AU718358B2 - Inhibitors of testosterone 5-alpha reductase activity - Google Patents

Description

P/00/01 1 Regulation 3.2

AUSTRALIA

Patents Act 1990

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT

I

.4 0 4 4.

44 4 .4 9* I I 4.4.

44I4 .44 I I 44 6 44 4 4.

4 94 44II** Invention Title: INHIBITORS OF TESTOSTERONE 5-ALPHA REDUCTASE

ACTIVITY

The following statement is a full description of this invention, including the best method of performing it known to me: GH REF: P21053-AU VNV:SY 1A INHIBITORS OF TESTOSTERONE 5a-REDUCTASE ACTIVI-TY Field of the Invention This invention relates to pharmaceutical compositions and methods for the treatment of androgen-related diseases, said compositions having novel inhibitors of testosterone activity. The inhibitors have a good combination of inhibitory effect on 5a-reductase activity, low or no androgenic activity and, in some embodiments, antiandrogenic activity. More particularly, certain embodiments of the invention relate to derivatives of 4-azaandrostanone or 4-aza-androstenone.

Background of the Invention Prior art inhibitors of 5a-reductase fail to provide an optimal combination of lack of inherent androgenic activity and ability to inhibit both of two different forms of testosterone 5a-reductase is an enzyme which catalyzes the conversion of the androgen, testosterone, to the much more potent androgen dihydrotestosterone DHT is the 20 more active androgen in many target organs (Anderson and Liao. Nature 219:277-279, 1968). The same enzyme catalyzes the conversion of androstenedione into androstanedione.

Inhibitors of 5a-reductase inhibit biosynthesis of the products whose formation is catalyzed by e 25 5a-reductase has been studied in different species (Liang et al. Endocrinology 117:571-579, 1985). Its isolation and structure, and the expression of cDNA encoding it have been described (Anderson and Russell. Proc. Natl.

Acad. Sci. 87:3640-3644, 1990).

Recent data have demonstrated the presence of at least two different genes expressing 5a-reductase in humans. Type I 5a-reductase (Andersson and Russell, Proc. Natl. Acad.

Sci. 87, 3640-3644, 1990) is expressed at a low level in the human prostate while type II 5a-reductase is the predominant -2 enzyme isoform expressed in this tissue (Andersson et la., Nature 354, 159-161, 1991).

The blockade of 5a-reductase has been intensively studied in view of developing pharmaceutical drugs for the therapy of diseases, such as prostate cancer. In European Patent Appln. No. EP 285 383 Ramusson et al. disclose the treatment of prostatic carcinoma with 1 7 p-N-monosubstitutedcarbamoyl-4-aza-5a-androst-l-en-3-ones). Diseases for which inhibitors are also being studied include acne, baldness (Rittmaster et al., J. Clin. Endocrinol. Metab.

65:188-193,1987) and benign prostatic hyperplasia (Metcalf et al., Tips 10:491-495, 1989).

The 4-aza-steroid N,N-diethyl-4-methyl-3-oxo-4-aza-5aandrostane-17p-carboxamide, 4-MA has proven useful in inhibiting the formation of DHT from testosterone in rat prostate in vitro and in vivo (Brooks et al., Endocrinology 109: 830-836, 1981), thus reducing the testosterone-induced increase in ventral prostate weight in these animals.

Another 4-aza-steroid, MK-906 (PROSCAR), has been found to S 20 cause a reduction in the intraprostatic concentration of DHT and a 25-30% reduction in prostatic size in men (Imperato- McGinley et al., Proc. 71st Ann. meet Endocr. Soc., p. 332, abst 1639, 1989). However, Proscar is reported to be a potent inhibitor of the type II enzyme but a weak inhibitor of the type I enzyme (Andersson et al., Nature 354, 159-161, 1991). Such a low inhibitory potency on type I probably explains why the highest doses of Proscar used in men generally fails to reduce serum dihydrotestosterone levels below 25 to 35% of control, thus leaving a highly 30 significant concentration of circulating androgens (Vermeulen et al., The Prostate 14, 45-53, 1989). The inhibitory effect of the drug on prostatic volume in men remains limited to 25 to 35% over a period of 6 months (Stoner, J. Steroid Biochem. Mol.Biol. 37, 375-378, 1990).

There is thus a need to develop compounds which can 3 efficiently inhibit both type I and type II 5a-reductase and thus cause a more complete inhibition of circulating dihydrotestosterone levels.

In U.S. Patent Appln. No. 4,317,817, Belgian Patent Appln. No. 883091 and British Patent Appln. No. 204 8888, Blohm and Metcalf discuss the use of certain diazo-steroids as steroid 5a-reductase inhibitors. Metcalf et al. describe the synthesis of related compounds in Tetrahedron Lett. 21, 15-18, 1980.

In EP Publication No. 343 954, EP Publication No. 375 347, U.S. Patent Appln. No. 4,882,319, U.S. Patent Appln.

No. 4,937,237 and J. Med. Chem. 33: 937-942, 1990, Holt et al. discuss the use of certain A-ring aryl steroid derivatives as steroid 5a-reductase inhibitors.

In EP Publication No. 289 327 and Publication No. 427 434, on one hand, and in J. Steroid Biochem. 34: 571-575, 1989 and Biochemistry 29: 2815-2824, 1990, on the other hand, Holt and Levy discuss, respectively, the use of androstene- and pregnene-3-carboxylate derivatives as steroid-5a-reductase inhibitors.

In EP Publication No. 375 351, Holt et al. discuss the preparation of phosphoric acid substituted steroids as testosterone 5a-reductase inhibitors.

In EP Publication No. 271 219, EP Publication No. 314 25 199, and EP Publication No. 155 096, Rasmusson and Reynolds discuss the preparation of 17p-substituted-4-aza-5aandrostenones as steroid-5a-reductase inhibitors.

Brooks, et al. (Steroid 47: 1-19, 1986; Prostate 9: 76, 1986) have reported 5a-reductase inhibiting and S* 30 androgen-blocking activities for some 4-aza-steroids.

Rasmusson et al. discuss certain aza-steroids as inhibitors of rat prostatic 5a-reductase (in J. Med. Chem.

27: 1690-1701, 1984; idem 29: 2298-2315, 1986, and J. Biol.

Chem. 259: 734-739, 1984).

4 In EP Publication No. 277 002, Holt et al. discuss 170substituted-4-aza-5a-androstane-3-ones.

In EP Publication No. 271 220, Carlin et al. discuss the preparation of 17p-(N-monosubstituted carbamoyl)-4-aza- 5a-androstane-3-ones.

In EP Publication No. 200 859, Cainelli et al. discuss the preparation of certain 4-aza-steroid derivatives which are stated to be steroidal 5a-reductase inhibitors.

In International Publication No. WO 91/12261, Panzeri et al. discuss the preparation of 17p-substituted-4-aza-5androstan-3-one derivatives.

In US 4,396,615, Steroids 38: 121-140, 1981 and J.

Steroid Biochem 19: 1491-1502, 1983, Petrow et al discuss certain 6-methylene progesterone derivatives stated to be inhibitors of steroid In U.S. 4,377,584 (see column 13), U.S. 4,220,775 and in EP Publication No. 414 490 and 414 491, Rasmusson et al. discuss certain 17p-substituted-4-aza-5a-androstanones (including acyl amino substitutions) as inhibitors.

In EP Publication No. 052799, Alig et al. discuss the use of certain D-homosteroids as steroid inhibitors.

In U.S. 4,191,759, Johnston and Arth discuss N- S 25 substituted-170-carbamoyl-androst-4-en-3-ones as steroid reductase inhibitors.

In Be 855 992, Benson and Blohm discuss steroidal inhibitors of testosterone 5a-reductase, for treating skin disorders.

In CA 970 692, Voight and Hsia discuss compounds inhibiting 5a-reductase activity.

In FR 1 465 544, Jolly and Warnant discuss 4-azaaromatic steroid derivatives as steroid 5 inhibitors.

In US 4,087,461, Robinson discuss certain allenic steroids as testosterone 5a-reductase inhibitors.

In EP Publication No. 414 529, Metcalf discuss certain 17-substituted steroidal acids as testosterone inhibitors (see e.g. the Abstract). See also, Holt, et al., EP Publication No. 427,434.

In EP Publication No. 298 652, Bhattacharya disclose the synthesis of 4-aza-Al-steroids.

In U.S. Patent Appln. No. 5,061,803 and 5,061,801, Williams discusses a method for the synthesis of 17palkanoyl-3-oxo-4-aza-5a-androst-l-enes and 3-oxo-4-azaandrost-1-ene 17p-ketones.

In U.S. Patent Appln. No. 5,061,802, Steinberg and Rasmusson discuss the preparation of 17P-aminobenzoyl-4-aza- 5a-androst-l-en-3-ones as benign prostatic hypertrophy agents.

Lan-Hargest et al. discuss the synthesis of bridged A ring steroids as 5a-reductase inhibitors (Tetrahedron Lett.

28:6117-6120, 1987).

Weintraub et al. (in J. Med. Chem. 28: 831-833, 1985) :discuss the preparation of 20-hydroxymethyl-4-methyl-4-aza- 2-oxa-5a-pregnan-3-one as inhibitors of testosterone reductase.

S 25 Kadohama et al. (Cancer Res. 44; 4947-4954, 1984) discuss sodium 4-methyl-3-oxo-4-aza-5a-pregnane-20 (S) a. carboxylate inhibition of prostatic tumor MacIndoe et al. in J. Steroid Biochem. 20, 1095-1100, 1984, discuss the 5a-reductase inhibiting effect, in MCF-7 human breast cancer cells and rat prostate, of certain 6methylene steroids.

Liang et al. Biol. Chem. 256:7998-8005, 19981) discuss 17B-N,N-diethylcarboxyamoyl-4-methyl-4-aza-5a- 6 androstan-3-one as a reversible inhibitor of Salomons and Doorenbos Pharm. Sci. 63:19-23, 1974) and Doorenbos et al. Pharm. Sci 60: 1234-1235, 1971; idem 62: 638-640, 1973; Chem. and Ind; 1322, 1970) discuss synthesis of 170 amino 4-aza-steroids.

Nakayama et al. Antibiotics XLII: 1221-1229, 1989; idem, 1230-1234, 1989, idem, 1235-1240, 1989) discuss the isolation of WS-9659 from Streptomyces and its inhibitory activity on testosterone In EP Publication No. 294 937 and EP Publication No.

294 035, Nakai et al. discuss the preparation, respectively, of cinnamoyl amide derivatives and (benzoylamino)phenoxy) butanoic acid derivatives, as inhibitors of U.S. Patent No. 5,026,882 and EP Publication No. 375 349 relates to certain steroid-3-phosphinic acid compounds for use as inhibitors of steroid 5a-reductase. These patents also summarize in their description of the related art, numerous compounds which are stated to be prior art reductase inhibitors. See, for example, Table 1 of U.S.

Patent 5,026,882 and the discussion in the prior art section of the patent.

EP Publication No. 435 321 relates to A-nor-steroid-3carboxylic acid derivatives, which reportedly exhibit reductase inhibition.

25 In International Publication No. WO 91/13060 and EP Publication No. 458,207, Okada et al. discuss the preparation of Indole derivatives as testosterone reductase inhibitors.

