FIELD OF THE INVENTION The present invention relates to certain novel diazo-phenanthrenone compounds, pharmaceutical compositions containing these compounds, methods for making these compounds, and methods for using these compounds to inhibit steroid 5-α-reductase.

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 hypertrophy 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-α- reduced androgens 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-α-reduced analog, in these tissues but not in others such as muscle and testes. Steroid 5-α-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 the discovery of a genetic steroid 5-α-reductase deficiency in male pseudohermaphrodites. Imperato-McGinley, J., et al., (1979), J. Steroid Biochem. 11:637-648.

Recognition of the importance of elevated DHT levels in various disease states has stimulated many efforts to synthesize inhibitors of this enzyme. A number of 5-α-reductase inhibiting compounds are known in the art. For example,

1. J. Steroid Biochem. (1989) Vol. 34 Nos. 1-6, pp. 571-575 , by M.A. Levy, et al., describes the interaction mechanism between rat prostatic steroid 5- alpha reductase and 3-carboxy-17β-substituted steroids;

2. J. Med. Chem. (1990) Vol. 33, pp. 937-942, by D.A. Holt, et al., describes the 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 5-α-reductase in benign prostatic hyperplasia, male pattern baldness and acne;

4. EPO Publn. No. 0 343 954 A3, to D. A. Holt, et al., (SmithKline Beckmann) describes steroidal 3-carboxylic acid derivatives as useful 5-α-reductase inhibitors; and

5. EPO Publn. No. 0 531 026 Al , to K.S. Hirsch, et al., describes hexahydrobenzo [f] quinolin-3 -ones as 5 -α-reductase inhibitors .

However, none of the above references specifically suggests that any of the novel diazo-phenanthrenone compounds of the present invention would have utility as potent testosterone 5-α-reductase inhibitors.

SUMMARY OF THE INVENTION

The present invention resides in the discovery that steroid 5-α-reductase is inhibited by certain diazo-phenanthrenone compounds. Presently preferred compounds of the invention, and compounds used in the invented pharmaceutical compositions and the invented methods, have the formula:

or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R! is hydrogen or Ci -Cg alkyl;

R2 is independently hydrogen; halogen; NO2; cyano; trifluoromethyl; C^- C alkyl; Cj-Cg alkoxy; carboxy; Ci -Cβ alkoxycarbonyl; amino; C1-C4 alkylamino; C1-C4 dialkylamino; amido; C1 -C4 alkylamido; C1-C4 dialkylamido; mercapto; -Cg alkylthio; C1-C6 alkylsulfmyl; C1-C6 alkylsulfonyl; -C(=O)R ³ where R ³ is C^-Cg linear or branched alkyl which can optionally be substituted with one or more of trifluoromethyl, halogen, -OH, -C(=O)OH, -O(Cι-C4 alkyl), -(CH2) _p OH where p is 1-4, -(eH2) _m COOH where m is 1-3, or a protected -OH group; aryl optionally substituted with one or more of trifluoromethyl, halogen,

-OH, -C(=O)OH, -O(Cι-C ₄ alkyl), -(CH ₂ ) _p OH where p is 1-4, -(CH ₂ ) _m C(=O)OH where m is 1-3, or a protected -OH group; or -R3-R4 where R ³ is Cj-Cg alkylene,

C2-C _β alkenylene or C2-Cg alkynylene, and R4 is halogen, hydroxy, trifluoromethyl, C^-Cg alkoxy, carboxy, C^-Cg alkoxycarbonyl, amino, C1-C4 alkylamino, C\-Ca dialkylamino, amido, C1-C4 alkylamido, C1-C4 dialkylamido, C1 -C4 alkylsulfonylamino, aminosulfonyl or C1-C4 alkylaminosulfonyl; and n is 1 or 2.

A presently preferred compound of the invention, and a compound used in the invented pharmaceutical compositions and invented methods, has the name: (+/-) ^{■ ■} trans- 1 -diazo-8-methoxy-4a-methyl- 1 ,2,3 ,4,4a,9, 10, 1 Oa- octahydrophenanthren-2-one. Another aspect of the invention is a pharmaceutical composition comprising a compound represented by the Formula (I), as defined previously, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

DETAILED DESCRIPTION OF THE INVENTION

DEFINITIONS As used herein, the term "alkyl" refers to a monovalent hydrocarbon radical having a straight or branched chain of carbon atoms, where the number of carbon atoms is selected from within a stated range. Exemplary alkyl radicals include methyl (-CH3), ethyl (-CH2CH3), «-propyl (-CH2CH2CH3), isopropyl (-CH(CH ₃ ) ), Λ-butyl (-CH2CH2CH2CH3), isobutyl (-CH ₂ CH(CH ₃ ) ₂ ), .yec-butyl (-CH(CH3)CH2CH3), t-butyl (-C(CH3)3) and, where indicated, higher homologs and isomers such as w-pentyl (-CH2CH2CH2CH2CH3), H-hexyl (-CH2CH2CH2CH2CH2CH3), 2-methylpentyl (-CH ₂ CH(CH ₃ )CH2CH3) and the like.

