OA10195A - Carbazole derivatives with 17 20-lyase-inhibiting activity - Google Patents

Abstract

This invention relates to substituted carbazole derivatives, to processes for their preparation, to pharmaceutical compositions containing them and to their use in medicine for the reduction of oestrogen and/or androgen levels. Particular compounds of the invention are compounds of formula (I) wherein R1 and R4 each independently represent a hydrogen atom or a C1-6alkyl group; each R2 may be the same or different and represents an electron-withdrawing group; each R3 may be the same or different and represents an electron-withdrawing group; R5 is a group of formula (a), (b) or (c); R6 is a halogen atom, a C1-6alkyl group or a C1-6alkoxy group; m is zero or an integer 1 to 4; n is zero or an integer 1 to 3; and p is zero, 1 or 2 and pharmaceutically acceptable salts and solvates thereof.

Description

1 010195

CARBAZOLE DERIVATIVES WITH 17,20-LYASE-INHIBITING ACTIVITY

This invçntion relates to substituted carbazole dérivatives, to processes for theirpréparation, to phaîmaceutical compositions containing them and to their use inmedicine for the réduction of oestrogen and/or androgen levels. 5 17,20-Lyase activity is responsible for the conversion of 17a- hydroxyprogesterone to androstenedione and of 17a-hydroxypregnenolone todéhydroépiandrostérone. This activity is critical for the biosynthesis ofandrogens and estrogens, and inhibition of 17,20-lyase by kétoconazole (aknown antifungal agent) has been shown to reduce testosterone levels in 10 animais (English, K.F. et al.. Cancer Res. 47, 38-42 (1986) and men (Pont, A. etal., Arch. Intern. Med. 142, 2137-2140(1982)'

High dose kétoconazole has been used in the treatment of prostate cancer(Trachtenberg, J., J. Urol. 132, 61-63 (1984) and Pont, A., et al.. Arch. Intern.Med. 145, 1429-1431 (1985)). However, doses of kétoconazole that produce 15 inhibition of testosterone production are significantly more toxic than antifungaldoses, and more sélective 17,20-lyase inhibitors would be expected to hâve amuch better therapeutic index.

Inhibition of 17,20-lyase, as an âpproach to inhibition of testosteronebiosynthesis, also has the advantage of blocking adrenal as well as testicular 20 derived androgens (English, H.F. et al.. Cancer Res. 46, 38-42 (1986)), andshould be a more effective treatment for prostate cancer than that afforded byLHRH agonists.

Thus the présent invention provides compounds of general formula (I) R, CRÛ, 0) chr4r5 25 wherein R-| and R4 each independently represent a hydrogen atom or aC-j-ealkyl group; 2 010195 each R2 may be the same or different and represents an electron-withdrawinggroup; each R3 may be the same or different and represents an electron-withdrawinggroup;

Rs is a group of formula

Rg is a halogen atom, a C^alkyl group or a Cv6 alkoxy group;m is zéro or an integer 1 to 4;n is zéro or an integer 1 to 3; andp is zéro, 1 or 2 and pharmaceutically acceptable salts and solvatés thereof.

The substituents R2 may each occupy any available position of the 5, 6, 7 or 8positions.

The substituents R3 may each occupy any available position of the 1, 2, 3 or 4positions, preferably the 1, 3 or 4 positions. r

The group —CHR4R5 may occupy any of the 1, 2, 3 or 4 positions, for example,the 1,2 or 3 positions. Preferably the group —CHR4R5 will be in the 2 position.

Thus, in one aspect the invention provides compounds of formula (la)

wherein R<, R2, R3, R4, R5. m and n are as hereinbefore defined. 3 010195

The substituent Rg may be attached to any carbon atom in the pyridyl ring, butpreferably is attached in the 3,4 or 5 positions.

It will be appreciated by those skilled in the art that the numbering of the ringSystem for individûal compounds within the scope of formula (I) will vary 5 according to the nature, number and position of substituents. For conveniencethe numbering of ring atoms adopted herein is that shown in formula (I) aboveexcept where individûal compound names are given.

It will be appreciated that the compounds of formula (I) may contain a chiralcentre. It is to be understood that formula (I) is intended to encompass ail 10 enantiomers and diastereoisomers of the compounds of the invention as well asmixtures thereof, including racemates.

Electron-withdrawing groups are well known to those skilled in the art and anysuch group may be employed. Such groups include halogen atoms, such asfluorine, chlorine and bromine atoms, nitrile groups, nitro groups, trifluoromethyl 15 groups, aldehydo groups, keto groups and carboxylic acid and ester groups andare preferably selected from fluorine atoms, chlorine atoms and nitrile groups.Particularly preferred as compounds of formula (I) are those wherein each of F?2and R3 represents a fluorine atom. r

Suitably R2 represents a fluorine atom and m is an integer 1 to 4. 20 When m is 1 R2 will preferably be in the 5 or 7 position, especially the 7-position. C-] _galkyl and C1_6alkoxy groups may contain straight or branched chain alkyl *groups, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, pentyl orhexyl groups, preferably Calkyl groups. Thus, for example, each of R-| and 25 R4 may be a hydrogen atom or a methyl, ethyl, propyl or butyl group. R-| and R4 are each preferably à hydrogen atom or a methyl group. 4 010195

In one preferred group of compounds of formula (I) R5 is a pyridin-3-yl orpyridin-4-yl group. f

Suitably represents a fluorine atom, a methyl group or a methoxy group.

In a preferred group of compounds of formula (I), R-| and R4 each represent ahydrogen atom or a methyl group, R2 and R3 are fluorine atoms, R6 is a fluorineatom, a methyl group or a methoxy group, m is an integer 1 to 4, n is zéro or aninteger 1 to 3, and p is zéro, 1 or 2.

In a particularly preferred group of compounds of formula (l), R1 and R4 eachrepresent a hydrogen atom, R2 and R3 are fluorine atoms, R6 is a fluorine atomor a methyl or methoxy group and m, n an3 p each independently representzéro, 1 or 2.

Spécifie compounds according to the invention include : 2-Fluoro-7-[1,2,4]triazol-1 ylmethyl-9H-carbazole 2-Fluoro-7-pyridin-3-ylmethyl-9H-carbazole 2-Fluoro-7-pyridin-4-ylmethyl-9H-carbazole 2-Fluoro-7-(3-fluoropyridin-4-ylmethyl)-9H-carbazole 2-Fluoro-7-(3-methylpyridin-4-ylmethyp-9H-carbazole 2-Fluoro-7-(3-methoxypyridin-4-ylmethyl)-9H-carbazole 1.7- Difluoro-2-[1,2,4]triazol-1 -ylmethyl-9H-carbazole 2,4-Diflûoro-7-[1,2,4]triazol-1 -ylmethyl-9H-carbazole 1.4.7- Trifluoro-2-[1,2,4]triazol-1-ylmethyl-9H-carbazole and pharmaceutically acceptable salts and solvatés thereof.

Suitable pharmaceutically acceptable salts of the compounds of formula (I) tinclude acid addition salts derived from inorganic and organic acids, such ashydrochlorides, hydrobromides, sulphates, phosphates, citrates, tartrates,maleates, fumarates, succinates, p-toluenesulphonates andmethanesulphonates. Other suitable salts will be readily apparent to one skilledin the art. Hydrochloride, sulphate and phosphate salts are especiallypreferred. Salts which are not pharmaceutically acceptable may be useful in the 5 010195 préparation of compounds of formula (I) and these form a further part of theinvention.

Compounds of the invention may be isolated in association with solventmolécules by crystallisation from or évaporation of an appropriate solvent. Suchsolvatés of the compounds of formula (I) are included within the scope of theprésent invention.

References hereinafter to a compound according to the invention includes boththe compounds of formula (I) and their pharmaceutically acceptable salts andsolvatés.

The compounds according to the invention are potent and sélective inhibitors ofthe enzyme steroidal 17,20-lyase, which is a key enzyme involved in theconversion of C2i-steroids (e.g. pregnjenolone) into androgens (e.g.testosterone) and oestrogens (e.g. oestradiol).

The 17,20-lyase-inhibiting activity of the compounds of formula (I) wasdemonstrated in vitro by their ability to inhibit the conversion of 17a-hydroxypregnenolone into déhydroépiandrostérone by human testicular 17,20-lyase and of 17a-hydroxyprogesterone into androstenedione by rat testicular17,20-lyase. These assays were conducted according to a method based onthat of Ayub and Level, J. Steroid Biochem, 1987, 28, 521.

In in vivo tests the compounds of the invention were tested for their ability tosuppress the élévation of testosterone levels produced in male rats whenstimulated with human chorionic gonadotrophin (hCG).

