous TFA and as the mobile phase. The appropriate fractions are collected and most of the solvents are eliminated by evaporation under reduced pressure, only retaining a small volume.
The concentrate is then lyoyphilized in order to produce the expected product.
Alternatively, a solution of 1-[2(R)-amino-3-mercaptopropyl]-2(S)-(2-mercaptoethyl)-4-(1-naphthoyl)-piperazine in aqueous CH3CN is agitated with air at a pI-1:
comprised between 6 and 8. In both cases, the reaction medium comprises a mixture of cyclized disulphide and its dimer in a proportion of approximately 4 to 1.
PHARMACOLOGICAL STUDY OF THE PRODUCTS OF THE INVENTION
As an example, the effect of treating an MCF-7 human cell fine with the compounds of formulae (I), (II), (XI), (XII), (GPI1) and (GPI2) described previously will be studied.
Electrophoresis will also be carried out on pancreatic cancer cens treated with the compounds of formulae (I) and (III). All these compounds are prepared casing the methods described in the patents mentioned or, in the case of (XII), according to the particular method described in Preparation 1 above.
Procedures Cell line The MCF-7 (human pleural cells, breast cancer) and Mia-PaCa2 (pancreatic cancer) cell fines were acquired from the American Tissue Culture Collection (Rockville, Maryland, USA).

