WO2000002558A1

WO2000002558A1 - Use of prenyltransferase inhibitors for preparing a medicine for treating pathologies resulting from heterotrimeric g protein membrane fixation

Info

Publication number: WO2000002558A1
Application number: PCT/FR1999/001611
Authority: WO
Inventors: Grégoire Prevost; Marie-Odile Lonchampt
Original assignee: Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.)
Priority date: 1998-07-08
Filing date: 1999-07-05
Publication date: 2000-01-20
Also published as: NO20010030L; NO20010030D0; EP1094810A1; JP2002520284A; CA2337261A1; AU4622499A; FR2780892A1; FR2780892B1

Abstract

The invention concerns the use of prenyltransferase inhibitors for preparing a medicine for treating pathologies resulting from prenylation of the η subunit of G protein. Said diseases comprise in particular diseases related to the following biological functions or disorders: smell, taste, light perception, neurotransmission, neurodegeneration, endocrine and exocrine gland functioning, autocrine and paracrine regulation, blood pressure, embryogenesis, viral infection, immunological functions, diabetes and obesity.

Description

USE OF PRENYLTRANSFERASE INHIBITORS FOR THE PREPARATION OF A MEDICINAL PRODUCT FOR TREATING CONDITIONS RESULTING FROM MEMBRANE FIXATION OF HETEROTRIMERIC PROTEIN

The present invention relates to the use of prenyltransferase inhibitors to prepare a medicament intended to treat pathologies which result from the membrane fixation of heterotimeric G protein. These diseases include in particular diseases linked to the following biological functions or disorders: smell, taste, perception of light, neurotransmission, neurodegeneration, functioning of the endocrine and exocrine glands, autocrine and paracrine regulation, blood pressure, embryogenesis, viral infection, functions immunology, diabetes and obesity.

The heterotrimeric G protein coupled to receptors with 7 trans-membrane domains is a mediator of extracellular information towards the interior of the cell. Several intracellular effectors of protein G are identified such as adenylate cyclase, phospholipase C or even ion channels (cf. Gilman, A.G., Biosci. Rep. 15, 65-97 (1995)).

The activity of adenylate cyclase is modulated by the various G proteins (Gs, Gi, Gq, Go) which thus regulate the biosynthesis of cyclic AMP (cAMP). So we know that for

15 modulate adenylate cyclase, it is necessary that the protein G is transiently in a heterotrimeric form, form in which the monomer constituted by an α subunit is associated with the dimer constituted by the β and γ subunits. It is also known that for protein G to be functional, it must be fixed by its γ subunit to the membrane, this fixing being possible when the γ subunit is prenylated. It is

20 only in this situation that the ligand, outside the cell, can, via the receptors with seven transmembrane domains, activate the α subunit of the protein G. The α subunit may after dissociation modulate the adenylate cyclase and modulate cAMP production.

The harmful effects of an abnormal level of cAMP are also known and take place in particular at the level of the following biological functions: smell, taste, perception of light, neurotransmission, neurodegeneration, functioning of the endocrine and exocrine glands, autocrine and paracrine regulation , blood pressure, embryogenesis, benign cell proliferation, oncogenesis, viral infection, immunological functions, 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 and coll., Cancer Re s. 55, 5302-5309 (1997)). The usefulness of prenyltransferase inhibitors in this type of treatment would come from their action which would prevent 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 Applicants have now discovered that the inhibitors of prenyltransferases can also be used to modulate the action of the heterotrimeric G protein, and thus treat all kinds of diseases linked to it.

The invention therefore relates first of all to the use of prenyltransferase inhibitors for preparing a medicament intended for treating pathologies which result from the membrane fixation of the heterotrimeric G protein. In particular, it relates to the use of said inhibitors for preparing medicaments intended for treating diseases linked to the following biological functions or disorders: smell, taste, perception of light, neurotransmission, neurodegeneration, functioning of the endocrine and exocrine glands, autocrine regulation and paracrine, blood pressure, embryogenesis, viral infection, immunological functions, diabetes and obesity.

More particularly, the invention relates to the use of prenyltransferase inhibitors to prepare a medicament intended to treat cholera, Acquired Immune Deficiency Syndrome (AIDS), traveller's diarrhea and male familial precocious puberty.

Among the prenyltransferase inhibitors which can be used for the invention, mention may in particular be made of those chosen from a group consisting of the following compounds: the compounds of general formula (GPI) defined below (in particular those described in patent application WO 97 / 21701, as the compound of formula (IV) defined below, and those described in patent application WO 97/16443, such as for example the compound of formula (VI) defined below), thiazole derivatives such as those described in patent application WO 98/00409 (for example the compound of formula (I) defined below), peptidomimetic derivatives of 4-aminobenzoic acid (for example the compounds of formulas (II) and (H) bis defined below) or else peptidomimetic analogs such as those described in patent application WO 96/21456 (for example the compound of formula (III) defined below), tricyclic amides such as those described in patent application WO 97/24378 (for example the compound of formula (V) defined below), polyacids such as those described in patent application WO 97/17321 (for example the compound of formula (VII) defined below), peptidomimetic derivatives of piperidines such as those described in the patent application WO 97/18813 (for example the compound of formula (VIII) defined below), peptidomimetic derivatives of benzodiazepines such as those described in patent US 5,532,359 (for example the compounds of formulas (IX) and (1X) bis defined below), tripeptide derivatives of phosphonic or phosphinic acids such as those described in US Patents 5,523,430 and 5,510,510 (for example the compounds of formulas (X) and (X) bis defined below), 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 below), and tetrapeptide analogs of the Cys-Al-A2-A3 type where Cys is a Cysteine , A1 and A2 are aliphatic amino acids and A3 any amino acid (such analogs being described for example in US Patent 5,627,202).

Preferably, the prenyltransferase inhibitors can be chosen from the group consisting of compounds corresponding to the general formula (GPI):

(GPI)

in which :

Ri represents OH, SH, NH ₂ or CH ₂ OH;

R ₂ represents a lower alkyl radical or COR ₅ ;

R ₃ and R ₄ independently represent a halogen atom or an OH radical;

R ₅ represents a lower alkyl radical, a substituted or unsubstituted carbocyclic or heterocyclic arylalkyl or aryl radical or a saturated heterocyclic radical having 5 to 6 members, said links being chosen from CH ₂ , O, NH and S; and

Y represents CO or CS or is absent; or one of the following formulas:

(I)

(ll) bis: RH

(V)

(IX): R = CH ₂ SCH ₃ (IX) bis: R = iPr

(X): X = CH ₂ (X) bis: X = O

(XII)

We can more particularly select the prenyltransferase inhibitors chosen from a group consisting of the following compounds: derivatives 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 formulas (II) and (ïl) bis), peptidomimetic analogs such as those described in patent application WO 96/21456 (for example the compound of formula (III) defined below), 1 - [(2R) - amino-3-mercaptopropyl] - (2S) - (2-mercaptoethyl) -4- (1-naphthoyl) -piperazine-1, 2-cyclodisulfide (XII), or also pseudodipeptides based on an N-carbonylpyrazine structure such as those described in patent application WO 95/00497 (for example the compound of formula (XI) defined). More preferably, the prenyltransferase inhibitors can be chosen from the group consisting of compounds corresponding to the general formula (GPI) represented below:

(GPI)

in which :

R _! represents OH, SH, NH ₂ or CH OH;

R represents a lower alkyl radical or COR ₅ ;