The term "alkenylene" refers to a divalent hydrocarbon radical having a straight or branched chain of carbon atoms, and a single carbon-carbon double bond (C=C), where the number of carbon atoms is selected from within a stated range. Exemplary alkenylene radicals include ethylene (-CH=CH-), l-propene-l,3-diyl (-CH ₂ -CH=CH- or -CH=CH-CH ₂ -), 2-butene-l,4-diyl (-CH ₂ -CH=CH-CH ₂ -), l-butene-l,4-diyl (-CH=CH-CH ₂ CH ₂ - or -CH ₂ CH ₂ CH=CH-) and the like.

The term "alkoxy" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, where the alkyl, alkenyl or alkynyl radical is bonded through an oxygen atom to the residue of a compound of Formula (I). Exemplary alkoxy radicals include methoxy (-OCH3),

ethoxy (-OCH2CH3), n-propoxy (-OCH2CH2CH3), isopropoxy (-OCH(CH3)2), propynyloxy (-OC^C≡CH) and the like.

The term "alkoxycarbonyl"refers to an alkyl, alkenyl or alkynyl radical, having a number of carbon atoms selected from within a stated range, where the alkyl, alkenyl or alkynyl radical is bonded successively through an oxygen atom (-O-) and a carbonyl group (-C(=O)-) to the residue of a compound of Formula (I). Exemplary alkoxycarbonyl radicals include -C(=O)-OCH3, -C(=O)-OCH(CH3)2, -C(=O)-OCH2CH ₂ CH ₂ CH=CH2 and the like.

The term "alkylamido"refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, where the alkyl, alkenyl or alkynyl radical is bonded to the nitrogen tom of an amide group (-C(=O)NH-) and the carbon atom of the amide group is bonded to the residue of a compound of Formula (I). Exemplary alkylamido radicals include methylamido (-(C=O)NH(CH ₃ )), isopropylamino (-(C=O)NH(CH(CH3)2)), 3-butenylamino (-(C=O)NH(CH ₂ CH ₂ CH=CH ₂ )) and the like.

The term "alkylamino" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, where the alkyl, alkenyl or alkynyl radical is bonded through an amine diradical (-NH-) to the residue of a compound of Formula (I). Exemplary alkylamino radicals include methylamino (-NH(CH3)), isopropylamino (-NH(CH(CH3)2)), 3-butenylamino (-NH(CH ₂ CH ₂ CH=CH2)) and the like.

The term "alkylaminosulfonyl" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, where the alkyl, alkenyl or alkynyl radical is bonded to the nitrogen atom of an aminosulfonyl group (-S(=O)2NH-), and the sulfur atom of the aminosulfonyl group is bonded to the residue of a compound of Formula (I). Exemplary alkylaminosulfonyl radicals include methylaminosulfonyl (-S(=O)2NHCH3), ethylaminosulfonyl (-S(=O)2NHCH ₂ CH ₃ ), isopropylaminosulfonyl (-S(=O) NHCH(CH ₃ )2), 2- methyl-3-butenylaminosulfonyl (-S(=O)2NHCH ₂ CH(CH3)CHCH ₂ ) and the like. The term "alkylene" refers to a divalent hydrocarbon radical having a branched or straight chain of carbon atoms, where the number of carbon atoms is selected from within a stated range. Exemplary alkylene radicals include methylene (-CH2-), 1,2-ethanediyl (-CH2CH2-), 3-methyl-l,5-pentanediyl (-CH2CH2CH(CH3)CH ₂ CH ₂ -) 1,6-hexanediyl (-CH2CH2CH2CH2CH2CH2-) and the like.

The term "alkylsulfinyl" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, and bonded to a sulfur atom of a sulfinyl group (-S(O)-), where the sulfur atom of the sulfmyl group is also bonded to the residue of a compound of Formula (I). Exemplary alkylsulfinyl radicals include -S(O)CH ₃ , -S(O)CH ₂ CH ₃ , -S(O)CH(CH ₃ ) ₂ and the like.

