AP412A

AP412A - 17a and 17B-Alkylketone-3-carboxyaromatic a ring steroidal compounds and pharmaceutical compositions containing these compounds.

Info

Publication number: AP412A
Application number: APAP/P/1993/000542A
Authority: AP
Inventors: Dennis Alan Holt; Mark Alan Levy
Original assignee: Smithkline Beecham Corp
Priority date: 1992-06-30
Filing date: 1993-06-30
Publication date: 1995-09-29
Also published as: KR950702118A; BG99313A; WO1994000125A1; GB9213835D0; EP0651643A1; EP0651643A4; MX9303970A; FI946189A; OA10122A; RU94046378A; AP9300542A0; SI9300350A; BR9306748A; FI946189A0; MA22927A1; SK159994A3; ZA934645B; CA2138956A1; NO945074L; IL106158A0

Abstract

Invented are

Description

SPECIFICATION

Be it known that I, DENNIS ALAN HOLT, residing at 4450A Gebhart School Road, Mohnton, Pennsylvania, 19540 a citizen of the United States of America; and MARK ALAN LEVY, residing at 115 Reveille Road, Wayne, Pennsylvania, 19087 a citizen of the United States of America; have invented a new and useful COMPOUNDS of which the following is a full, clear, and exact specification.

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- 1 c COMPOUNDS

FIELD OF THE INVENTION

The present invention relates to certain novel 17a and 17B-alkylketone-3carboxy aromatic A ring steroidal compounds, pharmaceutical compositions containing these compounds, and methods for using these compounds to inhibit steroid 5-a-reductase isozyme 1 and steroid 5-a-reductase isozyme 2. Also invented are novel intermediates and processes useful in preparing these compounds.

DESCRIPTION OF RELATED ART

The class of steroidal hormones known as androgens is responsible for the physical characteristics that differentiate males from females. Of the several organs that produce androgens, the testes produce these hormones in the greatest amounts. Centers in the brain exert primary control over the level of androgen production. Numerous physical manifestations and disease states result when ineffective control results in excessive androgen hormone production. For example, acne vulgaris, seborrhea, female hirsutism, male pattern baldness and prostate diseases such as benign prostatic hypertropy are correlated with elevated androgen levels. Additionally, the reduction of androgen levels has been shown to have a therapeutic effect on prostate cancer.

Testosterone is the principal androgen secreted by the testes and is the primary androgenic steroid in the plasma of males. It now is known that 5-areduced androgeos are the active hormones in some tissues such as the prostate and sebaceous gland. Circulating testosterone thus serves as a prohormone for dihydrotestosterone (DHT), its 5-a-reduced analogue, in these tissues but not in others such as muscle and testes. Steroid 5-a-reductase is a Nicotinamide Adenine dinucleotide Phosphate(NADPH)dependent enzyme that converts testosterone to DHT. The importance of this enzyme in male development was dramatically underscored by discovery of a genetic steroid 5-a-reductase deficiency in male pseudohermaphrodites. Imperator-McGinley, J., & al., (1979), J, Steroid Biochem. 11:637-648.

A number of 3-carboxy-estra 1,3,5(10) triene 5-a-reductase inhibitors are known in the art. For example, bad original

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-ΣΙ. J, Steroid Biochem.. Vol. 34, Nos. 1-6 pp. 571-575(1989), by M.A.

Levy, et al., describes the interaction mechanism between rat prostatic steroid 5alpha reductase and 3-carboxy-17B-substituted steroids;

2. J, Med, Chem. (1990) Vol. 33, pp. 937-942, by D.A. Holt, et al., describes a new steroid class of A ring aryl carboxylic acids;

3. TIPS (December 1989) Vol. 10, pp. 491-495, by B.W. Metcalf, et al., describes the effect of inhibitors of steroid 5a reductase in benign prostatic hyperplasia, male pattern baldness and acne.

In addition U.S. Patent No. 4,954,446 describes a group of 3-carboxy-estra l,3,5(10)triene-17B-substituted compounds which are disclosed as being useful as inhibitors of steroid 5-a-reductase. However, none of the cited references disclose or suggest that any of the novel 17B-alkylketone-3-carboxy aromatic A ring steroidal compounds of the present invention would have utility as highly potent inhibitors of both steroid-5-a-reductase isozyme 1 and steroid 5-a-reductase isozyme 2 (dual inhibitors).

SUMMARY OF THE INVENTION This invention relates to a compound of the formula I:

wherein Z is a or B O

II

-DR in which R is Cj_20 linear or branched, saturated or unsaturated alkyl and 25 pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

The invention also is a method for simultaneously inhibiting 5-a-reductase isozyme 1 and 5-a-reductase isozyme 2 activity in mammals, including humans, that comprises administering to a subject an effective amount of a presently invented 5-a-reductase inhibiting compounds. In a further aspect of the invention there are provided novel intermediates and novel processes useful in preparing the

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-3prescntly invented dual 5-a-reductase inhibiting compounds. Included in the present invention are pharmaceutical compositions comprising a pharmaceutical carrier and compounds useful in the methods of the invention. Also included in the present invention are methods of co-administering the presnetly invented dual 5-areductase inhibiting compounds with further active ingredients.

DETAILED DESCRIPTION OF THE INVENTION The compounds of this invention that inhibit both 5-a-reductase isozyme 1 and 5-a-reductase isozyme 2 have the following Formula (I):

wherein Z is a or β O

II

-C-R in which R is Cj.20 linear or branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

As used herein such compounds are referred to as dual inhibitors of steroid

5-a-reductase.

Preferred among the presently invented compounds are those having Formula (Π):

in which R is Ci_20 linear or branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

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-4Prefeired among the presently invented Formula Π compounds are those having Formula ΙΠ

in which R² is Cq.g linear or branched alkyl and pharmaceutically acceptable salts, hydrates solvates and esters thereof.

Preferred among Formula (III) compounds are those in which R² is methyl, ethyl, propyl, 3-methylbutyl, isopropyl, n-butyl, isobutyl, 1-methylpropyl, t-butyl, pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, octyl or 3,3-dimethylbutyl.

Particularly preferred among Formula (ΠΙ) compounds are those in which R² is 1-methylpropyl, n-butyl, isopropyl, n-pentyl, 3-methylbutyl, 2,2-dimethylpropyl, t-butyl, 1,1-dimethylpropyl, isobutyl n-octyl, tert-pentyl, n-propyl, methyl or 3-3-dimethylbutyl.

Particularly preferred among Formula (III) compound are:

17B-(isobutylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(octylcarbonyl)-estra-l ,3,5(10)-triene-3-carboxylic acid,

17B-(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(2,2-dimethylpropylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(propylcarbonyl)-estra-1,3,5(10)-triene-3-carboxylic acid,

17B-(methylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid and

17B-(3,3-dimethylbutylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

The term a, as used herein, follows standard chemical terminology and means down or that the corresponding substituent is attached below the plane of the paper.

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-5The term β, as used herein, follows standard chemical terminology and means up or that the corresponding substituent is attached above the plane of the paper.

By the term alkyl, Ci__nalkyl and derivatives thereof and in all carbon 5 chains as used herein, unless otherwise defined, is meant a Cj._n linear or branched carbon chain having 1 to n carbons. Examples of alkyl and derivatives thereof as used herein include: methyl, ethyl, propyl, 3-methylbutyl, isopropyl, n-butyl, isobutyl, 1-methylpropyl, t-butyl, n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, n-octyl, tert-pentyl and 3,3-dimethylbutyl.

By the term treating as used herein, is meant prophylatic or therapeutic therapy.