Measurement of tlze intracellular guantity of cyclic AMP for MCF 7 tells MCF-7 tells (2.104) seeded on a 24-well plate are cultured for 5 days in Dulbecco's modified Eagle medium ((Jibco-Brl, Cergy-Pontoise, France) completed with 10%
of foetal calf serum inactivated by heating (Gibco-Brl, Cergy-Pontoise, France ), units/1 of penicillïn and 50 mg/1 streptomycin (Gibco-Brl, Cergy-Pontoise, France), and 2 mM of glutamine (Gibco-Brl, Cergy-Pontoise, France). The culture medium is replaced alter two washes with a medium without serum completed or net completed with the specified agents for a tune indicated in the different figures.
Agents activating the production of cAMP are then added at 37 °C. The reaction is stopped alter 30 minutes by suppressing the medium and rapidly adding 200 pl of a O. l h1 solution of HCI. These extracts are frozen at -80 °C until they are used. The concentration of cAMP is measured using a commercial measurement kit (reference NEK033 from NEN, Les Ulis, France) following the manufacturer's instructions. The radioactivity is determined using a Gamma counter (Gamma Master-1277, LKB, Turku, Finland).
Cell proliferation tests 3000 MCF-7 tells placed in 80 pl of Dulbecco's modified Eagle medium (Gibco-Brl, Cergy-Pontoise, France) completed with 10% foetal calf serum inactivated by heating (Gibco-Brl, Cergy-Pontoise, France ), 50000 units/1 of penicillin and 50 mg/1 streptomycin (Gibco-Brl, Cergy-Pontoise, France), and 2 mM of glutamine (Gibco-Brl, Cergy-Pontoise, France) vvere seeded on a 96-well plate on day 0. The tells were treated on day 1 for 96 hours with increasing concentrations up to 50 pM of each of the compounds to be tested. At the end of this period, the quantification of tell proliferation is evaluated by a colorimetric test based on the cleavage of the tetrazolium sait by mitoclhondrial dehydrogenases in the viable tells leading to the formation of formazan (Boehringer Mannheim, Meylan, France). These tests are carned out in duplicata ~with 8 determinations per concentration tested. For each compound to be tested, thc: values included in the linear part of the sigmoid were used for linear regression analys;is and to estimate the ICSO inhibitory concentration.
Analysis of the subcellular localization of the,Qycomplex by the western-bloc method The Mia-PaCa2 tells (400,000) are seeded in Petri dishes (10 cm diameter) in Dulbecco's modified Eagl.e medium (Gibco-Brl, Cergy-Pontoise, France) completed with 10% foetal calf serurn inactivated by heating (Gibco-Brl, Cergy-Pontoise, France), 50,000 units/1 of penicillin and 50 mg/1 of streptomycin (Gibco-Brl, Cergy-Pontoise, France), and 2 mM of glutamine (Gibco-Brl, Cergy-Pontoise, France). The compounds are added on day 1 at a concentration of 30pM. The tells are harvested by scraping on day 3 alter washing twice with the phosphate buffer (PBS, Gibco-Brl, Cergy-Pontoise, France) at 4°C. The cells are separated by low speed centrifugation (800g, 5 minutes).
The cells are resuspended in an A buffer containing SOmM Hepes, pH 7.5, SmM
MgCl2, 1mM EDTA, pro~tease inhibitors (Cocktail tablets, no. 1836-170, Boehringer Mannheim, Meylan, France). The cells are lysed by three cycles of " freezing in liquid nitrogen - thawing at 4 °C' ". The lysate is centrifuged first at a low speed (1200g, 5 minutes, 4 °C) in order to eliminate the unlysed cens. The supernatant is centrifuged at high speed (200,OOOg, SU minutes, 4 °C) in order to separate the cytosolic fraction and the membrane fraction (pellet). This pellet is resuspended in the A buffer.
The concentration of proteins in each fraction is determined by Bradford colorimetric assay (Bio-Rad protein assay, Ivry, France). The proteins (20pg) are separated by electrophoresis on denatuong polyacrylamide gel (16% Gel-tricine, Novex, Prolabo, Fontenay sous Bois) following the manufacturer's recommendations. The proteins thus separated are transferred onto a nitrocellulose membrane (C-hybond, RPN 2020C, Amersham, Les Ulis, France) in a semi-dry transfer. The (3 protein is detected by the T20 antibody, sc378, Santa-Cruz, TEBU, Le Perray en Yvelines, France), itself recognized by the second antibody coupled with the peroxydase enzyme. The chemoluminescence is obtaïned with the ECL-Plus system (RPN2132, Amersham, Les Ulis, France). The signal is revealed on Kodak BioMax-light autoradiographie films, 237358, Sigma, St Quentin, France). The quantity of chemolurninescence is proportional to the quantity of proteins present.
Eguipment The vaso-intestinal peptide (VIP) and the mevastatin were acquired from Bachem (Voisins le Bretonneux, France) and TEBU (Le Perray en Yvelines, France) respectively. The comp~ounds of formulae (I), (II) and (III) were supplied by Biomeasure Inc. (Milford, MA, USA). All these compounds were used following their manufacturers' recommend~ations.
Results VIP has been presented as an extracellular ligand of a receptor coupled with protein G
which stimulates the synth.esis of cAMP in human breast cancer cens. Figure 1 shows that treatment of MCF-7 human breast cancer cens with VIP increases the intracellular quantity of cAMP in a concentration-dependent manner. A VIP concentration of nM, which offers quasi-optimum production of cAMP, will be used for the following tests. This concentration agrees with data already published concerning the human breast cancer cell fine T47D.