R ₃ and R ₄ independently represent a halogen atom or an OH radical;

Y represents CO or CS or is absent;

and compounds corresponding to one of the following formulas:

(I)

(XII) According to a preferred variant of the invention, the prenyltransferase inhibitors used for the invention will correspond to the general formula (GPI) and may in particular be chosen from the following compounds:

- 1 -acetyl-3- (3-chlorophenyl) -5- [(4-chlorophenyl) hydroxy- (1 -me thy 1- 1 H-imidazolyl) methyl] indole (GPIi);

- 1- (4-mo holinecarbamoyl) -3- (3-chlorophenyl) -5 - [(4-chlorophenyl) hydroxy- (1-methyl-1H-imidazolyl) methyl] indole (GPI ₂ );

- 4- (3-chlorophenyl) -6 - [(4-chlorophenyl) hydroxy] - (1-methyl-1H-imidazol-5-yl) methyl- 2 (1H) -quinolinone (GPI ₃ );

- 4- (3-cr-lorophenyl) -6 - [(4-chlorophenyl) hydroxy] - (1-methyl-1H-imidazol-5-yl) methyl- 2 (1H) -quinolinone (GPI ₄ ).

According to another preferred variant of the invention, the prenyltransferase inhibitors will be one of the compounds of formula (I), (II), (ïl) bis, (III) or (XII):

(I)

(XII)

Among the compounds mentioned above, it will be preferred to use as inhibitor of prenyltransferases one of the famesyltransferase inhibitors of formula (I), (II) or (ll) bis, or (XII):

(I)

(ll) bis: R = H

A compound particularly preferred for use according to the invention is the compound of formula (XII):

(XII)

The compounds of general formula (GPI) can be prepared according to the methods described in PCT patent application WO 97/21701 when Y represents CO or CS, or according to the methods described below when Y is absent. 1 - [(2R) -amino- 3-mercaptopropyl] - (2S) - (2-mercaptoethyl) -4- (1-naphthoyl) -piperazine- 1, 2-cyclodisulfide (XII) can be prepared as described in Experimental part.

The compounds of general formula (GPI) are described in patent application US 09/098141 dated June 16, 1998 as well as in a subsequent continuation-in-part of this application filed very recently, said application having been the subject of 'an international patent application No. PCT / US99 / 13303 not yet published on the date of this application. Part of the content of this PCT application is included in the experimental part. The pharmaceutical compositions comprising a compound of the invention can be in the form of solids, for example powders, granules, tablets, capsules, liposomes or suppositories. Suitable solid carriers can be, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, carboxymethyl cellulose sodium, polyvinylpyrrolidine and wax.

The pharmaceutical compositions comprising a compound of the invention can also be presented in liquid form, for example, solutions, emulsions, suspensions or syrups. Suitable liquid carriers 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 medicament according to the invention can be done by topical, oral, parenteral route, by injection (intramuscular, subcutaneous, intravenous, etc.), etc. The route of administration will of course depend on the type of disease to be treated.

The administration dose envisaged for a medicament according to the invention is between 0.1 mg and 10 g depending on the type of pathology to be treated.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as that commonly understood by an ordinary specialist in the field to which this invention belongs. Likewise, all publications, patent applications, all patents and all other references mentioned herein are incorporated by reference.

The following examples are presented to illustrate the above procedures and should in no way be taken as limiting the scope of the invention.

EXPERIMENTAL PART

Preparation of certain compounds used in the invention Preparation A:

Compounds of general formula (GPI) in which Y does not exist:

Said compounds are prepared according to the methods described in international patent application No. PCT / US99 / 13303, under priority of a patent application US 09/098141 dated June 16, 1998. Part of the protocols for preparing the products of this PCT application is repeated below. The invention (subject of PCT / US99 / 13303) relates to the compound of general formula (A)

(AT)

or a pharmaceutically acceptable salt 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 present in a compound of general formula (A);

m, n, p, and q are all independently 0 or 1;

Tj, T, T ₃ and T ₄ each time they occur are chosen from the group consisting of CR ₂₆ R ₂₇ , S, O, C (O), S (O) ₂ and NR ₂₈ ;

X is NY, O or S, Y being chosen from the group consisting of H, CR ₁₄ R ₁₅ R ₁₆ , S (O) R ₁₇ , S (O) 2R ₁₈ , C (O) R ₁₉ , C (O) NR ₂₀ R ₂ ι, C (S) NR ₂₂ R ₂ 3, C (O) OR ₂₄ , C (S) OR ₂₅ , S (O) NR ₂₉ R ₃ 0 and S (O) ₂ NR ₃₁ R ₃₂ ;

Z is chosen from the group consisting of H, hydroxy, alkoxy, aryloxy, cyano, halo, CR .4R 15R 16, S (O) R ₁₇₎ S (O) ₂ R _{1 8} , C (O) R ₁₉ , C (O) NR ₂₀ R ₂ ι, C (O) OR ₂₄ , C (S) NR ₂ 2R23, S (O) NR ₂₉ R ₃ o, C (S) OR ₂ s and S (O) ₂ NR ₃₁ R32 , it being understood that when the optional bond to Z is present then Z is an oxygen or sulfur atom;

Ri, R ₂ , R ₃ , R ₄ , R5, Rό. Rπ, R12. R13. R14. Ri5, Rie. ^R 26 and R ₂₇ , each time they occur, are each independently chosen from the group consisting of H, halo, hydroxy, thio and cyano, or an optionally substituted radical chosen from the group composed of alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, aryl, arylalkyl, alkyloxy, aryloxy, alkylthio, arylthio, alkylamino, arylamino and alkyl carbonyl amino radicals;

or Ri and R ₂ when they are in adjacent positions, or R ₄ and R ₅ , or Rπ and R ₁₂ when they are in adjacent positions, together form a bivalent radical chosen from the group consisting of -O-CH ₂ -O-, -O-CH ₂ -CH ₂ -O-, -O-CH = CH-, -O-CH ₂ - CH ₂ -, -O-CH2-CH2-CH2- and -CR ₃₃ = CR ₃₄ -CR3 ₅ = CR ₃ 6-;

R ₇ , R ₈ and R ₉ are each independently chosen from the group consisting of H, halo, amino, cyano, hydroxycarbonyl, or an optionally substituted radical chosen from the group consisting of aryl, alkyl, alkyloxy, alkylthio, aryloxy, arylthio radicals , mono- or di-alkylamino, alkoxycarbonyl, alkyl-S (O) -alkyl, alkyl-S (O) ₂ -alkyl, cyanoarylalkyl, and arylalkyl;

Rio is selected from the group consisting of H, amino, azido, hydroxy, halo, alkyl, substituted alkyl, cyano, hydroxycarbonyl, mono- or di-alkylaminoalkyl, mono- or di-alkylamino, alkyloxy, alkylcarbonylalkyl, alkyloxy-carbonylalkyl, carboxyalkyl , cycloalkyle, cycloalkylamino, cycloalkyloxy, imidazolyle, substituted imidazolyle, aminocarbonylalkyle, aryloxy, thio, alkylthio, arylthio, OS (O) ₂ Rι ₈ , OC (O) Ri ₉ , OC (O) NR ₂ oR ₂ ι, OC (S ) NR ₂₂ R23, OS (O) NR ₂₉ R ₃ o and OS (O) 2NR ₃₁ R ₃ 2;

