CA1154008A - STEROID 5.alpha.-REDUCTASE INHIBITORS - Google Patents

Abstract

ABSTRACT Compound of the following general formula are testosterone 5c-reductase inhibitors rendering the com-pounds useful in the treatment of acne and oily skin and benign prostatic hypertrophy: wherein R is: -O, -OH, -OCO-alkylC: 5, -COOH, -CH2CH, -COOalkylC1-?, -COCH3, , , and the 20? isomers of , , or .

Description

M-90~ CIP
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NOVEL STEROID 5~-REDUCTASE INHIBITORS

FIELD OF INVENTION
This invention relates to the use o~ certain steroid compounds for treatment of acne and oily skin, baldness and of prostate hypertrophy through inhibition of tes-tosterone 5c-reductase.
BACKGROUND OF INVENTION
It is known that skin rèsponds to androgens and is an active site of androgen metabolism. The androgen testosterone is metabolized in the skin to dihydrotestos-terone (DHT) which is a more potent androgen than tes-tosterone. As set out in Arch.J Oermatol. 111, 1496 (1975) there is considerable evidence that DHT is involved in the pathogenesis of acne as well as other androgen associated conditions. Studies in the hamster flank organ, which is an androgen dependent sebaceous structure, indicate that DHT stimulates the growth of this structure.
It has been found that acne-bearing skin produces from 2 to 20 times more DHT than normal skin. Therefore, it is believed that agents capable of blocking the formation of DHT would be effective in the treatment of an acne condition. Also, many studies indicate that prostatic hypertrophy may be treated by administering an agent that prevents the formation of DHT from testosterone, that is, a testosterone 5c-reductase inhibitor. Testosterone is converted to DHT by the enzymatic action of testos-terone 5c-reductase. One pOS5i ble means of blocking the ~ 8 ~ C3 CIP

formation of DHT is to inhibit the activity of testos-terone 5c-reductase. More desirably, in the treatment of acne the activity of the 5~-reductase enzymes is inhibited locally, that is in the region of the acne-bearing skin.
Many workers in the art have believed that steroidcompounds effective for testosterone 5a-reductase inhibi-tion purposes required a Q4-3-keto conflguration and inferentially, at 1east presence of other ring A sub- -lO stituents would add little, and might even detract from the usefulness of a compound for inhibition purposes.
Thus, th~ numerous compounds discussed by Voigt and Hsia in their U.S. patent 3,917,829 and by Benson and slohm in their U.S. patent 4,088,760 all contain the ring A ~ 4 -3-one structure.
It may be noted that the ~4-3 keto steroids hereto-fore suggested for inhibition of the 5c-reductase act through competitive inhibition and their effectiveness depends upon maintaining a significant, perhaps substan-tial, concentration of inhibitor in the target organJ
e.g., in the patient's skin or in the prostate. Mani-fes;ly competitive inhibitors offer a lesser degree of effectiveness than an irreversible or quasi~irreversible inhibitors.
A theory for irreversible inhibition of 5a-reductase and ~5-3-keto isomerase has been offered in U.S. patent ~,087,461. Briefly the ~heory calls for employment of a steroid containing a potentially reactive group, which ~roup becomes reactive when the target enzyme carries 3 out its transformation. At that time a chemical reaction ensues, e.g., an alkylation, directly at the active site of the enzyme or so closely adjacent thereto as to irreversibly inhibit the enzvme. U.S. Patent 4,087,461 suggests allenic seco-steroids fbr irreversible inhibi-tion of Q5-3-keto steroid isomerases and of testosterone ., ~.
.

~lS4(~ 8 5~-reductase. U.S. patent 4,087,461 describes the enzyme catalyzed transformation as involving reaction at the C-4 position.
The present invention relates to use of certain 4-substituted steroids which inhibit the activity of 5a-redurtase rendering said compounds useful in treating acne or oily skin. It is not known whether the inhibition is irreversible or is near to irreversible, i e., quasi-irreversible, because the 4-substituted steroid does not readily decouple from the enzyme.
SUMMARY OF_THE INVENTION
Compounds of the following formulas have been found to be at least quasi-irreversible inhibitors of the 5~-reductase. R

~ b wherein R is:
=0, -OH, -OCO-Alkyl 1-5, -CQOH, -CH20H, -CHO, -COO alkyl 1-6, -COCH3, -CH-COOH, -CH-COO alkyl 1-6, ~ ~ and the 20R i somers of -CH-CH20H, -CH-CHO, -CH-COOH or -CH-COO-alkyl C~
It is believed that the inhibition attributable tothe presence of the N2 substituent at C-4 is cummulative of the competitive inhibition attributable to the presence of a preferred 17 substituent. By and large, the 17 substitutents, i.e., R, are those described in the prior art, as for example by patents 3~917J829 and 4,o88,760 for inhibition of the 5~-reductase.
A preferred set of R substituents are:
CH3 CH3 CH3 < ~
20R-CH-CH20H, 20R-CH-CHO, 20R-CH-COOH) and --/

~2 ~,~, LV.