Salle, et al., "17-acylurea Derivatives of 4- Azasteroids as Inhibitors of Testosterone Sa-Reductase" relates to studies regarding the effectiveness on reductase of a new series of 17P-acylurea substituted derivatives.

U.S. Patent 5,053,403 describes the use of certain 7 androgen receptor blocking agents together with certain reductase enzyme inhibitor in the treatment or prevention of sebaceous gland hypertrophy, hirsutism and male-pattern baldness.

Prior art inhibitors of 5a-reductase are not believed to fully inhibit both forms of 5a-reductase without exhibiting or causing undesirable androgenic or other hormonal activity.

SUMMARY OF THE INVENTION During treatment of certain diseases whose progress is stimulated by the activation of androgen receptors, it is desirable to reduce activation of those receptors. This may be accomplished by reducing the availability of "agonists", natural androgens and other compounds capable of activating the receptors or by reducing the availability of receptors and/or by blocking access to the receptors by compounds which would otherwise activate them. The latter function may be achieved by administering an "antagonist", a compound with affinity for a receptor which binds the 20 receptor and blocks access by agonists. In the case of androgen receptors, an androgen antagonist ("an antiandrogen") may desirably bind the androgen receptor i. without activating the receptor. Its physical presence blocks access to the receptor by natural or other androgens which, given access to the receptor, could bind and activate the receptor.

In accordance with the present invention, novel testosterone 5a-reductase inhibitors are used in the treatment of androgen-sensitive diseases whose progress can 30 be slowed by inhibiting activation of androgen receptors.

Compounds of the invention inhibit the activity of reductase which catalyzes the synthesis of the potent androgen, dihydrotestosterone. Thus availability of dihydrotestosterone to activate androgen receptors is desirably reduced.

8 It is important to achieve this desirable reduction of activity without causing adverse effects on the ultimate goal of inhibiting activation of androgen receptors. Hence, even if a compound effectively inhibits Sa-reductase activity, its therapeutic effect is reduced if the inhibitor itself has inherent androgenic properties such that the inhibitor activates the very receptors whose activation it is intended to reduce. Likewise, the inhibitor should resist being converted in vivo into an androgenic compound.

*00* 0 00 0 0 0 00 9 Conversely, however, a 5a-reductase inhibitor having antiandrogenic properties, displays two, rather than one, desirable effect on the treatment of androgen-related diseases. First, it inhibits enzymatic conversion of testosterone to dihydrotestosterone, thus reducing the amount of dihydrotestosterone available to activate androgen receptors. Second, it antagonistically blocks androgen receptors, shielding them from activation by any available androgens, including any dihydrotestosterone which may have been synthesized inspite of the inhibitor.

Thus, inhibitors of 5a-reductase activity preferably display a combination of desirable qualities, including an ability to effectively inhibit activity (preferably both types of 5a-reductase); and 15 a substantial lack of androgenic activity (and resistance to being converted in vivo into an androgen).

It is also desirable that the inhibitors have antiandrogenic properties. In order to eliminate undesirable side effects, preferred 20 inhibitors also substantially lack glucocorticoid characteristics.

Accordingly, the present invention provides pharmaceutical compositions having 5a-reductase inhibitors possessing little intrinsic androgenic activity and little propensity to be converted in vivo to another compound possessing intrinsic androgenic activity.

The present invention further provides methods for inhibiting Sa-reductase activity more effectively, and preferably methods for reducing the activity of both known Stypes of human S:21053AF 10 The invention further provides methods for the treatment of androgen-related diseases whose progress is aided by activation of androgen receptors. Such diseases include, for example, prostate cancer, prostatic hyperplasia and sexual deviance and may be treated by the methods of reducing 5o-reductase activity provide herein.

Methods of treatment are provided which utilize inhibitors, either alone or in combination with another active ingredient, an antiandrogen, as part of a combination therapy.

Pharmaceutical compositions are provided comprising the 5a-reductase inhibitors disclosed herein together with pharmaceutically acceptable carriers or diluents.

These pharmaceutical compositions are administered to a S 15 patient afflicted with a disease such as those discussed above, whose progress is aided by activation of androgen receptors.

a a.

••ooo S:21053AF 11 In one embodiment of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of an inhibitor of testosterone Sa-reductase having the molecular formula: R17a R1 7P

R

7 R4 R6 wherein the dotted line is an optional pi bond; S 1 5 wherein R 4 is hydrogen or methyl; wherein R 6 is a hydrogen or Ci-C 3 saturated or unsaturated hydrocarbon; .6 7 wherein R 7 is hydrogen; and wherein R 17 a is hydrogen or a C 1

-C

8 alkyl; and wherein R 17 0 is tertiary amino or acyl tertiary amino; and when R 6 is Ci-C 3 saturated or unsaturated hydrocarbon, then R17 can also be selected from the group consisting of acyl, carboxamide, tertiary amino and acyl tertiary amino; or wherein

R

17 a is selected from the group consisting of Ci-C 6 alkyl, Ci-C 6 hydroxyalkyl, Ci-C 6 haloalkyl,

C

2

-C

6 carbonylalkyl,

C

3

-C

6 cyclopropylalkyl,

C

3

C

6 epoxyalkyl and unsaturated analogs of the foregoing; and wherein R17 is hydrogen, hydroxy or a moiety converted to hydroxy in vivo.

30952DESCRIPTION.DOC 12 In another embodiment of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of an inhibitor of testosterone 5cL-reductase selected from the group consisting of: 0

CH

3 173- (N-n-amyl-N-formamido)-4mty one;

S

S

S S

*SSS

S

S

S

S

555* S. S

SS

S.

SeSe

S

*SSS

S.

S S S S

S.

S

0)N~

IH

l73- (N-n-butyl-N-formamido) 4 -methyl-4-aza-5(x-androstan-3one; 30952DESCRIPTION.DOC 13 0 H N--

OH

3 73- (N-n-hexyl-N-formamido) -4-mrethyl-4-aza-5-androstan-3one;

S

S

S

S.

S S S S

S

**SS

S

S

S

S. S S S

S

S S S S

S

17p-lly-17-hydoxy4-mthyl4-aa-SX-anrosan--one; 30952DESCRLPTION.DOC

UH

3 l 7 a-propyl-7-hydroxy4methy4aza-5(xandrostan-3-one; and 9

C.

C C a C C C a

S.C.

Sn.

CC**

C

C

S. C

C.

C

S C

C

Ca C

C

17x- (4-lodobutynyl) -l7p-hydroxy-4-methyl-4-aza5x androstane-3-one.

In another embodiment of the invention, there is provided the use of the pharmaceutical composition described above for the preparation of a medicament for the treatment of androgen-related diseases.

30952DESCRIPTION.DOC In another embodiment of the invention, there is provided the use of the pharmaceutical composition described above for the preparation of a medicament for reducing testosterone 5a-reductase activity.

In another embodiment of the invention, there is provided a method of reducing testosterone activity comprising administering to a patient in need of such reduction a therapeutically effective amount of a pharmaceutical composition described above.

In another embodiment of the invention, there is provided a method of treating androgen-related diseases comprising administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition described above.

15 Capsules having the 5a-reductase inhibitors discussed herein may also be utilized. The inhibitors and .:compositions containing them are utilized in accordance with the invention in methods for reducing activity, and in the treatment of diseases when progress is aided by activation of androgen receptors, e.g.

prostrate cancer, benign prostatic hyperplasia, acne, seborrhea, hirsuitism, androgenic alopecia and the like.

C

30952DESCRIPTION.DOC 16 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In some, but not all, embodiments, an azasteroidal nucleus has a substituent other than hydrogen), on at least one of the 4, 6 or 7 positions, 4-methyl and/or 6-lower alkyl and/or 7-lower alkyl.

In some embodiments, the R 17 0 substituent is a tertiary amine such as -N(R 19

(R

20 where R 19 is lower alkyl or haloalkyl, and R 20 is lower alkyl. In other embodiments R 17 0 is a tertiary amido substituent, acylamino substituents such as -N(R 25

)C(O)R

2 6 wherein R 26 is hydrogen or a lower alkyl and R 25 is a CI-C 6 saturated or unsaturated hydrocarbon such as cyclopropyl, cyclohexyl, butyl or isobutyl.

As used herein, the terms "tertiary amino" or "tertiary amido" refer to amino or amido substituents wherein the amino or amido nitrogen is not hydrogen substituted.

Preferred substituents for the nitrogen include but are not limited to acyl and lower alkyl.

In order to avoid steric interaction between the R 17 0 20 and R'17, it is preferred that at least one of these two

S

substituents be hydrogen, hydroxy or a substituent which is converted to hydroxy in vivo benzoyloxy, acetoxy).

Hydrocarbon substituents may be saturated or unsaturated. Unsaturated substituents are believed to be 25 especially useful at the R 7 and R 17 positions. In some

S.

embodiments, R 7 is a C 2

-C

6 alkyl, alkenyl or alkynyl

*S*

substituent.

In some embodiments, especially when R 17P is hydrogen, hydroxyl (or an ester derivative thereof), R 17 is Ci-C6 S 30 alkyl, Ci-Cs hydroxyalkyl,

C

1 -C haloalkyl,

C

2

-C

6 carbonylalkyl, C 3

-C

6 epoxyalkyl or an unsaturated analog of the foregoing. Preferred unsaturated analogs include, for example, halo or hydroxy alkynyl or alkenyl substituents, especially where the halo or hydroxy group is at the end of the substituent, farthest from the aza-steroidal 17 D-ring. An unsaturated at the 1,2 or 3 position of the 17(x substituent is also preferred.

Except where otherwise specified, substituents may have c or f stereochemistry. Optional pi bonds denoted by dotted lines in a molecular structure are independent of any other optional bonds appearing in that structure, the presence of one not being dependent on the presence or absence of another, unless valence requires interpendency. Compounds discussed herein may be formulated as salts thereof. Atoms of the azasteroidal nucleus for which no substituent is shown may optionally be further substituted (as valence permits) so long as such substitution does not adversely affect the compound's ability to inhibit activity, and does not render the compounds substantially more androgenic.

As used herein, the term "lower" when describing a chemical moiety means a moiety having 8 or fewer atoms. For instance, a "lower alkyl" means a C 1 to C 8 alkyl. Any moiety of more than two atoms may be straight- or branch-chained unless otherwise specified.

As discussed in more details below, carriers or diluents include solids and liquids. The novel pharmaceutical compositions of the invention may be used in the treatment of androgen-related diseases. When 25 administered systemically by injection, for treatment of prostate cancer, benign prostatic hyperplasia, and other diseases not primarily effecting the skin, conventional diluents or carriers which are known in the art to be pharmaceutically acceptable for systemic use are used, e.g., 9 o9 saline, water, aqueous ethanol and oil. When the inhibitors of the invention are utilized for the treatment of androgen related diseases such as acne, seborrhea, hirsutism, androgenic alopecia, the inhibitors are preferably administered together with a conventional topical carrier or diluent such as a mixture of ethanol and propylene glycol.