Rπ, Ris, R19, R ₂₀ , R21, R22. R23. R24 »R25, R28> R29, R30> R31. ^R 32 and R37, each time they are involved, are each independently chosen from the group consisting of H and an optionally substituted radical chosen from the group consisting of alkyl, alkenyl, cycloalkyl, aryl and arylalkyl radicals;

or R ₂₀ and R ₂₁ , or R ₂₂ and R ₂₃ , or R ₂₉ and R ₃₀ , or R ₃₁ and R ₃₂ together form a bivalent radical chosen from the group consisting of the radicals - (CH ₂ ) r-NR ₃₇ - ( CH ₂ ) _s -, - (CH ₂ ) _r - O- (CH ₂ ) _s - and - (CR ₃₈ R ₃₉ ) _t -, in which r and s independently represent integers from 1 to 3 and t represents an integer from 2 to 6;

and R ₃₃ , R ₃₄ , R ₃₅ , R ₃₆ , R ₃₈ and R ₃₉ are each independently selected from the group consisting of H, amino, halo, cyano, alkyl, substituted alkyl, aryl, substituted aryl, alkyloxy, aryloxy, alkylthio , arylthio, mono- or di-alkylamino, arylamino, 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 at a time in a compound. When the optional bond directly attached to the variable Z is present, then Z is an oxygen or sulfur atom.

- The term "alkyl" refers to linear or branched unsubstituted hydrocarbon chains having 1 to 20 carbon atoms, and preferably 1 to 7 carbon atoms.

- The term "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, amino, alkylamino, arylamino, aralkylamino, disubstituted amines in which the 2 substituents of the amino function are chosen from alkyl, aryl or aralkyl; alkanoylamino, aroylamino, aralkanoylamino, alkanoylamino, substituted arylamino, substituted aralkanoylamino substituted thiol, alkylthio, arylthio, aralkylthio, alkylthiono, arylthiono, aralkylthiono, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfonamido, eg SO ₂ NH _2, substituted sulfonamido, nitro, cyano, carboxy, carbamyl, for example CONH ₂ , substituted carbamyl for example CONH alkyl, CONH aryl, CONH aralkyl or in the cases in which there are two substituents on the nitrogen these are chosen from alkyl, aryl and aralkyl; alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocycles, such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like. When it is indicated that the substituent is itself substituted, it will be by an alkyl, alkoxy, aryl or aralkyl radical.

- The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

- The term "aryl" refers to monocyclic or bicyclic aromatic groups having from 3 to 12 carbon atoms and from 0 to 2 nitrogen atoms and from 0 to 1 sulfur atom in their cyclic part such as phenyl, naphthyl, biphenyl , imidazoyl, pyridyl and diphenyl, each of which may be substituted.

- The term "arylalkyl" refers to an aryl group directly linked by an alkyl group, such as benzyl.

- The term "substituted aryl" refers to an aryl group substituted by, for example, one to five substituents such as alkyl; substituted alkyl, halo, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, arylalkylamino, arylalkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, ureido, nitro, cyano, carboxy, carboxythoxy, carboxyalkyl, carboxyalkyl, carboxyalkyl, carboxyalkyl , alkylsulfonyl, sulfonamido, aryloxy and the like. The substituent may itself be substituted by hydroxy, alkyl, alkoxy, aryl, substituted aryl, substituted alkyl, or arylalkyl. - The term "alkenyl" refers to unsubstituted linear or branched hydrocarbon chains having 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and more preferably 2 to 8 carbon atoms, having one to four double bonds.

- The term "substituted alkenyl" 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 , alkylsulfonyl, sulfonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl, guanidino, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl pyridyl, pyrimidyl and the like.

- The term "alkynyl" refers to unsubstituted linear or branched hydrocarbon chains having 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and more preferably 2 to 8 carbon atoms, having one to four triple bonds.

- The term "substituted alkynyl" refers to an alkenyl group substituted by one to four substituents, for example, a substituent, such as, halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, thiol, alkylthio , alkylthiono, alkylsulfonyl, sulfonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl, guanidino and heterocyclo, for example imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like.

- The term "cycloalkyl" refers to optionally substituted saturated hydrocarbon rings or saturated hydrocarbon ring systems, preferably comprising from 1 to 3 rings and from 3 to 7 carbon atoms per ring which can themselves be fused with a unsaturated C ₃ -C ₇ carbocyclic ring. Such groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyle, cyclododecyl and adamantyl. The substituents include, by way of example, one or more of an alkyl group as described above, or one or more of a group described above as a substituent of an alkyl group.

- The terms "heterocycle", "heterocyclic" and "heterocyclo" refer to an optionally substituted, aromatic or non-aromatic and saturated or unsaturated cyclic group, which is, for example, a monocyclic system having 4 to 7 atoms, a a bicyclic system having 7 to 11 atoms, or a tricyclic system having 10 to 15 atoms, which has at least one heteroatom in a ring comprising at least one carbon atom. Each cycle of the heterocyclic group comprising a heteroatom can comprise 1, 2 or 3 heteroatoms chosen from nitrogen, oxygen and sulfur, oxygen and sulfur atoms which can optionally be oxidized and the nitrogen atoms which can optionally be in the form of quaternary cations. The heterocyclic group can be attached to any carbon atom or hetero atom.

- Monocyclic heterocyclic groups include, for example, pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiazolyl, thiazadylyl, thiazadyl) , thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl, thiamorpholinyl, thiamorpholinyl and tetrahydro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl, thiiranyl, triazinyl, and triazolyl, and the like.

- Bicyclic heterocyclic groups include, for example, benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, quinolinyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzolurinyl, benzofuryl, chromonininyl, quinolurinyl, quinofuryl , furopyridinyl (such as furo [2,3-c] pyridinyl, furo [3, lb] pyridinyl) or furo [2,3-b] pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4-oxo -quinazolinyle), benzisothiazolyl, benzisoxaxolyle, benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl, benzpyrazolyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolinyl, thienofuryl , thienopyridyl, thienothienyl, and the like.

- The substituents for heterocyclic systems include, for example, one or more of an alkyl group as described above, or one or more of a group described above as a substituent of an alkyl group. Also included are smaller heterocycles, such as epoxides and aziridines.

- The term "heteroatom" includes oxygen, sulfur and nitrogen.

The compounds of formula (A) can be prepared according to the methods described below or by analogy to these methods: Example 1: (±) -3- (3-chlorophenyl) -5-r (4-chlorophenyl) hvdroxy (1-methyl-1H- imidazol-5-yllmethyll indole

To a solution of 1-methylimidazole (53 mg) in anhydrous THF (3 ml) is added dropwise a solution of butyllithium in hexane (1.6 M, 430 μl) at approximately -78 ° C. The mixture is stirred at about -78 ° C for about 15 minutes. To the solution is added dropwise a solution of chlorotriethylsilane in THF (1.0 M; 660 μl). The mixture is gradually brought to room temperature and then stirred at room temperature for one hour. The mixture is cooled to approximately -78 ° C and a solution of butyllithium in hexane (1.6 M; 430 μl) is added dropwise. The solution is stirred at approximately -78 ° C for approximately 1 hour and, over the next 15 minutes, brought to a temperature of approximately -15 ° C. The solution is cooled to approximately -78 ° C. A solution of 1 -methylsulfonyl-3- (3-chlorophenyl) -5- (4-chlorobenzoyl) indole (95 mg, see preparation 7) in THF (2 ml) is added dropwise. The mixture is gradually brought to room temperature and then stirred for about 2 hours. The solution is cooled to about 0 ° C and methanol and water are added. The mixture is stirred 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 MgSθ ₄ , filtered and concentrated by evaporation under reduced pressure. The crude product is purified by column chromatography on silica, eluting with a DCM / MeOH 95: 5 mixture to give the expected compound (60 mg; yield 63%). R _f = 0.20 (silica, DCM / MeOH 9: 1). MS (ES) 447.2 (Cale. MM = 447.4).