By the term isobutyl as used herein, is meant -CH2CH(CH3)2·

By the term metal-catalyzed coupling reaction as used herein is meant that the prepared 3-trifluoromethyl sulfonate or 3-fluorosulfonate compound is reacted in a suitable organic solvent, preferably toluene, dimethylformamide or THF with a base, preferably a tertiaiyamine base such as triethyiamine, pyridine or tributylamine, a phosphine such as bis(diphenylphosphino)alkane, preferably 1,3 bis(diphenylphosphino)propane or tri-o-tolyphosphine, or a Cj.galkOH, and a metal catalyst, preferably a palladium catalyst such as palladium (II) acetate, palladium (Π) chloride or bis(triphenylphosphine) palladium II acetate, and a coupling reagent.

By the term coupling reagent as used herein is meant a compound which is capable of reacting with an aryl radical to form a carboxylic acid substituent. Carbon monoxide is a preferred coupling reagnet, which when added to the metal25 catalyzed coupling reaction, as described herein, yields the desired carboxylic acid group.

Compounds of Formula (I) and compounds of formula (IV) are included in the pharmaceutical compositions of the invention and used in the methods of the invention. Where a -COOH group is present, pharmaceutically acceptable esters can be employed, for example methyl, ethyl, pivaloyloxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodnig formulations.

The term α-receptor antagonist, as used herein, refers to a known class of alpha-andrenergic receptor antagonist comounds, such as described in Lafferty, et al. U.S. Patent No. 4,963,547, which are utilized in treating vascular disorders such

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-6as diabetes, cardiovascular disease, benign prostatic hypertrophy and ocular hypertension.

Preferred alpha-andrenergic receptor antagonists for use in the compositions and methods of the invention include amsulosin, terazosin, doxazosin, alfuzosin, indoramin, prazosin and 7-chloro-2-ethyl-3,4,5,6-tetrahydro-4-methylthieno[4,3,2ef|[3]-benzapine.

By the term amsulosin as used herein is meant a compound of the structure

and salts, hydrates and solvates thereof.

Chemically, amsulosin is designated as (-)-(R)-5-[2-[[2-(Oethoxyphenoxy)ethyl]amino]propyl]-2-methoxybenzenesulfonamide.

Amsulosin is disclosed in U.S. Patent Number 4,703,063 and claimed in

U.S. Patent Number 4,987,125 as being useful in treating lower urinary tract dysfunction.

By the term terazosin as used herein is meant a compound of the structure

and salts, hydrates and solvates thereof.

Chemically, terazosin is designated as l-(4-amino-6,7-dimethoxy-2 quinazolinyl)-4-[(tetrahydro-2-furoyl)carbonyl]piperazine. Terazosin is disclosed 25 in U.S. Patent Number 4,251,532.

By the term doxazosin as used herein is meant a compound of the structure

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and salts, hydrates and solvates thereof.

Chemically doxazosin is designated as l-(4-amino-6,7-dimethoxy-2quinazolinyl)-4-[(2,3-dihydro-1,4-benzodioxin-2-yl)carbonyl]-piperazine. Doxazosin is discolsed in U.S. Patent Number 4,188,390.

By the term alfuzosin as used herein is meant a compound of the structure

and salts, hydrates and solvates thereof.

Chemically alfuzosin is designated as N-[3-[(4-amino-6,7-dimethoxy-210 quinazolinyl)methylamino]propyl]tetrahydro-2-furancarboxamide.

Alfuzosin is disclosed in U.S. Patent Number 4,315,007.

By the term indoramin as used herein is meant a compound fo the structure /-γ ^f_/⁼\ ch₂ch₂ n \-nhc—6 y and salts, hydrates and solvates thereof.

Chemically indoramin as designated N-[[ 1-(2-( lH-indol-3-yl)ethyl]-4piperidinyljbenzamine.

Indoramin is disclosed in U.S. Patent Number 3,527,761.

By the term prazosin as used herein is meant a compound of the structure

and salts, hydrates and solvates thereof.

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Chemically prazosin is designated as l-(4-amino-6,7-dimethoxy-2quinazolinyl)-4-(2-furanylcarbonyl)piperazine.

Prazosin is disclosed in U.S. Patent Number 3,511,836.

7-chloro-2-eAyl-3,4,5,6-tetrahydro-4-meAylAieno[4,3,2-ef][3]benzazepine as used herein is meant a compound of the structure

-8S XH₂CH₃ and salts, hydrates and solvates thereof.

7-chloro-2-eAyl-3,4,5,6-tetrahydro-4-meAylAieno[4,3,2-ef]-[3]benzazepine is disclosed in U.S. Patent No. 5,006,521. Additionally, all compounds disclosed in U.S. Patent No. 5,006,521 as alpha-adrenergic receptor antagonist are preferred alpha-adrenergic receptor antagonist as used herein.

Persons skilled in the art can readily determine if a compound other than one specifically referred to herein is a alpha-andrenergic receptor antagonist by utilizing Ae assay described in Lafferty I. Thus, all such compounds are included wiAin Ae scope of Ae term alpha-andrenergic receptor antagonist as used herein.

By Ae term minoxidil as used herein is meant the compound of Ae structure:

Z ^H2^N\ /N /NH chemically minoxidil is designated as 2,4-pyrimidineadiamine, 6-(1-piperidinyl)-,3oxide. Minoxidil is Ae active ingredient in Rogaine® which is sold as topical solution for stimulating hair growA by Ae Upjohn Company, Kalamazoo, Michigan.

The term aromatase inhibitor, as used herein, refers to a known class of compounds, steroidal and non-steroidal, which prevent Ae conversion of androgens to estrogens, such as described in Gormley et al. International Publication Number WO 92/18132. Aromatase inhibitors are disclosed in Gormley et al. as having____

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-9utility in treating benign prostatic hyperplasia when used in combination with a 5α-reductase inhibitor.

A preferred aromatase inhibitor for use in the compositions and methods of the invention 4-(5,6,7,8-tetrahydroimidazo-[l,5-a]pyridin-5-yl)benzonitrile (fadrazole). Fadrazole is disclosed in U.S. Patent No. 4,728,645. Additionally, all compounds disclosed in Gormley, et al. International Publication No. WO 92/18132 as having aromatase inhibiting activity are preferred aromatase inhibitors as used herein.

As used herein, when a dual inhibitor of 5-a-reductase, as described herein and a further active ingredient or ingredients are utilized together, said 5-areductase inhibitor can be co-administered with said further active ingredient or ingredients.

By the term co-administering and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of a 5-a-reductase inhibiting compound, as described herein, and a further active ingredient or ingredients, such as other compounds known to treat the disease states of acne vulgaris, seborrhea, female hirsutism, male pattern baldness, benign prostate hypertrophy or prostatic adenocarcinoma or compounds known to have utility when used in combination with 5-a-reductase inhibitors. Preferably, if the administration is not simultaneous, the compounds are administered in a close time proximity to each other. Furthermore, it does not matter if the compounds are administered in the same dosage form, e.g. one compound may be administered topically and another compound may be administered orally.

The novel compounds of Formula (Π) of the present invention can be prepared by methods outlined in schemes 1-4 below and in the Examples from known and readily available estrone which has the formula:

R or from the 17B-carboxylic acid analogue of estrone, which is known and readily available.

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- 10. Scheme I

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- 11 Ο

II

?

(β) ?

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- 12Scheme I outlines formation of Formula Π compounds. As used in scheme I compound (b) is prepared from compound (a) according to the procedure of Baldwin, et al., J. Chem. Soc. (c), 1968,2283-2289.

Compound (b) is then stirred in an appropriate organic solvent, preferably 5 methanol, with a base, preferably sodium hydroxide, and then acidified to yield compound (c). Compound (c) is next treated with a Grignard reagent, described hereinbelow, or a lithium reagent in an appropriate organic solvent, preferably tetrahydrofuran or diethylether solvent, preferably at reflux temperature to yield formula (d) compounds.

