OA12973A - 1-N-phenylamino-1H-imidazole derivatives as aromatase inhibitors. - Google Patents

Abstract

The present invention relates to imidazole derivatives of the formula (I) in which R1, R2,R3, R4, R5, R6 and n are as defined in the specification. The invention also relates to pharmaceutical compositions containing these derivatives, and to the uses thereof.

Description

1 012973 1-N-phenylamino-lH-imidazole dérivatives as aromatase inhibitors

The présent invention relates to 1-N-phenylamino-lH-imidazole dérivatives asaromatase inhibitors and to pharmaceutical compositions containing them.Aromatase is the physiological enzyme responsible for the spécifie conversion ofandrogens such as androstenedione or testosterone, into estrogens such as estroneand estradiol, respectively (Simpson ER et al., Endocrine Reviews, 1994, 15 : 342-355). Inhibition of aromatase is, therefore, a strategy of choice to interfère withnormal or pathological estrogen-induced or estrogen-dependent biological processessuch as female sexual différentiation, ovulation, implantation, pregnancy, breast andendométrial cell prolifération as well as régulation of spermatogenesis or prostatecell prolifération in male or of non-reproductive fonctions such as bone formation orimmune T cell and cytokine balance (Simpson ER et al., Recent Progress inHormone Research, 1997, 52 : 185-213 and the whole issues of Endocrine RelatedCancer (1999, volume 6, n° 2) and Breast Cancer Research Treatment (1998,volume 49, supplément n° 1)). A large number of azole dérivatives are known as antifungal agents. Some imidazoleor triazole dérivatives hâve already been described as inhibitors of the enzymearomatase. Generally, the imidazolyl or the triazolyl group is associated witharomatic rings as found in letrozole (EP-A-236 940; Lamb HM and Adkins JC, Drugs,1998, 56 : 1125-1140) :

or anastrozole (EP-A-296 749; Wiseman LR and Adkins JC, Drugs Aging, 1998, 13 :321-332) : 2 012973

Imidazoles or triazoles linked via a methylene group to a benzotriazole are describedin EP-A-293 978 :

5 Diterbutyl phénols having a N-amino-imidazole moiety in the para position aredescribed in US patent 4,908,363 and are presented as having inflammation-inhibiting and oedema-inhibiting properties :

More recently, M. OKADA et al. (Chem. Pharm. Bull., 44 (10), 1996, 1871-1879)10 described a sériés of [4-(bromophenylmethyl)-4-(cyanophenyl)amino]-azoles and their azine analogs : 3 012973 10 10 15 15

CN

Br YM 511

It has now been found that imidazole dérivatives which invariably contain a 1-[N-phenylamino] group demonstrate an unexpectedly high potency to inhibitaromatase.

Accordingly, one object of this invention is to provide l-[N-phenylamino]imidazoledérivatives which are potent aromatase inhibitors.

Another object of this invention is to provide a pharmaceutical compositioncontaining, as active ingrédient, a l-[N-phenylamino]imidazole dérivative asdepicted below or a pharmaceutically acceptable acid addition sait thereof. A further object of this invention is to provide the use of a l-[N-phenylamino]-imidazole dérivative in the manufacture of a médicament intended for treating orpreventing various diseases and for managing reproductive fonctions in women, inmen as well as in female and male wild or domestic animais.

The l-[N-phenylamino]imidazole dérivatives of this invention are represented by thefollowing general formula (I) : (I) and acid addition salts, solvatés and stereoisomeric forms thereof, wherein : 4 012973 • Ri and R2 are each independently hydrogen, a (Ci-C6)atkyl or a (C3-C8)cycloalkyl ; • n = 0, 1/ 2 ; • R3, R4, Rs and Rg are each independently hydrogen, or a (Ci-C6)alkyl,halogen, cyano, (Ci-ÇOaIkoxy, trifluoromethyl, (CrC6)alkylthio, (Ci-C6)alkylsulfonyl, sulfonamido, acyl, (Ci-C6)alkoxycarbonyl, orcarboxamido group ; • R3 and Rg together with the phenyl ring bearing them can also form abenzofurane or a N-methylbenzotriazole.

In the description and daims, the term "(CrC6)alkyl" is understood as meaning alinear or branched hydrocarbon chain having 1 to 6 carbon atoms. A (Ci-C6)alkylradical is for example a methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl,pentyl, isopentyl or hexyl radical.

The term "halogen" is understood as meaning a chlorine, bromine, iodine or fluorineatom.

The term "(C3-C8)cycloalkyl" is understood as meaning a saturated monocyclichydrocarbon having 3 to 8 carbon atoms. A (C3-C8)cycloalkyl radical is for example acydopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl radical.

The term "(Ci-C6)alkoxy" is understood as meaning a group OR in which R is a (CrCe)alkyl as defined above. A (Ci-C6)alkoxy radical is for example a methoxy, ethoxy,propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, n-pentyloxy or isopentyloxyradical.

The term "acyl" is understood as meaning a group R'—C in which R' is hydrogen or a

O (CrC6)alkyl as defined above.

Compounds of formula (I) form acid addition salts, for example with inorganic acidssuch as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoricacid and the like or with organic carboxylic acids such as acetic acid, propionic acid,glycolic acid, pyruvic acid, oxalic acid, malic acid, fumaric acid, tartaric acid, citricacid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid and the like.Preferred compounds of formula (I) are those wherein : • n is 0 or 1 ; • Ri and R2 are each independently hydrogen or (Ci-C6)alkyl ; 5 012973 • R3 is cyano or trifluoromethyl ; • Rj is hydrogen, (Ci-C6)alkyl, halogen, cyano, (Ci-C6)alkoxy,trifluoromethyl, (CrQalkylthio, (Ci-C6)alkylsulfonyl or (CrC6)alkoxycarbonyl ; • R5 is hydrogen, halogen, (CrCOalkoxy or trifluoromethyl ; • Ré is hydrogen ; • or R3 and Re together with the phenyl ring form a N-methylbenzotriazole.Also preferred are the compounds of formula (I) wherein : • n is 0 or 1 ; • Ri, R2 and Re are each hydrogen ; • R4 is halogen, cyano or trifluoromethyl.

Especially preferred compounds of formula (I) are those wherein R3 is cyano ; thosewherein R5 is hydrogen or trifluoromethyl ; and those wherein n is 1.

Valuable compounds are selected from the group consisting of: 4-[N-(lK-imidazol-l-yl)-N-(4-trifluoromethylphenylmethyl)amino]benzonitrile 4-[N-(lH-imidazol-l-yl)-N-(4-chlorophenylmethyl)amino]benzonitrile, 4-[N-(lH-imidazol-l-yl)-N-(4-cyanophenylmethyl)amino]benzonitrile, 4,4'-[N-(lH-imidazol-l-yl)amino]bis-benzonitrile, 4-[N-(lH-imidazol-l-yl)-N-(4-fluorophenylmethyl)amino]benzonitrile, 4-[N-(lH-imidazol-l-yl)-N-(3,4-difluorophenylmethyl)amino]benzonitrile, andthe acid addition salts, solvatés or stereoisomeric forms thereof.

By virtue of their capability to inhibit aromatase, and thus to exhaust ail sources ofendogenous estrogens, the compounds of the présent invention can be used aloneor in combination with other active ingrédients for the treatment or the préventionof any estrogen-dependent disorder or for the management of estrogen-regulatedreproductive functions, in humans as well as in wild or domestic animais.