Alternatively, the compound of Example 1, the (±) -3- (3-chlorophenyl) - 5 - [(4-chlorophenyl) hydroxy (1-methyl-1H-imidazol-5-yl) methyl] indole can be synthesized according to the procedure below. To a solution of 1-methylimidazole (53 mg) in anhydrous THF (3 ml) at approximately -78 ° C is added dropwise a solution of butyllithium in hexane (1.6 M, 430 μl). The mixture is stirred at about -78 ° C for about 15 minutes. To the solution is added dropwise a solution of chlorotriethylsilane in THF (1.0 M; 660 μl). The mixture is gradually brought to room temperature and then stirred at room temperature for one hour. The mixture is cooled to approximately -78 ° C and a solution of butyllithium in hexane (1.6 M; 430 μl) is added dropwise. The solution is stirred at approximately -78 ° C for approximately 1 hour and, over the next 15 minutes, brought to a temperature of approximately -15 ° C. The solution is cooled to approximately -78 ° C. A solution of 1 -methylsulfonyl-3- (3-chlorophenyl) -5- (4-chlorobenzoyl) indoline (95 mg, see preparation 6) in THF (2 ml) is added dropwise. The mixture is brought back gradually at room temperature and then stirred for approximately 2 hours. The solution is cooled to about 0 ° C and methanol and water are added. The mixture is stirred 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 MgSO ₄ , filtered and concentrated by evaporation under reduced pressure. The crude product is purified by column chromatography on silica, eluting with a DCM / MeOH 95: 5 mixture to give the expected compound.

The enantiomers of the compound of Example 1 can be separated using techniques known to those skilled in the art, such as preparative HPLC on a chiral column.

Example 2: (+) - 1 -methylsulfonyl-3- (3-chlorophenyl V

5-r (4-chlorophenyl) hyolroxy -methyl-1H-irnidazol-5-yl) methyl1indole

To a solution of 1-methylimidazole (88 mg) in anhydrous THF (3 ml) at approximately -78 ° C is added dropwise a solution of butyllithium in hexane (1.6 M; 694 μl). The mixture is stirred at about -78 ° C for about 15 minutes. To the solution is added dropwise a solution of chlorotriethylsilane in THF (1.0 M; 1.08 ml). The mixture is gradually brought to room temperature and then stirred at room temperature for one hour. The mixture is cooled to approximately -78 ° C and a solution of butyllithium in hexane (1.6 M, 694 μl) is added dropwise. The solution is stirred at approximately -78 ° C for approximately 1 hour and then brought to a temperature of approximately -15 ° C. It is stirred at approximately -15 ° C for approximately 15 minutes. The solution is cooled to approximately -78 ° C. A solution of 1-methylsulfonyl-3- (3-chlorophenyl) -5- (4-chlorobenzoyl) indoline (150 mg, see preparation 6) in THF (2 ml) is added drop by drop. The mixture is gradually brought to room temperature and then stirred for about 2 hours. The solution is cooled to about -78 ° C. Methanesulfonyl chloride (116 mg) is added dropwise. The solution is brought slowly to room temperature and stirred overnight. The solution is cooled to about 0 ° C. Water is added and the mixture is stirred for about 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 CHCl ₃ / MeOH 95: 5 mixture to give the expected compound in the form of a solid (30 mg; yield 17%). MS (ES): 525.1 (Cale. MM = 525.5). Alternatively, the compound of Example 2 can be prepared as follows. To a solution of 1-methylimidozole (88 mg) in anhydrous THF (1.5 ml) at about -78 ° C is added dropwise a solution of butyllithium in hexane (1.6 M; 694 μl) . The mixture is stirred at about -78 ° C for about 30 minutes. To the solution is added dropwise a solution of chlorotriethylsilane in THF (1.0 M; 1.11 ml). The mixture is gradually brought to room temperature and then stirred at room temperature for one hour. The mixture is cooled to approximately -78 ° C and a solution of butyllithium in hexane (1.6 M, 694 μl) is added dropwise. The mixture is stirred at approximately -78 ° C for approximately 1 hour and then brought to approximately -15 ° C for the next 30 minutes. The solution is cooled to approximately -78 ° C. A solution of 1-methylsulfonyl-3- (3-chlorophenyl) -5- (4-chlorobenzoyl) indole (150 mg, see preparation 7) in THF (1 ml) is added drop by drop. The mixture is gradually brought to room temperature and then stirred at room temperature for 19 hours. The solution is cooled to approximately -78 ° C. Methanesulfonyl chloride (163 mg) is added dropwise, then diisopropylethylamine (87 mg). The solution is brought to room temperature in one hour and stirred at room temperature for 3 hours. To the solution are added 4 ml aqueous solution of IN HCl and 4 ml of THF. The solution is stirred at 0 ° C for an hour and a half. The organic solvent is removed by evaporation under reduced pressure. The aqueous solution is brought to pH 8 by addition at 0 ° C of an aqueous solution of KOH 6N. The aqueous solution is extracted twice with DCM. The combined organic phases are washed once with brine, dried over MgSO ₄ , filtered and reduced by evaporation under reduced pressure. The residue is purified by column chromatography on silica, eluting with a CH ₃ Cl / MeOH / Et ₃ N mixture, 98: 2: 0.1. The expected compound is obtained in the form of a solid (44 mg; yield 25%). MS (ES) 525.2 (Cale. MM = 525.3).

The enantiomers of the compound of Example 2 can be separated using techniques known to those skilled in the art, such as preparative HPLC on a chiral column.

Example 3: (± Vl-fN.N-dimethylcarbamoyl ') - 3- (3-chlorophenyl') - 5-r ('4-chlorophenyl) hydroxyC 1 -methyl- 1 H-iιnidazol-5-yDmethyllindole

This compound is synthesized analogously to the first method described for preparing the compound of Example 2, using dimethylcarbamyl chloride in place of methanesulfonyl chloride. MS (electrospray): 518.2 (Cale. MM: 518.5). Example 4: (+) - 1 -methylsulfonyl-3- (3-chlorophenyl) -5-raminoC4-chlorophenyl) (1-methyl-1H-imidazol-5-y methvn indole

To a solution of (±) - 1 -methylsulfonyl-3- (3-chlorophenyl) -5 - [(4-chlorophenyl) hydroxy (1-methyl-1H-imidazol-5-yl) methyl] indole (100 mg ) and triethylamine (58 mg) in anhydrous DCM (2 ml) is added methanesulfonyl chloride (66 mg) at -78 ° C. The solution is brought slowly to room temperature and then stirred at room temperature for 19 hours. The solution is diluted by adding DCM and washed twice with brine, dried over MgSθ, 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 about 60 ° C overnight. The solvent is removed by evaporation under reduced pressure. The residue is dissolved in water and DCM. The organic phase is separated, washed once with brine, dried over MgSO ₄ , filtered and reduced by evaporation under reduced pressure. The residue is dissolved in an EtOAc / ΗOAc 9: 1 mixture. 200 mg of palladium on carbon (Pd / C 10%) are added. The mixture is stirred under Η ₂ (40 psi) for about 3 hours. The solvent is removed by evaporation under reduced pressure. The residue is purified by column chromatography on silica, eluting with a CH ₃ Cl / MeOH / Et ₃ N mixture, 98: 2: 0.1, to give the expected compound.