Inhibitors of 17,20-lyase reduce circulating and local levels of androgens andoestrogens. The compounds of the invention can thus be used in the treatment *of androgen- and/or oestrogen-dependant diseases such as malignant andbenign diseases of the breast, endometrium, ovary, prostate and pancréas.These diseases include cancer of the prostate, breast and endometrium,prostatic hypertrophy and hyperplasia, fibrocystic breast disease, endometriosisand polycystic ovarian disease. The compounds of formula (I) are also useful inthe treatment of Cushing's syndrome, gynecomastia, prématuré labour,precocious puberty, female hirsutism, premenstrual syndrome, male pattern 6 010195 baldness and acné. The compounds of the invention will be particularly usefulin the treatment of prostate cancer.

The invention thus further provides compounds of formula (I) and theirpharmaceutically acceptable salts and solvatés for use as active therapeuticagents in particular for the treatment of conditions whose underlying aetiology isassociated with elevated androgen and/or oestrogen levels in animais(especially humans).

In a particular aspect of the présent invention there is provided a compound offormula (I) or a pharmaceutically acceptable sait or solvaté thereof for use in thetreatment of prostate cancer.

In a further or alternative aspect there is provided a method for the lowering ofthe levels of androgens and/or oestrogens .in a mammal including a humancomprising administration of an effective amount of a compound of formula (I) ora pharmaceutically acceptable sait or solvaté thereof.

There is also provided in a further or alternative aspect use of a compound offormula (I) or a pharmaceutically acceptable sait or solvaté thereof for themanufacture of a médicament for the lowering of levels of androgens and/oroestrogens. Λ

It will be appreciated by those skilled in the art that reference herein totreatment extends to prophylaxis as well as the treatment of establishedsymptoms.

While it is possible that, for use in therapy, a compound of the invention may beadministered to a patient as the raw Chemical, it is préférable to présent theactive ingrédient as a pharmaceutical formulation.

The invention accordingly provides a pharmaceutical formulation comprising acompound of formula (I) or a pharmaceutically acceptable sait or solvaté thereoftogether with one or more pharmaceutically acceptable carriers or excipientsand, optionally, other therapeutic and/or prophylactic ingrédients. The carriersmust be "acceptable" in the sense of being compatible with the other ingrédientsof the formulation and not deleterious to the récipient thereof. 7 010195

Pharmaceutical formulations include those suitable for oral, rectal, nasal, topical, implant or parentéral (including intramuscular, subcutaneous and intravenous) administration or in a form suitable for administration by inhalation« or insufflation. Tbe formulations may, where appropriate, be convenientlypresented in discrète dosage units and may be prepared by any of the methodswell known in the art of pharmacy. Ail methods include the step of bringing intoassociation the active compound with liquid carriers or finely divided solidcarriers or both and then, if necessary, shaping the product into the desiredformulation.

For oral administration, the pharmaceutical compositions may take the form of,for example, tablets or capsules prepared by conventional means withpharmaceutically acceptable excipients such as binding agents (e.g. pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropyl · »· methylcellulose); fillers (e.g. lactose, microcrystalline cellulose or calciumphosphate); lubricants (e.g. magnésium stéarate, talc or silica); disintegrants(e.g. potato starch or sodium starch glycollate); or wetting agents (e.g. sodiumlauryl sulphate). The tablets may be coated by methods well known in the art.

Liquid préparations for oral administration may take the form of, for example,solutions, syrups or suspensions, or they may be presented as a dry product forconstitution with water or other suifable vehicle before use. Such liquidpréparations may be prepared by conventional means with pharmaceuticallyacceptable additives such as suspending agents (e.g. sorbitol syrup, methylcellulose or hydrogenated edible fats); emulsifying agents (e.g. lecithin oracacia); non-aqueous vehicles (e.g. almond oil, oily esters or ethyl alcohol); andpreservatives (e.g. methyl or propyl-p-hydroxybenzoates or sorbic acid).

For topical administration in the mouth, the pharmaceutical compositions may *take the form of buccal or sub-lingual tablets, drops or lozenges formulated inconventional manner.

For topical administration to the epidermis the compounds of the invention maybe formulated as creams, gels, ointments or lotions or as transdermal patches.Such compositions may, for example, be formulated with an aqueous or oily 8 010195 base with the addition of suitable thickening, gelling, emulsifying, stabilising,dispersing, suspending, and/or colouring agents.

The compounds of the invention may also be formulated as depot préparations.Such long acting formulations may be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus, forexample, the compounds may be formulated with suitable polymeric orhydrophobie materials (for example as an émulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble dérivatives, for example as a sparinglysoluble sait.

The compounds of the invention may be formulated for parentéral administrationby injection, conveniently intravenous, intrarrtyscular or subeutaneous injection,for example by bolus injection or continuous intravenous infusion. Formulationsfor injection may be presented in unit dosage form e.g. in ampoules or in multi-dose containers, with an added preservative. The compositions may take suchforms as suspensions, solutions or émulsions in oily or aqueous vehicles, andmay contain formulatory agents such as suspending, stabilising and/ordispersing agents. Alternatively, the active ingrédient may be in powder form forconstitution with a suitable vehicle, e.g. stérile pyrogen-free water, before use.

The compounds of the invention may ^Iso be formulated in rectal compositionssuch as suppositories or rétention enemas, e.g. containing conventionalsuppository bases such as cocoa butter or other glyceride.

For intranasal administration the compounds of the invention may be used, forexample, as a liquid spray, as a powder or in the form of drops.

For administration by inhalation the compounds according to the invention areconveniently delivered in the form of an aérosol spray présentation frompressurised packs or a nebuliser, with the use of a suitable propellant, e.g.dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane,1,1,1,2-tetrafluoroethane, carbon dioxide or other suitable gas. In the case of apressurised aérosol the dosage unit may be determined by providing a valve todeliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an 010195 inhaler or insufflator may be formulated containing a powder mix of a compoundof the invention and a suitable powder base such as lactose or starch.

Any of the pharmaceutical compositions described above may be presented in aconventional manner associated with controlled release forms.

Preferably the pharmaceutical compositions according to the invention aresuitable for oral, rectal or topical administration. A convenient unit dose formulation contains the active ingrédient in an amountof from 0.1 to 200mg.

It will be appreciated that the amount of a compound of formula (I) required foruse in treatment will vary not only with the'particular compound selected, but also with the route of administration, the nature of the condition being treated «- and the âge, weight and condition of the patient and will ultimately be at thediscrétion of the attendant physicien or veterinarian. In general, however, asuitable dose will be in the range of from about 1 to about 500mg per day,preferably in the range of 20 to 200mg per day, most preferably in the range of50 to 120mg per day. A suitable daily dose for use in prophylaxie will generally be in the range of0.1 mg to 50mg. '

The desired dose may conveniently be presented in a single dose or as divideddoses administered at appropriate intervals, for example as two, three, four ormore sub-doses per day. The compound is conveniently administered in unitdosage form.

The compounds of the présent invention may also be used in combination with «other therapeutic agents, for example, other androgen and/or oestrogenlowering agents, or anticancer agents. In particular the compounds of theinvention may be employed together with known anticancer agents.

The invention thus provides, in a further aspect, a combination comprising acompound of formula (I) as defined herein together with another therapeuticallyactive agent, in particular an anticancer agent. 10 010195

The combination referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a pharmaceutically acceptable carrier therefor comprise a further aspect of the invention. »

When compounds of formula (I) are used in combination with a secondtherapeutic agent, the compounds may be administered either sequentially orsimultaneously by any of the routes described above.

Suitable therapeutic agents for use in the combinations defined above include,for example cyproterone acetate, flutamide and anandron.

When compounds of formula (I) are used in combination with a secondtherapeutic agent effective to reduce levels of androgens and/or oestrogens in amammal including a human the dose of each compound may vary from thatwhen the compound is used atone. Thus when compounds of formula (I) areused together with a second therapeutic agent the dose of each compound may - be the same or different to that employed when the compound is used alone.Appropriate doses will be readily appreciated by those skilled in the art.

The compounds according to the invention may be prepared by any processknown in the art for the préparation of compounds of analogous structure. Inthe following description R-|, R2, R3, R4, R5, Rg, ni, n and p are as defined forgeneral formula (I) unless otherwise specified.

In one general process (A), a compound of general formula (I) wherein R-|represents a hydrogen atom may be prepared from an intermediate of formula (II)

N 3

(II) by cyclisation. The reaction is conveniently effected in the presence of a suitable solvent, such as a hydrocarbon solvent, for example dodecane, or a 11 010195 halogenated solvent, such as dichlorobenzene, preferably at elevatedtempérature, for example 100 to 300θ0, preferably 150 to 220θ0. Generalprocess (A) is particularly useful for the préparation of compounds of formula (I) t wherein R5 is a triazole group. » 5 Intermediates of formula (II) may be prepared from the corresponding amines of

by treatment with sodium nitrite in the presence of a minerai acid, e.g. sulphuricacid, followed by sodium azide. The reaction is conveniently effected in 10 aqueous solution.