When used topically, it is preferred that the diluent or 18 carrier does not promote transdermal penetration of the active ingredients into the blood stream or other tissues where they might cause unwanted systemic effects. When a composition is prepared other than for immediate use, an art recognized preservative is typically included benzyl alcohol).

When the compound is administered in a cutaneous or topical carrier the carrier may be any known carrier in the cosmetic and medical arts, any gel, cream, lotion, ointment, liquid or non liquid carrier, emulsifier, solvent, liquid diluent or other similar vehicle which does not exert deleterious effect on the skin or other living animal tissue. Examples of suitable topical carriers include, but are not limited to liquid alcohols, liquid glycols, liquid polyalkylene glycols, water, liquid amides, liquid esters, liquid lanolin and lanolin derivatives and similar materials. Alcohols include mono and polyhydric alcohols, including ethanol, glycerol, sorbitol, isopropanol, diethylene glycol, propylene glycol, ethylene glycol, hexylene glycol, mannitol and methoxyethanol. Typical carriers may also include esters, diethyl and dipropyl ether, methoxypolyoxyethylenes, carbowaxes, polyethyleneglycerols, polyoxyethylenes and sorbitols.

Usually, the topical carrier includes both water and alcohol S 25 in order to maximize the hydrophylic and lipophylic solubility. A typical carrier will comprise 75% ethanol or oo: isopropanol and 15% water.

The topical carrier may also include various ingredients commonly used in ointments and lotions and well 30 known in the cosmetic and medical arts. For example, fragrances, antioxidants, perfumes, gelling agents, thickening agents such as carboxymethylcellulose, I surfactants, stabilizers, emollients, colouring agents and other similar agents may be present.

As illustrated by the examples which follow, the compositions of the present invention may contain well known and currently used ingredients to form creams, lotions, gels and ointments which are dermatologically 19 acceptable and non toxic. The composition may be applied as a gel, a cream, an ointment, a lotion or the like.

A dry delivery system, as described in U.S .Patent Nos.

3,742,951, 3,797,494 or 4,568,343 may be used.

Solvents or devices as described in U.S. Patent Nos.

5,064,654, 5,071,644 or 5,071,657 can also be used to facilitate transdermal penetration when systemic effects are desired.

The compound can also be administered by the oral route. The compound in the present invention can be typically formulated with conventional pharmaceutical excipients, e.g. spray dried lactose and magnesium stearate into tablets or capsules for oral administration. Of course, taste improving substances can be added in the case of oral administration forms. When capsules for oral ingestion are desired, any pharmaceutical capsules known in the art may be filled with the 5a-reductase inhibitors of the invention, with or without additional diluents and other additives discussed herein.

The active substance can be worked into tablets or dragee cores by being mixed with solid, pulverulent carrier oo. substances, such as sodium citrate, calcium carbonate or I: dicalcium phosphate, and binders such as polyvinyl pyrrolidone, gelatin or cellulose derivatives, possibly by 25 adding also lubricants such as magnesium stearate, sodium lauryl sulfate, "Carbowax" or polyethylene glycol.

As further forms, one can use plug capsules, e.g. of hard gelatin, as well as closed solf-gelatin capsules comprising a softener or plasticizer, e.g. glycerine. The 000 30 plug capsules contain the active substance preferably in the form of granulate, e.g. in mixture with fillers, such as lactose, saccharose, mannitol, starches, such as potato starch or amylpectin, cellulose derivatives or highly dispersed silicic acids. In solf-gelatin capsules, the active substance 20 is preferably dissolved or suspended in suitable liquids, such as vegetable oils or liquid polyethylene glycols.

The following non-limiting examples describe the preparation of a typical cream, lotion, gel and ointment, respectively. In addition to these vehicles, one skilled in the art may choose other vehicles in order to adapt to specific dermatologic needs.

EXAMPLE A. A typical lotion contains 5% active compound, 15% propylene glycol and 75% ethanol and water EXAMPLE B. A typical gel contains 5% active compound, 5% propylene glycol, 0.2% Carbomer 940 (available as Carbopol 940 R from B.F. Goodrich), 40% water, 0.2% triethanolamine, 2% PPG-12Buteh-16 (available as UconR fluid from Union Carbide), 1% hydroxypropyl and 46.8% ethanol (95% ethanol-5% water).

EXAMPLE C. A typical ointment contains 5% active compound, 13% propylene glycol, 79% petrolatum, 2.9% glycerylmonostearate and 0.1% polylparaben.

EXAMPLE D. A typical cream contains 5% active compound, 0.2% propylparaben, 5% lanolin oil, 7.5% sesame oil, 5% cetyl alcohol, 2% glyceryl monostearate, 1% triethanolamine, 5% propylene glycol, 0.1% Carbomer 9 4 0 R and 69.2% water.

The 5a-reductase inhibitors of the invention are 25 preferably formulated into pharmaceutical compositions at conventional concentrations for 5a-reductase inhibitors.

The attending clinician may elect to modify the concentration and/or dosage in order to adjust the dose to the particular response of each patient.

30 When 5a-reductase activity inhibitors are administered S* in accordance with the invention, they are preferably administered orally or parenterally. Dosage preferably ranges from about 1 mg to about 1000 mg of active 21 expedient, i.e. 5a-reductase inhibitor(s), per day per 50 kg of body weight, most preferably from about 2.5 mg to about 500 mg per day per 50 kg of body weight.

Concentration of active expedient varies in a known manner depending upon the method of administering the pharmaceutical composition. A composition suitable for oral administration may preferably include at least one inhibitor of 5a-reductase activity wherein the total concentration of all such inhibitors in said pharmaceutical composition is from about 1% to 95% of the composition (by weight), and preferably from about 5% to about 20%. The pharmaceutically acceptable diluent is preferably starch or lactose (with or without tartrazine) When prepared for parental injection, the inhibitor is preferably added at a concentration between about 2.0 mg/ml and about 50 mg/ml (preferably about 5.0 mg/ml to about mg/ml) into a carrier preferably selected from the group consisting of saline, water, aqueous ethanol and oil.

In certain alternative embodiments, the pharmaceutical composition of the invention may be formulated for sustained release in accordance with known techniques. The sustained release formulations are preferably prepared in a known •S manner appropriate for either oral, intramuscular or

S*

o subcutaneous administration.

25 When the pharmaceutical composition is for topical use, the 5a-reductase inhibitor(s) is preferably formulated together with a carrier selected from the group consisting :of propylene glycol, ethanol, isopropanol and water at a concentration ranging from 0.5% to 10% by total weight of the pharmaceutical composition. The composition for topical use may be formulated, for example, as an ointment, a gel, a cream or a lotion, to be applied to the affected areas of S: the skin in need of treatment twice daily.

22 In some embodiments of the invention, the Sa-reductase inhibitors of the invention are used in combination with another active ingredient as part of a combination therapy.

For example, the novel inhibitors may be utilized together with a separate antiandrogen which may be incorporated into the same pharmaceutical composition as the inhibitor, or which may be separately administered. An active compound may possess both antiandrogenic and reductase inhibiting activity, and may be supplemented with another compound to reinforce either or both of these activities another antiandrogen or another inhibitor of 5a-reductase). Combination therapy could also include treatment with one or more compounds which inhibit the production of testosterone or its precursors.

When antiandrogen is used in combination therapy in addition to the 5a-reductase inhibitors of the invention, the antiandrogen may be, for example:

OH

H CC U(CH 2 2

C

i.

CF

3 Flutamide (systemic) EM 248 (topic) *o *o -23 The antiandrogen is formulated at conventional concentrations and administered at conventional dosages, at the same concentrations and dosages set forth above for the 5ax-reductase inhibitor.

The antiandrogen Flutamide is commercially available from Schering Corp. (New Jersey). The antiandrogen EM-248 may be synthesized as follows: OH1AO 0 ab 0 O 0"

T~

SS

04 CSC~O

OHO

OJSS 0 0c~ C F. EM 248 24 Compound b To a solution of testosterone 1 (288.43 g, 1.0 mole) in glacial acetic acid (3.5 ethanedithiol (85 ml, 1.01 mole) and boron trifluoride (800 mol)were added at 10 0

C.

The mixture was stirred at this temperature for 1 hour and poured over ice (2 kg). From this aqueous phase a white solid separated and was collected by filtration, washed with water (2x 2L) and air dried. Crystallization from methanol gave the pure compound b. Yield: 328.28 g Compound c A solution of b (182.3 g. 0.5 mole) in dry dichloromethane (1.5 L) was added dropwise to a solution of pyridinium chlorochromate (150 g, 0.7 mole), molecular sieves 3A (200 g) and sodium acetate (25 g) at room temperature with mechanical stirring. After the addition was completed, the mixture was stirred for 16 hours and then diluted with diethyl ether (2 L) and filtered through silica gel in a fritted funnel. The filtrate was concentrated in vacuo and the resulting solid was crystallized from methanol to give the pure compound c. Yield: 158.7 g, Compound d 2(3-Butynyloxy)tetrahydro-2H-pyran (112.5 g, 0.729 mole) was added dropwise to a solution of methyllithium (500 ml of MeLi 1.4M in ether, 70 mole) in 1 L of anhydrous THF 25 at -30 0 C under argon atmosphere in a 5 L round bottom flask.

After the addition was completed, the cooling bath was removed and the solution was allowed to stand at room temperature for 4 hours. The solution was cooled again at 30° and a solution of c (75 g, .207 mole) in 2.5 L of S 30 anhydrous THF was added dropwise. After the addition, the cooling bath was removed and the mixture was allowed to stand at room temperature for 16 hours. To this mixture, **t «t 25 100 ml of brine was added and the solution was diluted with ethyl acetate washed with brine and dried with anhydrous MgSO 4 The solvent was evaporated and a solid crystallized after a short period of time. Hexane was added to complete the precipitation. The solid was then filtered and washed with hexane. The compound was used in the next step without further purification. Yield: 95.8 g Compound e A mixture of compound d (30 g, 058 mole) and methyliodide (65 ml, 1 mole) in methanol 96% (750 ml) was heated under reflux for 16 hours. The solvent was then removed in vacuo and the crude mixture was diluted with ethyl acetate (1 The organic phase was washed with NaOH 3% (3 x 500 ml) and dried over MgS04. After the evaporation of the solvent the solid was washed with diethyl ether, filtered on a fritted funnel and washed again with diethyl ether. This compound can be used without further purification in the last step. Yield 17- (chlorobutynyl)-17p-hydroxy 4-androsten-3-one (f, EM 248) A mixture of compound d (15 g, 0.4 mole), triphenylphosphine (21 g, .08 mole) and carbon tetrachloride 06 mole) was heated under reflux in 1 L of anhyrous dichloromethane for 10 hours. After the evaoporation of the 25 solvent the crude mixture was adsorbed on silica gel and chromatographied on silica gel (flash) with diethyl ether: hexane (70:30). The compound was further purified by crystallization in diethyl ether. Yield A combination therapy involving 5a-reductase inhibitor 30 and antiandrogen has the beneficial effect of inhibiting activation of androgen receptors by two different mechanisms without significantly reducing testosterone levels, the reduction of which may cause undesirable side effects 26 is some patients. In appropriate cases, i.e. where prostate cancer or another androgen related disease is not responding acceptably to treatment, a concurrent therapy designed to decrease testosterone levels may also be utilized surgical or chemical castration, for example, by administering a LHRH agonists or antagonists known in the art) oo o a.

a.

a.