.. .

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As set forth hereinabove there is evidence that dihydrotestosterone (DHT) which is a metabolite of tes-tosterone has a stimulatory effect on sebaceous glands and thereby is involved in the pathogenesis of acne, and agents which inhibit the formation of DHT would be useful in the treatment of acne. The compounds employed in the present invention have been found to be inhibitors of testosterone 5~-reductase, the enzyme which transforms testosterone to the more active androgen DHT. HenceJ
the compounds employed in the present invention, that is, the compounds of the formulas set out above, being inhibi-tors of tes~osterone 5~-reductase are useful in the treat-ment of acne and oily skin conditions. The compounds are also useful for treatment of benign prostatic hyper-trophy and male pattern baldness.
The potency of an enzyme inhibitor is often expressedin terms of its inhibition constant, Ki, which is deFined mathematically in terms of established relationships of enzyme-catalyzed reaction rates to inhibitor and substrate concentrations. The derivation of these relationships and the mathematical definition of Ki are found in standard texts on enzymology. Simply stated, the Ki expresses quantitatively the combining power of the inhibitor for the enzyme; the lower the value of Ki, the greater the combining power (affinity). In the case of the substrate, the corresponding value is known as the Km, the relation-ship to combining power being roughly the same. The fol-lowing comparison exhibits the increase in combining power for 5~-reductase obtained in the compounds of ~his 3 series, over that of the substrate (testosterone) or one of the most potent prevlously known inhibitors.

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~4~ 8 Ki QH
1.0 x 10 ~M (Km) testosterone ~ 2.2 x 10 7M
0~

I~J 2.0 x 10 8M
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It can be seen that the representative compound of the present invention possesses 50 times the affinity of the substrate for the enzyme, and is at least 10 times as potent as the previous inhibitor.
The utility of the compounds employed in the present invention can be demonstrated by the ability of the com-pounds to inhibit the activity of 5a-reductase isolated from rat prostate gland. For example, using prostate microsomes containing testosterone 5a-reductase in an amount equivalent to 180 mg of fresh tissue, and 4-14C-testosterone at a concentration of 2 x 10 ~M, at 3 x 10 ~M (5a,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one was found to inhibit the conversion of testosterone to DHT and androstanediol (ADIOL) overall by 49~. Under the same conditions, a concentration of 3 x 10 7M
inhibited the conversion of testosterone to 5a-reduced products by 75~.

~1~4~08 M-903 CIP

To achieve the desired anti-acne or anti-seborrheic effect ~he compounds employed in the present invention can be administered orally, parenterally, for example, intra-muscularly and subcutaneously, and topically to a patient in need of treatment. Topical administration is pre-ferred. As used herein in association with the treatment of acne or oily skin the term patient is taken to mean a warm-blooded mammal, for example, primates, human males and females having an acne condition or an oily skin condition in need of treatment. The compounds of the invention can be administered alone or suitably admixed in the form of a pharmaceutical preparation to the patient being treated. The amount of compound administered will vary with the severity of the acne condition or oily skin condition and repetitive treatment may be desired. For oral and parenteral administration the amount of compound administered, that is, the anti-acne or anti-seborrheic effective amount, is from 0.1 to 50 mg/kg of body weight per day and preferably from 1 to 30 mg/kg of body weight per day. Unit dosages for oral or parenteral adminis-tration may contain, for example, from 5 to 200 mg of the active ingredient. For topical administration the anti-acne or anti-seborrheic effective amount of the com-pounds of the invention on a percent basis can vary from ~ ', .:
' ' ` - ' ' , :

.:

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0.001~ to 5~ and preferably from 0.005% to 1%. For topical administration the formulated active ingredient, that is, a compound of the invention can be applied directly to the site requiring treatment or can be applied to the oral or nasal mucosa. Applicator sticks carrying the formulation may be employed in administering the compounds.
In the treatment of benign prostatic hypertrophy (bph) the compounds of the invention may be administered in various manners to the patient being treated to achieve the desired effect. As used herein in the treat-ment of bph the term patient is taken to mean male warm blooded anima1s, such as male rats, male dogs and human males. The compounds can be administered alone or in combination with one another. Also, the compounds can be administered in the form of a pharmaceutical preparation.
The compounds may be administered orally, parenterally, for example, intravenously, intraperitoneally, intra-muscularly or subcutaneously, including injection of the ~ 20 active ingredient directly into the prostate. The amount of compound administered will vary over a wide range and can be any effective amount. Depending on the patient to be treated, the condition being treated and the mode of administration, the effective amount of compound adminis-