A formula (d) compound and a base, preferably 2,5-di-t-butyl-3-methylpyridine in an appropriate organic solvent, preferably dichloromethane, is cooled to -20°C to 20°C, preferably 0°C, and reacted with a trihaloalkyl sulfonic anhydride, preferably trifluoromethanesulfonic anhydride to form compounds (e).

Formula (f) compounds are prepared by reacting a formula(e) compound in a metal catalyzed coupling reaction. Preferably a formula (e) compound dissolved in dimethyl formide (DMF) an organic base preferably, triethylamine, a phosphine, preferably bis(diphenylphosphino)propane, a palladium(II) compound, preferably, palladium(II) acetate, and a Cj.galkyl alcohol (Ci.galkOH), followed by addition of carbon monoxide (CO). Compounds (f) next are reacted with a suitable base, preferably potassium carbonate, and acidified to yield compounds (g).

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-132£ΗΕΜΕΠ

Ο

FOjSO

ο

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(9)

Scheme Π outlines formation of Formula Π compounds. The starting materials in Scheme Π are formula (d) compounds prepared as described in Scheme

I.

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As used in Scheme Π, a formula (d) compound and a base, preferably 2,5di-t-butyl-3-methyl-pyridine in an appropriate organic solvent, preferably dichloromethane, is cooled to -20°C to 20°C, preferably 0°, and reacted with fluorosulfonic anhydride to form compounds (h). Formula (f) compounds are prepared by reacting a Formula (h) compound in a metal-catalyzed coupling reaction. Preferably a Formula (h) compound is dissolved in dimethylformide (DMF) an organic base preferably triethylamine, a phosphine preferably bis (diphenylphospine)propane, a palladium(II) compound, preferably, palladium(II) acetate, and a Cj.galkyl alcohol (Cj.^alkOH), followed by addition of carbon monoxide (CO). Compounds (f) next are reacted with a suitable base, preferably potassium carbonate, and acidified to yield compounds (g).

_z z' z’ /

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-15Scheme PI %

_ -COH

Scheme ΠΙ outlines formation of Formula Π compounds. As used in

Scheme ΠΙ R.1 is CF3O2SO- or FO2SO-. As used in scheme III in the alkylation process (step C), the pyridylthio ester is reacted with an LiR or an XMgR (X=C1, Br) Grignard reagent (as described hereinbelow), preferably isobutylmagnesium bromide, n-octylmagnesium chloride, tert-pentylmagnesium chloride or 2,2 dimethylpropylmagnesium bromide, preferably in tetrahydrofuran to form the desired product, preferably 17B-(isobutylcarbonyl)-estra-l,3,5(10)-triene-3carboxylic acid, 17B-(octylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid, 17B(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid, or 178-(2,2-

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- 16dimethylpropylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid respectively in one or two steps.

In Route 1, the 3-hydroxyl acid (i) is converted to the 3trifluoromethylsulfonylate or 3-fluorosulfonylate derivative (j) (step A) by treating (i) with trifluoromethylsulfonyl anhydride or fluorosulfonic anhydride and an amine base, such as pyridine, preferably 2,5 di-t-butyl-3-methyl-pyridine, in an appropriate organic solvent, preferably dichloromethane at about -20°C to 20°C, preferably 0°.

The activated ester (k) is produced (step B) by treating (j) with 2,2dithiopyridyl and triphenylphosphine in an appropriate organic solvent solution preferably, tetrahydrofuran/toluene at room temperature for about 8-14 hours.

The 17-acyl derivative (1) is produced (step C) by treating (k) with a Grignard reagent, described hereinbelow, in tetrahydrofuran or diethyl ether solvent, at a temperature of about -50 to -70°C, for 1-16 hours.

The 3-alkyl ester (f) is produced (step D) by treating (1) under carbonylation conditions, preferably by bubbling carbon monoxide gas through a solution of (1) in an appropriate organic solvent, preferably methanol, containing palladium acetate catalyst, triphenylphosphine, and a tertiary organic amine preferably triethylamine at about room temperature for 1-16 hours. Compounds (f) next are reacted with a suitable base, preferably potassium carbonate and acidified to yield compounds (g).

Compounds (g) can also be produced (step G) by treating (1) under carboxylation conditions, preferably by bubbling carbon monoxide gas through a solution of 0) in an an appropriate non-alcoholic solvent, preferably DMSO, containing a palladium catalyst, preferably palladium (Π) diacetate and 1,1Bis(diphenylphosphino)ferrocene (DPPF); and a base, preferably potassium acetate, preferably at increased temperatures.

Route 2 involves converting the starting steroidal acid (i) to the 3trifluoromethylsulfonylate or the 3-fluorosulfonylate derivative (j) by the abovedescribed step A; carbonylating (j) to (m) by step D; forming the activated 2pyridylthio ester (n) by step B; forming the 17-acyl compound (f) by step C; and hydrolyzing the 3-ester to the 3 acid final product (g) by step F.

Route 3 involves converting the starting acid (i) to the activated ester (o) by the above-described step B; forming the 17-acyl compound (d) by reacting (o) by the above described step C; converting (d) to the 3-trifluoromethylsulfonylate or 3fluorosulfonylate derivative (1) by the above-described step A; and converting 0) to

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AP Ο Ο Ο 4 1 2 the final product (g) by the above described step G or by the above-described step

D followed by the above described step F.

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(Grignard Reagent) (r)

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Scheme IV outlines formation of Formula Π compounds.

As used in Scheme IV in the alkylation process (to prepare compounds of Formula (s)), the carboxaldehyde is reacted with an Li-R or an XMgR (x=Cl, Br) Grignard reagent (as described hereinbelow), preferably propylmagnesium bromide, methylmagnesium bromide or 3,3-dimethylbutyl magnesium chloride, preferably in tetrahydrofuran to form the desired product, preferably 17B(propylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid, 17B-(methylcarbonyl)estra-1,3,5 (10)-triene-3-carboxylic acid, 176-(3,3-dimethylbutylcarbonyl)-estral,3,5(10)-triene-3-carboxylic acid respectively, in one or two steps.

The starting material in Scheme IV is Formula (c) compound prepared as in Scheme I.

As used in Scheme IV, a Formula (c) compound and a base, preferably 2,5di-t-butyl-3-methyl-pyridine in an appropriate organic solvent, preferably dichloromethane, is cooled to -20°C to 20°C, preferably 0°C, and reacted with a trihaloalkyl sulfonic anhydride, preferably trifluoromethanesulfonic anhydride to form compounds (p).

Formula (q) compounds are prepared by reacting a Formula (p) compound in a metal catalyzed coupling reaction. Preferably a Formula (p) compound dissolved in dimethylformide (DMF) and organic base preferably, triethylamine, a

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- 19phosphine, preferably bis(diphenylphosphino)propane, a palladium(II) compound, preferably, palladium(II) acetate, and a C i-Chalky 1 alcohol (CpCgalkOH), followed by addition of carbon monoxide (CO). Formula (q) compounds are reacted with a reducing agent, preferably diisobutylaluminum hydride, to yield

Formula (r) compounds.

Formula (s) compounds are produced by treating Formula (r) compounds with a grignard reagent (as described in Scheme III) or a lithium reagent in tetrahydrofuran or diethylether solvent, at a temperature of about -50° to -70°C, for 1-16 hours.

Formula (g) compounds are prepared by oxidation of Formula (s) compounds. Preferably said oxidation will utilize a Jones reagent or ¢-% tetrapropylamonium perruthenate followed by sodium chlorite.

Grignard reagents of the type, XMgR, (where R is Cj_20 linear or branched, saturated or unsaturated alkyl) for use in preparing all of the species included within the scope of this invention, are available or can be made readily by one skilled in the art.

Formula I compounds in which Z is in the a position are prepared from compounds which contain the corresponding β substituent by the General Method below.