27 Inhibitors of testosterone 5ci-reductase of the formula: 0 R -pN I1

P

3 cH3 are not limited to those include but below.

set forth in Table 1 TA13LE 1 Inhibitor R 3 316 336 337 347 401 402 405 407 422 423 424 436

CH

3 Cyclo C 3

H

5 CYClo C 6

H

11

C

4 H9

C

5

H

1 j

C

6 H1 3 ISO amyl Ethyipropyl ISO C 4

H

9

C

3 jH 7

C

4

H

9

CH

2

C

6

H

5

H

H

H

H

H

H

H

H

H

H

CH

3

H

5ax-reductase inhibition K i (nM) 29 11.2 11.9 2.6 1.8 7.2 2.2 10.6 7.3 5.1 11.8 5.6 S

S

S

9 9* 0 S S

S.

28 Testosterone 5crx-reductase inhibitors of the formula: 0 NHI2 include but are not limited to those set forth in Table II below.

TABLE II Compound R, R2 S-reductase inhibition Ki (nM) EM 373 Cyclo C 3

H

5

CH

3 EM 374 Cyclo C 3

H

5

C

6

H

5 2.3 EM 390 Cyclo C 3

H

5 C3H7 7.1 EM 392 Cyclo C 3

H

5

C

2 H5 13.5 EM 394 Cyclo C 3

H

5

C

4

H

9 3.3 EM 396 Cyclo C 3

H

5

ISOC

3

H

7 14.0 EM 397 Cyclo C 3

H

5 Cyclo C 6

H

1 I 3.2 EM 408 GCl 3

C

6

H

5 8.2 4.

4 4 4 4 44 .4 4 4 4 4 4* 4 4 4 4 44 4 4 4 0 4.

44 4* 44 44 4 4 4 4 29 Testosterone 5c-reductase inhibitors of the formula: I Rl" include but are not limited to those set forth in Tables 1 and II as well as those compounds set forth below in Table 111.

TA13LE III Compound Steroid R' R RI R1 7 (x R 17

P

nucleus EM 316 Nil CH-, H H H N (CH 3

CHO

EM 336 Nil CH H H H N (C 3

H

5

CHO

EM 337 Nil CH3 H H H N (C 6 HI) CHO EM 347 Nil eli 3 H H H N (C 4

H

9

CHO

EM 401 Nil CH 3 H H H N (C 5 HI) CHO EM 402 Nil CH 3 H H H NC 6

H

1 3

CHO

EM 405 Nil CH_ 3 H H H N(iso-amyl)CHO EM 407 Nil CH 3 H H H N(ethylpropyl)CHC EM 422 Nil CH 3 H H H N (iSO-C 4

H

9

CHO

9 9e 9*

S

0

S

S

55 30 EM 423 Nil CH 3 H H H N (C 3

CHO

EM 424 Nil CH 3 H H H N (C 4

H

9

COCH

3 EM 436 Nil CH 3 H H H N (CH 2

C

6

H

5

CHO

EM 373 Nil CH 3 H H H N (CYClo-C 3

H

5

CONHCH

3 EM 374 Nil CH 3 H H H N (CYClo-C 3

H

5 CONHCcH EM 390 Nil CH 3 H H H N (CYClo-C 3

H

5

CONHC

3

H

7 EM 392 Nil CR 3 H H H N (CYClo-C 3

H

5

CONHC

2

H

EM 394 Nil CH 3 H H H N (CYClo-C 3

H

5

CONHC

4 Hq EM 396 Nil CH 3 H H H N (CYClO-C 3

H

5 CONH (I SO-C 3

H

7 EM 397 Nil CH 3 H H H N (CYCl10- C 3

H

5 CONH (C yco CH 1 EM 408 Nil CH 3 H H H N (CH3) CONHC 6

H

EM 409 Nil CH 3 H H H N (CH 3

CONHCH

3 EM 322 Nil CH 3 H H C3C 5

OH

EM 441 Nil H H H CH OH EM 378 Nil CR 3 H H C. H 7

OH

EM 352 Nil H R H C, 1 H- OH EM 450 Nil CH 3 H H H N (C 4

H

9

COCH

2 Cl EM 314 A' CH 3 H H H N (CHA)CHO EM 420 A' CH-, H H H N (CYClo-CH 5

CONHC

2

H

EM 346 A' H H H H N (CH 3 CR0 EM 448 Nil H H H C-C (CH 2 2 T1 OH a S

S*

S@

S S S S 55 5 S S

S.

S S a

S.

*S*S

S S S S a. S S

S

S

.5 S S @5 31 EM 465 Nil H H H C-C (CH2)Br OH EM 358 Nil H H H C-C (CH 2 3 Br OH EM 471 Nil H H H C=C (CH 2 3 1 OH EM 321 Nil CH3 H H C-C (CH 2 Br OH EM 320 Nil CH 3 H H C-C (CH 2 I OH EM 501 Nil H H H C-C (CH 2 2 C1 OH EM 502 Nil CH 3 I H H C-C (CH 2 2 C1 OCOC6H EM 503 Nil CH 3 H H C-C (CH 2 2 C1 OH EM 548 Nil CH 3

CH

3 H H N (C 4

H

9

CHO

EM 561 Nil CH 3 H C 4

H

8 OH H N (C 4

H

9

CHO

EM 572 Nil H H H H N (C 4

H

9

COCH

3 EM 591 Nil CH 3 H H H N (C 4

H

9 S0 2

-CH

3 EM 621 Nil CH 3 H H H N (C 4

H

9 P0 3

(CHA)

2 EM 435 Nil CH 3 H H H N (ter-C 6

HIA)CHO

U U 0 9* U U U U U U

U

U.

0*SU

U

U U. Us U U 0

U

32 Except where otherwise indicated, inhibition is measured by the following method. Plates having 24 dishes per unit are used. Into each dish were placed 100,000 cells from the human carcinoma metastic prostatic cell line DU 145 (ATCC HTB 81) in MEM medium containing 2% dextran charcoal-treated calf serum, 1% penicillin, 1% streptomycin and 1% non essential amino acids. After 24 h, the medium was removed and replaced by ml of MEM medium containing 2% dextran charcoal-treated calf serum, 10 nM of tritiated 4-androsten-3,20-dione and 1% of ethanol. Each dish had different concentrations of the Sa-reductase inhibitor being tested. The cells were incubated for 24 h at 37'C under an atmosphere of 5% CO 2 saturated with water. The extracellular medium was then removed, centrifuged at 1000 RPM for 10 min., and decanted in test tubes. To these test tubes was added 100 p1 of ethanol containing 25 pg of 4-androstanedione, 25 ig of dihydrotestosterone and 25 tg of testosterone. The steroids were then extracted from the mixture by two extractions with 20 2 ml of diethyl ether. Phase separation was achieved by freezing of the aqueous phase. The organic phase was then evaporated, thus leaving a steroid residue which was :r dissolved in a few drops of methylene chloride and spotted on TLC plates (Whatman WH 4420222). After developing twice with a mixture of benzene-acetone the spots were visualized by U.V. and gaseous iodine, cut, put into 0* separate vials for each steroid and extracted for 15 min with 1 ml of ethanol. After addition of 10 ml of scintillation cocktail (NEN 989), the vials were shaked and 30 counted.

The 5a-reductase activity is the sum of the transformation of 4-androstenedione to androstanedione and the transformation of testosterone to dihydrotestosterone.

Conversion of tritiated 4-androsten-3,20-dione to other steroids in the present of different concentrations of 33 inhibitor and a constant concentration or radioactive 4androstenedione is measured, and IC 5 0 the concentration of inhibitor required to inhibit 50% of activity) values are then used to calculate Ki values of inhibition for each compound according to Cheng and Prusoff (Biochem. Pharmacol 22, 3099-3108, 1973).

Set forth below are non-limiting examples of methods of synthesizing Sa-reductase inhibitors for use in the present invention. Those of skill in the art may readily alter these syntheses by known conventional techniques to produce other 5-reductase inhibitors of the invention.

EXAMPLES OF SYNTHESIS OF PREFERRED INHIBITORS OF STEROID 5-REDUCTASE ACTIVITY EXAMPLE 1 Preparation of 173-(N-n-butyl-N-formamido)-4-methyl-4aza-5a-androstan-3-one (11, Ri=C 4 Hg, EM 347) Synthesis described in scheme 1 Preparation of 17p-hydroxy-5-oxo-A-nor-3,5seconandrostan-3-oic Acid To a stirred mixture of the 20 testosterone acetate (Steraloids Inc. Wilton HN USA) (200 g, 0.605 mol) in tert-butyl alcohol (2 L) was added a solution of sodium carbonate (96.3 g, 0.908 mol) in 460 mL of water.

The mixture was brought to reflux and a solution of sodium 04O0 periodate (893.8 g, 4.176 mol) and potassium permanganate (70.8 g, 0.448 mol) in water (75°C) was added gradually (1 h) while the reflux temperature was maintained. The reaction was cooled to 30 0 C, and after 15 min the solids were removed by filtration. The solid was washed with 800 mL of water, and the combined filtrates were concentrated 30 under reduced pressure to remove most of tert-butyl alcohol (final volume 1.0 The aqueous residue was cooled and acidified to pH 3.0 with concentrated hydrogen chloride solution. The aqueous solution was extracted with 34 methylene chloride (4 X 800 mL) and the combined organic phase was washed with water, dried and concentrated to solid. Thus the solid obtained was subjected to acetate hydrolysis by refluxing with NaOH (34.3 g, 0.857 mol) in methanol (2.0 L) for 12 h. The reaction mixture was concentrated to 400 mL, diluted with water (600 mL) and acidified to pH 3. The solid was filtered, washed with water and dried. The filtrate was extracted with methylene chloride (3 X 1.0 and the combined organic phases were concentrated to syrup. Both the precipitates and the syrup were swished with boiling EtOAc and cooled at 0°C for overnight to give 125 g (67% yield) of colorless crystals; mp 205-207 0

C.

Preparation of 17P-hydroxy-4-methyl-4-aza-androst-5ene-3-one In a Schlenk tube, MeNH 2 was bubbled till saturation to a mixture of the seco acid 1 (8.0 g, 25.974 mmol) in ethylene glycol (80 mL) at room temperature. The clear yellowish solution was heated gradually (3 OC/min) up to 180 °C and held at this temperature for 1 h. The 20 reaction mixture was cooled to 10 °C and water (80 mL) was added with stirring. The solid was filtered, washed with water (20 mL) and dried to give 6.1 g of 3 mp 181-183 0C.

Preparation of 17p-hydroxy-4-methyl-4-aza-5a-androstan- 3-one A solution of the compound 3 (6 g, 20.7 mmol) in acetic acid 130 mL) was hydrogenated in the presence of platinium oxide (600 mg) at 45 starting at room temperature and heated to 60 °C over 12 h. The reaction mixture was cooled and filtered. The catalyst was washed 30 with acetic acid (30 mL), and the combined filtrates were concentrated to a solid (5.5 g, mp 178-180 oC.