Example 21: (±) -1-acetyl-3- (3-chlorophenylV5-r (4-chlorophenyl) hydroxy (1-methyl-1 H-imidazol-5-yl) methyl] indole

This compound is synthesized analogously to the first method described to prepare the compound of Example 2, using acetic anhydride in place of methanesulfonyl chloride. MS (electrospray): 489.2 (Cale. MM: 489.4).

The following compounds can 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 those skilled in the art. The compounds of Examples 5, 6, 8, 10, 12, 16, 18, 20 and 22 can be synthesized analogously to the compound of Example 4. Examples 7, 9, 11, 15, 17 and 19 can be synthesized analogously to the compound of Example 2.

Example 5

PREPARATION 1: 2-f3-chlorophenylVN-methoxy-N-methyl-acetamide

A solution of 3-chlorophenylacetic acid (5.00 g; 29.3 mmol), 1- (3-dimethyl-aminopropyl) -3-ethylcarbodiimide hydrochloride (6.18 g; 32.2 mmol), and 1-hydroxybenzotriazole (HOBt; 4.00 g; 29.3 mmol) in dichloromethane (DCM; 40 ml) is stirred at room temperature for about 10 minutes. The solution is cooled to about 0 ° C. N, O-dimethylhydroxylamine hydrochloride (2.86 g; 29.0 mmol) and diisopropylethylamine (DIEA; 3.80 g, 29.3 mmol) are added. The reaction mixture is brought to ambient temperature and stirred for approximately 5 hours. The solution is diluted with 100 ml of DCM and washed with a saturated aqueous solution of NaHCO ₃ (2 times), an aqueous solution of HC1 IN (2 times) and brine (2 times), dried over anhydrous MgSO ₄ , filtered and concentrated by evaporation under reduced pressure. The liquid obtained is purified by chromatography on a silica column, eluting with a 1: 1 EtOAc / hexane mixture. The expected compound is obtained in the form of a colorless liquid (5.60 g, 89%). R _f = 0.44 (silica, EtOAc / hexane 1: 1).

¹ H NMR (300 MHz, CDC1 ₃ ): 7.18-7.34 (m, 4H); 3.76 (S, 2H); 3.66 (S, 3H); 3.22 (S, 3H).

PREPARATION 2; 2-f3-chlorophenylVacetaldehyde

A suspension of LiAlUt (1.90 g, 51 mmol) in anhydrous ether (250 ml) is stirred at room temperature under a nitrogen atmosphere for about 1 hour. The suspension is cooled to approximately -45 ° C. A solution of 2- (3-chlorophenyl) -N-methoxy-N-methyl-acetamide (8.19 g; 38.3 mmol) is added dropwise. ) in 10 ml of anhydrous tetrahydrofuran (THF). The mixture is brought to about 0 ° C and stirred for about 3 hours. The solution is then cooled to approximately -45 ° C. To this solution is slowly added a solution of KHSO ₄ (13 g) in water (approximately 30 ml). The resulting mixture is filtered through CELITE. The filtrate is concentrated by evaporation under reduced pressure, and the resulting solution is diluted with DCM and washed with an aqueous HC1 IN solution (2 times), and brine (2 times), dried over anhydrous MgSO ₄ , filtered and concentrated by evaporation under reduced pressure. The expected compound compound is obtained in the form of a liquid (5.80 g), which is used immediately in the additional step without further purification. R _f = 0.71 (silica, EtOAc / hexane 1: 3).

PREPARATION 3; 3- (3-chlorophenv 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 in nitrogen by passing N ₂ through the solution. The solution is then brought to reflux for approximately 2.5 hours. The solvent is removed by evaporation under reduced pressure and the residue obtained is dissolved in DCM and washed with an aqueous solution of HC1 IN (2 times), an aqueous solution saturated with NaHCO ₃ (2 times) and brine (2 times) , dried over anhydrous MgSθ ₄ , filtered and concentrated by evaporation under reduced pressure. The crude product is purified by chromatography on a silica column, eluting with an EtOAc / hexane mixture 1: 6. The expected compound is obtained in the form of a reddish oil (5.30 g; 62%). R _f = 0.26 (silica, EtOAc / hexane 1: 4)

PREPARATION 4: 3-G-cMorophenylVindoline

3- (3-chlorophenyl) indole (5.30 g; 23.3 mmol) was dissolved in 50 ml of 1M BH ₃ in THF solution. The mixture is cooled to approximately 0 ° C. Trifluoroacetic acid (TFA, 50 ml) is then added slowly. After the addition, the solution is stirred for about 10 minutes. To the solution is slowly added a solution of ₃ M BH in THF (40 ml). The mixture is stirred for approximately 5 minutes and then concentrated by evaporation under reduced pressure. The residue is purified by chromatography on a silica column, eluting with an EtOAc / hexane mixture 1: 6. The expected compound is obtained in the form of an oil (3.93 g; 74%). R _f = 0.20 (Silica, EtOAc / hexane 1: 4). MS (ES): 229.1 (Wed. MM = 229.7).

PREPARATION 5: 1-methylsulfonyl-3-f3-chlorophenvnindoline

To a solution of 3- (3-chlorophenyl) indoline (3.88 g; 16.9 mmol) and DIEA (2.40 g; 18.6 mmol) in DCM (40 ml) at approximately 0 ° C is added dropwise methanesulfonyl chloride (2.13 g; 18.6 mmol). The mixture is stirred for about an hour and a half. The solution is diluted with DCM and washed with a saturated aqueous solution of NaHCO ₃ (2 times), an aqueous HC1 IN solution (2 times) and brine (2 times) then dried over anhydrous MgSO ₄ , filtered and concentrated by evaporation under reduced pressure. The crude product is purified by chromatography on a column of silica eluting with an EtOAc / hexane mixture 1: 4. The expected compound is obtained in the form of an oil (4.40 g; 85%). R _f = 0.41 (silica, EtOAc / hexane 1: 2).

NMR ^! H (300 MHz, CDC1 ₃ ): 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).

PREPARATION 6: l-methvIsrifonyl-3-f3-cMorophenyl) -

5- (4-chlorobenzoyDindoline

To a solution of 1-methylsulfonyl-3- (3-chlorophenyl) indoline (4.40 g; 14.3 mmol, see Preparation 5) and 4-chlorobenzoyl chloride (3.25 g; 18.6 mmol) at approximately 0 ° C in CS ₂ (25 ml) is added in portions A1C1 ₃ (7.62 g; 57.2 mmol). A brown precipitate is immediately formed. The mixture is stirred for approximately 2 hours. To this mixture are slowly added 100 ml of cold water containing 3 ml of concentrated HCl. The solution is diluted with DCM and the organic phase is separated and washed with an aqueous solution of HC1 IN (2 times), an aqueous solution saturated with NaHCO ₃ (2 times) and brine (2 times), then dried over MgSθ ₄ anhydrous, filtered and concentrated by evaporation under reduced pressure. The crude product is purified by chromatography on a silica column, 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 (Cale. MM = 445.4).

NMR 'H (300 MHz, CDC1 _3): 7.67 to 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).