Compounds of formula (III) may be prepared from the corresponding nitrocompounds of formula (IV)

(IV) by réduction using hydrogen or a hydrogen-donor, e.g. ammonium formate, in15 the presence of a catalyst, such as a noble métal catalyst, e.g. platinum, « palladium, platinum oxide or rhodium, which may be supported, e.g. oncharcoal. The réduction may be carried out in a solvent such as an alcohol e.g.methanol or éthanol (which may be aqueous), acetic acid, aqueous acetic acid,an ether e.g. dioxan, an ester e.g. ethyl acetate or an amide e.g. 20 dimethylformamide, and conveniently at a température of from '10 to +50θΰ,preferably 20 to 30°C. 12 010195

Compounds of formula (IV) may be prepared from compounds of formula (V)

by treatment with a compound of formula HR5 or the sodium sait thereof. Thereaction is conveniently effected in a suitable solvent, e.g dimethylformamide.

Intermediates of formula (V) may be prepared from compounds of formula (VI)

by free radical bromination, for example using N-bromosuccinimide in thepresence of an initiator, such as a peroxide and/or ultra-violet light. Thereaction is conveniently effected in a'non-polar solvent, such as a halogenated 10 solvent, e.g. chloroform or tetrachloromethane, at a température of 20 to 80^C.Compounds of formula (VI) may be prepared from compounds of formula (VII)

(wherein L represents a readily displaceable atom or group) by reaction with acompound of formula (VIII)

B(OH)2 (VIII) 15 R^c 13 010195 in the presence of a suitable palladium(O) catalyst such astetrakis(triphenylphosphine)palladiurn(O) and a base, e.g. sodium carbonate, ina suitable aqueous solvent such as an alcohol, e.g. éthanol, an aromatichydrocarbon, e.g. benzene, or an ether, e.g. dimethoxyethane, or an aqueous 5 mixture of solvents. Suitable atoms or groups represented by L include halogenatoms, e.g. bromine or iodine atoms, or a triflate group.

In another general process (B), a compound of formula (I) may be preparedfrom a compound of formula (IX) .(R,), 3'a «y. (IX) C(OH)R4R5 10 by deoxygenation. The deoxygenation reaction is effected using a suitablereducing agent such as hydrogen in the presence of a catalyst, such as a noblemétal catalyst, e.g. platinum, palladium, platinum oxide or rhodium, which maybe supported, e.g. on charcoal. The reaction may conveniently be carried out ina solvent such as an alcohol, e.g. methanol or éthanol, which may be aqueous, 15 in the presence of an acid, e.g. hydrochloric acid, preferably at elevatedtempérature, e.g. at the reflux température of the solvent or at elevatedpressure. General process (B) is particularly useful for the préparation ofcompounds of formula (I) wherein R5 is a pyridyl group.

Intermediates of formula (IX) may be prepared from compounds of formula (X)

by reaction with compounds of formula (XI)Hal - Rs (XI) 14 010195 in the presence of a suitable base, such as an alkyllithium, e.g. n-butyllithium.The reaction is conveniently effected in the presence of a suitable solvent suchas an ether, e.g. diethyl ether, dimethoxyethane or tetrahydrofuran, or a mixtureof solvents, suitably at low température, e.g. -90 to - 50θ, preferably about 5 -70°C.

Compounds of formula (X) may be prepared from compounds of formula (XII)

by oxidation. Suitable oxidising agents will be readily apparent to one skilled inthe art and include pyridinium chlorochromate, potassium dichromate in 10 sulphuric acid and barium manganate. The reaction may conveniently beeffected in the presence of a solvent, e.g. a halogenated solvent such asdichloromethane.

Compounds of formula (XII) may be prepared from compounds of formula (XIII)

15 by cyclisation. The reaction is conveniently effected in the presence of asuitable solvent, such as a hydrocarbon solvent, e.g. dodecane, or ahalogenated solvent, e.g. dichloromethane, preferably at elevated température,e.g. 100 to 300°C, preferably 150 to 220°C.

Compounds of formula (XIII) may be prepared from compounds of formula (XIV) (XIV) 010195

by treatment with sodium nitrite in the presence of a minerai acid, e.g. sulphuricacid, followed by sodium azide. The reaction is conveniently effected inaqueous solution.

Compounds of formula (XIV) may be prepared from compounds of formula (XV)

(wherein G represents a hydroxy protecting group) by réduction using hydrogenor a hydrogen-donor, e.g. ammonium formate, in the presence of a catalyst,such as a noble métal catalyst, e.g. platinum, palladium, platinum oxide or 10 rhodium, whicb may be supported, e.g. on charcoal and subséquent removal ofthe protecting group G. The réduction may conveniently be carried out in asolvent such as an alcohol, e.g. methanol or éthanol, which may be aqueous,optionally in the presence of an acid, e.g. hydrochloric acid. Suitable hydroxyprotecting groups are described hereinafter. 15

Compounds of formula (XV) may be prepared from compounds of formula (XVI) (R,)

L NO2 « (XVI) (wherein L represents a readily displaceable atom or group) by reaction with a compound of formula (XVII) 16 B(OH)2 (XVII) 010195

in the presence of a suitable patladium(O) catalyst such astetrakis(triphenylphosphine)palladium(O) and a base, e.g. sodium carbonate, ina suitable aqueous solvent such as an alcohol, e.g. éthanol, an aromatichydrocarbon, e.g. benzene, or an ether, e.g. dimethoxyethane, or an aqueousmixture of solvents preferably at elevated température. Suitable atoms orgroups represented by L include a halogen atom, e.g. a bromine or iodine atom,and a triflate group.

Alternative synthetic routes to the intermediates of formula (X) will be readilyapparent to those skilled in the art.

In another general process (C) a compound of formula (I) according to theinvention may be converted into another compound of the invention usingconventional procedures.

According to one embodiment of general process (C) a compound of formula (I)wherein R-| represents a hydrogen atom may be alkylated using conventionaltechniques. The reaction may be effected using a suitable alkylating agentsuch as an alkyl halide, alkyl tosylate or dialkylsulphate. The reaction mayconveniently be carried out in an inert organic solvent such as an amide, e.g.dimethylformamide, or an ether, e.g. tetrahydrofuran, preferably in the presenceof a base. Suitable bases include, for example, alkali métal hydrides, e.g.sodium hydride, alkali métal carbonates, e.g. sodium carbonate, or alkali métalalkoxides, e.g. sodium or potassium, methoxide, ethoxide or t-butoxide. The talkylation reaction is conveniently effected at a température of 25 to 100θ0.

According to another general process (D), a compound of formula (I) accordingto the invention, or a sait thereof may be prepared by subjecting a protecteddérivative of formula (I) or a sait thereof to reaction to remove the protectinggroup or groups. Thus, at an earlier stage in the préparation of a compound offormula (I) or a sait thereof it may hâve been necessary and/or désirable to 010195 17

• I protect one or more sensitive groups in the molécule to prevent undesirableside reactions. Such protection may be effected in conventional manner, forexample as described in ’Protective Groups in Organic Chemistry' Ed.J.F.W.McOmie (Plénum Press 1973) or ’Protective Groups in Organic Synthesis’ byT W Greene (John Wiley and Sons 1981).

In compounds of formula (I) wherein R-j représente hydrogen the group NR-|may be protected for example with a conventional amino protecting group.Such groups may include for example aralkyl groups, such as benzyl,diphenylmethyl or triphenylmethyl groups; and acyl groups such as tosyl, N-benzyloxycarbonyl or t-butoxycarbonyl.

Removal of any amino protecting groups. présent may be achieved byconventional procedures. Thus an aralkyl group such as benzyl, may becleaved by hydrogenolysis in the presence'of a catalyst (e.g. palladium oncharcoal); an acyl group such as t-butoxycarbonyl may be removed by cleavagewith, for example, hydrogen chloride in dioxan or sodium methoxide inmethanol.

As will be appreciated, in some of the general processes (A) to (C) describedabove it may be necessary or desired to protect any sensitive groups in themolécule as just described. Thus, a .reaction step involving deprotection of aprotected dérivative of general formula (I) or a sait thereof may be carried outsubséquent to any of the above described processes (A) to (C).