35 4-Methyl-4-aza-5a-androstan-3, 17-dione The following method is the representative. To a stirred solution of compound 5 (7.3 g, 25 mmol) in methylene chloride (260 mL) was added pyridinium chlorochromate (8.1 g, 37 mmol) and the mixture was stirred at room temperature for 3 h. The contents were passed through Florisil (30-60 mesh) to remove the precipitates and the filtrates were washed with water (2 X 200 mL) and dried. The resulting residue was purified by flash column chromatography to give the dione 7 (4.4 g, mp 126-128 OC.

Preparation of 17p-(N-butyl)-amino-4-methyl-4-aza-5aandrostan-3-one Ri=C 4 H) The following method is representative. To a mixture of dione 7 (0.150 g, 0.495 mmol) and n-butylamine (0.040 g, 0.54 mmol) in 1,2dichloroethane was added sodium triacetoxyborohydride (0.156 g, 0.74 mmol) followed by acetic acid (0.03 g, 0.49 mmol) under argon at room temperature. After 16 h, the reaction mixture was diluted with methylene chloride (15 mL) and washed with 1 N aqueous sodium hydroxide (2 X 20 mL), followed by brine (20 mL), dried and solvent removed to give •the crude product which was purified by flash column chromatography to provide the 17-N-butyl-derivative (0.110 g, 61 yield).

Preparation of 173-(N-n-butyl-N-formamido)-4-methyl-4- 25 aza-5a-androstan-3-one (11, Ri=C 4

H

9 EM 347) To a solution of formic acid (0.026 g, 0.556 mmol) in chloroform (1.5 mL) was added dropwise dicyclohexylcarbodiimide (DCC) (0.114 g, 0.56 mmol) in chloroform (1.5 mL) at 0°C. After 5 min the above solution was added to the compound 9 (0.10 g, 0.28 S 30 mmol) in pyridine (2 mL). The mixture was then stirred for S* 1 hour at room temperature. Evaporation of solvent was followed by addition of ether gave dicyclohexylurea which was removed via filtration and the solid was washed 36 with ether. The combined filtrate was concentrated and purified by flash column chromatography to give the product 11, (EM 347), (0.075 g, 70%) 1 H MR (CDCl 3 6 0. 66 3H) 0.74-1.42 (in, 11 1.2-2.08 (mn, 19 2.37-2.48 (mn, 2 H), 2.87 3 3.04 (dd, 1 H, J=4, 13 Hz), 3.2-3.4 (in, 3H), 8.12 0.8H), 8.24 0.2H); 1 3 C NMR (ODC1 3 6 170.56, 164.54, 162.97, 68.59, 65.62, 51.99, 51.71, 44.11, 36.85, 36.40, 34.12, 32.88, 30.64, 29.68, 29.04, 28.96, 25.20, 24.37, 22.86, 20.55, 20.21, 13.-77, 12.34: HRMS: calcd for

C

2 4

H

4 0

N

2 0 2 388.3089; found 388.3147.

37 SCHEME I OAc 0~

OH

F0 2

C

C0 2 P- H, 3 PR-H 4 P-H 5 R-CH3

P

1

NH

C

C C C

C.

C.

C C C C

C

CC..

CCC.

C C C C C S C C. CC C C

C

CC C C C

CC

6 P-H 7 P-CH 3 P 1 N- CHZO 8 A-H 9 P-OH 3 10 P-H 11 P-OH 3 13 R-HX-0 14 P-H,X-S

P-CHXCS

38 EXAMPLE 2 17p-(N-n-Amyl-N-formamido)-4-methyl-4-aza-5a-androstan- 3-one (11, Ri=C 5 Hl, EM 401) This synthesis is described in scheme 1.

Preparation of 17p-(N-n-amyl)-amino-4-methyl-4-aza-5aandrostan-3-one RI=C 5 To a mixture of dione 7 (3.4 g, 22.6 mmol) and n-pentylamine (1.07 g, 12.3 mmol) in 1,2dichloroethane was added sodium triacetoxyborohydride g, 16.7 mmol) followed by acetic acid (0.68 g, 11.2 mmol) under argon at room temperature. After 16 h, the reaction mixture was diluted with methylene chloride (150 mL) and washed with 1 N aqueous sodium hydroxide (2 X 200 mL), followed by brine (200 mL), dried and solvent removed to give the crude product which was purified by flash column chromatography to provide the 17-N-n-amyl-derivative (2.5 g, 61% yield).

Preparation of 17p(N-n-Amyl-N-formamido)-4-methyl-4aza-5a-androstan-3-one (11, Ri=C 5

HI

1 EM 401). To a solution of formic acid (0.504 mL, 13.36 mmol) in chloroform (37 mL) was added dropwise dicyclohexylcarbodiimide (DCC) (2.75 g, 13.36 mmol) in chloroform (37 mL) at 0°C. After 5 min, the above solution was added to the compound 9 (2.5 g, 6.68 m mol) in pyridine (20 mL). The mixture was then stirred for 1 hour at room temperature. Evaporation of solvent was 25 followed by addition of ether gave dicyclohexylurea which was removed via filtration and the solid was washed with ether. The combined filtrate was concentrated and purified by flash column chromatography to give the compound 11,

R

1

=C

5 H11 (EM-401) (2.5 g, The NMR spectroscopy o.

39 analysis gave a mixture of two conformers, M.P. 149- 151 0 C; I' MR (CDC1,): 6 0. 67 3H) 0. 86 3H) 0. 82- 1.19 (in, 6H), 1.21-1.42 (mn, 11H), 1.56-1.57 (mn, 2H), 1.69- 1.91 (mn, 4H), 1.92-1.97 (mn, 3H), 2.38-2.43 (mn, 2H), 2.89 (s, 3H), 3.0 (dd, 1H, J= 3 2 12.4 Hz), 3.21-3.28 (in, 3H), 8.14 0.8H), 8.2 0.2H); 1 3C MR (CDC1 3 6 170.56, 164 8, 162.97, 68.54, 65.58, 61.98, 51.93, 51.66, 51.24, 46.69, 44.28, 37.24, 36.8, 36.37, 34.07, 32.83, 32.12, 29.64, 29.13, 29.04, 28.8, 28.19, 25.15, 24.33, 23.22, 22.84, 22.36, 20.51, 13.94, 12.75, 12.32; HRMS: calcd for C2 5

H

42 0 2

N

2 402.3245, found 402.3242.

40 EXAMPLE 3 By analogous methods to those described in example 1, the following compounds were synthesized.

EM 316: 17p-(N-methyl-N-formamido)-4-methyl-4-aza-5aandrostan-3-one (11, R 1 The product was prepared in 78% yield and the NMR spectroscopy analysis gave a mixture of two conformers, M.P. 194-196 oC, H NMR (CDC 3 6 0.74 2.4 0.75 0.6 0.88 0.89 (s,2.4H), 0.78-1.14 3H), 1.26-1.47 6H), 1.60-1.90 7H), 2.01-2.07 2H), 2.41-2.46 2H), 2.90 3H), 2.92 (s, 3H), 3.02-3.07 (dd, 1H, J 12.58, 3.2 Hz), 3.32 0.8 H, J 9.6 Hz), 4.21 0.2 H, J 10 Hz), 8.15 0.8 H),8.18 0.2 13 CNMR (CDC13) 6 170.45, 164.20, 163.32, 68.95, 65.43, 61.36, 51.82, 51.37, 51.12, 45.50, 44.21, 37.10, 36.66, 36.26, 33.87, 33.58, 32.70, 29.59, 29.51, 28.92, 28.83, 25.04, 23.03, 22.84, 22.74, 21.49, 20.35, 13.20, 12.61, 12.21; HRMS: calcd for C 21

H

34

N

2 0 2 346.2620; found, 346.2645.

EM 336: 17p-(N-Cyclopropyl-N-formamido)-4-methyl-4-aza- 20 5a-androstan-3-one (11, R 1 =cyclo C 3

H

5 The product was prepared in 74% yield and the NMR spectroscopy analysis gave 99 a mixture of two conformers, M.P. 163-165 OC; H NMR (CDC13); 6 0.39-0.44 0.4 0.61-0.85 10H), 0.86-1.17 (m, 2H), 1.18-1.41 7H), 1.42-1.56 1H), 1.57-2.06 (m, 6H), 2.35 (dd, J 4.5, 9.4 Hz, 2 2.38-2.54 2H), 2.85 3H),2.97 (dd, J 3.4, 12.5 Hz, 1H), 3.24 J 8.7, 8.9 Hz, 0.4 4.0 J 9.2, 9.5 Hz, 0.6 8.27 0.4H), 8.33 0.6 H) "C NMR (CDC1 3 6 170.6, 165.4, 163.5, 69.9, 65.6, 64.4, 51.9, 51.8, 51.3, 45.7, 44.2, 37.9, 37.5, 36.4, 34.2, 29.7, 29.3, 29.1, 29.0, 28.8, 25.3, 22.2, 20.7, 13.6, 12.3, 9.9, 8.1, 6.4, 6.2. HRMS: calcd for

C

23

H

36

N

2 0 2 372.2796; found 372.2820.

41 EM 337: 17- (N-Cyclohexyl-N-formamido)-4-methyl-4-aza- 5a-androstan-3-one (11, R 1 =cycloC 6 H) The product was prepared in 52% yield and NMR spectro-scopy analysis gave a mixture of two conformers, M.P. 144-146 OC; 1H NMR (CDC13): 6 0.70 1.5H), 0.77 1.5H), 0.85 0.87 1.5H), 0.81-1.41 12H), 1.44-1.83 13H), 1.90-2.04 2H), 2.40 (dd, J 4.6, 9.3 Hz, 2H), 2.90 2.99 (dd, J 3.2, 12.4 Hz, 1H), 3.16 J 9.7, 9.9 Hz, 0.5H), 3.70-3.82 0.5H), 4.28 J 9.6, 9.9 Hz, 0.5H), 8.30 0.5H), 8.38 0.5H). 13C NMR (CDCI 3 6 170.7, 163.5, 163.1, 77.2, 66.3, 65.7, 61.9, 55.4, 54.4, 52.4, 52.1, 51.4, 44.7, 43.1, 37.2, 36.8, 36.5, 34.2, 34.1, 33.7, 32.9, 30.9, 30.6, 29.8, 29.7, 29.3, 29.1, 29.0, 27.9, 26.9, 26.2, 26.0, 25.4, 25.3, 23.2, 22.9, 20.7, 20.5, 13.0, 12.6, 12.4; HRMS: calcd for C 26

H

42

N

2 0 2 414.3246 found 414.3270.

EM 402: 17p-(N-n-Hexyl-N-formamido)-4-methyl-4-aza-5aandrostan-3-one (11, R 1 =CHi) The product was prepared in yield. The NMR analysis gave a mixture of two conformers, M.P. 101-103 0 C, 'H NMR (CDCl 3 6 0.68 3H), 0.86 3H), 0.77-1.09 6H), 1.22-1.42 10H), 1.43- 1.57 2H), 1.60-1.82 5H), 1.89-2.03 4H), 2.38-2.43 2H), 2.89 3H), 2.97-3.03 (dd, 1H, J 12.4, 3.2 Hz), 3.18-3.28 2.4H), 4.12 02H, J=10 Hz), 8.14 0.8H), S 25 8.2 0.2H); "C NMR (CDC13) 170.57, 164.49, 162.97, 68.55, 65.58, 61.99, 51.95, 51.66, 51.26, 46.72, 45.66, 44.32, 44.22, 37.24, 36.8, 36.37, 34.07, 32.83, 32.43, 31.32, 29.64, 29.19, 29.04, 28.95, 28.47, 26.64, 26.35, 25.15, 24.34, 23.23, 22.84, 22.49, 20.51, 13.92, 12.32; 30 HRMS: calcd for C 26

H

44 0 2

N

2 416.3382, found, 416.3355.