General Method A

To a stirrred solution of a substituted 17β steroidal dual 5a-reductase inhibiting compound of Formula (II) in an appropriate solvent, preferably ethylene glycol or dimethyl sulfoxide, is added a base such as a hydroxide or alkoxide base, preferably sodium hydroxide, potassium hydroxide or sodium methoxide, at a temperature over 100°C preferably at reflux temperatures to yield the corresponding a epimer, after isolation and work up.

In determining the appropriate solvent for conducting the epimerization, dimethyl sulfoxide or other non-reactive high boiling solvents are preferred when the starting 17B dual 5a-reductase inhibiting steroidal compound contains reactive substituents or reactive unsaturated bonds that are, for example, subject to nucleophilic attack and ethylene glycol, or other reactive high boiling solvents can be used when the reactivity of the substituents or any unsaturated bonds of the starting 17B dual 5a-reductase inhibiting steroidal compound is not a consideration.

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-20Also within the scope of the present invention are the ketone reduction products of I, the secondary alcohols of the formula (IV):

wherein Y is a or β

OH

I — C—R in which R is 0μ20 li^near °^r branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

Preferred among the presently invented ketone reduction products described above are the secondary alcohols wherein the OH

I — C—R substituent is in the β position.

Particularly preferred among the presently invented ketone reduction products described above are 17B-(l-hydroxyethyl)-estra-l,3,5(10)-triene-3_:carboxylic acid < and 17B-(l-hydroxybutyl)-estra-l,3,5(10)-triene-3-carboyxlic acid.

These compounds can be made by conventional sodium borohydride ( reduction of the carbonyl attached to R without epimerization of the 17 substituent or reducing the carboxyl in Ring A or the aromatic A ring.

The borohydride reduction can be carried out in e.g. water or aqueous methanol, at a temperature of room temperature to 50°C and the product then isolated and purified by conventional means. The compounds are also active as dual inhibitors of 5-alpha reductase.

By the term increased temperatures as used herein and in the claims is meant above 25°C, preferably at reflux temperatures.

By the term solvent or appropriate solvent as used herein and the in the claims is meant a solvent such as methylene chloride, ethylene chloride, chloroform, ethylene glycol, carbon tetrachloride, tetrahydrofuran (THF), ethyl ether, toluene, ethyl acetate, dimethylsulfoxide (DMSO), N,N'-dimethyl-N,N'-_

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-21 P50095-1 propylene urea, N-methyl-2-pyrrolidinone, methanol, isopropylalcohol, dimethylformamide (DMF), hexane, water, pyridine, quinoline or ethanol.

Pharmaceutically acceptable salts, hydrates and solvates of Formula (I) and Formula (TV) compounds are formed, where appropriate, by methods well known to those of skill in the art.

In preparing the presently invented compounds of Formula (I), novel intermediates of the Formula (V) are synthesized;

II

C-R (V) in which R is C4.20 linear or branched, saturated or unsaturated alkyl and

R⁴ is fluorosulfonyloxy.

Also preferred in synthesising the presently invented formula (I) compounds were novel intermediates of the formula (VI):

OH \

H<

(VI) in which R is as described in formula (II).

A preferred process for preparing a compound of Formula (Π)

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in which R is Cj_20 l'^near or branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof comprises reacting a compound of the formula

in which R is as described above with fluorosulfonic anhydride and a base, preferably, 2,5-t-butyl-3-methyl-pyridine, in a solvent, preferably dichloromethane, to form a compound of the formula

FQj,SO

in which R is as described above and subsequently reacting said compound in a metal-catalyzed coupling reaction in the presence of an approrpirate coupling reagent, preferably, carbon monoxide followed by an optional, if applicable, hydrolysis reaction to form a compound of formula U, and thereafter optionally forming a pharmaceutically acceptable salt, hydrate, solvate or ester thereof.

Because the presently invented pharmaceutically active compounds are potent dual inhibitors of steroid 5-a-reductase activity, they have therapeutic utility in treating diseases and conditions wherein decreases in DHT activity produces the desired therapeutic effect. Such diseases and conditions include acne vulgaris,__

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-23seborrhea, female hirsutism, male pattern baldness, prostate diseases such as benign prostatic hypertrophy, and prostatic adenocarcinoma.

In determining potency in inhibiting the human 5<x-reductase enzyme, the following procedure was employed:

Preparation of membrane particulates used as source for recombinant steroid

Sgjeductasg isozyme 1,

Chinese hamster ovary (CHO) cells containing expressed, recombinant human steroid 5a-reductase isoenzyme 1 (Andersson, S., Berman, D.M., Jenkins,

E.P., and Russell, D.W. (1991) Nature 354 159-161) were homogenized in 20 mM potassium phosphate, pH 6.5, buffer containing 0.33 M sucrose, 1 mM dithiothreitol, and 50 pM NADPH (buffer A) using a Dounce glass-to-glass hand homogenizer (Kontes Glass Co., Vineland, N.J.). Membrane particulates were isolated by centrifugation (100,000 x g at 4°C for 60 minutes) and resuspended in

20 mM potassium phosphate, pH 6.5, containing 20% glycerol, 1 mM dithiothreitol, and 50 μΜ NADPH (buffer B). The suspended particulate solution was stored at -80°C.

Preparation of prostatic membrane particulates used as source for steroid

5a-reductase isozyme 2.

Frozen human prostates were thawed and minced into small pieces ( Brinkmann Polytron (Sybron Corp., Westbury, New York). The solution was sonicated for 3 to 5 minutes with a Sonifier (Branson Sonic Power Co.) followed by hand homogenization in a Dounce hand homogenizer. Prostatic particles were obtained by differential centrifugation at 600 or 1000 x g for 20 minutes and 140,000 x g for 60 minutes at 4°C. The pellet obtained from the 140,000 x g centrifugation was washed with 5 to 10 tissue volumes of the buffer described above and centrifuged at 140,000 x g. The resulting pellet was suspended in buffer B and the particulate suspension was stored at -80°C.

Preparation of membrane particulates used as source for recombinant steroid 5-aredii£ias£js<2zym£_2x

Chinese hamster ovary (CHO) cells containing expressed, recombinant human steroid 5-a-reductase isozyme 2 were homogenized in 20 mM potassium phosphate, pH 6.5, buffer containing 0.33 M sucrose, ImM dithiothreitol, and 50 μΜ NADPH (buffer A) using a Douce hand homogenizer. Membrane particulates

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-24containing the recombinant human enzyme were isolated by centrifugation (100,000 x g at 4°C for 60 minutes) and resuspended in 20 mM potassium phosphate, pH 6.5 containing 20% glycerol, ImM dithiothreitol, and 50 pM NADPH (buffer B). The suspended particulate solution was stored at -80°C until used.

Assay for enzymes activities and inhibitors potency.

A constant amount of [ C]testosterone (50 to 55 mCi/mmol) in ethanol and varying amounts of potential inhibitor in ethanol were deposited in test tubes and concentrated to dryness in vacuo. To each tube was added buffer, 10 pL (recombinant isoenzyme 1 or isoenzyme 2) or 20 pL (isoenzyme 2 from human prostate tissue) of 10 mM NADPH and an aliquot of a steroid 5a-reductase preparation to a final volume of 0.5 mL. Assays for human steroid 5a-reductase isoenzyme 1 were conducted with a sample of the recombinant protein expressed in CHO cells in 50 mM phosphate buffer, pH 7.5 while assays of isoenzyme 2 were conducted with a suspension of human prostatic particulates and/or recombinant protein expressed in CHO cells in 50 mM citrate buffer at pH 5.0.