The term "alkylsulfonyl" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range, and bonded through a sulfonyl group (-S(=O)2-) to the residue of a compound of Formula (I). Exemplary alkylsulfonyl groups include methylsulfonyl (-S(=O)2CH3), ethylsulfonyl (-S(=O)2CH CH ₃ ), 3-butenylsulfonyl (-S(=O)2CH2CH CH=CH2), isopropylsulfonyl (-S(=O)2CH(CH3)2) and the like.

The term "alkylsulfonylamino" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from with a stated range, and bonded to the sulfur atom of a sulfonylamino (-NH-S(=O)2-) group, where the nitrogen atom of the sulfonylamino group is bonded to the residue of a compound of Formula (I). Exemplary alkylsulfonylamino radicals include methylsulfonylamino (-NHS(=O) ₂ CH ₃ ), ethylsulfonylamino (-NHS(=O) ₂ CH ₂ CH3), isopropylsulfonyla ino (-NHS(=O)2CH(CH3)2), 2-butenylsulfonylamio (-NHS(=O)2CH CH=CHCH ₃ ) and the like.

The term "alkylthio" refers to an alkyl, alkenyl or alkynyl radical having a number of carbon atoms selected from within a stated range,where the alkyl, alkenyl or alkynyl radical is bonded through a sulfur (S) atom to the residue of a compound of Formula (I). Exemplary alkylthio radicals include methylthio (-SCH3), ethylthio (-SCH2CH3), /.-propylthio (-SCH2CH2CH3), isopropylthio (-SCH(CH ₃ )2) and the like.

The term "alkynylene"refers to a divalent hydrocarbon radical having a branched or straight chain of carbon atoms, and a single carbon-carbon triple bond (C≡C), where the number of carbon atoms is selected from within a stated range. Exemplary alkynylene radicals include 1,2-acetylenediyl (-C≡C-), l-propyne-1,3- diyl (-C≡C-CH2-), 2-butyne-l,4-diyl (-CH2-C≡C-CH ₂ -) and the like. The term "amido" refers to the -C(=O)NH2 radical. The term "amino" refers to the -NH2 radical. The term "aminosulfonyl" refers to the -S(=O)2NH2 radical. The term "carboxy" refers to the -C(=O)OH radical, as well as the salt, hydrate and solvate forms thereof.

The term "cyano", also known as "nitrile", refers to the -C≡N radical. The terms "co-administering" or "co-administered" or "co-administer" etc. refer to either the simultaneous administration, or any manner of separate sequential administration, of a 5-α-reductase inhibiting compound, as described herein, and a further active ingredient or ingredients. Such further active ingredient or ingredients include compounds known to treat the disease states of acne vulgaris, seborrhea, female hirsutism, male pattern baldness, benign prostate hypertrophy, prostatic adenocarcinoma, or any other compound known to have utility when used in combination with 5-α-reductase inhibitors. Preferably, if the administration is not simultaneous, the compounds being co-administered 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 admimstered topically and another compound may be administered orally.

The term "dialkylamido" refers to a monovalent radical having two groups bonded to the nitrogen atom of an amide group (-C(=O)N), where the carbon atom of the amide group is bonded to the residue of a compound of Formula (I), and where the two group bonded to the nitrogen atom are selected from alkyl, alkenyl or alkynyl radicals, where each alkyl, alkenyl or alkynyl radical has a number of carbon atoms selected from within a stated range, and selected independently from where the other alkyl, alkenyl or alkynyl radical. Exemplary dialkylamido radicals include dimethylamido (-C(=O)N(CH ₃ ) ₂ ), methylethylamido (-C(=O)N(CH3)(CH ₂ CH ₃ )), isobutenylisopropylamido (-C(=O)N(CH2C(CH3)=CH2)(CH(CH3)2)) and the like.

The term "dialkylamino" refers to a radical having two groups bonded to a single nitrogen atom (N), where the nitrogen atom is also bonded to the residue of a compound of Formula (I), where the two groups bonded to the nitrogen atom are selected from alkyl, alkenyl or alkynyl radicals, wherein each alkyl, alkenyl or alkynyl radical has a number of carbon atoms selected from within a stated range and selected independently from the other alkyl, alkenyl or alkynyl radical. Exemplary dialkylamio radicals include dimethylamino (-N(CH3)2), ethylmethylamino (-N(CH3)(CH2CH3)), isobutenylisopropylamino (-N(CH2C(CH ₃ )CH2)(CH(CH ₃ )2)) and the like.