EM 405: 17- (N-iso-Amyl-N-formamido)-4-methyl-4-aza-5aandrostan-3-one (11, RI iso-CH 11 The product was prepared in 66% yield. The NMR analysis gave a (4:1) mixture of two conformers, M.P. 87-89 0 C, 1H NRM 42 (CDClI) 5 0.67 3H), 0.77-1.06 12H), 1.21-157 (m, 1.71-1.80 4H), 1.82-1.97 3H), 2.38-2.43 (m, 2H), 2.89 3H), 2.97-3.01 (dd, 1H, J 12.4, 3.2 Hz), 3.20-3.29 1.8H), 4.16 0.2H, J=10 Hz), 8.13 (s, 0.8H), 8.2 0.2H); 3 C NMR (CDC1 3 6 170.57, 164.46, 162.91, 68.49, 65.58, 51.92, 51.63, 44.19, 42.72, 37.19, 36.80, 36.35, 34.07, 32.81, 29.64, 29.19, 29.04, 28.93, 26.28, 25.92, 25.15, 24.27, 22.84, 22.43, 20.51, 12.77, 12.32; HRMS: calcd for C 2

.H

42 0 2

N

2 402.3245, found, 402.3230.

EM 407: 17p-(N-l-Ethylpropyl-N-formamido)-4-methyl-4aza-5c-androstan-3-one (11, R=iso-CsH 1 The product was prepared in 50% yield and the NMR analysis gave a (2.33:1) mixture of two conformers, M.P. 111-113 'H NMR (CDCI 3 6 0.74-1.2 15H), 1.24-1.84 17H), 1.95-2.1 2H), 2.41 -2.42 2H), 2.91 3H), 2.99-3.1 1.7H), 3.95 0.3H, J 10 Hz), 8.24 0.3H), 8.48 0.7H); 13C NMR (CDCl 3 5 170.60, 163.88, 163.46, 66.28, 65.74, 65.62, 63.81, 52.85, 52.02, 51.89, 51.77, 44.94, 43.17, 37.24, 36.39, 34.22, 32.89, 29.75, 29.66, 29.07, 29.01, 28.89, 28.56, 28.47, 26.19, 25.32, 25.22, 23.25, 23.05, 22.98, 20.78, 20.57, 13.25, 13.07, 12.36, 11.79, 11.44, 10.62; 0 HRMS: calcd for C 2 5H 42 0 2

N

2 402.3246, found 402.3265.

EM 422: 17p-(N-iso-Butyl-N-formamido)-4-methyl-4-aza-5- S 2 androstan-3-one (11, R 1 =iso-C 4

H

9 The product was prepared 25 in 90% yield and the NMR analysis gave a mixture of two conformers, M.P. 52-54 IH NMR (CDCI 3 6 0.67 (s, 3H), 0.85 3H), 0.68-1.15 9H), 1.21-1.38 6H), 1.5-1.79 6H), 1.81-1.99 3H), 2.38-2.42 2H), 2.88 3H), 2.94-3.02 (dd, 1H, J 12.4, 3.3 Hz), 3.1-3.15 (dd, 30 0.2H, J 13.2, 5.7 Hz), 3.23 0.8, J 0.8 3.32-3.39 (dd, 0.8H, J 13.2, 6.6. Hz), 4.05 0.2H, J 10 Hz), 00 8.15 0.2H), 8.29 0.8H); 13C NMR (CDCI 3 6 170.57, 164.66, 162.82, 69.18, 65.57, 52.26, 51.89, 51.83, 51.23, 45.99, 44.25, 37.16, 36.34, 32.81, 29.04, 28.92, 28.03 43 27.12, 26.76, 25.12, 24.94, 23.05, 22.81, 20.17, 19.96, 19.78, 12.84, 12.33; HRI4S: calod for C 2 4

H

4 0

N

2 0 2 388.3089, found, 388.3069.

EM 423: 173- (N-n-propyl-N-formamido) -4-methyl-4-aza-cLandrostan-3-one (11, RI=C: H 7 The product was prepared in 82% yield and the MR analysis gave a mixture of two conformers, M.P. 127-129 0 C; 'H NMR (CDCl 3 8 0.65 3H), 0.83 3H), 0.69-1.13 (in, 6H), 1.19-1.81 (mn, 13H), 1.86- (in, 3H), 2.35-2.4 (in, 2H), 2.86 3H), 2.94-3.0 (dd, 1H, J 12.4, 3.2 Hz), 3.10-3.28 (in, 1.82H), 4.1 0.18H, J 10 Hz), 8.11 0.82 8.17 0.18 13 C NMR (CDC1_,) 6 170.51, 164.43, 162.99, 68.51, 65.52, 51.89, 51.62, 48.30, 45.82, 44.17, 36.32, 34.03, 32.78, 29.58, 28.98, 28.89, 25.11, 24.31, 22.78, 21.61, 20.47, 12.27, 11.24; HPJ'S: calcd for C 2 3

H

3 8 0 2

N

2 374.2933, found 374.2903.

EM 436: 173- (N-Benzyl-N-formanido) -4-inethyl-4-aza-Standrostan-3-one (11, R 1

=CH

2

C

6

H,

5 The product was prepared in 80% yield. The MR analysis gave a mixture of two conformers m.p. 89-91'C, IR v cm 1 (KBr) 1640, 1610; 1 NMR (CDCL 3 6 0.74 3H) 0.85 3H), 2.37-2.42 (in, 2H), 2.87 3H), 2.95-3.0 (dd, 1H, J=12.5, 3.4 Hz), 3.28 0.8H, J=9.8 Hz), 4.18 0.2 H, J 9.7 Hz), 4.4 0.8H,J 15.5 Hz, 4.5 0.4 H, J 3.8 Hz) 4.79 (d, 0. 8 Hb, J=15.5 Hz) 7.1-7. 3 (in, 5H) 8.28 0.2H), 8.42 0.8H); 1 CMR(CDCl 3 170.38, 165.17, 162.93, 138.76, 137.38, 128.55, 128.34, 127.27, 126.93, 125.93, 68.01, 65.42, 62.67, 51.80, 51.63, 51.20, 50.36, 47.28, 45.81, 44.11, 37.39, 37.06, 36.29, 34.04, 32.75, 29.61, 29.49, 28.87, 25.04, 24.65, 23.29, 22.72, 20.54, 12.95, 12.41, 30 12.23; HRMS: calcd for C 2 7HI3 8

O

2

N

2 422.2933, found 422.2924.

44 EXAMPLE 4 Preparation of 17p-(N-methyl-N-formamido)-4-methyl-4aza-5a-androst-l-ene-3-one (17) Preparation described in scheme 2 Preparation of 17p-(N-methyl-N-formamido)-4-methyl-4aza-5a-androst-l-ene-3-one (17, R 1

=CH

3 EM 314). The following method is representative.

Bis(trimethylsilyl)trifluoroacetamide (0.305g, 1.185 mmol) was added to the mixture of formamide 11, Ri=CH 3 (EM 316) (0.10 g, 0.289 mmol) and 2,3-dichloro-5,6-dicyano-1,4benzoquinone (0.066g, 0.289 mmol) in dioxane (4mL) under nitrogen. After 4 h at 25'C, the contents were heated at 110 0 C for 24h. The resulting dark red solution was poured into the stirred mixture of methylene chloride (6 mL) and 1% aqueous sodium bisulphite solution (1.8 mL). The heterogeneous mixture was filtered. The dark red organic layer was washed with 2 mL of 2N HC1 followed by brine, dried and concentrated. The crude mixture was purified by column chromatography to give 41 mg of the product 17 RI=CH 3 20 (EM 314) 1 H NMR (CDC13) 0.72 3H 0.74 0.86-2.15 15H), 0.89 0.91 2.89 3H), 2.93 3H), 3.27-3.36 2H), 5.82-5.85 3H), 6.65 1H, J 11 Hz), 8.14-8.24 (m, 1H); HRMS: calcd for C 21

H

32

N

2 0 2 344.2463; found 344.2426.

45 SCHEME 2

R

1 N-ci-o

R-H

11R=H 16 R4{ 17

-H

RINL NHR 2

S

S.

S.

S S S S 5* S. S 5 0 55 5555 0e S S 55 S S

S

S

05 S5

S.

S

12 R-H 13 R.=01 3 18 R=H 19 R=4CH 3 46 EXAMPLE By analogous methods to those describe in example 4, the compounds 16 EM 346), 18 and 19 EM 420) were synthesized using the formamide 10 or the ureas 12 and 13 as starting material.

4 4*4444 4 44 4 4 4 .4 04 4 4 0 4* 4' S 4 4 4 4 4 *444 .4 4 4 4 4 .4 .4.4 .4 f~ 4.

4* 44 04 4 4 4 4 4, 0 4.

4.

4 4 47 EXAMPLE 6 Preparation of 17P-N-(1N-cyclopropyl-2N-phenylurea)-4methyl-4-aza-5a-androstan-3-one (13, Ri=C 3

H

5

R

2

=C

6

H

5 EM 374) Preparation described in scheme 1 Preparation of 17p-N-(1N-cyclopropyl-2N-phenylurea)-4methyl-4-aza-5a-androstan-3-one (13, Ri=cyclo C 3

H

5

R

2 EM 374). The following method is representative. To a solution of compound 9 (R 1 cyclo C 3

H

5 (220 mg, 0.64 mmol) in THF (100 mL) was added N-methylmorpholine (0.107 mL, 0.975 mmol) followed by phenylisocyanate (0.1 mL, 0.9 mmol) at 0°C under argon. The mixture was stirred overnight at 0 C to room temperature. The mixture was diluted with ethyl acetate and washed twice with 2N HC1, dried and solvent removed to give the crude product which was purified by column to give the pure product 13 (EM 374) (210 mg, 73%); SH NMR (CDCl 3 0.74-1.11 6H), 0.8 3H),0.86 (s, 3H), 1.23-2.06 14H), 2.34-2.65 4H), 2.9 3H), 2.99 (dd, 1H, J 4, 13 Hz), 4.06 1H, J= 9.5 Hz), 7.42 5H); 13 c NMR (CDC1) 6 170.72, 157.19, 139.09, 20 128.81, 122.77, 119.49, 65.63, 52.04, 51.10, 45.57, 38.08, 36.35, 34.22, 32.79, 29.70, 29.02, 27.79, 25.244, 23.43, 23.05, 20.72, 13.73, 12.35, 12.01, 9.96.