After incubating the solution at 37°C for 20 or 30 minutes the reaction was quenched by the addition of 4 mL ethyl acetate and 025 pmol each of testosterone, 5a-dihydrotestosterone, androstanediol, and androstanedione as carriers. The organic layer was removed to a second test tube and evaporated to dryness in a Speed Vac. The residue was dissolved in 40 pL chloroform, spotted on an individual lane of a 20 x 20 cm prechannelled silica gel TLC plate (Si 250F-PA, Baker Chemical) and developed twice with acetone:chloroform (1:9). The radiochemical content in the bands of the substrate and the products was determined with a BIOSCAN Imaging Scanner (Bioscan Inc., Washington, D.C.). The percent of recovered radiolabel converted to product was calculated, from which enzyme activity was determined. All incubations were conducted such that no more than 20% of the substrate (testosterone) was consumed.

The experimentally obtained data was computer fit to a linear function by plotting the reciprocal of the enzyme activity (1/velocity) against the variable inhibitor concentration; apparent inhibition constants (Kj _app) were determined by the Dixon analysis (Dixon, M. (1953).

The value for the inhibition constant (Ki) was calculated from known procedures (Levy, M. (1989), Biochemistry, 22:2815-2824).

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-25Α11 of the compounds within the scope of this invention are useful in inhibiting steroid 5-a-reductase isozyme 1 and steroid 5-a-reductase isozyme 2 in a mammal, including humans, in need thereof.

Compounds within the scope of this invention have been tested and have shown an activity of from 15 Ki(nM) to 180 Ki(nM) against isozyme 1 and an activity of from 0.5 Ki(nM) to 30 Ki(nM) against isozyme 2. Particularly preferred among the compounds of the invention and the compounds used in the invented pharmaceutical compositions and invented methods are

176-(isobutylcarbonyl)-estra-1,3,5(10)-triene-3-carboxylic acid,

176-(octylcarbonyl)-estra-l,3,5( 10)-triene-3-carboxylic acid,

176-(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

176-(2,2-dimethylpropylcarbonyl)-estra-1,3,5(10)-triene-3-carboxylic acid,

176-(propylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(methylcarbonyl)-estra-l ,3,5( 10)-triene-3-carboxylic acid,

176-(3,3-dimethylbutylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

176-(1-hydroxyethyl)-estra-1,3,5(10)-triene-3-carboxylic acid and

176-(1 -hydroxybutyl)-estra-1,3,5( 10)-triene-3-carboxylic acid.

The pharmaceutically active compounds of the present invention are incorporated into convenient dosage forms preferably as capsules, tablets, or injectable preparations. Solid or liquid pharmaceutical carriers are employed.

Solid carriers include, starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, saline, and water. Similarly, the carrier or diluent may include any prolonged release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies widely but, preferably, will be from about 25 mg to about 1 g per dosage unit. When a liquid carrier is used, the preparation will peferably be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampoule, or an aqueous or nonaqueous liquid suspension.

The pharmaceutical preparations are made following conventional techniques of a pharmaceutical chemist involving mixing, granulating, and compressing, when necessary, forlablet forms, or mixing, filling and dissolving the ingreidents, as appropriate, to give the desired oral or parenteral products.

Doses of the presently invented pharmaceutically active compounds in a pharmaceutical dosage unit as described above will be an efficacious, nontoxic

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-26quantity preferably selected from the range of

0.1 - 1000 mg/kg of active compound, preferably 1-100 mg/kg. When treating a human patient in need of steroid 5-a-reductase inhibition, the selected dose is administered preferably from 1-6 times daily, orally or parenterally. Preferred forms of parenteral administration include topically, rectally, transdermally, by injection and continuously by infusion. Oral dosage units for human administration preferably contain from 1 to 500 mg of active compound. Oral administration, which uses lower dosages is preferred. Parenteral administration, at high dosages, however, also can be used when safe and convenient for the patient.

The method of this invention of inhibiting steroid 5-a-reductase isozyme 1 and steroid 5-a-reductase isozyme 2 activity in mammals, including humans, comprises administering to a subject in need of such inhibition an effective dual inhibiting amount of a compound of the present invention.

The invention also provides for the use of a compound of Formula (I) or a compound of Formula (TV) in the manufacture of a medicament for use in the dual inhibition of steroid 5-a-reductase.

The invention also provides for a pharmaceutical composition for use in the treatment of benign prostate hypertrophy which comprises a compound of Formula I or a compound of Formula (IV) and a pharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use in the treatment of prostatic adenocarcinoma which comprises a compound of Formula I or a compound of Formula (IV) and a pharmaceutically acceptable carrier.

The invention also provides for a process for preparing a pharmaceutical composition containing a pharmaceutically acceptable carrier or diluent and a compound of Formula I or a compound of Formula (IV) which comprises bringing the compound of Formula I or the compound of Formula (IV) into association with the pharmaceutically acceptable carrier or diluent.

No unacceptable toxicological effects are expected when compounds of the invention are administered in accordance with the present invention.

In addition, the pharmaceutically active compounds of the present invention can be co-administered with further active ingredients, such as other compounds known to treat the disease states of acne vulgaris, seborrhea, female hirsutism, male pattern baldness, benign prostate hypertrophy or prostatic adenocarcinoma or compounds known to have utility when used in combination with 5-a-reductase inhibitors. Particularly preferred is the co-administration of a dual 5-a-reductase inhibitor, as disclosed herein, and minoxidil for use in the treatment of male pattern _____

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-27baldness. Particularly preferred is the co-administration of a dual 5a-reductase inhibitor, as disclosed herein, and a α-receptor antagonist for use in Ae treatment of benign prostatic hypertrophy. Preferred is Ae co-administration of a dual 5-areductase inhibitor, as disclosed herein, and an aromatase inhibitor for use in Ae treatment of benign prostatic hypertrophy. Preferred is Ae co-administration of a dual 5-a-reductase inhibitor, as disclosed herein, a a -receptor antagonist and an aromatase inhibitor for use in Ae treatment of benign prostatic hypertrophy.

WiAout further elaboration, it is believed Aat one skilled in Ae art can, using Ae preceding description, utilize Ae present invention to its fullest extent.

The following Examples are, therefore, to be construed as merely illustrative and not a limitation of Ae scope of Ae present invention in any way.

EXAMPLE 1-corresponding to Scheme ΠΙ UB-Isoburylcarbonvl-estra-l.B.SnOi-triene-B-carboxvlic Acid (i) . S-(2-pyridyl)-3-hydroxy-estra-l,3,5(10)-triene-17B-Aiocarboxylate.

A mixture of 3-hydroxy-estra-l,3,5(10)-triene-17B-carboxylic acid (0.11 g, 0.37 mmol), 2,2'-diipyridyl disulfide (0.163 g, 0.74 mmol), triphenylphosphine (0.19 g, 0.74 mmol) and dichloromethand (20 ml) was stirred at ambient temperature under argon for 4 hours. The resulting solution was concentrated and Ae residue was chromatographed (silica gel, eluting wiA 25 % eAyl acetate in hexane) to provide 0.13 g of Ae title compound as a white solid, mp 195-196°C (recrystallized from eAyl acetate - meAanol).

(ii) . 17B-Isobutylcarbonyl-3-hydroxy-estra-l,3,5(10)-triene.

Isobutylmagnesium bromide (2.3 mL; 2M in dieAyl ether) was added slowly to a solution of S-(2-pyridyl)-3-hydroxy-estra-l,3,5(10)-triene-17BAiocarboxylate (0.60 g, 1.53 mmol) in tetrahydrofuran (20 mL) at -78°C. After 1 hour, Ae mixture was quenched with saturated aqueous NH4CI and extracted with eAyl acetate. The organic extract was washed with brine, dried, and concentrated. The resulting residue was chromatographed (silica gel, eluting with 15% ethyl acetate in hexane) to give a foam. Trituration wiA diethyl ether and hexane gave a white solid (0.27 g).

(iii) . 17B-Isobutylcarbonyl-estra-1,3,5( 10)-triene-3-trifluoromethylsulfonate.