The term "diazo-phenanthrenone" refers to compound having the basic structure

where the carbons of the diazo-phenanthrenone nucleus of the invention are numbered as shown above. -

The term "halogen" or "halo" refers to any of fluoro, chloro, bromo or iodo.

The term "hydroxy" or "hydroxyl" refers to the -OH radical.

The term "mercapto", also known as "thio", refers to the -SH radical.

The term "alpha-receptor antagonist" or "α-receptor antagonist" refers to a known class of alpha-andrenergic receptor antagonist compounds, such as described in Lafferty, et al. U.S. Patent No. 4,963,547, which are utilized in treating vascular disorders such as 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, 7-chloro-2-ethyl-3,4,5,6-tetrahydro-4- methylthieno[4,3,2-ef][3]-benzazepine and 8-{3-[4-(2-methoxyphenyl)-l- piperazinyl]-propylcarbamoyl}-3-methyl-4-oxo-2-phenyl-4H-l-b enzopyran.

The term "amsulosin" refers to a compound of the structure

and salts, hydrates and solvates thereof. Chemically, amsulosin is designated as (-)- (i?)-5-(2-((2-(O-ethoxyphenoxy)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.

The term "terazosin" refers to a compound of the structure

cH ₃ o .N

NH ₂ and salts, hydrates and solvates thereof. Chemically, terazosin is designated as 1- (4-amino-6,7-dimethoxy-2 quinazolinyl)-4-((tetrahydro-2- furoyl)carbonyl)piperazine. Terazosin is disclosed in U.S. Patent Number 4,251,532.

The term "doxazosin" refers to a compound of the structure

and salts, hydrates and solvates thereof. Chemically doxazosin is designated as 1- (4-aπ-ino-6,7-dimethoxy-2-quinazolinyl)-4-((2,3-dihydro-l,4 -benzodioxin-2- yl)carbonyl)piperazine. Doxazosin is disclosed in U.S. Patent Number 4,188,390. The term "alfuzosin" refers to a compound of the structure

and salts, hydrates and solvates thereof. Chemically, alfuzosin is designated as N- (3-((4-ammo-6,7-dime oxy-2-quinazolinyl)memylamino)propyl)tetrahydro-2- furancarboxamide. Alfuzosin is disclosed in U.S. Patent Number 4,315,007. The term "indoramin" refers to a compound of the structure

H

and salts, hydrates and solvates thereof. Chemically indoramin as designated N-((l- (2-(lH-indol-3-yl)ethyl)-4-piperidinyl)benzamine. Indoramin is disclosed in U.S. Patent Number 3,527,761.

The term "prazosin" refers to a compound of the structure

and salts, hydrates and solvates thereof. Chemically, prazosin is designated as l-(4- - nino-6,7-dimethoxy-2-quinazolinyl)-4-(2-furanylcarbonyl)pipe razine. Prazosin is disclosed in U.S. Patent Number 3,511,836.

The term "7-chloro-2-ethyl-3,4,5,6-tetrahydro-4-methylthieno[4,3,2-ef ]- [3]benzazepine" refers to a compound of the structure

and salts, hydrates and solvates thereof. 7-chloro-2-ethyl-3,4,5,6-tetrahydro-4- methylthieno[4,3,2-ef]-[3]benzazepine is disclosed in U.S. Patent No. 5,006,521. All compounds disclosed in U.S. Patent No. 5,006,521 as alpha-andrenergic receptor antagonist are preferred alpha-andrenergic receptor antagonists as used herein.

The term "8-{3-[4-(2-methoxyphenyl)-l-piperazinyl]-propylcarbamoyl}-3 - methyl-4-oxo-2-phenyl-4H-l-benzopyran refers to a compound of the structure

and salts, hydrates and solvates thereof. 8-{3-[4-(2-methoxyphenyl)-l-piperazinyl]- propylcarbamoyl}-3-methyl-4-oxo-2-phenyl-4H-l-benzopyran is disclosed in EPO Publn. No. 0558245 Al, to Leonardi, et al., (Recordata S.A.). Additionally, all compounds disclosed in EPO Publn. No. 0558245 Al, as alpha-adrenergic receptor antagonists are preferred alpha-adrenergic receptor antagonists as used herein.

Persons skilled in the art can readily determine if a compound other than one specifically referred to herein is an alpha-andrenergic receptor antagonist by utilizing the assay described in U.S. Patent No. 4,963,547: all such compounds are

included within the scope of the term "alpha-andrenergic receptor antagonist" as used herein.