While post-transcriptional prenylation of G proteins is a necessary stage to allow it to fonction, the addition of prenyltransferase inhibitors should alter it significantly. The compound of formula (1) is known to be a powerful specific inhibitor of farnesyltransferases, capable of inhibiting the farnesylation of ras at concentrations in the region of a nanomole. Figure 2 shows that pre-treatment for 24 hours of cens taken from in vitro cultures with the compound of formula (I) inhibited, in a concentration-dependent marner, the accumulation of cAMP stimulated by the VIP.
Almost complete inhibition was obtained at a concentration of 30 pM of the compound of formula (I). These results clearly show that treatment with a specific inhibitor of farnesyltransferases is sufficient to block the transduction of the signal l:he route of which uses the heterotrimeric G proteins as medïators.
Figure 3 shows that treatment for 1 hour with the compound of formula (I) is rot sufficient to modify the re;sponse to the VIP. Despite a slight but reproducible effect alter treatment for 8 hours,, only treatment for 24 hours clearly inhibits the stimulation of the VIP. The action kinetics observed agrees with that expected for a prenyltransferase inhibitor allowing the appearance of non-prenylated forms of sub-units of the G protein.
The ïnhibition of the stinnulation by the VIP is rot restricted to compounds with a structure analogous to that of the compound of formula (I) but tan be extended to other prenyltransferase inhibitors, as shown by the results obtained in particular for the compound of formula (II) which is also a powerful inhibitor of farnesyltransferases capable of inhibiting the farnesylation of H-ras of human tumours at concentrations of the order of a nanomole (Table I).
Moreover, Figure 4 shows that the pretreatment of tells for 24 hours by the compound of formula (I) does rot modify the production of cAMP induced by the direct activator of the adenylate cyclase, forskolin. This shows that the adenylate cyclase itself is rot modified by treatment witlh the compound of formula (I).
Figure 5 shows that the pre-treatment of tells for 24 hours with the compound of formula (I) decreases the production of cAMP induced by the direct activator of the a sub-unit, the choleric toxin. Now this toxin tan only fix itself to the heterotrimeric complex (Warner et al., C'ell Signal, 8, 43-53 (1996)). This therefore suggests that pretreatment with the compound of formula (I) prevents the formation of the heterotrimeric complex.
The ~3y dimeric complex is very stable and tan be identified in a proteinic extract with an antibody directed against the (3 sub-unit. Figure 6 shows the results of a 'western-blot carried out on untreated cens, cens treated for 48 hours with mevastatin (a mevalonate synthesis inhibitor), wïth a geranyl-geranyl transferase inhibitor, the compound of formula (III), or with a pure inhibitor of farnesyltransferase, the compound of formula (I). The proteins of the cyosolic and membrane fractions of the control or treated cens are separated by denaturing electrophoresis on polyacrylamide gel before being transferred onto a semi-solid membrane allowing identification with an antibody (western-blot) directed against all forms of (3 proteins.
The quantity of the (3y c;amplex detected on the cell membrane of untreated cells (control) decreases greatly in cens treated for 48 hours with mevastatin, with a geranyl-geranyl transferase inhibitor (compound of formula (III)) or also with a pure inhibitor of farnesyl-transferase (compound of formula (I)). It can therefore be concluded that alter treatment with a prc~nyltransferase inhibitor, the y sub-unit therefbre no longer plays its rote in anchoring the ~iy complex to the membrane.
Finally, Table Il shows the in vitro values for the proliferation inhibition activities of the compounds of formula.e (I) and (II) with respect to MCF-7 human tumoral cens not carrying a Ras oncogen which has been subject to a mutation.

Compound Dose (pM) Inhibition of n the G
rotein (I) 10 52 3 II) 100 69 2 XI) 30 27 5 (XII 10 53 2 (GPI1 ) 10 22 1 (GPI2) 10 44 3 Table I
Effects of compounds incubated for 24 hours on the production of cyclic AMP
stimulated by VIP in MC.'F7 cells.
The cells are incubated for 24 hours in the presence of the tested compounds at the doses indicated and then stimulated by 10-g M of VIP. The quantification of cyclic AMP is determined by radioimmunoassay.
(~om ounds ICso (NM) Compound 19.4 (I) Com ound 35.8 II

Table II
Proliferation inhibition of the compounds of formulae (I) and (II) with respect to MCF-7 human tumoral tells not carrying a Ras oncogen having been subjected to a mutation.

Claims

Revendications 1. Utilisation d'inhibiteurs de prényltransférases pour priparer un médicament destiné à
bloquer ta transduction des signaux utilisant les proteines G
hétérotrimériques comme médiateurs.

2. Utilisation d'inhibiteurs de prényltransférases pour préparer un médicament destiné à
traiter les pathologies qui résultent de la fixation membranaire de ta protéine G
hétérotrimérique, à l'exception des maladies prolifcrativcs bénignes ou malignes et des infections virales.