Where it is desired to isolate a compound of the invention as a sait, for exampleas an acid addition sait, this may be achieved by treating the free base ofgeneral formula (I) with an appropriate acid, preferably with an équivalentamount. Solvatés of the compounds of the invention may be prepared by *crystallisation from or évaporation of an appropriate solvent solution of thecompounds of formula (I). Séparation of enantiomers of formula (I) may becarried out in conventional manner, for example by resolution of racemicmixtures e.g. using chiral HPLC techniques or by stereospecific synthesis fromisomerically pure starting materiel or any convenient intermediate, for exampleas described in Stereochemistry of Carbon Compounds by E.L. Eliel (McGrawHill, 1962) and Tables of Resolving Agents by S.H. Wilen. 010195 18

I

Thus, according to a further aspect of the invention, the following reactions may,if necessary and/or desired be carried out in any appropriate sequencesubséquent to any of the processes (A) to (C): (i) removal of any protecting groups; (ii) conversion of a compound of formula (I) or a sait or solvaté thereof into apharmaceutically acceptable sait or solvaté (for example, hydrate) thereof; (iii) séparation of a racemic mixture into individual enantiomers of formula (I).

As well as being employed as the last main step in the préparative sequence,the general methods indicated above for the préparation of the compounds ofthe invention may also be used for the introduction of the desired groups at anintermediate stage in the préparation of the required compound. It shouldtherefore be appreciated that in such multi-stage processes, the sequence ofreactions should be chosen in order that the reaction conditions do not affectgroups présent in the molécule which are desired in the final product.

As indicated above the compounds of the invention are useful as 17,20-lyaseinhibitors. 17,20-Lyase inhibition may be demonstrated by the following tests: ACTIVITY in vitro z

Biological activîty in vitro was determined by measuring the inhibition of 17,20-lyase activity in a microsomal préparation from human testes. A range ofconcentrations of each compound was incubated with the microsomalpréparation and 17-a-hydroxy (21 -14C) pregnenolone as substrate for 60 minsat 37°C. Radioactive product was assayed and enzyme inhibition determinedby comparison with uninhibited control samples. ACTIVITY in vivo

Biological activity in vivo was determined in a rat model of testosteronebiosynthesis. Compounds were administered p.o. at a dose of 3mg/kg. Onehour later the rats were given human chorionic gonadotrophin (hCG) s.c. tostimulate testosterone synthesis and two hours after hCG administration, bloodwas taken and sérum testosterone concentrations measured. Inhibitory activitywas determined by comparison to values from rats receiving vehicle only. 010195 19 «

The results of the in vitro and in vivo assays for the compounds of the followingexamples are shown in Table 1. The results for the known 17,20-lyase inhibitorkétoconazole are included for comparison.

T

Table 1

Example In vitro assav ICS0 concentration (nM) In vivo assav% Inhibition of sérumtestosterone Kétoconazole 85 56* 1 97 50 2 6 84 3 5 74 4 8 · 40 5 5 78 6 19 71 7 41 14 8 12 36 9 25 5 5 * The dose of kétoconazole was 30mg/kg.

Acute toxicity studies indicate that single doses of 3mg/kg p.o. of the compoundof Example 2 are well tolerated in rats and no overt toxicity was observed.

The invention is further illustrated by the following non-limiting examples. Ailtempératures are in °C. Chromatography was performed on silica gel. Dried 10 means dried over anhydrous magnésium sulphate. DMSO means dimethylsulphoxide. DMF means dimethylformamide. THF means tetrahydrofuran.DME means dimethoxyethane. IPE means isopropyl ether.

Intermediate 1 4-(t-Butyldimethvlsilvloxvmethyl)phenylboronic acid 15 (i) 4-(t-Butvldimethylsilyloxymethyl)-bromobenzene A solution of t-butyldimethylsilyl chloride (21.1g) in DMF (25ml) was treateddropwise with a solution of 4-bromo-benzyl alcohol (25g) and imidazole (18.2g)in DMF (100ml). Resulting solution was left standing at room température for 20 010195 72h. Reaction mixture was added to ethyl acetate (2000ml) and organic layerwas washed with 2N HCl (2 x 250ml), water (4 x 250ml), brine (250ml), driedand evaporated to give a yellow oil. Chromatography on silica gel (1kg), elutingwith cyclohexane/ethyl acetate (10.1 v/v) gave the product as a pale yellow oil. 5 (ii) 4(t-Butyldimethvlsilvloxymethyl)phenvlboronic acid A solution of (i) (32.37g) in dry tetrahydrofuran (250ml) was cooled to -70°under N2 n-Butyllithium (70ml of a 1.6M solution in hexane) was addeddropwise over 15 minutes maintaining température below -65°C. Resulting 10 yellow solution was stirred below -70°C for 2h, then triisopropyl borate (75ml)was added dropwise, maintaining température below -65°. When addition wascomplété, the reaction was allowed to warm to room température, thenquenched with water (550ml) and then ether (250ml).

The organic layer was separated off and the aqueous phase extracted with 15 ether (2 x 500ml). The combined organic layers were washed with brine(200ml), dried and evaporated to give the product as a pale yellowcrystalline solid.

For the avoidance of doubt, in the compounds of Examples 1 to 7 and 9, R2 is inthe 7-position of formula (I), in Example 8 R2 is in the 5 and 7 positions, in 20 Example 7 R3 is in the 1-position and Γη Example 9 R3 is in the 1 and 4 positionsof formula (I). Individual compound names are in accordance with IUPACnomenclature.

Example 1 25 2-Fluoro-7-f 1.2,4ltriazol-1 -ylmethyl-9H-carbazole 1 (i) 4-Fluoro-4,-methvl-2-nitro-biphenyl * A solution of 2-bromo-5-fluoronitrobenzene(15g) in benzene (60ml) was rapidlystirred under N2, and treated with tetrakistriphenylphosphine palladium (0)(1.13g), aq. 2M sodium carbonate (35.7ml) and a solution of p-tolylboronic acid 30 (10.2g) in 95% éthanol (25ml). Mixture was heated at 95°C and rapidly stirred under N2 for 16h. Reaction mixture was extracted with ethyl acetate (400ml)then (2x 100ml). The combined extracts were washed with water (100ml), brine(100ml), dried (MgSO4) and evaporated to give a brown oil. Chromatography 21 010195 on silica gel (500g), eluting with cyclohexane/ethyl acetate (25:1v/v) gave thetitle compound. « 1 (ii) 4'-Bromomethyl-4-fluoro-2-nitro-biphenyl5 A solution of 1 i) (2.04g) in carbon tetrachloride (40ml) was treated with a trace of benzoyl peroxide, and N-bromosuccinimide (1.81g). Mixture was refluxedover a 150W bulb for 1h, then the precipitate of succinimide was filtered off, andwashed with CCI4 (2 x 5ml). The filtrate and washings were evaporated to givethe title compound as a yellow oil. 10 1(iii) l-i^-Fluoro-Z-nitro-biphenyM-vImethvD-IH-fl, 2, 4)triazoleA solution of 1,2,4-triazole (0.613g) in DMF (5ml) was treated with sodiumhydride (0.266g of an 80% dispersion, in oil). After 5 minutes when aileffervescence had ceased, mixture was treated with a solution of 1 ii) (3.1g) in 15 DMF(3ml). Resulting pale brown solution was stirred at room température for20h. Mixture was added to ethyl acetate (500ml) and the organic phase waswashed with water (4 x 50ml) then extracted with 2N HCl (6 x 80ml), thenwashed with brine (100ml), dried (MgSO4) and evaporated. The residue waspurified by chromatography on silica gel (150g) eluting with CHCI3 to give a 20 yellow crystalline solid. Elution with CHCI3/MeOH (19:1 v/v) gave the titlecompound as a yellow gum. r 1 (iv) 4-Fluoro-4’-f1,2,4ltriazol-1 -ylmethvl-biphenyl-2-vlamineA solution of 1 iii) (1.20g) in acetic acid (5ml) was treated with 10% Pd/C catalyst 25 (400 mg) and stirred under H2 for 3h. The catalyst was filtered off through a

Kieselguhr pad, and washed with acetic acid (2x5 ml), then the filtrate andwashings were evaporated to a yellow gum. This was dissolved indichloromethane (100ml) and was carefully washed with sat. aq. NaHCO3 *(100ml), then the organic layer was dried (MgSO4) and evaporated to give the 30 title compound as a brownish crystalline solid. 1 (v) 1 -(2'-Azido-4,-fluoro-biphenvl-4-ylmethvl)-1 H-f 1.2.41triazoleA solution of 1iv) (970mg) in water (9ml) and conc. sulphuric acid (0.75ml) wascooled in an ice-bath. A solution of sodium nitrite (261 mg) in water (3ml) was 35 added dropwise. Resulting yellow solution was stirred at ice-bath température 010195 22 4 for 30 minutes. A solution of sodium azide (243mg) in water (6ml) was addeddropwise. Rapid effervescence of N2 gas was observed. After 30 minutes themixture was neutralized with sat. aq. NaHCO3, then extracted withdichloromethane (4x 50ml). The combined extracts were dried (MgSO4), andevaporated to give the title compound as a pale brown crystalline solid. 1 (vi) 2-Fluoro-7-f1,2.41triazol-1 -ylmethyl-9H-carbazole A solution of 1v) (200mg) in 1,2-dichlorobenzene (5ml) was heated at 170°C for4h. The reaction mixture was diluted with ethyl acetate (200ml) and extractedwith 2N HCl (3 x 60ml). The organic layer was then washed with brine (50ml),dried (MgSO4) and evaporated to give the impure product plusdichlorobenzene. Impure product was added to cyclohexane (100ml) and thesolid filtered off and dried to give crystals of the title compound. NMR δ (DMSO-d6) 5.57 s (2H), 6.99 t (1 H),.7.12 d (1H), 7.25 dd (1H), 7.38 s(1H), 8.00 s (1H), 8.05 d (1H), 8.10 dd (1H), 8.70 s (1H), 11.42 s (1H).