The term "aromatase inhibitor" refers to a known class of compounds, steroidal and non-steroidal, which prevent the 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 utility in treating benign prostatic hyperplasia when used in combination with a 5-α- reductase inhibitor. All compounds disclosed in Gormley et al. as having aromatase inhibiting activity are preferred aromatase inhibitors as used herein. Another preferred aromatase inhibitor for use in the compositions and methods of the present invention is 4-(5,6,7,8-tetrahydroimidazo-[l,5-α]pyridin-5-yl)benzonitri le, which is also known as fadrazole. Fadrazole is disclosed in U.S. Patent Number 4,728,645.

The term "minoxidil" refers to a compound of the structure

O

Chemically, minoxidil is designated as 2,4-pyrimidineadiamine-6-(l-piperidinyl)-3- oxide. Minoxidil is the active ingredient in Rogaine® which is sold as a topical solution for stimulating hair growth by the Upjohn Company, Kalamazoo, Michigan.

It has been discovered that the novel diazo-phenanthrenone compounds of Formula (I) can inhibit 5-α-reductase, and thus provide an attractive means for regulating DHT levels in mammals. Compounds of Formula (I) have the structure:

or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R ¹ is hydrogen or Ci -Cg alkyl; R is independently hydrogen; halogen; NO2; cyano; trifluoromethyl; C\-

C alkyl; Ci-Cg alkoxy; carboxy; Ci -Cg alkoxycarbonyl; amino; C1-C4

alkylamino; C1-C4 dialkylamino; amido; C1 -C4 alkylamido; C1 -C4 dialkylamido; mercapto; Ci-Cg alkylthio; Ci-Cg alkylsulfinyl; C\-C alkylsulfonyl; -C(=O)R ³ where R ³ is Cj-Cό linear or branched alkyl which can optionally be substituted with one or more of trifluoromethyl, halogen, -OH, -C(=O)OH, -O(Cι-C alkyl), -(CH2) _p OH where p is 1 -4, -(CH2) _m COOH where m is 1 -3 , or a protected -OH group; aryl optionally substituted with one or more of trifluoromethyl, halogen, -OH, -C(=O)OH, -O(Cι-C alkyl), -(CH ₂ ) _p OH where p is 1-4, -(CH ₂ ) _m C(=O)OH where m is 1-3, and a protected -OH group; or -R ³ -R4 where R ³ is Cj-Cg alkylene, C2-C6 alkenylene or C2-C6 alkynylene, and R^ is halogen, hydroxy, trifluoromethyl, Ci-Cβ alkoxy, carboxy, C\-C alkoxycarbonyl, amino, C1 -C4 alkylamino, C1-C4 dialkylamino, amido, C1-C4 alkylamido, C1 -C4 dialkylamido, C1-C4 alkylsulfonylamino, aminosulfonyl or C1-C4 alkylaminosulfonyl; and n is 1 or 2.

Preferred among the presently invented Formula (I) compounds are compounds wherein

R! is hydrogen or methyl;

R2 is independently hydrogen, halogen, cyano, Ci-C _β alkyl, \-C alkoxy, -C(=O)R ³ where R ³ is Cj-Cg linear or branched alkyl which can optionally be substituted with one or more of trifluoromethyl, halogen, -OH, -C(=O)OH, -O(C 1 -C4 alkyl), -(CH2) _p OH where p is 1 -4, -(CH2) _m C(=O)OH where m is 1 -3, or a protected -OH group; aryl optionally substituted with one or more of trifluoromethyl, halogen, -OH, -C(=O)OH, -O(Cι-C4 alkyl), -(CH2) _p OH where p is 1-4, -(CH2) _m C(=O)OH where m is 1-3, or a protected -OH group; and n is 1 or 2. More preferred among the presently invented Formula (I) compounds are compounds wherein R^ is methyl; R^ is methoxy; and n is 1.

SYNTHETIC METHODS The novel compounds of Formula (I) of the present invention can be prepared by the method outlined in SCHEME I below, and in the Example, from the starting material described in SCHEME I. Analogous compounds of the invention can be prepared according to similar methods from the same or alternative starting materials.

SCHEME I

SCHEME I shows a synthesis of a diazo-phenanthrenone compound of

Formula (I) in which R! is methyl, R^ is methoxy and n is 1. A compound comprising an α,β-unsaturated ketone is a convenient starting material for the preparation of the dizao-phenanthrenone compounds of the invention, and a representative α,β-unsaturated ketone compound can be prepared according to the method of Comforth, J.W.; Robinson, R. J. Chem. Soc. 1949, 1855. An α,β- unsaturated ketone compound can be converted to a compound readily susceptible to functionalization at the α-carbon, as shown by reaction 'b' in SCHEME I, which shows the conversion of an α,β-unsaturated ketone to a silyl enol ether

A silyl enol ether containing compound, including the trimethylsilyl enol ether compound shown in SCHEME I, can be reacted with an acylating agent to provide a 1,3-diketone which serves as a convenient precursor to the diazo- phenanthrenone compounds of Formula (I). As shown as reaction 'c' in SCHEME I, benzoyl chloride is a suitable acylating agent, which will react with a trimethylsilyl enol ether to provide a 1,3-diketone compound.