*S

S 0 9@

O

05

S

48 EXAMPLE 7 By analogous methods to those described in example 6, the following compounds were synthesized.

EM 373: 17J3-N (1N-cyclopropyl-2N-methylurea) -4-methyl-4aza-5cx-androstan-3-one (13, Rj=cyclo C 3

H

5

R

2

=CH

3 .The product was prepared in 68% yield, 1H NMR (CDCl 3 :60.71 (s, 3H), 0.82 3H), 1.94-198 (dd, 1H, J=12.6, 3.4 Hz), 2.28-2.33 (mn, 1H), 2.35-2.40 (dd, 2H, J=9.5, 4.8 Hz), 2.53 1H, J= 10.36 Hz), 2.76 3H, =J 4.8 Hz), 2.87 3H), 2.95-3.01, (dd, 1H, J 12.5, 3.4 Hz), 3.9 1H, J =10 Hz), 5.29 1H, J=5Hz); 13 C NMR (CDCl1 3 5 17 0. 66, 160.86, 67.74, 65.58, 52.01, 51.05, 45.16, 37.98, 36.31, 34.16, 32.75, 29.66, 28.96, 27.37, 25.21, 23.32, 22.94, 20.68,13.61, 12.29, 11.43, 9.51; MS: m e rel. int.) 344 (W-57).

EM 392: 17f-N(N-cyclopropyl-2N-ethylurea) -4-methyl-4aza-5ct-androstan-3-one (13, R 1 =cyclo-C 3

H

5

R

2

=C

2

H

5 .The product was prepared in 78% yield, 1 H NMR (CDCl 3 :60.70 (s, 3H),0.81 3H), 1.08 3H, J 7 Hz), 1.94 (dd, 1H, J too*: 20 12.4, 3.2 Hz), 2.25-2.31 (in, 1H), 2.36 (dd, 1H, J 9.5, 4.8 Hz), 2.42-2.57 1H, J=10 Hz), 2.86 3H), 2.95-3.19 (dd, 1H, J=12.5, 3.4 Hz), 3.20-3.23 (in, 2H), 3.9 1H,J 00.=10 Hz), 5.27 1H, J 5 Hz); 13C NMR (CDCl 3 8 170. *06: 160.03, 67.50, 65.55, 51.95, 50.96, 45.31, 37.95, 36.26, 35.27, 34.10, 32.70, 29.61, 28.92, 27.18, 25.17, 23.28, 22.90, 20.64, 15.47, 13.58, 12.26, 11.64, 9.57; HBMS: calcd for C 2 5

H

4 1

N

3 0 2 415.319, found 415.318.

EM 408: 17f3-N (1N-methyl-2N-phenylurea) -4-inethyl-4-aza- Sc-androstan-3-one (13, R 1

=CH

3

R

2

=C

6

H

5 .The product was obtained in 87% yield, 1 H NMR (CDCl 3 :60.73 3H) 0. 86 3H), 2.4 (dd, 2HJ 9.5, 4.7 Hz), 2.90 49 3H) 2. 96 3H) 2. 96-3. 0 (dd, 1H, J =12. 5, 4 Hz) 4. 24 1H, J 10 Hz) 6. 53 1H) 6. 98 1H, J= -7 Hz), 7.24 2H, J 7 Hz), 7.36 2H, J= 7 Hz); l-C NNR (CDC1 3 :6 170. 66, 156. 37, 139.24, 128.69, 122.70, 119.79, 65.55, 63.96, 51.92, 51.17, 44.91, 37.35, 36.34, 33.92, 32.75, 31.58, 29.66, 29.03, 28.95, 25.19, 23.11, 20.50, 13.10, 12.30; HPRMS: calcd for C 27

H

3 9

N

3 0 2 437.2804, found, 437 .2823.

5o EXAMPLE 8 Preparation of l7J3-N- (1N-cyclopropyl-2Nmethylthiourea) -4-methyl--4-aza-5c-androstafl3-ofe (15, EM 379) The preparation is analogous to the preparation of compounds 13 in example 6 (see scheme but using phenylisothiocyanate as reagent in the step converting compound 9 to compound 51 EXAMPLE 9 Preparation of 17a-allyl-17-hydroxy-4-methyl-4-aza-5aandrostan-3-one (21, R 3 =C3H5, EM 322).

Preparation described in scheme 3 Preparation of 17a-allyl-17p-hydroxy-4-methyl-4-aza-5androstan-3-one (21, R3=C 3

H

5 EM 322). The following method is representative. To a solution of compound 7 (see scheme 1) (0.1 g, 0.328 mmol) in THF (10 mL) was added allylmagnesium bromide (394 pL, 0.394 mmol) at -78°C. After addition the contents were stirred for 1 h and workup as usual to provide the crude product which was purified by column chromatography to give the pure product EM 322 (77 mg, 'H NMR (CDC1: 3 6 0.68-1.0 2H), 0.84 6H), 1.14-1.66 11H), 1.77-2.06 5H), 2.14-2.2 1H), 2.28-2.36 1H), 2.42-2.48 2H), 2.86 3H), 2.94- 2.99 (dd, 1H, J 4, 13 Hz), 5.14 (dd, 1H, J 17, 2 Hz), 5.2 (dd, 1H, 13, 2 Hz), 5.91-6.06 1H); "C NMR (CDCIs): 6 170.70, 134.85, 118.99, 82.13, 65.70, 51.91, 50.13, 46.33, 41.75, 36.45, 35.14, 34.78, 32.94, 31.63, 29.95, 29.03, 25.31, 23.55, 20.77, 14.55, 12.37; HRMS: calcd for C22H3 5 NO2, 345.2668; found, 345.2658.

a a* a.

ao *i 52 SCHEME3 6 R=H 7 R=CH 3 H20

R-H

21 fl=CHk

I

I.

I I I I

I.

II

II

*1 I I I I

I.

I

I I I. II I I

I

I I

I

*1 53 EXAMPLE Preparation of l7c-propyl-17j-hydroxy-4-methyl-4-aza- 5c-androstan-3-one (21, R 3

=C

3

H

7 EM 378) Same preparation as the preparation of compounds 21 in example 9, but using propylmagnesium bromide instead of allylmagnesium. bromide as reagent.

EXAMPLE 11 Preparation of 17ax-allyl-17f3-hydroxy-4-aza-5acandrostan-3-one (20, R 3 =C3H5, EM 441).

Same preparation than the preparation of compounds 21 in example 9 in using the compound 6 as starting material.

OV9 54 EXAMPLE 12 Synthesis of 17a-(4-bromo-butynyl)-17p-hydroxy-4-aza- 5a-androstane-3-one (24, x=2, P=Br, EM 465) (scheme 4).

Preparation described in scheme 1 and scheme 4.

Preparation of 17p-hydroxy-4-aza-androst-5-ene-3-one Referring first to scheme 1, in a pressure apparatus, NH: was bubbled till saturation to a mixture of the seco acid 1 (6.0 g, 18 mmol) in ethylene glycol (60 mL) at room temperature. The clear yellowish solution was heated gradually (3 "C/min) up to 180 0 C and held at this temperatue for 1 h. The reaction mixture was cooled to 10 0 C and water mL) was added with stirring. The solid was filtered, washed with water (20 mL) and dried to give 4.5 g of 2.

Preparation of 4-aza-5a-androstan-3,17-dione A solution of the compound 2 (160 g, 0.5 mol) in acetic acid (500 mL) was hydrogenated at 60 psi using PtO2 (15 g) as catalyst at 60 0 C for 60 min. After cooling and filtering, the catalyst was washed with acetic acid and the solvent removed. The residue dissolved in methylene chloride was washed with 1N sulfuric acid, brine, saturated sodium bicarbonate, and brine. The organic phase was dried, filtered, and evaporated. The residue was chromatographed on silica gel. Elution with ethyl acetate/hexane gave crystalline compound which was treated with 3% methanolic 25 sodium hydroxide solution at reflux for 90 min. After usual work up the obtained residue was dissolved in acetone (500 mL) cooled to 0°C and Jones' reagent was added (8N-chromic acid solution, 65 mL). After 15 min, isopropanol was added and the mixture was concentrated

C

55 under vacuo. Water was added and the mixture was extracted with ethyl acetate. The organic layers were washed with brine, dried and evaporated to dryness. The chromatography on silica-gel of the residue with ethyl acetate/hexane as eluent gave the aza-ketone 6 (152 g) of which structure was determined by spectroscopic mean.

17a-(4-tetrahydropyranyloxy-butynyl)-17p-hydroxy-4-aza- 5-androstan-3-one (22, Referring now to scheme 4, to anhydrous THF (140 mL) at -60°C was added methyl lithium (1.4 M, 100 mL) and a solution of 4tetrahydropyranyloxybutyne (21.6 g, 140 mmol). To this mixture warmed up to room temperature, stirred for 2 h, and cooled to -60 0 C was added dropwise a solution of above azaketone 6 (9.6 g, 30 mmol) in THF (350 mL), and the mixture was warmed up to room temperature and stirred for 16h.

After usual work up, the compound 22 was purified by silica-gel chromatography and its structure determined by spectroscopic mean. In a similar fashion, 17a-(4tetrahydropyranyloxybutynyl)-17p-hydroxy-4-methyl-4-aza-5aandrostan-3-one (23, x=2) was prepared.

17a-(4-bromo-butynyl) -17-hydroxy-4-aza-5a-androstane- 3-one (24, P=Br, x=2, EM 465). To a solution of aza-diol 24, (P=OH) (179 mg, 0.5 mmol) obtained by acidic hydrolysis of the compound 22, and triphenylphosphine (262 mg, 1 mmol) 25 in methylene chloride (15 mL) at 0 C was added CBr 4 249 mg, 1 mmol) and the mixture was stirred for 2h at room S. temperature, the solvent was removed and the compound 24 (P=Br, x=2 EM 465) was purified by flash silica-gel chromatography and this structure determined by spectroscopic mean. In a similar fashion, 17a-(4-bromobutynyl)- Q o 56 17J-hydroxy-4-methyl-4-aza-5ca-androstane-3-one (25, x=2, P=Br, EM 321) was prepared.

Preparation of 17c- (4-lodobutynyl) -17j3-hydroxy-4methyl-4-aza-5at-androstan-3-one (25, x=2, P=I, EM 320) To a mixture of l7cL- (4-bromobutynyl) -17p-hydroxy-4-methy1-4aza-5oc-androstan-3-one (25, x=2, P=Br, EM 321) (200 mg, 0.471 rnmol) in acetone (16 mL) was added sodium iodide (92 mg, 0.6132 rnmol) and the mixture was refluxed for 12 h.

Removal of acetone and usual workup provided the crude product which was purified by column chromatography to give the pure product (EM 320) (137 mg, 60%) 1 H NMR (CDCl 3 0.83 3H), 0.88 3H), 2.41-2.46 (in, 2H), 2.83 2H, J 6.84 Hz), 2.91 3H), 3.0-3.06 (dd, 1H, J=3.

4 12.5 Hz) 3.24 2H, 1 Hz); HPMS: calcd for C 2 3

H

3 4 1N0 2 483.1635; found, 483.1634.

S. U 57 SCHEME 4 F~4OOT HPu 6A'H 7 P=CH 3

P

22 R=H 23 R=C1I 3

H

24 fl-H

P=CH

3 *4 0 4 4 *44* 4044 4 4 *4 0

S.