3. Utilisation selon la revendication 1 ou 2, caractérisée en ce que les inhibiteurs de prényltransférases sont des inhibiteurs de farnésyltanstérases.

4. Utilisation selon la revendication 1 ou 2, caractérisée en ce que les inhibiteurs de prényltransférases sont choisis parmi un groupe constitué des composés suivants : des dérivés de thiazoles, des dérivés peptidomimétiques de l'acide 4-aminobenzoïque, des dérivés de quinolinone, des amides tricycliques, des polyacides, des dérivés peptidomimétiques de pipéridines, des dérivés peptidomimétiques de benzodiazépines, pics dérives tripeptidiques d'acides phosphoniques ou phosphiniques, des pseudodipeptides basés sur une strucaure N-carbonylpyrazine, et des analogues ~étrapeptidiques du type Cys-A1-A2-A3 où Cys est une Cystéine, A1 et A2 sont des acides aminés aliphatiques et A3 un acide aminé quelconque 5. Utilisation selon la revendication 4, caractcricée cn ce que les inhibiteurs de prényltranaférases sont choisis parmi un groupe constitué des composés correspondant à
formule générale (GPi) représentée ci-dessous dans laquelle:
R1 représente OH, SH ou NH Z;

R2 représente un radical alkyle inférieur ou COR5 ;

R3 et R4 représentent indépendamment un atome halogéne ou un radical OH ;
R5 représente un radical alkyle inférieur, un radical arylalkyle ou aryle carbocyclique ou hétérocyclique substitué ou non substitué ou un radical hétérocyclique saturé comptant de 5 à 6 chaînons, lesdits chaînons étant choisis parmi CH2, O, NH et S ; et y représente CO ou CS ou est absent;

et des composés correspondant aux formules suivantes.

6. Utilisation selon la revendication 5, caractérisée en ce que les inhibiteurs de prényltransférases sont des composes de formule générale (GPI).
7. Utilisation selon la revendication 6, caractérisée en ce que les inhibiteurs de prényltransférases sont choisis parmi un groupe constitué des composés suivants:
1-acétyl-3-(3-chlorophényl)-5-[(4-chlorophényl)hydroxy-(I-méthyl-1H-imidazolyl)méthyl]indole (GPI1);

1-(4-morpholinecarbamoyl)-3-(3-chlorophenyl)-5-[(4-chlorophényl)hydroxy-(1-méthyl-1H-imidazolyl)méthyl]indole (GPI2);
4-(3-chlorophényl)-6-[(4-chlorophényl)hydroxy]-(1-méthyl-1H-imidazol-5-yl)méthyl-2(1H)-quinolinone (GPI3);
- 4-(3-chlorophényl)-6-[(4-chlorophényl)hydroxy]-(1-méthyl-1H-imidazol-5-yl)méthyl-2(1H)-quinolinone (GPI4).
8. Utilisation selon la revendication 5, caractérisée en ce que l'inhibiteur de prényltransférases est choisi parmi le groupe constitué des composés de formule (I), (II), (II)bis, (III) et (XII):
2. Utilisation selon la revendication 8, caractérisée en ce que l'inhibiteur de prényltransférases est le composé de formule générale (XII) 10. Utilisation selon l'une des revendications 1 à 9, caractérisée en ce que la pathologie à
traiter est choisie parmi des pathologies liées aux fonctions biologiques ou désordres suivants: odorat, goût, perception de la lumière, neurotransmission, neurodégénérescence, fonctionnement des glandes endocrines et exocrines, régulation autocrine et paracrine, tension artérielle, embryogénèse, fonctions immunologiques, diabète et obésité.

11. Utilisation selon l'une des revendications 1 à 9, caractérisée en ce que la pathologie à
traiter est choisie parmi les pathologies suivantes le choléra, le Syndrome immuno-Déficitaire Acquis (SIDA), la diarrhée du voyageur et la puberté
précoce familiale masculine.