Example 2 2-Fluoro-7-pyridin-3-ylmethvl-9H-carbazole 2(i) 4-t-Butvldimethvlsilvloxvmethvl-4'-fluoro-2‘-nitrodiphenyl A solution of 4-(t-butyldimethylsilyloxymethy!)-phenylboronic acid(Jntermediate 1) (25g) and 2-bronio-5-fluoro-nitrobenzene (15.5g) in drydimethoxyethane (120ml) was treated with tetrakistriphenylphosphine palladium(O) (1.1g) and 2M aq sodium carbonate (40ml). Resulting mixture was refluxedunder N2 for 16h. Reaction mixture was diluted with ethyl acetate (600ml) andwashed with water (2 x 100ml), brine (100ml), dried (MgSO4) and evaporated togive a brown oil. Chromatography on silica gel (750g), eluting with cyclohexane-> cyclohexane/ethyl acetate (15:1 ) gave the title compound as a yellow oil. 2(ii) 2-Amino~4'-t-butvldimethvlsilyloxvmethvl-4-fluorodiphenvl A solution of 2(i) (21g) in methanol (300ml) was treated with 10% Pd/C catalyst.(3.4g) and dry ammonium formate (16.48g). Mixture was stirred under N2 atroom température for 24h. The catalyst was filtered off, washed with methanol(2 x 25ml), under a blanket of N2, then the nitrate and washings evaporated togive a brownish residue. This was dissolved in water (200ml) and extracted 010195 23 « with dichloromethane (3 x 200ml). Combined extracts were dried (MgSO4) andevaporated to give the title compound as a brown oil. « 2(iii) (2,-Azido-4'-fltforo-biphenvl-4-vl)-methanol A solution of 2(ii) (18.408g) in dioxan (200ml) and water (200ml) was treatedwith conc.sulphuric acid (11.47ml) then cooled to 5°C. A solution of sodiumnitrite (4.00g) in water (15ml) was added dropwise. The resulting orangesolution was stirred at 5°C for 30 minutes, then a solution of sodium azide(3.75g) in water (20ml) was added carefully, avoiding over-effervescence of N2gas. When addition was complété, reaction was stirred at 5°C for 30 minutes,then carefully treated with sat. aq NaHCO3 solution until neutral. Mixture wasextracted with dichloromethane (5 x 200ml) and the extracts were dried(MgSO4), and evaporated to give the title compound as a brown oil. » 2(iv) (7-Fluoro-9H-carbazol-2-vh-methanol A solution of 2(iii) (13.9g) in 1,2-dichlorobenzene (200ml) was refluxed under N2for 3h. * Mixture was allowed to cool, and then was added to cyclohexane(100ml). Solid material was filtered off, and washed with cyclohexane (2 x100ml), then dried in vacuo to give the title compound as a pale brown solid. 2(v) 2-Fluoro-7-formyl-carbazole-9-caFboxvlic acid tert-butyl ester A solution of 2(iv) (7.18g) in dichloromethane (1500ml) was treated with bariummanganate (54.5g) and stirred under N2 for 5h. The solid was filtered off andwashed with dichloromethane (4 x 100ml). Filtrate and washings wereevaporated to give a green-brown solid. This was suspended indichloromethane (100ml) and treated with 4-dimethylaminopyridine (3.78g) anddi-'butyldicarbonate (7.43g). Resulting brown solution was stood at roomtempérature for 30 minutes, then diluted with ethyl acetate. Organic solution *was washed with 2N HCl (2 x 400ml), water (200ml), brine (2 x 200ml) thendried (MgSO4) and evaporated to give impure product. This was purified bychromatography, on silica gel (500g), eluting with cyclohexane/ethyl acetate(15:1 v/v) to give the title compound as a pale cream solid. 2(vi)2-Fluoro-7-(hvdroxv-pyridin-3-vl-methyl)-carbazole-9-carboxvlic acid tert- butyl ester 010195 24 < A solution of 3-bromopyridine (0.306ml) in ether (10ml) was cooled to -45°Cunder N2. A solution of "butyllithium (2ml of 1.6M sol. in hexane) was addeddropwise. Resulting brownish suspension was warmed to 0°C, then cooledagain to -45°C and left for 15 minutes. A sol. of 2(v) (0.5g) in dry THF (10ml)was added. Mixture was stirred at -40°C for 1h, then quenched with ammoniumchloride solution (20ml). Mixture was extracted with ethyl acetate (3 x 30ml) andthe combined extracts were washed with brine (200ml), dried (MgSO4) andevaporated to give the title compound as a yellow foam. 2(vii) 2-Fluoro-7-pyridin-3-vlmethvl-9H-carbazole A solution of 2(vi) (670mg) in methanol (20ml) was treated under N2 withpalladium black catalyst (300mg) and conc. HCl (0.8ml). Mixture was stirredunder H2 for 3h at 50°C, then a further 300mg of catalyst was added andreaction continued for 3h. Catalyst was filtered off, and washed with methanol(2 x 10ml), then the filtrate and washings were evaporated .to give a stickyyellow foam. Chromatography on silica gel (100g), eluting with CHCI3/IPE (50:1-> 20:1 v/v) gave the title compound. NMR 5 (DMSO-d6) 4.12 s (2H), 6.97 t (1H), 7.08 d (1H), 7.22 d (1H), 7.32 m(1H), 7.68 d(1H), 8.05 m (2H), 8.41 d(1H), 8.58 s (1H), 11.31 s(1H).

Example 3 2-Fluoro-7-pyridin-4-vlmethvl-9H-carbazole i) 2-Fluoro-7-(hvdroxv-pvridin-4-vl-methyl)-carbazole-9-carboxvlic acid, tert butyl ester A solution of 4-bromopyridine (253mg) in ether (10ml) was cooled to -75° underN2. A solution of "butyllithium (1ml of 1.6M sol. in hexane) was added. Afterstirring for 10 minutes at -75° a solution of 2(v) (0.5g) in dry THF (5ml) wasadded. After stirring the reaction at -70e for 1h, a further mixture of 4- 4bromopyridine (506mg) and 2ml of "butyllithium (2ml of 1.6M sol.) in ether(10ml) was added (premixed at -70e). After a further 1h at -70°, aqueousammonium chloride (10ml) was added. The mixture was extracted with ether (4x 30ml). The combined ether extracts were then extracted with 2N HCl (3 x30ml). These combined acid extracts were treated with 40% aqueous sodiumhydroxide to pH 11, then extracted with dichloromethane (4 x 40ml), then the 010195 25 « combined extracts were dried (MgSO4) and evaporated to give a yellow solid.Préparative t.l.c. gave the title compound as a yellow solid. ii) 2-Fluôro-7-pyridin-4-ylmethyl-9H-carbazole A solution of 3i) (164mg) in methanol (5ml) was treated under N2 with palladium black catalyst (40mg) and conc. HCl (0.2ml). Mixture was stirred under H2 for 1,5h at 50°, then a further charge of catalyst (80mg) was added and then the reaction stirred under H2 at 50° for 3h. The catalyst was filtered off , washed with methanol (2 x 5ml) then the filtrate and washings evaporated to a gum.

This was dissolved in 4M HCl in dioxan (5ml) for 30 minutes then evaporated to give a yellow gum. This was dissolved in water (10ml), treated with 40% aqueous sodium hydroxide to pH 11 then the mixture was extracted with dichloromethane (4 x 20ml). The combined* extracts were dried (MgSO4) and evaporated to give a yellow solid. Préparative t.l.c. gave the title compound as* » a yellow solid. NMR (DMSOd6). 4.125 s (2H), 6.9751 (1 H), 7.065 d (1 H), 7.2-7.45 m (4H), 8.0-8.15 m(2H), 8.465 d (1 H), 11.335 s (1 H).