PREPARATION 7: 1-methylsulfonyl-3-f3-chlorophenylV5- (4-chlorobenzovnindole

A solution of 1-methylsulfonyl-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 brought to reflux under a nitrogen atmosphere for approximately 6 hours and then heated to approximately 95 ° C. overnight. The solvent is removed and the residue purified by chromatography on a silica column, 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 (hold. MM = 443.4). Rf = 0.38 (silica, EtOAc / hexane, 1: 2). Preparation B:

1 - [(2R) -amino-3-mercaptopropyl] - (2S) - (2-mercaptoethyl) -4- (l-naphthoyl) -piperazine-1,2-cyclodisulfide (XII):

a) Synthesis of 1-benzyl-3 (SVbenzyloxycarbonylmethyl piperazine-2.5-dione:

To a solution cooled by an ice bath of β-benzyl ester BOC-aspartic acid (10 g), hydroxybenzotriazole (HOBT, 4.2 g) and ethyl ester of N-benzylglycine (6.4 g) in 80 ml of CH ₂ CI ₂ is added a cold solution of dicyclohexylcarbodiimide (DCC, 7.1 g) in 20 ml of CH ₂ CI ₂ . The reaction medium is stirred for approximately 1 hour at a temperature of 0 to 5 ° C., then overnight at room temperature. The precipitate is removed by filtration and the filtrate is evaporated to dryness under reduced pressure. The residue is partitioned between ethyl acetate and water. The organic phase is washed with 100 ml of aqueous NaHCO ₃ , then water, and dried (MgSθ4). The solvent is removed by dry evaporation under reduced pressure to give 16 g of a solid. Thin layer chromatography (TLC) (silica gel: CHCl ₃ / acetone = 9: 1, R _f = 0.55).

The product obtained is then treated with trifluoroacetic acid 50% in CHC1 ₃ (40 ml) for approximately 1 hour and the volatile substances are eliminated by evaporation to dryness under reduced pressure. The residue is partitioned between ethyl acetate and a saturated aqueous solution of NaHCO ₃ . The organic phase is dried (MgSO ₄ ) and the solvent is removed by evaporation to dryness under reduced pressure to give 10 g. TLC (silica gel: CHCl / acetone = 9: 1, R _f = 0.14).

b) Synthesis of 4-benzyl-1-tert-butoxycarbonyl-2fS1- (2-hyciroxyethyD piperazine:

To a solution cooled by an ice bath of the product of step a) (9.73 g) in 200 ml of tetrahydrofuran (THF) is added in portions a mineral dispersion of lithium aluminum hydride (LAH) 50% ( 12.5 g) under a nitrogen atmosphere. The reaction medium is left at reflux overnight. After cooling in an ice bath, a saturated aqueous solution of Na ₂ S0 ₄ is added dropwise to decompose the excess of LAH and the white molasses obtained is eliminated by filtration. The filtrate is evaporated to dryness under reduced pressure and dissolved in dichloromethane (55 ml), treated with tert-butyl dicarbonate (5.9 g), and stirred for about 1 hour. A saturated aqueous solution of NaHCO ₃ (25 ml) is added and the mixture stirred for approximately 2 hours. The organic phase is washed with saturated aqueous sodium chloride solution and dried (MgSO ₄ ). After evaporation of the solvent, the residue is chromatographed on silica gel (160 g) using a CHCl ₃ / MeOH 19: 1 mixture as eluent. The appropriate fractions are combined, and the solvents are removed by evaporation to dryness under reduced pressure to give 10 g of a glass. TLC (silica gel: CHCl ₃ / MeOH = 9: 1, R _f = 0.56).

c) Synthesis of l-tert-butoxycarbonyl-2- (S) - (2-hydroxyethyl) piperazine:

The product from step b (8.7 g) is dissolved in ethanol (35 ml) and treated with Pd (OH) ₂ on charcoal (0.8 g) and acetic acid (3 ml) . The hydrogenation is then carried out under a pressure of 30 psi overnight. The reaction mixture is filtered and the solvent removed by evaporation to dryness under reduced pressure to give the expected product.

d) Synthesis of l-tert-butoxycarbonyl-2 (Sy (2-hydroxyethyl) -4-π-naphthoyD piperazine:

To a solution of the product of step c) (8.4 g) in acetonitrile (40 ml) are added 110 ml of aqueous solution of NaOH IN and then a solution of 1-naphthoyl chloride (5.14 g ) in acetonitrile (20 ml). After approximately 3 hours with stirring, a large part of the acetonitrile eliminated by evaporation under reduced pressure and the residual mixture is extracted with chloroform. The extract is dried (MgSO ₄ ) and the solvent removed by evaporation to dryness under reduced pressure to give 8.12 g of the expected product. TLC (silica gel: CHCl ₃ / MeOH = 9: 1, R _f = 0.64).

e) Synthesis of 1 -tert-butoxycarbonyl-2 ('S') - ( ^' 2-triphenylmethylthioethyl - 4-O-naphthoylVpiperazine:

To a solution cooled by an ice bath of triphenylphosphine (0.53 g) in 5 ml of anhydrous THF is added dropwise a solution of diethylazodicarboxylate (DEAD, 0.25 g) in 2 ml of THF. After stirring for approximately 30 minutes at a temperature of 0 to 5 ° C., a solution of the product from step d) (0.4 g) and triphenylmercaptan (0.55 g) in 10 ml of THF is added dropwise. drop. The mixture is stirred for about 1 hour at a temperature of 0 to 5 ° C and for about 1 hour at room temperature. The solvent is removed by evaporation to dryness under reduced pressure and the residue is chromatographed on silica gel (40 g) using CHC1 ₃ as eluent. The appropriate fractions are combined and the solvent is removed by evaporation to dryness under reduced pressure to give 420 mg of a pale yellow foam. Mass spectrometry (MS) (Electrospray) 665.2 (643 + 23 (sodium)). TLC (silica gel: CHCl ₃ / acetone = 9: 1, R _f = 0.53) f) Synthesis of 2 (S) - (2-triphenylmethylthioethyl -4- (l -naphthoyl) piperazine:

To a stirred solution of the product of step e) (2.2 g) in 30 ml of CH ₂ CI ₂ are added 10 ml of trifluoroacetic acid (TFA). The mixture is stirred for approximately 30 minutes. The volatile substances are removed by dry evaporation under reduced pressure. The residue is dissolved in CHC1 ₃ (50 ml) and treated with excess triemylamine (4 ml). The mixture is washed with water, then dried (MgSO ₄ ) and the volatiles are removed by evaporation to dryness under reduced pressure to give 2.1 g of a pale yellow glass. TLC (silica gel: CΗC _-3 / MeOH = 9: 1, R _f = 0.63)

g) Synthesis of l-r2 (R) -N-tert-butoxycarbonylamino-3-triphenylmethylthiopropyll- 2f S (2-triphenylmethyl-thioethy -4- (1 -naphthoyl Vpiperazine):