To a cooled (0°C) solution of 17B-Isobutylcarbonyl-3-hydroxy-estral,3,5(10)-triene (0.28 g, 0.82 mmol) and 2,6-di-tert-butyl-4-methylpyridine (0.17

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Ρ50095-1 g,0.83 mmol) in dichloromethane (20 ml) was slowly added trifluoromethane sulfonic anhydride (0.23 g, 0.82 mmol). The resulting solution was stirred at 0°C for 1 hour and then at ambient temperature of 30 minutes. The reaction mixture was washed with dilute HCl, water, dilute NaHCCfy brine dried and concentrated. The resulting residue was chromatographed (silica gel, 7% ethyl acetate in hexane) to provide 0.13 g of oil.

(iv) . Methyl- 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylate.

A mixture of 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3trifluoromethylsulfonate (0.13 g 0.28 mmol), palladium (II) acetate (4.2 mg, 0.0187 mmol), l,3-bis(diphenylphosphino)propane (dppp, 7.5 mg, 0.0182 mmol), triethylamine (0.08 mL), methanol (0.6 mL), 1,2-dichloroethane (0.32 mL), and DMSO (1 mL) was heated at 70-75°C under an atmosphere of CO overnight. The cooled reaction mixture was then diluted with dichloromethane, washed with water, dilute HCl, dilute NaHCC>3 brine, dried and concentrated. The residue was chromatographed (silica gel, eluting with 10% ethyl acetate in hexane) to provide 70 mg of the title compound.

(v) . 17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylic acid.

A mixture of methyl-17B-isobutylcarbonyl-estra-1,3,5( 10)-triene-3carboxylate (70 mg, 0.18 mmol), K2CO3 (0.12 g, 0.87 mmol), water (1.5 mL) and methanol (10 mL) was heated at reflux overnight. The reaction mixture was then concentrated. The residue was diluted with water, acidified with dilute HCl and extracted with ethyl acetate. The organic extract was washed with brine, dried, and concentrated. The title compound purified by HPLC. mp 202-206°C.

EXAMPLE 2-corresponding to Scheme ΠΙ 17 B-Isobutylcarbonyl-estra-1.3.5( 10)-triene-3-carboxyIic Acid (i). 17 β-Isobutylcarbonyl-estra-1,3,5( 10)-triene-3-fluorosulfonate.

A solution of 17|3-isobutylcarbonyl-estra-l,3,5(10)-trien-3-ol, prepared according to Example 1 (i-ii), in dichloromethane at 0°C is treated with 2,5-di-tbutyl-3-methyl-pyridine followed after 10 min with fluorosulfonic anhydride. The resulting mixture is stirred for 2 h and then diluted with dichloromethane. The organic layer is washed with saturated aqueous NaHCO3 and brine, dried over MgSO4, and evaporated to dryness. Chromatography on silica gel yields the title compound.

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-29(ii) . Methyl 173-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylate.

A mixture of 17|3-isobutylcarbonyl-estra-l,3,5(10)-triene-3-fluorosulfonate l,3-bis(diphenylphosphino)propane, palladium diacetate, triethylamine, methanol, DMSO and 1,2-dichloroethane is heated and vigorously stirred for 5 h at 80°C under an atmosphere of carbon monoxide. After cooling to ambient temperature, the resulting mixture is diluted with dichioromethane. The organic phase is thoroughly washed with water, dried (MgSO4) and evaporated to dryness. Chromatography on silica gel yields the title compound.

(iii) . 17J3-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylic Acid.

A mixture of methyl 17p-isobutylcarbonyl-estra-l,3,5(10)-triene-3carboxylate, K2CO3, water and methanol is heated at reflux for 5 h. The volatiles are then removed at reduced pressure and the residue is diluted with water, acidified with dilute aqueous HCl, and extracted with EtOAc. The organic extract is washed with water and brine, dried, and evaporated to yield the title compound.

EXAMPLE-3-corresponding to Scheme III 17B-Isobutvlcarbonvl-estr-1.3.5(10)-triene-3-carboxylic acid (i) . 3-(trifluoromethanesulfonyloxy)-estra-l,3,5(10)-triene-17B-carboxylic acid

A solution of 3-hydroxy-estra-l,3,5(10)-17B-carboxylic acid, 2,6-di-t-butyl4-methyl pyridine and trifuloromethane sulfonic anhydride in methylene chloride is stirred at 5°C for 20 hours. The organic solvent is evaporated and the residue is dissolved in tetrahydrofuran water (99.5.Ό.5) with 4-dimethylaminopyridine which upon acidification with hydrochloric acid followed by conventional workup yields title compound.

(ii) . S-(2-pyridyl)-3-(triflurormethanesulfonyloxy)-estra-1,3,5(10)-triene-17Bthiocarboxylate

A solution of 3-(trifluoromethanesulfonyloxy)-estra-l,3,5(10)-triene-17Bcarboxylic acid, triphenylphosphine and, 2,2'-dipyridyl disulfide in toluene is stirred under nitrogen for 20 hours. The reaction mixture is concentrated and the residue is passed directly through silica gel and appropriate fractions evaporated to yield title comopund.

(iii) . 17B-Isobutylcarbonyl-estea-l₂3,5(10)-triene-3-trifluoromethane sulfonate

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-30Το a solution of S-(2-pyridyl)-3-(trifluoromethanesulfonyloxy)-estral,3,5(10)-triene-17B-thiocarboxylate in tetrahydrofuran at about -50°C is added isobutylmagnesium bromide. The reaction mixture is warmed to about -10°C, and diluted with a saturated aqueous ammonium chloride solution. Conventional workup with subsequent isolation by column chromatography yields title compound.

(iv) . Methyl 17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylate

A solution of 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3trifluoromethane sulfonate, triphenyl phosphine, palladium II acetate, triethylamine, methanol and dimethyl formamide is stirred vigorously under a carbon monoxide atmosphere for 20 hours. Conventional workup with subsequent isolation by column chromatography yields title compound.

(v) . 17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylic acid

A mixture of methyl 176-isobutylcarbonyl-estra-l,3,5(10)-triene-3carboyxlate, K2CO3, water and methanol is heated at reflux for about 5 hours. Acidifications followed by conventional workup yields title compound.

EXAMPLE 4-corresponding to Scheme III 17B-Isobutylcarbonyl-estr-1.3.5(10)-triene-3-carboxylic acid (i) . 3-(fluorosulfonyloxy)-estra-l,3,5(10)-triene-17B-carboxylic acid

A solution of 3-hydroxy-estra-l,3,5(10)-17B-carboxylic acid, 2,6-di-t-butyl4-methyl pyridine and fluorosulfonic anhydride in methylene chloride is stirred at 5°C for 20 hours. The reaction mixture is washed with aqueous hydrochloric acid and water. The organic phase is concentrated and the resulting residue is purified by column chromatography to yield the title compound.

(ii) . S-(2-pyridyl)-3-(fluorosulfonyloxy)-estra-l,3,5(10)-triene-17B-thiocarboxylate

A solution of 3-(fluorosulfonyloxy)estra-l,3,5(10)-triene-17B-carboxylic acid, triphenylphosphine and, 2,2'-dipyridyl disulfide in toluene is stirred under nitrogen for 20 hours. The reaction mixture is concentrated and the residue is passed directly through silica gel and appropriate fractions evaporated to yield title compound.

(iii) . 17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-fluorosulfonate bad original 4

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-31 To a solution of S-(2-pyridyl)-3-(fluorosulfonyloxy)-estra-l,3,5(10)-triene17B-thiocarboxylate in tetrahydrofuran at about -50°C is added isobutylmagnesium bromide. The reaction mixture is warmed to about -10°C and diluted with a saturated aqueous ammonium chloride solution. Conventional workup with subsequent isolation by column chromatography yields title compound.