The 1,3-diketone compound prepared according to reaction 'c' can be converted to a diazoketone compound of Formula (I) by reaction with an azide compound such as tosylazide, as shown as reaction 'd' in SCHEME I.

The synthetic sequence shown in SCHEME I is amenable to the preparation of all of the compounds of Formula (I), if suitable starting materials are chosen.

UTILITY Because the pharmaceutically active compounds of the invention inhibit steroid 5-α-reductase activity, they have therapeutic utility in treating diseases and conditions wherein decreases in DHT activity produce the desired therapeutic effect. Such diseases and conditions include acne vulgaris, seborrhea, female hirsutism, male pattern baldness, prostate diseases such as benign prostatic hypertrophy, and prostatic adenocarcinoma.

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

Preparation of membrane particulates used as source for recombinant steroid 5- ft-redugta§e jsQzyme 1,

Chinese hamster ovary (CHO) cells containing expressed, recombinant human steroid 5-α-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 μM 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 μM NADPH (buffer B). The suspended particulate solution was stored at -80°C.

Preparation of membrane particulates used as source for recombinant steroid 5-α- reductase isozvme 2.

CHO cells containing expressed, recombinant human steroid 5-α-reductase isozyme 2 were homogenized in buffer A using a Dounce hand homogenizer. Membrane particulates containing the recombinant human enzyme were isolated by centrifugation (100,000 x g at 4°C for 60 minutes) and resuspended in buffer B. The suspended particulate solution was stored at -80°C until used.

Assay for enzvmes activities and inhibitors potency. A constant amount of [ ¹⁴ 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 μL (recombinant isoenzyme 1 or isoenzyme 2) or 20 μL (isoenzyme 2 from human prostate tissue) of 10 mM NADPH and an aliquot of a steroid 5-α-reductase preparation to a final volume of 0.5 mL. Assays for human steroid 5-α-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 0.25 μmol each of testosterone, 5α-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 μL 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 and used to determine enzyme activity. 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. The value for the inhibition constant (Ki) was calculated according to Levy, M. (1989) Biochemistry 29:2815-2824.

Compounds within the scope of this invention have been tested and have shown an activity of from 0.5 Ki(nM) to 120 Ki(nM) against isozyme 1 and an activity of >2000 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 is (+/-)-trα«5-l-diazo-8- methoxy-4a-methyl- 1 ,2,3 ,4,4a,9, 10,1 Oa-octahydrophenanthren-2-one.

All of the compounds within the scope of this invention are useful in inhibiting steroid 5-α-reductase in a mammal, including humans, in need thereof. When a 5-α-reductase inhibitor, as described herein, and a further active ingredient or ingredients are utilized together, said 5-α-reductase inhibitor can be co- administered with said further active ingredient or ingredients.

The pharmaceutically active compounds of the present invention are preferably incorporated into convenient dosage forms such as capsules, tablets, or injectable preparations. Solid or liquid pharmaceutical carriers may be 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 preferably be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid such as maintained in 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, for tablet forms, or mixing, filling and dissolving the ingredients, 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 quantity preferably selected from the range of 0.01 - 1000 mg/kg of active compound, preferably 0.1 - 100 mg/kg. When treating a human patient in need of steroid 5-α-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-α-reductase activity in mammals, including humans, comprises administering to a subject in need of such inhibition an effective steroid 5-α-reductase inhibiting amount of a pharmaceutically active compound of the present invention.

The invention also provides for the use of a compound of Formula (I) in the manufacture of a medicament for use in the inhibition of steroid 5-α-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) 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) 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) which comprises bringing the compound of Formula (I) 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-α-reductase inhibitors. Particularly preferred is the co-administration of a 5-α-reductase inhibitor, as disclosed herein, and minoxidil for use in the treatment of male pattern baldness. Particularly preferred is the co-aclministration of a 5-α-reductase inhibitor, as disclosed herein, and a α-receptor antagonist for use in the treatment of benign prostatic hypertrophy. Preferred is the co-administration of a 5-α-reductase inhibitor, as disclosed herein, and an aromatase inhibitor for use in the treatment of benign prostatic hypertrophy. Preferred is the co-administration of a 5-α-reductase inhibitor, as disclosed herein, an alpha-receptor antagonist and an aromatase inhibitor for use in the treatment of benign prostatic hypertrophy. The invention also provides a compound of Formula (I) for use in therapy and for use in the manufacture of a medicament for use as a steroid 5-α-reductase inhibitor.