The breasts being sensitive targets of estrogen-stimulated prolifération and/ordifférentiation, inhibitors of aromatase are especially useful in the treatment orprévention of benign breast diseases in women, gynecomastia in men and in benignor malignant breast tumors with or without metastasis both in men and women(Brodie AM and Njar VC, Steroids, 2000, 65 : 171-179; Pritchard Kl, Cancer, 2000,85, suppl 12 : 3065-3072), or in male or female domestic animais. 6 012973

Due to the învolvement of estrogens in the mechanisms of ovulation, implantationand pregnancy, inhibitors of aromatase according to the invention can be used,respectively, for contraceptive, contragestive or abortive purposes in women (NjarVC and Brodie AM, Drugs, 1999, 58 : 233-255) as well as in females of wild ordomestic animal species.

The utérus is another reproductive organ responsive to estrogenic stimulation andinhibition of aromatase is therefore useful to treat or prevent endometriosis, benignuterine diseases or benign or malignant uterine tumors with or without metastasis inwomen (Njar VC and Brodie AM, Drugs, 1999, 58 : 233-255) or in female domesticanimais.

The ovary being the physiological source of estrogen, inhibitors of aromatase can beused to treat abnormal or untimely ovarian estrogen production such as polycysticovary syndrome or precocious puberty, respectively (Bulun et al., J Steroid BiochemMol Biol, 1997, 61 : 133-139). Ovarian as well as non-ovarian but estrogen-producing benign or malignant tumors with or without metastasis (Sasano H andHarada N, Endocrine Reviews, 1998, 19 : 593-607) may also benefit from treatmentwith aromatase inhibitors according to the invention.

In males, prostate and testicular tissues are also responsive to estrogenicstimulation (Abney TO, Steroids, 1999, 64 : 610-617; Carreau S et al., Int J Androl, 1999, 22 : 133-138). Therefore, aromatase inhibitors can be used to treat or toprevent benign (Sciarra F and Toscano V, Archiv Androl, 2000, 44 : 213-220) ormalignant prostate tumors with or without metastasis (Auclerc G et al., Oncologist, 2000, 5 : 36-44) or to treat, prevent or control spermatogenesis functions ormalfunctions, in men as well as in male wild or domestic animais.

Estrogens are also known to be implicated in the régulation of bone turnover ;therefore, aromatase inhibitors may be useful, alone or in combination with otherantiresorbtive or proosteogenic agents, in the treatment or prévention of bonedisorders according to appropriate therapeutic sequences or regimens.

In addition, estrogens are involved in the régulation of the balance between Thi andTh2 prédominant immune functions and may therefore be useful in the treatment orprévention of gender-dependent auto-immune diseases such as lupus, multiplesclerosis, rheumatoid arthritis and the like. 7 012973

When the compounds of formula (I) are administered for the treatment orprévention of estrogen-dependent disorders, they can be combined with one orseveral other sexual endocrine therapeutic agents. In the case of the control ormanagement of reproductive fonctions such as male or female fertility, pregnancy,abortion or delivery, the compounds of formula (I) can be combined with forexample a LH-RH agonist or antagonist, an estroprogestative contraceptive, aprogestin, an anti-progestin or a prostaglandin. When the compounds of formula (I)are intended for the treatment or prévention of benign or malignant diseases of thebreast, the utérus or the ovary, they can be combined with e.g. an anti-estrogen, aprogestin or a LH-RH agonist or antagonist. In the case of the treatment orprévention of benign or malignant diseases of the prostate or the testis, thecompounds of formula (I) can be combined with for example an antiandrogen, aprogestin, a lyase inhibitor or a LH-RH agonist or antagonist.

The term "combined" refers herein to any protocol for co-administration of acompound of formula (I) and one or more other pharmaceutical substances,irrespective of the nature of the time of administration and the variation of doseover time of any of the substances. The co-administration can for example beparallel or sequential.

The invention thus also relates to a method of treating or preventing the above-mentioned diseases, comprising the administration to a subject in need thereof of atherapeutically effective amount of a compound of formula (I) or a pharmaceuticallyacceptable acid addition sait thereof, optionally in combination with another activeingrédient.

For the treatment/prevention of any of these diseases, the compounds of formula(I) may be administered, for example, orally, topically, parenterally, in dosage unitformulations containing conventional non-toxic pharmaceutically acceptable carriers,adjuvants and vehides. These dosage forms are given as examples, but otherdosage forms may be developed by those skilled in the art of formulation, for theadministration of the compounds of formula (I). The term parentéral as used hereinincludes subcutaneous injections, intravenous, intramuscular, intrasternal injectionor infusion techniques. In addition to the treatment of humans, the compounds ofthe invention are effective in the treatment of warm-blooded animais such as mice,rats, horses, cattle sheep, dogs, cats, etc. 8 012973

The pharmaceutical compositions containing the active ingrédient may be in a formsuitable for oral use, for example, as tablets, troches, lozenges, aqueous or oilysuspensions, dispersible powders or granules, émulsions, hard or soft capsules, orsyrups or élixirs. Compositions intended for oral use may be prepared according toany method known to the art for the manufacture of pharmaceutical compositionsand such compositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents and preservingagents in order to provide pharmaceutically élégant and palatable préparations.Tablets contain the active ingrédient in admixture with non-toxic pharmaceuticallyacceptable excipients which are suitable for the manufacture of tablets. Theseexcipients may be for example, inert diluents, such as calcium carbonate, sodiumcarbonate, lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid; binding agents, forexample starch, gelatin or acacia, and lubricating agents, for example, magnésiumstéarate, stearic acid or talc. The tablets may be uncoated or they may be coated byknown techniques to delay disintegration and absorption in the gastrointestinal tractand thereby provide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate may beemployed.

They may also be coated by the technique described in U.S. Patents 4,256,108;4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.Formulations for oral use may also be presented as hard gelatin capsules whereinthe active ingrédient is mixed with an inert solid diluent, for example, calciumcarbonate, calcium phosphate, or kaolin, or as soft gelatin capsules wherein theactive ingrédient is mixed with water or an oil medium, for example peanut oil,liquid paraffin, or olive oil.

Aqueous suspensions contain the active ingrédient in admixture with excipientssuitable for the manufacture of aqueous suspensions. Such excipients aresuspending agents, for example sodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gumtragacanth and gum acacia; dispersing or wetting agents, for example a naturally-occurring phosphatide, such as iecithin, or condensation products of an alkyleneoxide with fatty acids, for example polyoxyethylene stéarate, or condensation 9 012973

Products of ethylene oxide with long Chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxide withpartial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitolmonooleate, or condensation products of ethylene oxide with partial esters derived 5 from fatty acids and hexitol anhydrides, for example polyethylene sorbitanmonooleate. The aqueous suspensions may also contain one or more preservatives,for example ethyl, or n-propyl, p-hydroxybenzoate, one or more coloring agents,one or more flavoring agents, and one or more sweetening agents, such as sucrose,saccharin or aspartame. 10 Oily suspensions may be formulated by suspending the active ingrédient in avegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or inminerai oil such as liquid paraffin. The oily suspensions may contain a thickeningagent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents suchas those set forth above, and flavoring agents may be added to provide a palatable 15 oral préparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for préparation of an aqueous suspensionby the addition of water provide the active ingrédient in admixture with a dispersingor wetting agent, suspending agent and one or more preservatives. Suitable 20 dispersing or wetting agents and suspending agents are exemplified by thosealready mentioned above. Additional excipients, for example sweetening, flavoringand coloring agents, may also be présent. The pharmaceutical compositions of theinvention may also be in the form of an oil-in-water émulsion. The oily phase maybe a vegetable oil, for example olive oil or arachis oil, or a minerai oil, for example 25 liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occuring phosphatides, for example soy bean, lecithin, and esters or partial estersderived from fatty acids and hexitol anhydrides, for example sorbitan monooleate,and condensation products of the said partial esters with ethylene oxide, forexample polyoxyethylene sorbitan monooleate. The émulsions may also contain 30 sweetening and flavouring agents.