To a solution of the product of step f) (0.9 g) and of 2 (R) -N-tert-butoxycarbonylamino- 3-triphenylmethylthiopropan-al (1.2 g), prepared according to the procedure of OP Goel, et al., (Org. Syn. 1988, 67, 69-75), in CH ₂ C1 ₂ (20 ml) containing 1% acetic acid, 4 g of 4A molecular sieves are added and then in portions of Na ( OAc) ₃ BH (1 g) over a period of 30 minutes. After stirring for approximately 2 hours, the mixture is filtered and the filtrate is washed with water, 5% aqueous NaHCO ₃ solution, water, then dried (MgSU ₄ ). The solvent is removed by evaporation to dryness under reduced pressure, and the residue is chromatographed on silica gel (60 g) using CHC1 ₃ as eluent. The appropriate fractions are combined and the solvent is removed by evaporation to dryness under reduced pressure to give 0.6 g of a white foam. TLC (silica gel: CHCl ₃ / acetone = 9: 1; R _f = 0.55); MS (Electro Spray) 974.3.

h) Synthesis of l -r2 (R) -amino-3-mercaptopropyπ-2 (S) -2-mercaptoethyl) - 4- (l -naphthoyl Vpiperazine):

The product of step g) (450 mg) is treated for about 30 minutes with 50% TFA in CH ₂ C1 ₂ (10 ml) containing 1 ml of triethylsilane. The volatile substances are removed by dry evaporation under reduced pressure. The residue is triturated with ether, filtered, then dried to give 280 mg of 1- [2 (R) -amino-3-mercaptopropyl] -2 (S) - (2-mercaptoethyl) -4- (l- naphthoyl) -piperazine. MS (electrospray) 390.3.

i) Cclclisation of l-r2 (R) -amino-3-mercaptopropyll-2 (S) -2-mercaptoethyl) - 4- (l -naphthoyl Vpiperazine to form l-r2CRVamino-3-mercaptopropyll-2 (S) -

2-mercaptoethyl V4- (1-naphthoyl Vpiperazine-1.2-cyclodisulfide (XII):

100 mg of the product of step a) are dissolved in 10 ml of aqueous CH ₃ CN solution (H ₂ O / CH ₃ CN = 7: 3), and treated with 3 g of EKATHIOX ™ resin (0.34 mmol / g). The reaction medium is stirred for about 6 hours at room temperature. The mixture is then filtered, the resin is washed with an aqueous methanol solution (methanol water 1: 3), and most of the organic solvent is removed by evaporation under reduced pressure to keep only a small volume. The concentrate is subjected to preparative HPLC chromatography using 0.1% aqueous TFA and CH ₃ CN as the mobile phase. The appropriate fractions are combined and most of the solvents are removed by evaporation under reduced pressure to keep only a small volume. The concentrate is then lyophilized to give the expected product.

Alternatively, a solution of 1- [2 (R) -amino-3-mercaptopropyl] -2 (S) - (2-mercaptoethyl) -4- (1-naphthoyl) -piperazine in aqueous CH ₃ CN is stirred with air at pH between 6 and 8. In both cases, the reaction medium comprises a mixture of cyclized disulfide and the dimer of the latter in a proportion of approximately 4 to 1.

PHARMACOLOGICAL STUDY OF THE PRODUCTS OF THE INVENTION

By way of example, we will study the effect of a treatment of a human cell line MCF-7 with the compounds of formulas (I), (II), (XI), (XII), (GPIi) and (GPI ₂ ) described above. An electrophoresis of pancreatic cancer cells treated with the compounds of formulas (I) and (III) will also be carried out. All these compounds are prepared by the methods described in the cited patents or, for (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 lines were acquired from the American Tissue Culture Collection (Rockville, Maryland, USA).

Measurement of the intracellular amount of cyclic AMP for MCF-7 cells

MCF-7 cells (2.10) seeded on a 24-well plate are cultured for 5 days in the modified Eagle medium from Dulbecco (Gibco-Brl, Cergy-Pontoise, France) supplemented with 10% inactivated fetal calf serum 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 culture medium is replaced after two washes with a serum-free medium, whether or not supplemented with the agents specified for a time indicated in the various figures. Agents activating the production of cAMP are then added at 37 ° C. The reaction is stopped after 30 minutes by removing the medium and rapidly adding 200 μl of 0.1N HCl solution. 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) by following the manufacturer's instructions. Radioactivity is determined by a Gamma counter (Gamma Master-1277, LKB, Turku, Finland).

Cell proliferation tests

3000 MCF-7 cells placed in 80 μl of modified Eagle medium from Dulbecco (Gibco-Brl, Cergy-Pontoise, France) supplemented with 10% fetal calf serum 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) were seeded on a 96-well plate on day 0. The cells were treated on day 1 for 96 hours with increasing concentrations up to 50 μM of each of the compounds to be tested. At the end of this period, the quantification of cell proliferation is evaluated by colorimetric test based on the cleavage of the tetrazolium salt WST1 by mitochondrial dehydrogenases in viable cells leading to the formation of formazan (Boehringer Mannheim, Meylan, France ). These tests are carried out in duplicate with 8 determinations per concentration tested. For each compound to be tested, the values included in the linear part of the sigmoid were used for a linear regression analysis and used to estimate the inhibitory concentration IC ₅₀ .

Analysis of the subcellular localization of the βγ complex by the western-blot method The Mia-PaCa2 cells (400,000) are seeded in Petri dishes (diameter 10 cm) in the modified Eagle medium from Dulbecco (Gibco-Brl, Cergy-Pontoise, France ) supplemented with 10% fetal calf serum 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 glutamine (Gibco-Brl, Cergy-Pontoise, France). The compounds are added on day 1 at a concentration of 30 μM. The cells are harvested by scraping on day 3 after two washes with phosphate buffer (PBS, Gibco-Brl, Cergy-Pontoise, France) at 4 ° C. The cells are pelleted by centrifugation at low speed (800 g, 5 minutes) . The cells are resuspended in buffer A containing Hepes 50 mM pH 7.5, MgCl2 5 mM, EDTA 1 mM, Protease inhibitors (Cocktail tablets, n ° 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 a first time at low speed (1200 g, 5 minutes, 4 ° C) to remove the non-lysed cells. The supernatant is centrifuged at high speed (200,000 g, 50 minutes, 4 ° C) to separate the cytosolic fraction and the membrane fraction (pellet). The latter is resuspended in buffer A. The protein concentration in each fraction is determined by Bradford colorimetric assay (Bio-Rad protein assay, Ivry, France). The proteins (20 μg) are separated by electrophoresis in denaturing polyacrylamide gel (16% gel-tricine, Novex, Prolabo, Fontenay sous Bois) according to the manufacturer's recommendations. The proteins thus separated are transferred onto a nitrocellulose membrane (C-hybond, RPN 2020C, Amersham, Les Ulis, France) in semi-dry transfer. The β protein is detected by the antibody T20, sc378, Santa-Cruz, TEBU, Le Perray en Yvelines, France), itself recognized by the second antibody coupled to the peroxidase enzyme. Chemiluminescence is obtained with the ECL-Plus system (RPN2132, Amersham, Les Ulis, France). The signal is revealed on Kodak BioMax-light autoradiographic films, Z37358, Sigma, St Quentin, France). The amount of chemiluminescence is proportional to the amount of protein present. Equipment

The vaso-intestinal peptide (VIP) and mevastatin were acquired from Bachem (Voisins le Bretonneux, France) and TEBU (Le Perray en Yvelines, France) respectively. The compounds of formulas (I), (II) and (III) were supplied by Biomeasure Inc. (Milford, MA, USA). All these compounds were used following the recommendations of their manufacturers.