Example 4 2-Fluoro-7-(3-fluoropvridin-4-vl-methyl)-9H-carbazole i) 2-Fluoro-7-(hydroxv-(3-fluoropvridin-4-vl)-methvl)-carbazole-9-carboxylic acid, tert butyl ester ' A solution of Ν,Ν,Ν',Ν'-tetramethylethylenediamine (0.48ml) in dry THF (5ml)was cooled to -70e under N2. A solution of nbutyllithium (2ml of 1.6M sol. inhexane) was added. The resulting solution was stirred at -20° for 1h thencooled to -70°. A solution of 3-fluoropyrïdine (0.273ml) in dry THF (3ml) wasadded. The resulting suspension was stirred at -40° for 1 h, then a solution of2(v) (500mg) in dry THF (10ml) was added. over 2 minutes. The reaction wasstirred at -70° for 1h then quenched with aqueous ammonium chloride (20ml).The mixture was extracted with ethyl acetate (3 x 30ml), then the combinedextracts were washed with water (4 x 10ml ) and brine (10ml), then dried(MgSO4) and evaporated to give a yellow solid. Chromatography on silica gel(100g), eluting with chloroform/isopropanol (50:1 -> 20:1 v/v) gave the titlecompound as a white solid. ii) 2-Fluoro-7-(3-fluoropvridin-4-yl-methyl)-9H-carbazole 010195 26 « A solution of 4i) (328mg) in methanol (10ml) was treated with palladium blackcatalyst (320mg) and conc. HCl (0.4ml). Mixture was stirred under H2 for 6h at50°. The catalyst was filtered off and washed with methanol (3 x 5ml). Thefiltrate and washings were evaporated to give a yellow solid. This wasdissolved in water (20ml) and treated with 40% aqueous sodium hydroxide topH 11. The mixture was extracted with dichloromethane (4 x 30ml), then thecombined extracts were dried (MgSO4) and evaporated to give a cream solid.Chromatography on silica gel (50g), eluting with chlorofomrdsopropanol (50:1 ->20:1 v/v) gave the title compound as a white solid. NMR (DMSOd6). 4.19δ s (2H), 6.9751 (1H), 7.075 d (1H), 7.235 d (1H), 7.405 m(2H), 8.055 m (2H), 8.355 d (1 H), 8.525 s (1 H), 11.325 s (1 H).

Example 5 2-Fluoro-7-(3-methylpyridin-4-vl-methvl)-9H-carbazole i) 2-Fluoro-7-(hvdroxv-(3-methylpyridin-4-yl)-methyl)-carbazole-9-carboxvlic acid, tert butyl ester A solution of 4-bromo-3-methylpyridine (549mg) in ether (10ml) was cooled to-70° under N2. A solution of nbutyllithium (3.2ml of 1.6M sol. in hexane) wasadded. The resulting suspension was stirred at -70° for 30 minutes, then asolution of 2(v) (500mg)in dry THF (10ml) was added. The solution was stirredat -70e for 1h then quenched with aqueous ammonium chloride (20ml). Themixture was extracted with ethyl acetate (3 x 30ml) then the combined extractswere washed with brine (10ml), dried (MgSO4) and evaporated to give a yellowgum. Chromatography on silica gel (90g), eluting with chloroform:isopropanol(50:1 -» 20:1 v/v) gave the title compound as a pale yellow foam. ii) 2-Fluoro-7-(3-methylpvridin-4-yl-methyl)-9H-carbazole A solution of 5i) (267mg) in methanol (5ml) was treated with palladium black *catalyst (100mg) and conc. HCl (0.25ml). Mixture was stirred under H2 for 14hwith periodic additions of further batches of catalyst. The catalyst was filteredoff and washed with methanol (3 x 2ml) and acetic acid (2 x 1ml). The filtrateand washings were evaporated to give a pale yellow gum. This was dissolvedin water (10ml) and treated with 40% aqueous sodium hydroxide to pH 11. Themixture was extracted with dichloromethane (4 x 15ml), then the combinedextracts were dried (MgSO4) and evaporated to give a pale cream foam. 010195 27 <

Chromatography on silica gel (50g), eluting with chloroform:isopropanol (50:1 ->20:1 v/v) gave the title compound as a white solid. NMR (DMSOd6). 2.255 s (3H), 4.145 s (2H), 6.975 t (1 H), 7.015 d (1 H). 7.135 d(1 h). 7.235 m (2H),-8.0-8.15 m (2H), 8.335 m (2H), 11.275 s (1H).

Example 6 2-Fluoro-7-(3-methoxvpyridin-4-vl-methyl)-9H-carbazole i) 2-Fluoro-7-(hydroxy-(3-methoxvpvridin-4-vl)-methvl)-carbazole-9-carboxylic acid, tert butyl ester A solution of 4-bromo-3-methoxypyridine (530mg) in ether (10ml) was cooled to-70° under N2. A solution of nbutyllithium (1.76ml of 1.6M sol. in hexane) wasadded. The resulting suspension was stirred at -70° for 30 minutes, then asolution of 2(v) (442mg)in dry THF (10ml) was added. The solution was stirredat -70° for 1h then quenched with aqueous. ammonium chloride (20ml). Themixture was extracted with ethyl acetate (3 x 50ml) then the combined extractswere washed with water (30ml), brine (30ml), dried (MgSO4) and evaporated togive a yellow solid. Chromatography on silica gel (90g), eluting withchloroform.isopropanol (50:1 -> 20:1 v/v) gave the title compound as a whitesolid. ii) 2-Fluoro-7-(3-methoxvpvridin-4-yl-n»ethyl)-9H-carbazole A solution of 6i) (200mg) in methanol (5ml) was treated under N2 with palladiumblack catalyst (100mg) and conc. HCl (0.25ml). Mixture was stirred under H2 for8h at 50°. The catalyst was filtered off, washed with methanol and acetic acidthen the filtrate and washings evaporated to a white solid. This was dissolvedin 4M HCl in dioxan (15ml) and dichloromethane (50ml) and stirred at roomtempérature for 6h then evaporated to give a gum. This was dissolved in water(10ml), treated with 40% aqueous sodium hydroxide to pH 11 then the mixture *was extracted with dichloromethane (4 x 20ml). The combined extracts weredried (MgSO4) and evaporated to give a yellow solid. Chromatography on silicagel (50g), eluting with chloroform:isopropanol (50:1 -> 20:1 v/v) gave the titlecompound as a white solid. NMR (DMSOd6). 3.935 s (3H), 4.075 s (2H), 6.965 t (1H), 7.045 d (1H), 7.135 d(1 H), 7.215 d (1 H), 7.295 s (1 H), 7.95-8.155 m (3H), 8.335 s (1 H), 11.285 s (1 H). 28 010195

Examples 7 to 9

The following compounds were prepared by methods analogous to those described in Examples 1 to 6: « 7. 1,7-Difluoro-2-M ,'2,4ltriazol-1-ylmethvl-9H-carbazole NMR δ (DMSO-d6) 5.63 s (2H), 7.10 m (2H), 7.26 dd (1 H), 7.92 d (1 H). 7.98s (1 H), 8.15 dd (1H), 8.69 s (1 H), 11.90 s (1H). 8. 2,4-Difluoro-7-f 1,2,41triazol-1 -ylmethyl-9H-carbazole NMR δ (DMSO-d6) 5.59 s (2H), 6.96 t (1H), 7.19 m (2H), 7.44 s (1H), 7.99 m(2H), 8.72 s (1 H). 9. 1.4.7-Trifluoro-2-f1,2.41triazol-1-vlmethvl-9H-carbazole. NMR δ (DMSO-d6) 5.62 s (2H), 6.96 dd (1 H), 7.11 t (1 H), 7.31 dd (1 H), 8.02m (2H), 8.72 s (1H), 12.25 s (1H).

Example 10 i) 4-Fluorophenyl boronic acid A solution of 1-bromo-4-fluorobenzene (318.2g) in THF (3000ml) was cooled to-52° under N2. "Butyllithium (2.5M in hexane, 800ml) was added by cannulaover 5 minutes, température rising to —44°. After addition was complété, thereaction was allowed to store at -52° for 15 minutes, then tri-isopropyl borate

Z (1240ml) was added over 10 minutes. Cooling was removed and the reactionallowed to warm to -15e. It was quenched by adding water (2000ml). Themixture was then partitioned between a further 2000ml of water, and ethylacetate. (2000ml). The gelatinous white material remaining in the flask wasdiscarded.