The invention further provides a compound of Formula (I) for use in the manufacture of a medicament for use in treatment to reduce prostate size or the treatment of prostatic adenocarcinoma.

The invention also provides for a composition comprising a compound represented by Formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, for use in therapy or for use in the manufacture of a medicament for use as a steroid 5-α-reductase inhibitor. The invention further provides for a composition comprising a compound represented by Formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, for use in treatment to reduce prostatic size, or for use in treatment of prostatic adenocarcinoma.

The invention also provides a process for the preparation of

(a) a pharmaceutically acceptable carrier, diluent or excipient and

(b) an effective amount of a compound of the Formula (I) or a pharmaceutically acceptable salt, hydrate, solvate or ester thereof, which process comprises bringing component (a) into association with component (b).

The invention further provides for the use of a compound of Formula (I) in the manufacture of a medicament for use in inhibiting steroid 5-α-reductase. The invention also provides for a method of inhibiting steroid 5-α-reductase in mammals which comprises the administration to a mammal in need of such inhibition, an effective amount of a compound of Formula (I).

The invention further provides for the use of a compound of Formula (I) and an alpha-receptor antagonist compound as an active therapeutic substance, which use comprises co-administration of a compound of Formula (I) and an alpha-receptor antagonist compound. The invention also provides for the use of a compound of Formula (I) and an alpha-receptor antagonist compound, in the manufacture of medicament for use in the treatment of benign prostatic hypertrophy which use comprises co-administration of a compound of Formula (I) and an alpha-receptor antagonist compound. The invention also provides for the use of a compound of Formula (I) and minoxidil as an active therapeutic substance which use comprises co-administration of a compound of Formula (I) and minoxidil. The invention further provides for the use of a compound of Formula (I) and minoxidil, in the manufacture of a medicament for use in the treatment of male pattern baldness which use comprises co-administration of a compound of Formula (I) and minoxidil.

The invention also provides for the use of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, in the manufacture of a medicament for use in inhibiting steroid 5-α-reductase. The invention further provides for a method of inhibiting steroid 5-α-reductase in mammals which comprises the administration to a mammal in need of such inhibition, an effective amount of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient. The invention also provides for the use of composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and an alpha-receptor antagonist compound as an active therapeutic substance which use consist of co- administration of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and an alpha-receptor antagonist compound.

The invention also provides for the use of composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and an alpha- receptor antagonist compound in the manufacture of medicament for use in the treatment of benign prostatic hypertrophy which use comprises co-administration of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and an alpha-receptor antagonist compound. The invention also provides for the use of composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and minoxidil as an active therapeutic substance which use comprises co-administration of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient and minoxidil.

The invention also provides for the use of a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient and minoxidil in the manufacture of a medicament for use in the treatment of male pattern baldness which use comprises co-administration of a composition comprising a compound of

Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, diluent or excipient, and minoxidil.

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following Examples are, therefore, to be construed as merely illustrative and not a limitation of the scope of the present invention in any way.

EXAMPLE 1 Corresponding to SCHEME T: Preparation of (+IA-trans-\ -riiazo-8-methoxv-4a- methvl- 1.2. .4.4a.9.1 .1 Oa-octahvdro-phenanthren-2-one