The pharmaceutical compositions may be in the form of a stérile injectable aqueousor oleagenous suspension. This suspension may be formulated according to theknown art using those suitable dispersing or wetting agents and suspending agents 10 012973 which hâve been mentioned above. The stérile injectable préparation may also be astérile injectable solution or suspension in a non-toxic parenterally-acceptablediluent or solvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water, Ringer's solution 5 and isotonie sodium chloride solution. In addition, stérile, fixed oils areconventionally employed as a solvent or suspending medium. For this purpose anybland fixed oil may be employed including synthetic mono- or diglycerides. Inaddition, fatty acids such as oleic acid flnd use in the préparation of injectables.Dosage levels of the order of from about 0.0001 mg to about 20 mg/kg of body 10 weight per day are useful in the treatment of the above-indicated conditions, oraltematively about 0.1 mg to about 2000 mg per patient per day.

The amount of active ingrédient that may be combined with the carrier matériels toproduce a single dosage form will vary depending upon the host treated and theparticular mode of administration. Dosage unit forms will generally contain between 15 from about 0.1 mg to about 400 mg, preferably from about 1 mg to about 100 mg,of active ingrédient, typically 0.1 mg, 1 mg, 2 mg, 5 mg, 10 mg, 20 mg, 40 mg, 50mg, 60 mg, 80 mg, 100 mg or 400 mg.

It will be understood, however, that the spécifie dose level for any particular patientwill dépend upon a variety of factors including the âge, body weight, general health, 20 sex, diet, time of administration, route of administration, rate of excrétion, drugcombination and the severity of the particular disease undergoing therapy.

The 1-N-phenyl-amino-lH-imidazole dérivatives of formula (I) of the invention andtheir acid addition salts can be prepared following the general scheme 1. 11 012973

(l) 9 8 12 012973

According to scheme 1 aniline dérivative (1) is condensed with the aldéhyde offormula (2) and the imine intermediate is reduced with sodium borohydride orhydrogenated using palladium or platinum oxide as catalyst to afford the N,N-disubstituted aniline (3). Said aniline (3) can also be prepared by reaction of ahalogeno dérivative (8) with an aniline of formula (1).

The Ν,Ν-disubstituted aniline (3) is converted to its nitroso dérivative using standardconditions, then reduced to afford the 1,1-disubstituted hydrazine of formula (4).Alternative^, the 1,1-disubstituted hydrazine (4) can be prepared by sélective N-alkylation of a compound of formula (7) with a compound of formula (8) using theconditions described by U. LERCH and J. KONIG (Synthesis, 1983, 2, 157-8).

Then, condensation of (4) with a dialkyloxy-alkyl-isothiocyanate dérivative or anethylenedioxy-alkyl-isocyanate dérivative, affords the thiosemicarbazide (5) which istransformed to the l-amino-imidazole-2-thione (6) by treatment with an acid likeacetic acid or sulphuric acid.

Desulfurization of (6) in acetic acid, following the conditions described by S.GRIVASand É.RONNE in Acta Chemica Scandinavia, 1995, 49, 225-229, gives the final 1-N-phenylamino-lH-imidazole dérivative (I), which is optionally converted to one of itspharmaceutically acceptable acid addition salts. Alternatively compound (I) where R3or Re is an electron-withdrawing group can be obtained by condensation of the N-imidazoloaniline (9) with the halogeno dérivative (8).

The following examples and tests are intended to illustrate the invention withouthowever implying a limitation. PREPARATION OF THE N-ALKYLANILINES (3) EXAMPLE 1 N-(4-chlorophenylmethyl)-4-cyanoaniline

To a solution of 4-chlorobenzaldehyde (35.69 g, 0.253 mol) in absolute éthanol (250ml) was added portionwise 4-aminobenzonitrile (30 g, 0.253 mol). The reactionmixture was stirred at room température for 3 h, the precipitate was filtered,washed with ether and poured into a 1/1 mixture of THF/ethanol (250 ml). Theresulting suspension was ice-cooled, NaBH4 (4.8 g, 0.127 mol) was addedportionwise and the reaction mixture was stirred at room température for 0.75 h.After addition of acetic acid (3 ml) and water (500 ml), the precipitate was filtered,washed with water and dried to give a white solid (51.27 g, 84 %), mp 130°C. 13 012973 ^-NMR (CDCI3) : 4.35 (d, 2H), 4.75 (s, 1H), 6.55 (d, 2H), 7.20-7.50 (m, 6H)

Using the same procedure but replacing the 4-chlorobenzaldehyde by - 4-methoxybenzaldehyde, - 3-fluoro-4-methoxybenzaldehyde, - 4-cyano-N-(4-methoxyphenylmethyl)aniline (mp 109°C), and - 4-cyano-N-(3-fluoro-4-methoxyphenylmethyl)aniline (mp 108°C)were respectively obtained. EXAMPLE 2 4-cyano-N-(4-methylphenylmethyl)aniline A mixture of p-tolualdehyde (40.68 g, 0.338 moi), 4-aminobenzonitrile (40 g, 0.338mol) and 5 % Pd/C (4g) in absolute éthanol (300 ml) was hydrogenated at roomtempérature overnight. The reaction mixture was diluted with methylene chloride,fïltered on celite and concentrated to dryness. Crystallization from éthanol affordeda white solid (74.9 g, 99 %), mp 100°C. ^-NMR (CDCI3) : 2.35 (s, 3H), 4.3 (d, 2H), 4.7 (s, 1H), 6.55 (d, 2H), 7.10-7.30 (m,4H), 7.35 (d, 2K)

Using the same procedure but replacing the p-tolualdehyde by - 4-cyanobenzaldehyde, - 3,4-dimethoxybenzaldehyde, - 4-cyano-N-(4-cyanophenylmethyl)aniline (mp 156°C), and - 4-cyano-N-(3, 4-dimethoxyphenylmethyl)aniline (mp 150°C)were respectively obtained. PREPARATION OF N, N-DISUBSTITUTED HYDRAZINES (4)

These are generally prepared according to the procedure described in Tetrahedron1982, 38(3) : 419-423 and Organic Fonctional Group Préparations 1968, 1 : 374-376. EXAMPLE 3 N1-(4-chlorophenylmethyl)-N1-(4-cyanophenyl)hydrazine

To an ice-cooled suspension of 4-cyano-N-(4-chlorophenylmethyl)aniline (30 g,0.125 mol) in 2 N H2SO4 (150 ml) was added a solution of sodium nitrite (9.48 g,0.137 mol) in water (30 ml). The réaction mixture was stirred at room température 14 012973 for 2 h. A solution of sodium nitrite (9.48 g, 0.137 mol) in water (30 ml) was added,and the reaction was stirred overnight. A solution of sodium nitrite (6.6 g, 0.095mol) in water (20 ml) was added and the reaction was stirred for 1 h. Afterextraction with ethyl acetate, the organic layer was washed successively with asaturated sodium hydrogen carbonate solution, water, brine, dried over sodiumsulfate and evaporated under vacuum to give a white solid (30.5 g). To asuspension of the resulting solid in a mixture of ether (60 ml), AcOH (60 ml) andwater (60 ml), was added zinc powder (24.5 g, 0.375 mol) at such a rate as to keepthe température below 35° C. The mixture was stirred for 2 h, AcOH (60 ml), water(60ml) and zinc (6 g) were added and stirring was maintained for 0.5 h. Afteraddition of ether (200 ml), the reaction mixture was filtered, the inorganic materialwas washed with ethyl acetate, the product was extracted with ethyl acetate andthe organic layer was washed with water, brine and dried over sodium sulfate. Thesolvents were concentrated under vacuum and crystallization from diisopropyl etheryielded a solid (18.78 g, 58 %), mp 90° C. iH-NMR (CDCI3) : 3.75 (s, 2H), 4.69 (s, 2H), 7.05 (d, 2H), 7.15 (d, 2H), 7.35 (d,2H), 7.49 (d, 2H).