44.0 4.

4e 44 S. S 4 *4 4 58 EXAMPLE 13 In a similar fashion, to Example 12, the following compounds described in Table IV are prepared using different tetrahydropyranyloxy-alkynes and different carbon tetrahalides.

TABLE IV

I

R x P EM 501 H H 2 Cl.

EM 502 CH 3

COC

6

H

5 2 Cl EM 503 CH H 2 Cl EM 448 H H 21 EM 320 CH 3 H 21 471 H H 3 I .*EM 358 H H 3 Br 005 59 EXAMPLE 14 Preparation of 17p-(N-n-butyl-N-formamido)-4, 6dimethyl-4-aza-5a-androstan-3-one (31, EM 548).

Preparation described in Scheme Preparation of 17p-acetoxy-6a-methyl-4-androsten-3-one Testosterone (50 g) (Schering A.G. Germany) was treated in an apparatus equipped with a Dean-Stark, distillation apparatus, by diethylene glycol in toluene in the presence of a catalytic amount of p-toluenesulfonic acid at reflux for 16 h. The resulting ketal 26 was oxidized by monoperoxyphtalic acid, magnesium salt (Aldrich Chem. Comp.

Inc. Milwaukee Wis USA) in iso-propanol at 50 0 C for lh.

After removal of the solvent and crystallization the mixture of epoxyde 27 was heated at reflux with large excess methyl magnesium iodide in tetrahydrofuran for 18h. The ketal 28 was deprotected by standing overnight at room temperature with a mixture of acetone/water The hydroxy ketone 29 thus obtained was mono dehydrated by heating a mixture of 0.1 N sodium hydroxide in methanol and the 17p-hydroxyl was acetylated by usual manner (acetic anhydride/pyridine). The 6-methyl enone 30 was thus obtained (27 g) and characterized by spectroscopy.

17p-(N-n-butyl-N-formamido)-4, 6-dimethyl-4-aza-5aandrostan-3-one (31, EM 548). The 6-methyl enone 30 is transformed into 17-(N-n-butyl-N-formamido)-4,6-dimethyl-4aza-5a-androstan-3-one (31, EM 548) by a process analogous to the process described in example 1 for conversion of compound testosterone acetate to compound 11.

60 04 testosterone OR OH

S

S.

S S S

S@

S.

S

S

S

S.

S S S S *5

OSSS

S S S S

S.

S. 55 S S

S

S. S S S

S.

H c

OH

3

CI:-

31 61 EXAMPLE Preparation of 17p-(N-alkyl-N-formamido)-4-methyl-7ahydroxyalkyl-4-aza-5a-androstan-3-one (34) Synthesis is described in scheme 6 The following method is representative. The commercial 17p-acetoxy-4, 6-androstadien-3-one 32 (Steraloids Inc.

Wilton, NH, USA) is treated by an 1.5 excess of

TBDMSO(CH

2 )xCu(CN)Li (prepared from TBDMSO(CH 2 )xI, t-BuLi and CuCN) in ether and tetrahydrofuran in presence of trimethylsilyl chloride at -78 0 C. The mixture is heated to room temperature and usual work up is made. The resulting compound 33 is transformed into 17p-(N-alkyl-N-formamido)-4methyl-7a-hydroxyalkyl-4-aza-5a-androstan-3-one (34) by a process similar to the process described in example 1. The last step is the deprotection of the silyl group.

o• 62 SGHEME 6 32

-IN-

34 63 EXAMPLE 16 Preparation of alkylamide, alkylsulfamide and alkylphosphite derivatives of 17p-N-alkyl-4-methyl-4-aza-5aandrostan-3-one.

Synthesis is described in scheme 7.

The compounds 8 or 9 prepared in accordance with scheme 1 are treated at room temperature by acyl chloride in tetrahydrofuran using a 2 fold excess of K 2 CO3 powder of base. After usual work up, the compounds 35 or 36 are obtained. The use of sulfonyl chloride instead of acyl chloride gives in the same conditions respectively the compounds 37 or 38, and the use of dialkyl chloro phosphate gives respectively the compounds 39 or EM 424: 17p-(N-n-butyl-N-acetamido)-4-methyl-4-aza-5aandrostan-3-one. The product was prepared in 85% yield.

The NMR analysis gave a (1.85:1) mixture of two conformers, M.P. 152-154°C, 1 H NMR (CDC~) 6 0.67 1.95 0.74 (s, 1.05 0.80-1.0 9H), 1.07-1.69 12H), 1.71-1.91 (m, 5H), 1.99-2.07 (m 1H), 2.12 1.05H), 2.14 1.95H), 2.42-2.45 2H), 2.84-3.0 0.35H), 2.91 3H), S. 3.05 (dd, 1H, J 12.3, 3.2 Hz), 3.11-3.18 0.65H), 2.24- 2.29 (m,0.65H), 3.68-3.71 0.7H), 4.49 0.65H, J 9.9 Hz); "C NMR (CDC1 3 6 171.43, 171.14, 170.54, 67.29, 65.53, 25 62.27, 51.05, 46.36, 45.55, 44.55, 44.34, 37.10, 36.31, 33.96, 33.01, 32.75, 30.87, 29.67, 29.22, 28.98, 28.94, 25.18, 24.59, 23.65, 23.17, 22.70, 22.33, 20.49, 20.34, 4 S 20.04, 13.66, 12.94, 12.73, 12.29; HRMS: calcd for C 25

H

42

N

2 0 2 402.3246, found, 402.3234.

402.3246, found, 402.3234.

9 64 SCHEME 7

PI

1

NH

p a &Rl=H R 35 R=H 9 R,=GH- 3 36r IL=O

R

1 N- GO 3 0

R

1 -N-P(0R 3 2 a 4 4 a a 4* 44 4 a 9 a.

0* 4 4 C C 4 4 *0 @4 4 @4 4444 A 44 C 4S 4. 44 *4 0 4 S 4 37 R=H 38RC13 39 R-H 40 R=GHi 4 4 65 Additional examples of pharmaceutical compositions are set forth below: EXAMPLE 17 Composition suitable for injection Ingredient by Weight of Total Composition EM 378 (a 5c-reductase inhibitor) 0.4 Ethanol 6.4 NaCl 0.8 Water 91 Benzyl alcohol 0.9 EXAMPLE 18 Topical Lotion Ingredient by Weight of Total Composition EM 402 (a 5c-reductase inhibitor) Ethanol 47.5 Propylene glycol 47.5 0 000000

C

C.

C

C

CC

0S C S 0 C 0e

CS

0 0 S C 0000

C

5505 0 S C '9 0 0@

S.C.

SC

OC

C.

00 50 C S

S

S C 55 C 0 S S *5 66 EXAMPLE 19 A Tablet

T

Ingredient by Weight of Total Composition 46.5 EM 378 (a Sa-reductase inhibitor) Flutamide (an antiandrogen available from Schering Corp., New Jersey) Gelatin 46.5 4 4e 4 4 4 9 *449 4. 9 .4 9 4*

S

Lactose Starch EXAMPLE 20 A Topical Lotion Ingredient by Weight of Total Composition EM 402 (a 5c-reductase inhibitor) EM 248 (an antiandrogen) Ethanol 45.0 Propylene glycol 45.0 The terms and descriptions used herein are preferred embodiments set forth by way of illustration only, and are not intended as limitations on the many variations which 10 those of skill in the art will recognize to be possible in practicing the present invention as defined by the claims.

In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the words "comprises" and "comprising" are used in the sense of "includes" and "including", i.e. the features specified may be associated with further features in various embodiments of the invention.

Claims (12)

1. A pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of an inhibitor of testosterone 5a-reductase having the molecular formula: R17P 10 C D A B R 7 S4 R wherein the dotted line is an optional pi bond; wherein R 4 is hydrogen or methyl; wherein R is a hydrogen or Ci-C 3 saturated or unsaturated hydrocarbon; wherein R 7 is hydrogen; and wherein R 17 i is hydrogen or a C 1 -Cs alkyl; and wherein R 17 is tertiary amino or acyl tertiary amino; and when R 6 is C 1 -C 3 saturated or unsaturated S. hydrocarbon, then R 17 can also be selected from the group a consisting of acyl, carboxamide, tertiary amino and acyl tertiary amino; or wherein R 17 is selected from the group consisting of C 1 -C 6 alkyl, Ci-C 6 hydroxyalkyl, Ci-C 6 haloalkyl, C

2 -C 6 carbonylalkyl, C

3 -C 6 cyclopropylalkyl, C 3 C 6 epoxyalkyl and unsaturated analogs of the foregoing; and

30952.DOC 68 wherein R 17 is hydrogen, hydroxy or a moiety converted to hydroxy in vivo. 2. A pharmaceutical composition comprising a pharmaceutically acceptable diluent or carrier and a therapeutically effective amount of an inhibitor of testosterone 5cix-reductase selected from the group consisting of: 0 0A 5 OH 3 173- (N-n-amyl-N-formamido) -4-methyl-4-aza-5cx-androstan-3 one; S S. S S C. C S S S S S SCSS C C S.C. C S SC S S. C S C C *SCSCS S S 0 AN'-- 173- (N-n-butyl-N-formamido) -4-methyl-4-aza-5x-androstan-3- one; 30952.DOC 69 73- (N-n--hexyl-N-fornam-ido) -4-methyl-4-aza-5cx-androstan-3 one; S S S S S. S. S S S S S S 5.55 S S S 55.. S S. S S S .5 *555 S 55 S S S. S 55 S 17p-lly-17-hydoxy4-mthyl4-aa-5x-anrosan--one; 30952.DOC 70 OH l 7 ax-propyl-17-hydroxy-4methy4aza5xandrostan-3-one; and 9* 9 4 4 9 4449 4 9 *4 9. 49 4 9.44 4 4*4* *4 9 4 94 *49499 9 17x- (4-lodobutynyl) -17J-hydroxy-4-methyl-4-aza-5Sa- androstane-3 -one. 3. The use of the pharmaceutical composition according to claim 1 or claim 2 for the preparation of a medicament for the treatment of androgen-related diseases. 30952.DOC -71-

4. The use of the pharmaceutical composition according to claim 1 or claim 2 for the preparation of a medicament for reducing testosterone activity.

5. A method of reducing testosterone activity comprising administering to a patient in need of such reduction a therapeutically effective amount of a pharmaceutical composition of either claim 1 or claim 2.

6. A method of treating androgen-related diseases comprising administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition of either claim 1 or claim 2.

7. A pharmaceutical composition of claim 1 or 15 claim 2 when used in reducing testosterone activity.

8. A pharmaceutical composition of claim 1 or claim 2 when used in treating androgen-related diseases.

9. Use of a pharmaceutical composition of claim 1 or claim 2 in reducing testosterone 5a-reductase activity.

10. Use of a pharmaceutical composition of claim 1 or claim 2 in treating androgen-related diseases.

11. A pharmaceutical composition according to claim 1 substantially as herein described with reference to any one of Examples 17 to

12. A method according to claim 5 or 6 substantially as herein described. Dated this 18th day of February 2000 ENDORECHERCHE INC. By their Patent Attorneys GRIFFITH HACK 30952.DOC