A. (+/-)-trαr75-8-Methoxy-4a-methyl- 1 -trimethylsilyl- 1 ,2,3 ,4,4a,9, 10, 1 Oa-octahydro- phenanthren-2-one

A solution (+/-)-8-methoxy-4a-methyl-2,3,4,4a-9,10-hexahydrophenanthren - 2-one (1.0 g, 4.1 mmol, prepared by the method of Comforth, J. W.; Robinson, R. J. Chem. Soc. 1949, 1855) and aniline (27 μL, 2.9 mmol) in tetrahydrofuran (25 mL) was added dropwise to a solution of lithium (65 mg, 9.4 mmol) in doubly distilled ammonia (100 mL) at -78 °C. The resulting mixture was stirred at -78 °C for 2 h and then treated with dry isoprene until the blue color was discharged. The volatiles were removed by slowly warming the mixture to room temperature under an argon flow, followed by evaporation in vacuo. The residue was dissolved in tetrahydrofuran (25 mL) and cooled in an ice bath. A filtered solution of trimethylsilylchloride (0.785 mL, 1.5 equiv) and triethylamine (0.862 mL, 1.5 equiv) was added slowly with stirring. After stirring at ice temperature for a further 18 h. the solvent was evaporated in vacuo and the residue partitioned between diethyl ether and cold saturated aqueous sodium bicarbonate solution. The separated ether phase was washed with cold saturated sodium chloride solution, dried over MgSO ₄ , filtered, and concentrated in vacuo to afford a yellow oil which crystallized on standing. The oil was purified by flash chromatography (silica, 33% dichloromethane/hexane) to yield the titled compound (815 mg, 63 %). mp 95- 96 °C. Anal. (Cι ₉ H ₂ 8θ ₂ Si) calcd: C, 72.10; H, 8.92. found: C, 71.87; H, 8.65.

B. (+l-)-trans- 1 -Benzoyl-8-methoxy-4a-methyl- 1 ,2,3,4,4a,9, 10, 1 Oa-octahydro- phenanthren-2-one (+/-)-tr «5-8-Methoxy-4a-methyl- 1 -trimethylsilyl- 1 ,2,3,4,4a,9, 10, 10a- octahydro-phenanthren-2-one (500 mg, 1.58 mmol) in diethyl ether (10 mL) was

added dropwise with stirring to a solution of methyl lithium (1.2 mL of a 1.4 M solution in diethyl ether, 1.69 mmol) in diethyl ether (15 mL) at room temperature. The solution was stirred for 45 min, cooled to -78 °C under argon and added to a solution of benzoyl chloride (238 mg, 1.69 mmol) in diethyl ether (10 mL) at -78 °C. After 30 min at -78 °C, and a further 30 min at room temperature, the solution was diluted with diethyl ether (25 mL) and water (30 mL). The ether phase was separated, dried over MgSO ₄ , filtered, and concentrated in vacuo to afford an oil. Purification by preparative chromatography (silica, 15% ethyl acetate/hexane) and recrystallization (chloroform hexane) gave the titled compound (305 mg, 56 %). mp 143-144 ^O C. Anal. (C23H24O3) calcd: C, 79.28; H, 6.94. found: C, 79.17; H, 6.90.

C. (+/-)-trans- 1 -Diazo-8-methoxy-4a-methyl- 1 ,2,3 ,4,4a,9, 10, 10a- octahydrophenanthren-2-one

(+l-)-trans- 1 -Benzoyl-8-methoxy-4a-methyl- 1 ,2,3 ,4,4a-9, 10,1 Oa-octahydro- phenanthren-2-one (270 mg, 0.76 mmol) in tetrahydrofuran (2 mL) was added dropwise to a stirred suspension of sodium hydride (60 % dispersion in oil, 65 mg, 1.63 mmol) in tetrahydrofuran (3 mL). The mixture was cooled in a water bath and tosylazide (250 mg, 1.27 mmol) was added dropwise under argon. After 45 min stirring at room temperature, the mixture was filtered and concentrated in vacuo. Dichloromethane (30 mL) was added and the mixture was washed with water (20 mL), dried over MgSO ₄ , filtered, and concentrated in vacuo to afford a brown oil which crystallized on standing. Purification by preparative chromatography (silica, 30% ethyl acetate/hexane) gave the titled compound (95 mg, 46 %). mp 131- 133 °C. MS (DEI). (Ci6Hi8N ₂ 0 ₂ ) calcd: 270.1378. found: 270.1368.

EXAMPLE 2 An oral dosage form for administering Formula (I) compounds is produced by screening, mixing, and filling into hard gelatin capsules the ingredients in the proportions shown in Table 1 , below.

able I Ingredients. Amounts

(+/-)-trørts-8-methoxy-4a-methyl-l- 50 mg trimethylsilyl- 1 ,2,3,4,4a,9, 10, 10a- octahydro-phenanthren-2-one magnesium stearate 5 mg lactose 75 mg

EXAMPLE 3 The sucrose, calcium sulfate dihydrate and Formula (I) compound shown in Table II 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.

Table II

Ingredients Amounts

(+/-)-tranj-8-methoxy-4a-methyl- 1 - 100 mg trimethylsilyl- 1 ,2,3 ,4,4a,9, 10, 10a- octahydro-phenanthren-2-one calcium sulfate dihydrate 150 mg sucrose 20 mg starch 10 mg talc 5 mg stearic acid 3 mg

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 instrumentalities herein disclosed and that the right to all modifications coming within the scope of the following claims is reserved.