Using the same procedure but replacing the 4-cyano-N-(4- chlorophenylmethyl)aniline by - 4-cyano-N-(4-methoxyphenylmethyl)aniline, - 4-cyano-N-(3-fluoro-4-methoxyphenylmethyl)aniline, - 4-cyano-N-(4-methylphenylmethyl)aniline, - 4-cyano-N-(4-cyanophenylmethyl)aniline, - 4-cyano-N-(3, 4-dimethoxyphenylmethyl)aniline, - 4-bromo-N-(4-cyanophenylmethyl)aniline, - N1-(4-methoxyphenylmethyl)-N1-(4-cyanophenyl)hydrazine (mp 74°C), - N1-(3-fluoro-4-methoxyphenyimethyl)-N1-(4-cyanophenyl)hydrazine (mp102°C), - N1-(4-methylphenylmethyl )-N1-(4-cyanophenyl)hydrazine (mp 74°C), - N1-(4-cyanophenylmethyl )-N1-(4-cyanophenyl)hydrazine (mp 215°C), -1^-(3,4-dimethoxyphenylmethyl )-N1-(4-cyanophenyl)hydrazine (mp 15 012973 134°C), and - N1-(4-cyanophenylmethyl )-N1-(4-bromophenyl)hydrazine (mp 114°C)were respectively obtained. EXAMPLE 4 N1-(4-trifluoromethylphenylmethyl)- N1-(4-cyanophenyl)hydrazine

In a nitrogen atmosphère, powdered sodium amide (95%, 4.8 g, 0.117 mol) wasintroduced with stirring into a flask containing THF (100 ml). The solution was ice-cooled and 4-cyano-phenylhydrazine hydrochloride (prepared following JoséLCastro et al. J.Med.Chem. 1994, 37, 3023-3032) (10 g, 0.058 mol) was addedportionwise. The ice bath was removed and a stream of nitrogen was passedthrough the orange suspension over a period of 1 h to remove most of the dissolvedammonia. With ice-cooling, 4-trifluoromethylbenzyl chloride (12 g, 0.062 mol) wasadded, then the reaction mixture was stirred at room température for 1.5 h andpoured into water (100 ml). After extraction with ethyl acetate, the organic layerwas washed with water, dried over sodium sulfate and evaporated under vacuum.Trituration from diisopropyl ether gave a yellow solid (7.5 g, 43 %), mp 98°C.JH-NMR (CDCI3) : 3.8 (s, 2H), 4.8 (s, 2H), 7.05 (d, 2H), 7.33 (d, 2H), 7.5 (d, 2H), 7.6 (d, 2H)

Using the same procedure but replacing the 4-trifluoromethylbenzyl chloride by : - 4-fluorobenzonitrile, - 4-fluorobenzyl bromide, - 4-methylthiobenzyl chloride - 3,4-difluorobenzyl chloride, -2,4-difluorobenzyl chloride, -3,5-difluorobenzyl chloride, - 4-bromobenzyl bromide, - N1-bis-(4-cyanophenyl)hydrazine (mp 222°C), - N1-(4-fluorophenylmethyl)- N1-(4-cyanophenyl)hydrazine (mp 114-115°C), - N1-(4-methylthiophenylmethyl)-N1-(4-cyanophenyl)hydrazine (mp 72°C), - N1-(3,4-difluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine (mp 72°C), - N1-(2,4-difluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine (mp 70°C), 16 012973 - 1^-(3,S-difluorophenylmethyQ-fP-^-cyanophenyOhydrazine (mp 124°C),and - N^-bromophenylmethyO-N^-cyanophenyOhydrazine (mp 90°C)were respectively obtained.

EXAMPLE 4A N1-(4-cyanophenylmethyl)-N1-(4-methoxyphenyl)hydrazine

Chloromethylbenzonitrile (25 g, 164.90 mmol) was introduced with stirring into aflask containing toluene (200 ml) and triethylamine (46.40 ml, 329.80 mmol). 4-cyano-phenylhydrazine hydrochloride (prepared following José L. Castro et al., J.Med.Chem. 1994, 37, 3023-3032) (28.80 g, 164.90 mmol) was added portionwiseand the reaction mixture was stirred 3 h at reflux. After cooling, the mixture wasfiltered, washed with toluene (50 ml) and with water (200 ml) to give a white solid(27.20 g, 65%), mp: 115°C. ‘H-NMR (DMSO d6) : 3.65 (s, 3H), 4.30 (s, 2H), 4.57 (s, 2K), 6.77 (d, 2H), 6.94 (d,2H), 7.48 (d, 2K), 7.76 (d, 2H). PREPARATION OF THE IMIDAZOLES OF FORMULA (I) EXAMPLE 5 4-[N-(lH-imidazol-l-yl)-N-(4-trifluoromethylphenyl)amino]benzonitrile

To a cold solution (10-15°C) of tBuOK (1.06 g, 0.00895 mol) in DMSO (18 ml) wasadded portionwise N-(lH-imidazol-l-yl)-4-trifluoromethylaniline (1.85 g, 0.00814mol) (prepared by desulfurization from the corresponding 2,3-dihydro-lH-imidazol- 2-thione, J.G.Schantl, Heterocycles, 37(3), 1873, 1994). The reaction mixture wasstirred at room température for 1 h, then 4-fluorobenzonitrile (0.936 g, 0.00773mol) in DMSO (18 ml) was added, the reaction mixture was stirred for 2 h andpourred into water and concentrated sodium hydroxyde solution. The precipitatewas collected and dried under vacuum. Flash chromatography on silica gel (toluene! dioxane : 7/3) and crystallization from diisopropyl ether yielded a solid (1.6 g,60 %), mp 104 °C.

Analysis Calculated. : C : 62.2 ; H : 3.38 ; F : 17.36 ; N : 17.07Found : C : 62.22 ; H : 3.40 ; F : 17.3 ; N : 17.1 ^-NMR (DMSO d6) : 6.96 (d, 2H), 7.15 (s, 1H), 7.31 (d, 2H), 7.68 (s, 1H), 7.6-7.85(m, 4H), 7.87 (s, 1H). 17 012973 EXAMPLE 6 4-[N-(lH-imidazol-l-yl)-N-(4-trifluoromethylphenylmethyl)amino]- benzonitrile a) 4-[N-(2,3-dihydro-lH-imidazol-l-yl-2-thione)-N-(4-trifluoromethylphenyl5 methyl)amino]benzonitrile

To a suspension of N1-(4-trifluoromethylphenylmethyl)-N1-(4-cyanophenyl)hydrazine(7.5 g, 0.025 mol) in éthanol (100 mi) was added dropwise 2,2-dimethoxyethylisothiocyanate (4 g, 0.027 mol) and the reaction mixture was heatedto reflux for 2 h. After cooling thë solvent was evaporated under vacuum, the 10 resulting residue was poured into 2N H2SO4 (20 ml) and the suspension was heatedto reflux for 0.3 h. After extraction with ethyl acetate, the organic layer was washedwith water, dried over sodium sulfate and concentrated under vacuum. Flashchromatography on silica gel (toluene / dioxane : 8/2) and trituration fromdiisopropyl ether / éthanol afforded a yellow solid (2.7 g, 28 %), mp 200 °C. 15 XH-NMR (DMSO d6) : 5-5.4 (m, 2H), 6.65 (d, 2K), 6.95 (d, 1H), 7.15 (d, 1H), 7.7(m, 6H). b) 4-[N-(lH-imidazol-l-yl)-N-(4-trifluoromethylphenylméthyl)amino]benzonitrile 35 % hydrogen peroxide (1.1 ml, 0.035 mol) was added dropwise to an ice-cooledsuspension of 4-[N-(2,3-dihydro-lH-imidazol-l-yl-2-thione)-N-(4- 20 trifluoromethylphenylmethyl)amino]benzonitrile (2.7 g, 0.0072 mol) in acetic acid(20 ml). When TLC showed complété reaction, the reaction mixture was diluted withwater, adjusted to pH 11 with sodium hydroxide, treated with sodium hydrogensulfite and extracted with ethyl acetate. The organic layer was dried over sodiumsulfate and concentrated under vacuum. Flash chromatography on silica gel 25 (toluene / ethyl acetate : 7/3 then 6/4) and crystallization from diisopropyl etheryielded a solid (1 g, 41 %), mp 134 °C.