(iv) . Methyl 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylate

A solution of 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3-fluorosulfonate, triphenyl phosphine, palladium II acetate, triethylamine, methanol and dimethyl formamide is stirred vigorously under a carbon monoxide atmosphere for 20 hours. Conventional workup with subsequent isolation by column chromatography yields title compound.

(v) . 17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylic acid

A mixture of methyl 17B-isobutylcarbonyl-estra-l,3,5(10)-triene-3carboxylate, K2CO3, water and methanol is heated at reflux for about 5 hours. Acidification followed by conventional workup yields title compound.

Example 5 - Corresponding to Scheme ΠΙ 17B-(octvlcarbonyl)-estra-1.3.5flO)-triene-3-carboxyIic acid (i) 17P-(octylcarbonyl)-estra-1,3,5(10)-triene-3-trifluoromethylsulfonate.

The title compound was prepared according to Example 1 (i-iii) by substituting n-octylmagnesium chloride for isobutylmagnesium bromide in step (ii).

(ii) 17^-(octylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid.

A mixture of 17|3-(octylcarbonyl)-estra-l,3,5(10)-triene-3trifluoromethylsulfonate (0.38 g, 0.7 mmol), potassium acetate (0.27 g), palladium(II) diacetate (0.008 g, 0.036 mmol), 1.Γbis(diphenylphosphino)ferrocene(dppf; 0.08 g, 0.14 mmol), in DMSO (15 ml) was purged with carbon monoxide for 2 minutes and stirred under a CO balloon at 60°C overnight The reaction mixture was diluted with water, acidified with 0.5N hydrochloric acid and extracted with dichloromethane. The organic layer was washed with water, dried (MgSO4), and evaporated under vacuum. The residue was chromatographed (silica gel, eluting with 20% ethylacetate, 1% acetic acid in hexane) to give solid triturated with methanol-acetonitrile to yield the title compound 0.22 g (73%) mp 175°C.

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-32Exarople 6 - Corresponding to Scheme ΠΙ

17B-(tert-pentvlcarbonyl)-estra-1.3.5(10)-triene-3-carboxylic acid (i) Trifluoromethyl-17|3-(tert-pentylcarbonyl)-estra-1,3,5 (10)-triene-3sulfonate.

The tide compound was prepared according to Example 1 (i-iii) by substituting tert-pentylmagnesium chloride for isobutylmagnesium bromide in step (ii) .

(ii) 17P-(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid.

The title compound was prepared according to Example 5 (ii) by substituting trifluoromethyl-17|3-(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-sulfonate, as prepared in Example 6 (i), for trifluoromethyl-17β-(οοΐγ1θ3τ1κ>ηγ1)-65ϋ·3-1,3,5(10)triene-3-sulfonate. mp 199-200°C.

Example 7 - Corresponding to Scheme HI 17S-(2.2-dimethvlpropylcarbonyl)-estra-1.3.5(10)-triene-3-carboxylic acid (i) Trifluoromethyl-17()-(2,2-dimethylpropylcarbonyl)-estra-1,3,5(10)-triene-3sulfonate.

The title compound was prepared according to Example 1 (i-iii) by substituting 2,2-dimethylpropylmagnesium bromide for isobutylmagnesium bromide in step (ii).

(ii) 17(3-(2,2-dimethylpropylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid.

The title compound was prepared according to Example 5 (ii) by substituting trifluoromethyl-17(3-(2,2-dimethylpropylcarbonyl)-estra-l,3,5(10)-triene-3sulfonate, as prepared in Example 7 (i), for trifluoromethyl-17P-(octylcarbonyl)estra-l,3,5(10)-triene-3-sulfonate. mp 210°C.

I

Example 8 - Corresponding to Scheme IV 17&-(propylcarbonyl)-estra-1.3.5(10')-triene-3-carboxylic acid (i) 17P-cyano-estra-l,3,5(10)-triene-3-methancsulfonate.

The title compound is known and is prepared from estrone according to the procedure of Baldwin et al., J. Chem. Soc. (C), 1968,2283-2289.

(ii) 17f)-Cyano-estra-1,3,5 (10)-triene-3-ol.

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-33The title compound of 17P-cyano-estra-l,3,5(10)-triene-3-methanesulfonate (10 g) in methanol (100 mL) is added dropwise a solution of NaOH (42 mL of a 20% solution in 1:1 MeOH-water). The resulting mixture is heated to 40°C for 24 h after which time the mixture is cooled to 0°C, diluted with water (350 mL), and acidified with dilute HCl. The resulting white precipitate is isolated by vacuum filtration, washed with water, and dried under vacuum. Recrystallization from acetonitrile provides the title compound mp 249-250°C Dec..

(iii) 17fi-cyano-estra-l,3,5(10)-triene-3-trif]uoromethanesulfonate.

17p-cyano-estra-l,3,5(10)-triene-3-ol (4.2 g) and 2,6-di-t-butyl-4methylpyride (3.6 g) was dissolved in 50 ml of methylene chloride. The mixture was stirred at room temperature for 30 min. Triflic anhydride (4.2 ml) was added and the mixture was stirred another 40 mins, diluted with 50 ml CH2CI2, filtered, concentrated, and chromatographed on silica gel. Elution with 20% ethyl acetate in hexane gave 5.3 g (87%) of the title compound, mp 115-117°C.

(iv) Methyl 17J3-cyano-estra-l,3,5(10)-triene-3-carboxylate.

To a solution of 17|3-cyano-estra-l,3,5(10)triene-3-oI (10 g) in 77 ml of DMSO and 50 ml of MeOH was added 7 ml of triethylamine, 0.35 g of palladium acetate, 0.64 g of bis(diphenylphosphino)propane and 1,2-dichloroethane (26 ml). Carbon monoxide was bubbled through the solution and the reaction mixture was then stirred at 75°C overnight under 1 atm of CO (balloon). The mixture was diluted with EtOAc and washed with water (3x), dried, and concentrated. The residue was chromatographed (silica gel, eluting with 10% EtOAc in hexane) to provide 4.5 g of the title compound, mp 161-163°C.

(v) 3-Hydroxymethyl-estra-1,3,5(10)-triene-17 β-carboxaldehyde. Methyl-17fS-cyano-estra-l,3,5(10)-triene-3-carboxylate (0.8 g) was dissolved in 30 ml of toluene and treated with DIBAL (6 ml, IM). The mixture was stirred at room temp under argon for 2.5 hours. The mixture was then poured into 50 ml of 5% H2SO4 and the mixture was stirred for 1 hour, filited, dried, and concentrated. The residue was chromatographed (silica gel, eluting with 20% EtOAc in hexane) to provide 424 mg of the title compound, mp 146-150°C.

(vi) 17P-(l-hydroxybutyl)-estra-l,3,5(10)-triene-3-hydroxymethyl.

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-34Α solution of 3-hydroxymethyl-estra, l,3,5(10)-triene-17P-carboxaldehyde (75 mg in 2 ml of THF) was added slowly to a solution of propylmagnesium bromide (2 ml, 1.6 M). The mixture was stirred at room temp for 2 hours.

The mixture was quenched with saturated aqueous NH4CI and extracted with CH2CI2. The organic extract was washed with brine, dried, and concentrated. The resulting residue was chromatogrpahed (silica gel, eluting with 30% ethyl acetate in hexane) to give 57 mg of title compound.

(vii) 17(3-(propylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid.

17|3-(l-hydroxybutyl)-estra-l,3,5(10)-triene-3-hydroxymethyl (57 mg) was dissolved in acetone (5 ml) and treated with Jones reagent. The mixture was stirred for 1 hour and then quenched with 2-propanol, extracted with CH2CL2 and purified by chromatography (silica gel, eluting with 30% etase in hexane with 0.5% HOAc added) to give 47 mg of the title compound, mp 211-213°C.