Results

VIP has been presented as an extracellular ligand for a G protein-coupled receptor that stimulates cAMP synthesis in human breast cancer cells. Figure 1 shows that VIP treatment of human breast cancer cells MCF-7 increases the intracellular amount of cAMP in a concentration-dependent manner. The VIP concentration of 10 nM which provides a quasi-optimal cAMP production will be used for the following tests. This concentration is in agreement with the data already published concerning the human breast cancer cell line T47D.

If the post-transcriptional prenylation of G proteins is a necessary step to enable its function, the addition of prenyltransferase inhibitors should significantly alter it. The compound of formula (I) is known to be a powerful specific famesyltransferase inhibitor, capable of inhibiting farnesylation of ras at concentrations of the order of nanomoles. FIG. 2 shows that a 24-hour pre-treatment of MCF-7 cells from in vitro cultures with the compound of formula (I) inhibited, in a concentration-dependent manner, the accumulation of cAMP stimulated by the VIP. Almost complete inhibition was obtained at a concentration of 30 μM of the compound of formula (I). These results clearly show that treatment with a specific famesyltransferase inhibitor is sufficient to block the signal transduction, the pathway of which uses heterotimeric G proteins as mediators.

FIG. 3 shows that a treatment of 1 hour with the compound of formula (I) is not sufficient to modify the response to VIP. Despite a weak but reproducible effect after an 8 hour treatment, it is only a 24 hour treatment which gives a clear inhibition of VIP stimulation. The kinetics of action observed are in agreement with that which was expected for a prenyltransferase inhibitor allowing the appearance of non-prenylated forms of protein G subunits.

The inhibition of stimulation by VIP is not restricted to compounds of structure analogous to that of the compound of formula (I) but can be extended to other inhibitors of prenyltransferases, as shown by the results obtained in particular for compound of formula (H) which is also a powerful farnesyltransferase inhibitor, capable of inhibiting the farnesylation of H-ras of human tumors at concentrations of the order of nanomoles (Table I).

Furthermore, FIG. 4 shows that the pretreatment of the cells for 24 hours with the compound of formula (I) does not modify the production of cAMP induced by the direct activator of adenylate cyclase, forskolin. This shows that adenylate cyclase itself is not modified by treatment with the compound of formula (I).

FIG. 5 shows that the pretreatment of the cells for 24 hours with the compound of formula (I) decreases the production of cAMP induced by the direct activator of the α subunit, the cholera toxin. However, the latter can only be fixed on the heterotrimeric complex (Warner et al., Cell Signal, 8, 43-53 (1996)). This therefore suggests that the pretreatment with the compound of formula (I) prevents the formation of the heterotrimeric complex.

The βγ dimer complex is very stable and can be identified in a protein extract by an antibody directed against the β subunit. FIG. 6 shows the results of a western blot performed on untreated cells, cells treated for 48 hours with mevastatin (an inhibitor of mevalonate synthesis), with a geranyl-geranyl transferase inhibitor, the compound of formula (III), or by a pure faraesyltransferase inhibitor, the compound of formula (I). The proteins of the cytosolic and membrane fractions of the control or treated cells are separated by denaturing electrophoresis in polyacrylamide gel before being transferred to a semi-solid membrane allowing identification with an antibody (western-blot) directed against all forms of β proteins.

The amount of βγ complex detected on the cell membrane of untreated cells (control) decreases sharply in cells treated for 48 hours with mevastatin, by a geranyl-geranyl transferase inhibitor (compound of formula (IH)) or by a pure farnesyl transferase inhibitor (compound of formula (I)). We can therefore conclude that after treatment with a prenyltransferase inhibitor, the γ subunit therefore no longer fulfills its role in anchoring the βγ complex to the membrane.

Finally, table II shows the in vitro values of the proliferative inhibition activities of the compounds of formulas (I) and (II) with respect to human tumor cells MCF-7 not carrying a Ras oncogene which has undergone a mutation. .

Table I

Effects of compounds incubated for 24 hours on the production of VIP-stimulated cyclic AMP in MCF7 cells.

The cells are incubated for 24 hours in the presence of the compounds tested at the doses indicated and then stimulated with 10 ⁸ M of VIP. The quantification of cyclic AMP is made by radioimmunoassay.

Table II

Proliferative inhibition of the compounds of formulas (I) and (II) compared to human tumor cells MCF-7 not carrying a mutant Ras oncogene.

Claims

1. Use of prenyltransferase inhibitors to prepare a medicament intended to treat pathologies which result from the membrane fixation of the heterotimeric protein G.

2. Use according to claim 1, characterized in that the prenyltransferase inhibitors are chosen from a group consisting of the following compounds: thiazole derivatives, peptidomimetic derivatives of 4-aminobenzoic acid, quinolinone derivatives, tricyclic amides , polyacids, peptidomimetic derivatives of piperidines, peptidomimetic derivatives of benzodiazepines, tripeptide derivatives of phosphonic or phosphinic acids, pseudodipeptides based on an N-carbonylpyrazine structure, and tetrapeptide analogs of the Cys-Al-A2-A3 type where Cys is a Cysteine, A1 and A2 are aliphatic amino acids and A3 is any amino acid.

3. Use according to claim 1, characterized in that the prenyltransferase inhibitors are famesyltransferase inhibitors.

4. Use according to claim 1, characterized in that the prenyltransferase inhibitors are chosen from a group consisting of the compounds corresponding to general formula (GPI) represented below

(GPI)

in which :

Ri represents OH, SH or NH ₂ ;

R ₂ represents a lower alkyl radical or COR ₅ ; R ₃ and R ₄ independently represent a halogen atom or an OH radical;

Y represents CO or CS or is absent;

and compounds corresponding to the following formulas:

(D

(XII)

5. Use according to claim 4, characterized in that the prenyltransferase inhibitors are compounds of general formula (GPI).

6. Use according to claim 5, characterized in that the prenyltransferase inhibitors are chosen from a group consisting of the following compounds:

- 1-acetyl-3- (3-chlorophenyl) -5 - [(4-chlorophenyl) hydroxy- (1-methyl-1H-imidazolyl) methyl] indole (GPIi); - 1- (4-mo holinecarbamoyl) -3- (3-chlorophenyl) -5 - [(4-chlorophenyl) hydroxy- (1-methyl-1H-imidazolyl) methyl] indole (GPI ₂ );

- 4- (3-corophenyl) -6 - [(4-chlorophenyl) hydroxy] - (1-methyl-1H-imidazol-5-yl) methyl- 2 (1H) -quinolinone (GPI ₃ );

- 4- (3-corophenyl) -6 - [(4-corophenyl) hydroxy] - (1-memyl-1H-imidazol-5-yl) methyl- 2 (1H) -quinolinone (GPI ₄ ).

7. Use according to claim 4, characterized in that the prenyltransferase inhibitor is chosen from the group consisting of compounds of formula (I), (II), (ll) bis, (III) and (XII):

(D

(IH)

(XII)

8. Use according to claim 7, characterized in that the prenyltransferase inhibitor is the compound of general formula (XH):

(XII)

9. Use according to one of claims 1 to 7, characterized in that the pathology to be treated is chosen from pathologies linked to the following biological functions or disorders: smell, taste, perception of light, neurotransmission, neurodegeneration, functioning of the glands endocrines and exocrines, autocrine and paracrine regulation, blood pressure, embryogenesis, viral infection, immunological functions, diabetes and obesity.

10. Use according to one of claims 1 to 7, characterized in that the pathology to be treated is chosen from the following pathologies: cholera, Acquired Immunodeficiency Syndrome (AIDS), traveler's diarrhea and precocious familial puberty masculine.