Analysis Calculated. : C : 63.1 ; H : 3.8 ; N : 16.3

Found : C : 63.4 ; H : 3.58 ; N : 16.3 'H-NMR (DMSO d6) : 5.15 (s, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.4 (s, 1H), 7.5-7.9 (m, 30 7H). M+ = 342

Using the same procedure but replacing the N1-(4-trifluoromethylphenylmethyl)-N1-(4-cyanophenyl)hydrazine by : 18 012973 - N1-(4-chlorophenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(4-methoxyphenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(3-fluoro-4-methoxyphenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(4-methylphenylmethyl)-N1-(4-cyanophenyl)hydrazine, 5 - N1-(4-cyanophenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(3,4-dimethoxyphenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-bis-(4-cyanophenyl)hydrazine, - N1-(4-fluorophenylmethyl)-N1-(4-cyanôphenyl)hydrazine, - N1-(4-methylthiophenylmethyl)-N1-(4-cyanophenyl)hydrazine, 10 - N1-(3,4-difluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(2/4-difluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(3,5-difluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine/ - N1-(4-bromophenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(4-cyanophenylmethyl)-N1-(4-bromophenyl)hydrazine, 15 - N1-(4-cyanophenylmethyl)-N1-(l-methylbenzotriazol-6-yl)hydrazine/ - N1-(4-cyanophenylmethyl)-N1-(3-trifluoromethyl-4-cyanophenyl)hydrazine, - N1-(phenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(4-cyanophenyl)-N1-(3-trifluoromethyl-4-cyanophenyl)hydrazine, - N1-(3-fluorophenylmethyl)-N1-(4-cyanophenyl)hydrazine, 20 - N1-(4-methoxycarbonylphenylmethyl)-N1-(4-cyanophenyl)hydrazine, - N1-(4-cyanophenyl)-N1-(4-fluorophenyl)hydrazine, - N1-(3-trifluoromethylphenylmethyl)-N1-(4-cyanophenyl)hydrazine,the following compounds were respectively obtained : EXAMPLE 7 25 4-[N-(lH-imidazol-l-yl)-N-(4-chlorophenylmethyl)amino]benzonitnle,hydrochloride

mp 190°C

Analysis Calcuiated : C : 59.15 ; H : 4.09 ; Cl : 20.54 ; N : 16.23Found : C : 59.04 ; H : 3.99 ; Cl : 20.5 ; N : 16.2 30 'H-NMR (DMSO d6) : 5.15 (s, 2H), 6.95 (d, 2H), 7.4 (s, 4H), 7.75 (m, 3H), 8.10 (s,1H), 9.6 (s, 1H) EXAMPLE 8 19 012973 4-[N-(lH-imidazol-l-yl)-N-(4-methoxyphenylmethyl)amino]benzonitrile, hydrodiloride

mp 178°C

Analysis Calculated : C : 63.44 ; H : 5.03 ; Cl : 10.40 ; N : 16.44Found : C : 63.4 ; H : 5.01 ; Cl : 10.4 ; N : 16.6 ^-NMR (DMSO d6) : 3.70 (s, 3H), 5.05 (s, 2H), 6.85 (d, 2H), 7 (d, 2H), 7.25 (d,2H), 7.79 (s, 1H), 7.80 (d, 2H), 8.1 (s, 1H), 9.50 (s, 1H) EXAMPLE 9 4-[N-(lH-imidazol-l-yl)-N-(3-fluoro-4-methoxyphenylmethyl)-amino]benzonitrile, hydrochloride

mp 190°C ^-NMR (DMSO d6) : 3.80 (s, 3H), 5.05 (s, 2H), 7 (d, 2H), 7.05-7.15 (m, 2H), 7.30(d, 1H), 7.8 (d, 2H), 7.75 (s, 1H), 8.1 (s, 1H), 9.50 (s, 1H) EXAMPLE 10 4-[N-(lH-imidazol-l-yl)-N-(4-methylphenylmethyl)amino]benzonitrile

mp 156°C

Analysis Calculated : C : 74.98 ; H : 5.59 ; N : 19.43Found : C : 74.55 ; H : 5.56 ; N : 19.3 ’H-NMR (DMSO d6) : 2.25 (s, 3H), 4.95 (s, 2H), 6.65 (d, 2H), 6.98 (s, 1H), 7.10 (d,2H), 7.2 (d, 2H), 7.35 (s, 1H), 7.70 (m, 3H) EXAMPLE 11 4-[N-(lH-imidazol-l-yl)-N-(4-cyanophenylmethyl)amino]benzonitrile, hydrochloride

mp 195°C

Analysis : Calculated : C : 64.38 ; H : 4.2 ; Cl : 10.56 ; N : 20.86

Found : C : 64.31 ; H : 4.29 ; Cl : 10.6 ; N : 20.9 ‘H-NMR (DMSO d6) : 5.30 (s, 2H), 7.10 (d, 2H), 7.62 (d, 2H), 7.70-8 (m, 5H), 8.17(s, 1H), 9.7 (s, 1H) EXAMPLE 12 4-[N-(lH-imidazol-l-yl)-N-(3,4- dimethoxyphenylmethyl)amino]benzonitrile, hydrochloride

mp 167°C

Analysis Calculated : C : 61.54 ; H : 5.16 ; Cl : 9.56 ; N : 15.11 20 012973

Found : C : 61.39 ; H : 5.13 ; Cl : 9.57 ; N : 15.1 ^-NMR (DMSO d6) : 3.70 (s, 3H), 3.73 (s, 3H), 5 (s, 2H), 6.65-7.15 (m, 5H), 7.75(S, 1H), 7.80 (d, 2H), 8.10 (s, 1H), 9.5 (s, 1H) EXAMPLE 13 ^'-[Ν-ΙΙΗ-ϊιηΐάθζοΙ-Ι-γΟβηιίηοΙΜβ-^βηζοηΐϋΊΐβ, hydrochloride

mp 199°C

Analysis : Calculated : C : 63.46 ; H : 3.76 ; Cl : 11.02 ; N : 21.76

Found : C : 63.2 ; H : 3.81 ; Cl : 11.0 ; N : 21.9 ^-NMR (DMSO d6) : 7.26 (d, 4H), 7.9 (s, 1H), 7.92 (d, 4H), 8.30 (s, 1H), 9.80 (s,1H) EXAMPLÎ14 4-[N-(lH-imidazol-l-yl)-N-(4-fluorophenylmethyl)amino]benzonitrile,hydrochloride hemihydrate