Example 9 - Corresponding to Scheme IV 17fi-fmethylcarbonyn-estra-1.3.5(10)-triene-3-carboxylic acid The title compound was prepared according to Example 8 (i)-(vii) by substituting methylmagnesium bromide for propylmagnesium bromide in step (vi). mp 199-202°C.

Example 10 - Corresponding to Scheme IV 17fi-(3.3-dimethylbutylcarbonyl)-estra-1.3.5(10)-triene-3-carboxylic acid

The title compound was prepared according to Example 8 (i)-(vii) by substituting 3,3-dimethylbutylmagnesium chloride for propylmagnesium bromide in step (vi). mp 252-255°C.

Example 11 - Corresponding to Scheme IV

17p-ri-(R.SFhydroxyethyn-estra-1.3.5(10)-triene-3-carboxyIic acid (i) A solution of 17P-(methylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid (6 mg) - prepared according to Example 9 - in methanol was treated with NaBH4 (0.7 mg). The mixture was warmed to 45°C and stirred overnight. The solvent was removed by rotary evaporation and the residue was dissolved in dichloromethane and filtered. The title compound (1.6 mg) was purified by preparative thin layer chromatography, mp 202-204°C.___ bad original £ (ii) Pure (R) and (S) forms are obtained by separation techniques readily available and known to those of skill in the art.

AP ο Ο Ο 4 1 2

Ρ5ΟΟ95-1

-35Example 12 - Conesponding to Scheme IV

17Ml-(R.S)-hydroxybutyll-estra-1.3.5(10)-triene-3-carboxylic acid (i) The title compound was prepared according to Example 11 by substituting 17{5-(propylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid, prepared according to Example 8 (i)-(vii) for 17P-(methylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid, mp 208-211 °C.

(ii) Pure (R) and (S) forms are obtained by separation techniques readily available and known to those of skill in the art.

Example 13

An oral dosage form for administering Formula I comounds is produced by screening, mixing, and filling into hard gelatin capsules the ingredients in the proportions shown in Table 1, below.

Tahiti

Ingredients Amounts

17B-isobutylcarfionyl-estra-1,3,5(10)- 50 mg triene-3-carboxylic acid magnesium stearate 5 mg lactose 75 mg

EXAMPLE,,14

The sucrose, calcium sulfate dihydrate and Formula (I) compound shown in Table Π below, are mixed and granulated in the proportions shown with a 10% gelatin solution. The wet granules are screened, dried, mixed with the starch, talc and stearic acid, screened and compressed into a tablet.

BAD ORIGINAL

AF Ο ο ο 4 1 ζ

Ρ50095-1

-36Table II

Ingredients Amounts

17B-Isobutylcarbonyl-estra-l, 3,5(10)- 100 mg triene-3-carboxylic acid calcium sulfate dihydrate 150 mg sucrose 20 mg starch 10 mg talc 5 mg stearic acid 3 mg

Example. 15

17B-Isobutylcarbonyl-estra-l,3,5(10)-triene-3-carboxylic acid, 75 mg, is 5 dispursed in 25 ml of normal saline to prepare an injectable preparation.

While the preferred embodiments of the invention are illustrated by the above, it is to be understood that the invention is not limited to the precise instructions herein disclosed and that the right to all modifications comming within the scope of the following claims is reserved.

Claims

What is claimed is:

1. A compound represented by the formula:

wherein Z is a or β O

II

5 -C-R in which R is C]_20 linear or branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.
2. A compound of claim 1 having the following formula:

in which R^ is Cpg linear or branched alkyl and pharmaceutically acceptable salts, hydrates solvates and esters thereof.
3. A compound of claim 2 wherein R^ is methyl, ethyl, propyl, 315 methylbutyl, isopropyl, n-butyl, isobutyl, 1-methylpropyl, t-butyl, pentyl, 1,1dimethylpropyl, 2,2-dimethylpropyl, octyl or 3,3-dimethylbutyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.

wherein Y is a or β

BAD ORIGINAL

P50095-1C

AP Ο 00
4 1 g

-38OH

I — C—R in which R is Cj_20 linear or branched, saturated or unsaturated alkyl and pharmaceutically acceptable salts, hydrates, solvates and esters thereof.
5 5. A compound selected from the group consisting of

17B-(isobutylcarbonyl)-estra-1,3,5( 10)-triene-3-carboxylic acid, 17B-(octylcarbonyl)-estra-l ,3,5( 10)-triene-3-carboxylic acid, 17B-(tert-pentylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

176-(2,2dimethylpropylcarbonyl)-estra-1,3,5(10)-triene-3-carboxylic acid,

10 17B-(propylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(methylcarbonyl)-estra-l,3,5(10)-triene-3-carboxylic acid,

17B-(3,3-dimethylbutylcarbonyl)-estra-1,3,5(10)-triene-3-carboxylic acid, 17B-(hydroxyethyl)-estra-l,3,5(10)-triene-3-carboxylic acid and 17B-( 1 -hydroxybutyl)-estra-1,3,5(10)-triene-3-carboxylic acid.
6. A pharmaceutical composition comprising a compound according to anyone of claims 1 to 5 and a pharmaceutically acceptable carrier.
7. A compound according to any one of claims 1 to 5 for use in therapy.
8. A compound according to any one of claims 1 to 5 in the manufacture of a medicament for use as a steroid 5-a-reductase inhibitor.
9. A compound according to any one of claims 1 to 5 in the

25 manufacture of a medicament for use in treatment to reduce prostate size.
10. A compound according to any one of claims 1 to 5 in the manufacture of a medicament for use in treatment of prostatic adenocarcinoma.

30
11. An intermediate of the formula:

BAD ORIG^,nAL £

APO 00 4 1 2

P50095-1C in which R is Cj.20 linear or branched, saturated or unsaturated alkyl and R^ is fluorosulfonyloxy.

5
12. The use of a compound according to anyone of claims 1 to 5 and an alpha-receptor antagonist compound as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 to 5 and an alpha-receptor antagonist compound.

10
13. The use of a compound according to anyone of claims 1 to 5 and an alpha-receptor antagonist compound in the manufacture of medicament for use in the treatment of benign prostatic hypertrophy which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 to 5 and an alpha-receptor antagonist compound.
14. The use of a compound according to anyone of claims 1 to 5 and minoxidil as an active therapeutic substance which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 to 5 and minoxidil.
15. The use of a compound according to anyone of claims 1 to 5 and minoxidil in the manufacture of a medicament for use in the treatment of male pattern baldness which use consist of separate sequential or simultaneous administration of a compound according to anyone of claims 1 to 5 and minoxidil.
16. An intermediate of the formula:

Bad ORIGINAL d

AP 0 ϋ ο 4 1 2

P50095-1C in which R is Cj_20 l*^near °^r branced, saturated or unsaturated alkyl.
17. A process for the preparation of a compound of the Formula (Π) in which R is Cj_20 ^^near °^r branched, saturated or unsaturated alkyl, and pharmaceutically acceptable salts, hydrates, solvates and esters thereof which comprises either

-- 7

i) oxidation of a compound of the Formula in which R is described above, or (ii) reacting a compound of the Formula

BAD ORIGINAL ft

P50095-1C

AP Ο Ο Ο 4 1 2

-41 in which R is as described above, in a metal-catalyzed coupling reaction in the presence of an appropriate coupling reagent, preferably, carbon monoxide followed by an optional, if applicable, hydrolysis reaction, or

5 (iii) hydrolyzing a compound of the Formula in which R is as described above, or (iv) reacting a compound of the formula O

II in which R is as described above in a metal catalyzed coupling reaction in the presence of an appropriate coupling reagent, preferably, carbon monoxide followed by an optional, if applicable, hydrolysis reaction and thereafter optionally forming a pharmaceutically acceptable salt, hydrate, solvate or 15 ester thereof.
18. Use of a compound according to anyone of claims 1 to 5 in the manufacture of a medicament for use in inhibiting steroid 5-a-reductase.