The aqueous phase was further extracted with ethyl acetate (2000ml), then the tcombined ethyl acetate layers were washed with brine (1500ml), dried (MgSO4)and evaporated to give a pale cream solid, (228.12g). The solid wascrystallized from boiling water (1100ml) to give the title compound (152.18g). ii) m-Nitro-p-bromobenzaldehyde

Conc. sulphuric acid (3125ml) and conc. nitric acid (1250ml) were mixedtogether (conc. nitric acid added in ~100ml portions) and subsequently cooled 010195 29 to 5eC, in an ice/salt bath. p-Bromobenzaldehyde (500g) was then addedportion-wise, maintaining the température below 10°C. Once the addition wascomplété, the resulting mixture was stored at room température for -3 hrs (until t ail p-bromobenzaldehyde dissolved), then carefully poured over ice. The» 5 precipitate formed was collected by filtration and washed thoroughly with water,until washings ~pH6, then dried in vacuo at 40°C to give the title compound aswhite powder (607.63g). iii) 4,-Fluoro-2-nitrobiphenyl-4-carbaldehvde 10 A mixture of 10i) (300g), 10ii) (448g), tetratristriphenyl phosphine palladium (0)(15g) and aq. 2M sodium carbonate (1.07L) in DME (3.2L) was refluxed underN2 for 16h. After cooling, the reaction mixture was extracted with ethyl acetate(2 x 2000ml). The combined extracts were washed with brine (1000ml), dried(MgSO4) and evaporated. Before complété évaporation, the product 15 crystallized. The crystals were filtered off, washed with IPE, then dried to givethe title compound (244.3g).

The washings and mother liquors were concentrated. The residue crystallizedand was triturated with ether, then the solid filtered and dried to give a further 20 crop of the title compound (120g). f iv) 2-(4'-Fluoro-2-nttrobiphenvl-4-vl)-[1,31-dioxolane A solution of 10iii) (357.4g) in isopropyl acetate (4000ml) was treated with p-toluene sulphonic acid, monohydrate (27g) and ethylene glycol (750ml). The 25 mixture was refluxed with a Dean-Stark trap for 2 hours. After cooling, thesolution was washed with water (2000ml, sat aq. NaHCO3 (2000ml), water (2 x2000ml), brine (1000ml), then dried (MgSO4) and evaporated to give the titlecompound as a yellowish syrup (520.96g). 30 Toluene has also been found to be a suitable solvent for this reaction. v) 2-M .31-Dioxolan-2-yl-7-fluoro-9H-carbazole

Xylene (1250ml) and triethylphosphite (1250ml) were mechanically stirred underN2 and heated to reflux (~142°). A solution of 10iv) (514.76g) in xylene 35 (3750ml) was added over 2h, maintaining reflux. Reflux was continued for a 010195 30 < further 5%h, and then further triethylphosphite (500ml) was added dropwise.Reflux was continued for VAh, then reaction was left to cool to roomtempérature overnight. After cooling in ice/water for 30 minutes, crystals werefiltered off, washed-with xylene and cyclohexane, then dried in vacuo at 40° togive the title compound (171 75g).

Further crops were obtained from the cyclohexane washes and the rest of themother liquors.

Neat triethylphosphite has also been found to be a suitable solvent for thisreaction. vi) 7-Fluoro-9H-carbazole-2-carbaldehyde A suspension of 10v) (209.626g) in acetone,(5000ml) was treated with 2N HCl(1250ml) and the resulting yellow brine solution was stored for 30 minutes. Theacetone was evaporated off, and the aqueous layer and solid was extracted withethyl acetate (7000ml). The organic solution was washed with water (1000ml),dried (MgSO4) and evaporated to give the title compound as a paleapricot/mango solid (171.59g). vii) 2-Fluoro-7-formyl-carbazole-9-carboxylic acid tert-butyl ester A suspension of 10vi) (168.25g) in ethyl acetate (300ml) was stirred with 4-dimethylaminopyridine (96.41g) at room température under N2. A solution of di-butyldicarbonate (172.2g) in ethyl acetate (1000ml) was added over 30 minutes,so CO2 effervescence was only at a slow rate and then further BOC anhydride(25g) was added to complété reaction. A white solid crystallized out. Afterstirring for ~30 minutes, this was filtered off, washed with 2N HCl (2 x 500ml),then water until neutral, then dried in vacuo to give the title compound *(187.44g).

Further crops were obtained from the mother liquors. . viii) 2-Fluoro-7-(hvdroxv-pvridin-3-ylmethyl)-9H-carbazole A solution of 3-bromopyridine (16.6ml) in THF (500ml) was cooled to -70°Cunder N2. This solution was added dropwise over 10 minutes to a solution of n- 010195 31 butyllithium (220ml of a 1.6M solution in hexanes) that was cooled to -70eC.After 10 minutes, a solution of 10vi») (27g) in THF (700ml) that was cooled to-70°C was added dropwise over 20 minutes. The mixture was stirred at -70°Cfor 40 minutes, then quenched with ammonium chloride solution (300ml). Themixture was extracted with ethyl acetate (2 x 500ml), and the combined organiclayers washed with brine (500ml), dried (MgSO4) and evaporated to give amixture of 2-fluoro-7-(hydroxy-pyridin-3-ylmethyl)-carbazole-9-carboxylic acidtert-butyl ester and 2-fluoro-7-(hydroxy-pyridin-3-ylmethyl)-9H-carbazole as anamber oil.

This oil was dissolved in methanol (500ml) and treated with sodium methoxide(37g of a 25% solution in methanol). After stirring for 24 hours, the reaction wasconcentrated to 50ml, then water (250ml) was added and the mixture wasextracted with ethyl acetate (3 x 250ml). The combined organic layers wereextracted with HCl (20% aqueous solution, 3 x 250ml). The combined HCllayers were washed with ethyl acetate (250ml). The aqueous layer was thenneutralized to pH10 with NaOH pellets, then was extracted with methylenechloride (2 x 500ml). The combined methylene chloride layers were washedwith brine (500ml), dried (MgSO4), and evaporated to 100ml. Toluene (250ml)was added, and the suspension concentrated to 150ml. Collection of theresulting precipitate afforded, upon drying, the title compound as an off whitepowder. ix) 2-Fluoro-7-pyridin-3-ylmethyl-9H-carbazole A solution of 10viii) in 50% methanol/THF (200ml) was treated withconcentrated HCl (5ml) and palladium on carbon catalyst (10% active catalyst,50% water wet). The suspension was placed in a Parr hydrogénation bottle andtreated with hydrogen gas at 10 PSI, 25’C for 3 hours. The catalyst was filtered *off, the catalyst cake was washed with HCl (10%, 100ml), water (100ml), and10% NaOH (100ml). The filtrate was concentrated to 200ml, then neutralizedwith NaOH pellets to pH9. The solution was then extracted with ethyl acetate (2x 250ml), and the combined organic layers washed with brine (250ml), dried(Na2SO4) and concentrated to give a white solid. The crude product wasdissolved in methanol (400ml) and treated dropwise with water (200ml). The 32 010195 resulting precipitate was collected and dried to give the title compound as awhite crystalline solid.

Melting fjoint: 196-197°C.

Mass Spectrum (FAB+): 277

Elemental analysis:

Calculated:

Found: % c H N 78.24 4.74 10.13 78.25 4.80 10.18

The following examples illustrate pharmaceutical formulations according to theinvention containing 2-fluoro-7-pyridin-3-y!methyl-9H-carbazole as the activeingrédient. Other compounds of the invention may be formulated in a similar 10 manner.

Tablets for oral administration a) Direct Compression Tablet

Comportent Composition (%) Active ingrédient 20.0 Microcrystalline cellulose ' 50.0 Spray dried lactose 24.2 Sodium starch glycolate 5.0 Colloïdal Silicon dioxide 0.3 Magnésium stéarate 0.5

Ail materials except magnésium stéarate are blended until sufficiently mixed.Magnésium stéarate is screened and added to the mixture which is blended * 15 thoroughly. The résultant mix is compressed to predetermined tablet size andweight. 33 010195 b) Wet Granulation Tablet

Component Composition (%) Active ingredient 30.0 Microcrystalline cellulose 55.2 Starch 1500 6.0 Sodium starch glycolate 5.0 10% polyvinylpyrrolidone 3.0 in water Magnésium stéarate 0.5 Colloïdal Silicon dioxide 0.3

Ail of the ingrédients except for the polyvinylpyrrolidone solution andmagnésium stéarate are blended in a fluidized air bed. Polyvinylpyrrolidonesolution is added to the blended powders with constant mixing until uniformly 5 moist. After drying, the granules are milled to reduce particle size and increasesize uniformity and blended with the magnésium stéarate. The granules arethen compressed to predetermined tablet size and weight.

Tablets of other strengths may be prepared by altering the ratio of activeingrédient e.g. to lactose or altering the compression weight. f 10 The tablets may be film-coated with suitable film-forming materials such ashydroxypropyl methylcellulose using standard techniques. Altematively thetablets may be sugar coated or enteric coated.