mp 176-178 °C

Analysis : Calculated : C : 60.45 ; H : 4.48 ; F : 5.62 ; Cl : 10.5 ; N : 16.59

Found: C : 60.71 ; H : 4.64 ; F : 5.54 ; Cl : 10.6 ; N : 17.0 ’H-NMR (DMSO de) : 5.15 (s, 2H), 7.2 (t, 2H), 7.4 (q, 2H), 7.8 (s, 1H), 7.0-7.8 (AB,4H), 9.6 (s, 1H). EXAMPLE 15 4-[N-(lH-imidazol-l-yl)-N-(4-inethylthiophenylmethyl>amino]- benzonitrile

mp 128 °C

Analysis : Calculated : C : 67.56 ; H : 5.03 ; N : 17.5 ; S : 10.02

Found : C : 67.12 ; H : 4.90 ; N : 17.2 ; S : 9.51 ‘H-NMR (DMSO d6) : 2.4 (s, 3H), 5.05 (s, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.15-7.3 (m,4H), 7.35 (S, 1H), 7.7 (m, 3H). EXAMPLE 16 4-[N-(lH-imidazoM-yl)-N-(4-methylsulfonylphenylmethyl)amino]- benzonitrile

mp 190 °C ’H-NMR (DMSO de) : 3.2 (s, 3H), 5.2 (s, 2H), 6.6 (d, 2K), 7 (s, 1H), 7.45 (s, 1H), 7.6-7.9 (m, 7H) 21 012973 EXAMPLE 17 4-[N-(lH-imidazol-l-yl)-N-(3,4-difluorophenylmethyl)amino]benzonitrile

mp 132 °C

Analysis Calculated : C : 65.86 ; H : 3.9 ; N : 18.07Found : C : 65.47 ; H : 3.78 ; N : 18.1 XH-NMR (DMSO d6) : 5.0 (s, 2H), 6.7 (d, 2H), 6.9-7.8 (m, 8H) EXAMPLE 18 4-[N-(lH-imidazol-l-yl)-N--(2,4-diflùor<)phenylmethyQaniino]benzonitrile

mp 149 °C

Analysis Calculated : C : 65.86 ; H : 3.9 ; N : 18.07Found : C : 66.0 ; H : 3.84 ; N : 18.2 ‘H-NMR (DMSO d6) : 5.1 (s, 2H), 6.7 (d, 2H), 6.9-7.8 (m, 8H) EXAMPLE 19 4-[N-(lH-imidazol-l-yl)-N-(3/5-difluorophenylmethyl)amino]benzonitrile

mp 170 °C

Analysis Calculated : C : 65.86 ; H : 3.9 ; N : 18.07Found : C : 65.73 ; H : 3.8 ; N : 18.02 'H-NMR (DMSO d6) : 5.1 (s, 2H), 6.6 (d, 2H), 7-7.2 (m, 4H), 7.5 (s, 1H), 7.75 (d,2H), 7.9 (s, 1H) EXAMPLE 20 4-[N-(lH-imidazol-l-yl)-N-(4-bromophenylmethyl)amino]benzonitrile, hydrochloride

mp 125 °C 'H-NMR (CDCI3) : 5 (s, 2H), 6.8 (d, 2H), 7 (s, 1H), 7.25 (m, 3H), 7.35 (s, 1H), 7.4(d, 2H), 7.65 (d, 2H), 9.85 (s, 1H) EXAMPLE 21 4-[N-(lH-imidazol-l-yl)-N-(4-bromophenyl)aminomethyl]benzonitrile

mp 138 °C

Analysis : Calculated : C : 57.81 ; H : 3.71 ; N : 15.86 ; Br : 22.62

Found : C : 57.85 ; H : 3.7 ; N : 15.9 ; Br : 23.2 'H-NMR (DMSO d6) : 4.85 (s, 2H), 6.5 (d, 2H), 7 (s, 1H), 7.1 (s, 1H), 7.25-7.55 (m,5H), 7.6 (d, 2H) EXAMPLE 22 22 012973 ^[N-Cl-methylbenzotriazol-e-yO-N-ilH-imidazol·!- yl)aminomethyl]benzomtrile

mp 184 °C 'H-NMR (DMSO d6) : 4.20 (s, 3H), 5.1 (s, 2H), 6.5 (dd, 1H), 6.95 (s, 1H), 7.2 (s,1H), 7.4 (s, 1H), 7.57 (d, 2H), 7.7-7.85 (m, 3H), 7.9 (d, 1H) EXAMPLE 23 4-[N-(lH-imidazol-l-yl)-N-(4-cyanophenylmethyl)amino]-3- trifluoromethylbenzonitrile

mp 226°C

Analysis : Calculated : C : 62.18 ; H : 3.29 ; N : 19.08

Found : C : 61.89 ; H : 3.35 ; N : 18.9 'H-NMR (DMSO d6) : 5.25 (s, 2H), 6.8-7.05 (m, 3H), 7.45 (s, 1H), 7.55 (d, 2H), 7.8(d, 3H), 8.05 (d, 1H) EXAMPLE 24 4-[N-(lH-îmidazol-l-yl)-N-(phenylmethyl)amino]benzonitrile

mp 107 °C XH-NMR (DMSO de) : 5.05 (s, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.25-7.4 (m, 6H), 7.65-7.75 (m,3K) EXAMPLE 25 4-[N-(lH-imidazol-l-yl)-N-(4-cyanophenyl)amino]-3- trifluoromethylbenzonitrile

mp 166 °C XH-NMR (DMSO d6) : 7.2 (m, 3K), 7.3 (d, 2H), 7.75 (s, 1H), 7.95 (d, 2K), 8.15 (d,1H), 8.25 (s, 1H) EXAMPLE 26 4-[N-(lH-imidazol-l-yl)-N-(3-fluorophenylmethyl)amino]benzonitrile

mp 112 °C

Analysis Calculated : C : 69.9 ; K : 4.48 ; N : 19.18Found : C : 69.38 ; K : 4.35 ; N : 19.3 *H-NMR (DMSO d6) : 5.05 (s, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.05-7.25 (m, 3H), 7.3-7.45 (m, 2H), 7.7-7.85 (m, 3H) EXAMPLE 27 23 012973

Methyl 4-[N-(4-cyanophenyl)-N-(lH-imidazol-l-yl)aminomethyl]benzoate

mp 178 °C 'H-NMR (DMSO d6) : 3.85 (s, 3H), 5.15 (s, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.4 (s, 1H), 7.5 (d, 2H), 7.65-7.8 (m, 3H), 7.9 (d, 2H) EXAMPLE 28 4-[N-(lH-imidazol-l-yi)-N-(4-fliiorophenyl)amino]benzonitrile

mp 113 °C XH-NMR (DMSO d6) : 6.45 (d, 2H), 7.1 (s, 1H), 7.25-7.45 (m, 2H), 7.5-7.7 (m, 5H),8.2 (s, 1H) EXAMPLE 29 4-[N-(lH-imidazol-l-yl)-N-(3-trifIuoromethylphenyl- methyl)amino]benzonitrile

mp 122 °C 'H-NMR (DMSO d6) : 5.15 (s, 2H), 6.7 (d, 2H), 7 (s, 1H), 7.4 (s, 1H), 7.5-7.8 (m,7H)