Syrup for oral administration

Component Composition (%) Active ingredient, microhized 5.00 Magnésium aluminum silicate 0.50 Sodium carboxymethylcellulose 0.80 34 010195

Component Composition (%) Sodium lauryl sulphate 0.01 Sorbitol solution, USP 26.0 • Methylparaben 0.20 Propylparaben 0.04 Flavour 0.50 Purified water 66.95

The sodium carboxymethylcellulose and magnésium aluminium silicate ishydrated in a solution of sodium lauryl sulphate in water for 24 hours. Activeingrédient is suspended in the vehicle with the aid of a mixer. Thepreservatives are dissolved in the remaining yvater by heating and after cooling 5 to room température, the sorbitol solution is added. The solution is added to thesuspension, flavour mixed in and the pH ’adjusted as needed. The finalsuspension is mixed in a homogenizer.

Soft gelatin capsules for oral administration

Component Composition (%) Active ingrédient, micronized 5.0 Polyethylene glycol r 47.5 Propylene glycol 47.5

The glycols are blended with warming until homogeneous. Active ingrédient is10 added and the mixture homogenised and filled into an appropriate gelatin mass to give soft gelatin capsules containing the appropriate fill weight.

Suppository for rectal administration

Component Composition (%) Active ingrédient, micronized 2 Witepsol W32, hard fat 98 A slurry of the active ingrédient in a portion of molten Witepsol (approximately36°C) is prepared using a high speed mixer and is then evenly dispersed in the 010195 35 remaining molten hard fat. The suspension is filled, using suitable machinery,into 1 or 2g size suppository moulds and allowed to cool.

Transdermal System

Component Composition (%) Active ingrédient 5 Silicone fluid 90 Colloïdal Silicon dioxide 5

The silicone fluid and active ingrédient are mixed together and the colloïdal5 Silicon dioxide is added to increase viscosity. The material is then dosed into asubsequently heat sealed polymeric laminete comprised of the following:polyester release liner, skin contact adhesive composed of silicone or acrylicpolymers, a control membrane which is a polyolefin (e.g. polyethylene orpolyvinyl acetate) or polyuréthane, and an imperméable backing membrane 10 made of a polyester multilaminate. The laminated sheet is then eut intopatches.

Formulations for parentéral administration a) Intravenous solution '

Component Composition (%) Active ingrédient 5.0 Sodium chloride USP 0.9 Phosphate buffer (monobasicand dibasic potassiumphosphate) 7.0 Water for Injection USP 87.1

Active ingrédient is dissolved in the water with the remaining components and15 stérile filtered (0.22pm fi,ter). The solution is filled into glass vials, stoppered and sealed before autoclaving. . 36 010195 b) Lyophilised product

Component Composition (%) Active ingrédient 2.5 Mannitol 5.0 Phosphate buffer (monobasicand dibasic potassiumphosphate) 7.0 Water for Injection USP 85.5

Active ingrédient is dissolved in the water with the remaining components andstérile filtered (0.22pm filter). The solution is filled into glass vials, stopperedand lyophilised before sealing. The lyophilised product is reconstituted with 5 saline prior to administration. 4

Claims (10)

010195 37 « Claims

1.A compound of general formula (I)

wherein R-| and R4 each independently represent a hydrogen atom or a5 C-]_galkyl group; each R2 may be the same or different and represents an electron-withdrawinggroup; each R3 may be the same or different and represents an electron-withdrawinggroup; 10 R5 is a group of formula

Rg is a halogen atom, a C-j^alkyl group or a alkoxy group;m is zéro or an integer 1 to 4;n is zéro or an integer 1 to 3; and 15 p is zéro, 1 or 2 and pharmaceutically acceptable salts and solvatés thereof. *

2-Fluoro-7-{3-methoxypyridin-4-ylmethyl)-9H-carbazole 1.7- DifIuoro-2-[1,2,4]triazol-1-ylmethyl-9H-carbazole2,4-Difluoro-7-{1,2,4]triazol-1 -ylmethyl-9H-carbazole 1.4.7- Trifluoro-2-[1,2,4]triazol-1-ylmethyl-9H-carbazoleand pharmaceutically acceptable salts and solvatés thereof. 14. 2-Fluoro-7-pyridin-3-ylmethyl-9H-carbazole and pharmaceuticallyacceptable salts and solvatés thereof.

15. Compounds of formula (I) as claimed in any one of daims 1 to 14 for use asactive therapeutic agents.

16. A pharmaceutical composition comprising a compound of formula (I) asclaimed in any one of daims 1 to 15 or a pharmaceutically acceptable sait orsolvaté thereof as adive ingrédient together with one or more pharmaceuticallyacceptable carriers or excipients.

17. A pharmaceutical composition as claimed in daim 16 adapted for oral,redal or topical administration.

18. A pharmaceutical composition as claimed in daim 16 or daim 17 which isformulated in unit dosage form comprising 0.1 to 200mg active ingrédient.

19. Use of a compound as claimed in any one of daims 1 to 15 or apharmaceutically acceptable sait or solvaté thereof for the manufacture of amédicament for the treatment of a condition whose underlying aetiology isassociated with elevated androgen and/or oestrogen levels in a mammal,including a human.

20. A method for the treatment of a mammal, including human, comprisingadministration of an effective amount of a compound as claimed in any one ofdaims 1 to 15 or a pharmaceutically acceptable sait or solvaté thereof, to lowerthe levels of androgens and/or oestrogens. 40 010195

21. A method of treating prostate cancer comprising administration of aneffective amount of a compound as ciaimed in any one of daims 1 to Î5 or apharmaceutically acceptable sait or solvaté thereof. &amp;

22. A process for the préparation of a compound as ciaimed in any one of5 daims 1 to 15 or a pharmaceutically acceptable sait or solvaté thereof which comprises (A) for the préparation of a compound of formula (I) wherein R·, represents ahydrogen atom, cyclising a compound of formula (II)

2-Fluoro-7-(3-methylpyridin-4-ylmethyl)-9H-carbazole

2. A compound as claimed in claim 1 wherein the group -CHR4R5 is in the 1, 2or 3 position.

3. A compound as claimed in claim 1 or claim 2 wherein the group -CHR4R5 is20 in the 2-position. 010195 38 ♦

4. A compound as claimed in any one of cfaims 1 to 3 wherein R2 and R3,which may be the same or different, represent a halogen atom or a nitrite, nitro,trifluoromethyl, aldehydo, keto, carboxylic acid or carboxylic ester group.

5. A compound aé claimed in any one of daims 1 to 4 wherein R2 and R3,which may be the same or different, represent a fluorine atom, a chlorine atomor a nitrile group.

6. A compound as claimed in any one of daims 1 to 5 wherein R2 represents afluorine atom and m is an integer of 1 to 4.

7. A compound as claimed in any one of daims 1 to 6 wherein m is 1 and R2 isin the 5 or 7 position, preferably the 7-position.

8. A compound as claimed in any one of daims 1 to 7 wherein R., and R4 areeach independently a hydrogen atom or a methyl group.

9. A compound as claimed in any one of daims 1 to 8 wherein R5 is a pyridin-3-yl or pyridin-4-yl group.

10. A compound as claime din any one of daims 1 to 9 wherein R6 Representsa fluorine atom, a methyl group or a methoxy group. f

11. A compound as claimed in any one of daims 1 to 10 wherein R-j and R4each represent a hydrogen atom or a methyl group, R2 and R3 are fluorineatoms, R6 is a fluorine atom, a methyl group or a methoxy group, m is an integer1 to 4, n is zéro or an integer 1 to 3, and p is zéro, 1 or 2.

12. A compound as claimed in any one of daims 1 to 10 wherein R-, and R4each represent a hydrogen atom, R2 and R3 are fluorine atoms, R6 is a fluorineatom or a methyl or methoxy group and m, n and p each independentlyrepresent zéro, 1 or 2. 13. 2-Fluoro-7-[1,2,4]triazol-1 ylmethyl-9H-carbazole2-Fluoro-7-pyridin-3-ylmethyl-9H-carbazole2-Fluoro-7-pyridin-4-ylmethyl-9H-carbazole2-F!uoro-7-(3-fluoropyridin-4-ylmethyl)-9H-carbazole 0 î 0195 39 <

10 (B) deoxygenating a compound of formula (IX) (IX) F R. C(OH)R4R5 or a protected dérivative thereof; or (C) converting a compound of formula (I) into another compound of formula (I); and if necessary and/or desired subjecting the compound resulting from Step A, *15 B or C to one or more further reactions comprising: (i) removing any protecting groups; (ii) converting a compound of formula (I) or a sait or solvaté thereof into apharmaceutically acceptable sait or solvaté thereof; (iii) separating a racemic mixture into individual enantiomers of formula (I).