Using the same procedure of Example 6 but replacing the 2-isocyanato-acetaldehyde dimethylacetal by : - 2-isothiocyanatopropionaldehyde diethylacetal, - l-isothiocyanatopropan-2-one diethylacetal, - 2-isothiocyanatobutan-3-one diéthylacetal,the following compounds were obtained : EXAMPLE 30 4-[N-(4-methyl-lH-imidazol-l-yl)-N-(4-bromophenylmethyl)amino]- benzonitrile

mp 152 °C

Analysis : Calculated : C : 58.9 ; H : 4.11 ; N : 15.26

Found : C : 58.67 ; H : 4.16 ; N : 15.3 'H-NMR (DMSO d5) : 2.1 (s, 3H), 5 (s, 2H), 6.65 (d, 2H), 7.05 (s, 1H), 7.3 (d, 2H), 7.5 (d, 2H), 7.6 (s, 1H), 7.7 (d, 2H) M+ = 366 24 012973 EXAMPLE 31 4-[N-(5-methyl-lH-imidazol-l-yl)-N-(4-bromophenylmethyl)amino]- benzonitrile

mp 152 °C ^-NMR (CDCI3) : 2 (s, 3H), 4.9-5.2 (m, 2H), 6.65 (d, 2H), 7 (s, 1H), 7.25 (d, 2H), 7.5 (d, 2K), 7.6 (d, 2H), 9.25 (s, 1H) M+ = 366 EXAMPLE 32 4-[N-(4,5-dimethyl*lH-imidazol-l-yl)-N-(4-bromophenylmethyl)amino]- benzonitrile

mp 150 °C

Analysis Calculated : C : 59.8 ; H : 4.49 ; N : 14.7Found : C : 58.7 ; K : 4.41 ; N : 14.6 ^-NMR (DMSO de) : 1.9 (s, 3H), 2.1 (s, 3H), 4.95-5.25 (m, 2H), 6.6 (d, 2H), 7.3-7.8 (m, 7H) M+ = 380EXAMPLE 33 4-[N-(lH-imidazol-l-yl)-N-(4-methoxyphenyl)amino]methylbenzonitrile a) 4-[N-(2,3-dihydro-lH-imidazol-l-yl-2-thione)-N-(4-methoxyphenyl)amino]methylbenzonitrile

To a suspension of N1-(4-cyanophenylmethyl)-N1-(4-methoxyphenyl)hydrazine(27.10 g, 106.98 mmol) in éthanol (250 ml) was added dropwise 2,2-dimethoxyethylisothiocyanate (17.30 g, 117.67 mmol) and the reaction mixture washeated to reflux for 2 h. After cooling the solvent was evaporated under vacuum,the resulting oil was diluted into acetic add/water (9/1, 250 ml) and the suspensionwas heated to reflux for 1.5 h and at room température overnight. The resultingresidue was poured into water (1400 ml) and a brown precipitate was collected.After trituration from éthanol, the brown solid afforded a white solid (9.60 g, 27 %),mp: 150 °C. XH-NMR (DMSO d6) : 3.70 (s, 3H), 5.08 (s, 2H), 6.60 (d, 2H), 6.75-7.00 (m, 3H),7.20 (S, 1H), 7.80 (s, 4H). b) 4-[N-(lH-imidazol-l-yl)-N-(4-methoxyphenyl)amino]methylbenzonitrile 25 012973 35 % hydrogen peroxide (1.1 ml, 0.035 mol) was added dropwise to ap ice-cooledsuspension of 4-[N-(2,3-dihydro-lH-imidazol-l-yl-2-thione)-N-(4-methoxyphenyl)amino]methylbenzonitrile (9.50 g, 28.24 mmol) in acetic acid (50 ml). When TLCshowed complété reaction, the reaction mixture was diluted with water, adjusted to 5 pH 11 with sodium hydroxide, treated with sodium hydrogen sulfite and extractedwith ethyl acetate. The organic layer was dried over sodium sulfate andconcentrated under vacuum. Flash chromatography on silica gel (toluene / dioxane :6/4) yielded a pure orange oil (5.80 g, 67 %). ‘H-NMR (DMSO d6) : 3.70 (s, 3H), 4.90 (s, 2H), 6.60-7.00 (m, 5H), 7.40 (s, 1H), 10 7.55 (d, 2H), 7.70 (s, 1H), 7.78 (d, 2H).

Crystallization from hydrochloric éthanol yielded white crystals (5.70 g, 66 %)

Mp: 207°C ‘H-NMR (DMSO d6) : 3.70 (s, 3H), 4.97 (s, 2H), 6.93 (d, 2H), 7.13 (d, 2H), 7.45 (d,2H), 7.70 (s, 1H), 7.84 (d, 2H), 8.04 (s, 1H), 8.18 (s, 1H), 9.55 (s, 1H). 15 BIQLQGJÇAL TE§T RES.U.LT.5In vitro testing

The JEG-3 cell line, derived from a human placental choriocarcinoma, is intrinsicallyvery rich in human aromatase (Bahn RS et al., J Clin Endocrinol Metab, 1981, 52 :447-450) and, therefore, a useful practical biological System to screen and evaluate 20 putative aromatase ïnhibitors in vitro (Yue W and Brodie AM, J Steroid Biochem MolBiol, 1997, 63 : 317-328). Stocks of JEG-3 cells are grown up to 80 % of confluenceas monolayers in plastic flasks in minimum Eagle's medium with 1 g/l glucose and10 % decomplemented fêtai calf sérum at pH 7.4 and 37° C, under a 5 % CO2atmosphère. Then, 24 hours before aromatase activity measurement, JEG-3 cells 25 are distributed for seeding in 96-well microplates (60,000 to 100,000 viable cells in100 pl of culture medium per well) ; 24 hours later, microplates are rinsed and freshmedium containing the radioactive aromatase substrate (ip-3H-androstenedione,10 nM) is added together with test compounds dissolved with 1 % dimethylsulfoxidein a test concentration range between 10 “ and ΙΟ"4 M in a total volume of 150 pl. 30 Two hours after the beginning of the incubation, 100 pl of the supernatants aretransferred to homologous new 96-well micfoplates. A solution of dextran-coatedcharcoal (1 %) is added in each well (100 pl/well) ; after standing for 10 minutes onice, microplates are centrifuged (1500 g) for 10 more minutes at 4° C. Ail steroids, 26 012973 including the radioactive substrate and the newly biosynthesized estrogens, aretrapped complexed by the charcoal ; only 3H-water specifically formed duringaromatisation of lp-3H-androstenedione, involving a spécifie oxidative stepremoving the 1β-3Η, remains in the supernatant at this stage. Transferred to 5 another homologous 96-well microplate, the 100 μΙ supernatants received somescintillation counting liquid (200 μΙ/well) for β-radioactivity measurement by aMicrobeta Plus 1450 microplate Counter (Wallac, EG &amp; G).

In parallel, the aromatase reaction is stopped in the cell-containing microplates bydestruction and solubilization of the JEG-3 cells in a 10 mM ethylenediamine 10 tetraacetate solution at pH 12.3. Then, DNA is measured by a standard fluorimetricmethod using the Hoechst 33258 fluorochrome and a Victor2 (Wallac, EG &amp; G)microplate fluorimeter.

Finally, aromatase activity is expréssed in fmoles/pg DNA in 2 hours and aromataseinhibition in percentage of control incubation without inhibitors. A non linear fit 15 analysis of % of inhibition vs. concentration allows the détermination of 50 %inhibitory concentration (IC50) : the lowest IC50 correspond to the most potentinhibitors (Table A). 27 012973

Table A : Inhibition of human aromatase in vitro

Compound IC50 ± sem (nM) n Anastrozole 8.21 ± 1.27 3 Example 28 1.57 ± 0.74 3 Example 30 1.16 ± 0.55 3 Example 21 1.12 ± 0.55 3 Example 25 0.66 ± 0.14 3' Example 31 0.58 ± 0.18 3 Letrozole 0.56 ± 0.10 12 Example 5 0.54 ± 0.26 3 Example 23 0.48 ±0.13 3 Example 13 0.44 ± 0.04 3 Example 19 0.40 ±0.06 3 Example 16 0.35 ± 0.07 3 Example 6 0.31 ± 0.01 3 YM 511 0.30 ± 0.04 3 Example 17 0.24 ± 0.02 3 Example 20 0.22 ± 0.05 3 Example 15 0.21 ± 0.05 3 Example 22 0.20 ±0.05 3 Example 29 0.19 ±0.05 3 Example 11 0.18 ± 0.02 3 Example 24 0.18 ± 0.09 3 Example 9 0.16 ± 0.06 3 Example 14 0.16 ± 0.02 3 Example 7 0.14 ± 0.05 3 Example 8 0.14 ± 0.05 3 Example 10 0.14 ± 0.03 3 Example 18 0.14 ± 0.01 3 Example 26 0.13 ± 0.05 3 28 012973

INHIBITION OF AROMATASE IN VIVO

Aromatase is the steroidogenic enzyme responsible for the biosynthesis of estradiol,the main female sex hormone among estrogens. In the rat, estradiol is 5 physiologically synthesized to high circulating levels in a particular period during the4-day estrous cycle : it is the so-called preovulatory surge taking place in theafternoon of proestrus, just before ovulation which occurs during the night betweenproestrous and estrous phases. A sensitive physiological model for in vivo évaluationof aromatase inhibition was developped based on the inhibition of this preovulatory 10 surge of estradiol levels.

Adult female Wistar rats are monitored for regular 4-day estrous cyclicity by dailyvaginal smear examination ; after 2 or 3 regular cycles, animais are treated by asingle oral administration of the very low discriminative dose of 10 micrograms/kg ina 4 ml/kg volume around 04:00 PM on diestrus, i.e. on the day before proestrus. 15 Precisely 24 hours later, aortic blood sam pies are drawn under gaseous anesthésia.Plasma estradiol levels are measured by radioimmunoassay with commerciallyavailable kits (Diagnostic Systems Laboratories, Webster, Texas, U. S. A.). Controland test groups usually consist of 7 to 10 rats, depending on the number ofregularly cycling rats to distribute among study groups. Results are expressed in 20 pg/ml and then in % of inhibition taking the estradiol levels of control animaisreceiving only the oral vehicle as the 100 % référencé to allow comparisonsbetween different independent studies, since the preovulatory surge level may varyin each control group from one study to another between about 25 to 40 pg/ml. 29 012973

Table B : Inhibition of aromatase in vivo

Compound % inhibition at 10 pg/kg n Anastrozole 18 % - 33 % 2 YM 511 35 % 1 Example 7 47% 1 Example 14 49 % - 57 % 2 Example 6 57 % 1 Example 11 57 % - 59 % 2 Example 17 60 % 1

It can be seen that compounds Falling within the general formula (I) according to 5 the présent invention induce a slightly or markedly higher inhibition of estradiolbiosynthesis due to inhibition of aromatase in vivo than does YM 511, taken as astructural référencé of N-triazole (Okada et al., Chem Pharm Bull, 1996, 44 : 1871-1879), or anastrozole, a standard antiaromatase already used therapeutically. Thecompounds falling within the general formula (I) thus represent a substantial 10 improvement over the latter substances.

In vivo data (Table B) were not absolutely correlated with in vitro data (Table A)but, taken as a whole, both sets of biological results indicate that the sériés of 1-N-phenylamino-lH-imidazole dérivatives according to the présent invention yieldsnumerous nanomolar and subnanomolar inhibitors of aromatase among which 15 several exert effective in vivo inhibition of estrogen biosynthesis. These compoundsare therefore useful for counteracting or managing pathological or physiologicalestrogen-dependent mechanisms, preferentially in females (women or femaleanimais) but also in males (men or male animais). 20

Claims (13)

1. 30 012973 CLAIMS

   1. An imidazole dérivative of formula (I) :

   (I) and acid addition salts, solvatés and stereoisomeric forms thereof, wherein : • Ri and R2 are each independently hydrogen, (Ci-C6)alkyl or (C3- C8)cycloalkyl ; • n is 0,1 or 2 ; • R3, R4, R5 and Re are each independently hydrogen or a (CrC6)alkyl,halogen, cyano, (CrQalkoxy, trifluoromethyl, (Ci-C6)alkylthio, (Ct-C6)alkylsulfonyl, sulfonamido, acyl, (Ci-QJalkoxycarbonyl, or carboxamidogroup ; • R3 and R6 together with the phenyl ring bearing them can also form abenzofurane, or a N-methylbenzotriazole.
2. 2. A dérivative according to claim 1, wherein : • n is 0 or 1 ; • Rx and R2 are each independently hydrogen or (CrC5)alkyl ; • R3 is cyano or trifluoromethyl ; • R4 is hydrogen, (C1-C6)alkylz halogen, cyano, (Ci-Ce)alkoxy,trifluoromethyl, (CrÇQalkylthio, (Ci-C6)alkylsulfonyl or (CrC6)alkoxycarbonyl ; • R5 is hydrogen, halogen, (Ci-C6)alkoxy or trifluoromethyl ; • R6 is hydrogen ; 31 012973 • or R3 and Re together with the phenyl ring bearing them form a N-methylbenzotriazole ; and acid addition salts, solvatés and stereoisomeric forms thereof.
3. 3. A dérivative according to claim 1 or 2, wherein : • n is 0 or 1 ; • Rlf R2 and Re are each hydrogen ; • R4 is halogen, cyano or trifluoromethyl ; and acid addition salts, solvatés and stereoisomeric forms thereof.
4. 4. A dérivative according to one of daims 1 to 3, wherein R3 is cyano ;and acid addition salts, solvatés and stereoisomeric forms thereof.
5. 5. A dérivative according to one of daims 1 to 4, wherein R5 is hydrogen ortrifluoromethyl ; and acid additions salts, solvatés and stereoisomeric forms thereof.
6. 6. A dérivative according to one of daims 1 to 5, wherein n is 1 ; and acid addition salts, solvatés and stereoisomeric forms thereof.
7. 7. A pharmaceutical composition comprising a dérivative according to one ofdaims 1 to 6, or a pharmaceutically acceptable acid addition sait thereof, and apharmaceutical acceptable carrier.
8. 8. A pharmaceutical composition according to claim 7, which is an aromataseinhibitor composition.
9. 9. A pharmaceutical composition according to claim 7 or 8, comprising from 0.1to 400 mg of said dérivative.
10. 10. Use of a dérivative according to one of daims 1 to 6 or a pharmaceuticallyacceptable acid addition sait thereof in the manufacture of a médicament for the 32 012973 treatment or prévention of estrogen-dependent disorders, wherein said dérivative isoptionally combined with a sexual endocrine therapeutic agent.
11. 11. Use of a dérivative according to one of daims 1 to 6 or a pharmaceutically5 acceptable acid addition sait thereof in the manufacture of a médicament for the control or management of reproductive fonctions such as male or female fertility,pregnancy, abortion or delivery, wherein said dérivative is optionally combined witha LH-RH agonist or antagonist, an estroprogestative contraceptive, a progestin, ananti-progestin or a prostaglandin. 10
12. 12. Use of a dérivative according to one of daims 1 to 6 or a pharmaceuticallyacceptable acid addition sait thereof in the manufacture of a médicament for thetreatment or prévention of benign or malignant diseases of the breast, the utérus orthe ovary, wherein said dérivative is optionally combined with an anti-estrogen, a 15 progestin or a LH-RH agonist or antagonist.
13. 13. Use of a dérivative according to one of daims 1 to 6 or a pharmaceuticallyacceptable acid addition sait thereof in the manufacture of a médicament for thetreatment or prévention of benign or malignant diseases of the prostate or the 20 testis, wherein said dérivative is optionally combined with an antiandrogen, aprogestin, a lyase inhibitor or à LH-RH agonist or antagonist.