2~ tered will vary from about 0.1 to 50 mg~kg of body weight per day and preferably from 1 to 30 mg/kg of body weight per day. Unit dosages for oral or parenteral administra-tion may contain, for example, from 5 to 200 mg of a compound of the invention.
These dosage ranges represent the amount of compound that wtll be effecttve in reducing the size of the pros-tate, i.e., the amount of compound effective in treating bph. The compounds can be administered from onset of hypertrophy of the prostate to regression of the symptoms, and may be used as a preventive measure.
Topical formulation can be, for example, in the form of a solution, suspension, emulsion, gel or cream of )08 either the oil-in-water or water-in-oil type/ ointment~
paste, jelly, paint or powder. Suitable bases for the topical preparation may be of any conventional type such as oleaginous bases, for example, olive oil, cottonseed oil, petrolatum, white petrolatum, mineral oils, silicones, such as, dimethylpolysiloxane~ or methylphenylpolysiloxane, lanolines, polyethyleneglycol, glyceryl monvsterate, methylcellulose and hydroxymethylcellulose. The topical formulation may contain pharmaceutically acceptable sur-factants, wetting agents, dispersing agents, emulsifiers,penetrants, emollients, detergents, hardeners, preserva-tives~ fillers, anti-oxidants, perfumes, cooling agents, such as, menthol, soothing agents, such as, camphor, or coloring agents, such as, zinc oxide. Aerosol preparations of a solution, suspension or emulsion containing the active ingredient in the form of a finely ground powder can also be employed for topical administration. The aerosol may be packaged in a pressurized aerosol container together with a gaseous or liquified propellant, for example, dichlorodifluoromethane, dichlorodifluoromethane with di-chlorodifluoroethane~ carbon dioxide, nitrogen, or propane with the usual adjuvant such as cosolvent and wetting agents as may be necessary or desirable. The compounds may also be administered in a non-pressurized form such as in a nebulizer or atomizer.
Following are illustrative topical pharmaceutical formulations which may be employed in practicing the present invention:
SOLUTION
(5~-20R)-4-diazo-21-hydroxy-20-methyl- 0.85 9 pregnan-~-one Alcohol 78.9 ml Isopropyl myristate 5.0 9 Polyethylene glycol 400 10.02 Purified water qs ad 100 ml i~, ~- yv~
- ~154~VI~
g Combine the alcohol, isopropyl myristate and polyethylene glycol 400 and dissolve the drug substance therein. Add sufficient purified water to give 100 ml.
A GEL
5 ( a) ( 5a,20R ) -4-diazo-21-hydroxy-20- 0.85 g methylpregnan-3-one (b) A lcohol 78.9 ml tc) Isopropyl myristate 5.0 9 (d) Polyethylene glycol 400 10.0 9 (e) Carbopol 940 (Carboxypolymethylene) .75 9 10 (f) Triethylamine qs (g) Purified water qs ad 85 9 Disperse the Carbopol 9~0 in the isopropyl myristate. To 38 ml of alcohol add 7 ml of purified water and the poly-ethylene glycol 400 and mix. Combine the two phases and 15 mix until well dispersed. Add sufficient triethylamine to give a neutral pH. Dissolve the drug substance in the balance of the alcohol and mix well into the batch.
Add and mix sufficient purified water to provide 85 9 of finished product.

(a) (5,20R)-2~-diazo-21-hydroxy-20- 0.85 9 methylpregnan-~-one (b) Absolute alcohol 75 ml (c) Polyethylene glycol 400 10.0 9 (d) Isopropyl myrista~e 5~0 9 25 (e) Stearic acid 4 3 9 (f) Sodium hydroxide 0-55 9 (g) Purified water qs ad 85 9 Combine the absolute alcohol, polyethylene glycol 400 and isopropyl myristate. Add the stearic acid and heat the 30 mixture to about 65C. Dissolve the sodium hydroxide in a small amount of water, add and mix. Add sufficient water to provide 85 9 of finished product. After formation of the applicator stick base the drug substance is suspended therein immediately before depositing and solidifying 35 the formulation.

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* Trade Mark ~ - 9v~
S~008 AEROSOL FOAM
(a) (5a~20R)-4-diazo-21-hydroxy-20- 1.0 9 methylpregnan-3-one (b) Propylene glycol 96.o 9 (c) Emulsifying Wax NF XIV3.0 9 5 (d) Dichlorodifluoromethane:cryfluorane 6.9 9 Disso1ve the drug substance in the propylene glycol. Add the emulsifying wax and heat to approximately 70C. Stir while cooling to room temperature. Charge a suitable aerosol unit with this concentrate and 6.9 9 of dichloro-difluoromethane:cryofluorane (20:80).
TOPICAL CREAM, VANISHING o/w _ _ _ _ .
(a) (5a,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one (b) Stearyl alcohol 15 (c) Sorbitan Monostearate 2 15 (d) Polyoxyethylene Sorbitan Monostearate 2.3 (e) Propylene glycol 5 (f) Methylparaben 0.025%
(g) Propylparaben 0.015%
(h) Purified Water qs POWDER
(a) (5a,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one (b) Silicon dioxide, anhydrous 0.5 (c) Cornstarch, lactose, fine powder qs OLEAG ! NOUS OlNTMENT
(a) ( 5G, 2oR)-4-dlazo-2l-hydroxy-2o- 1 Methylpregnan-3-one (b) White wax 5 (c) White petrolatum qs 100 ABSORPTION OINTMENT BASE
(a) (5a~2oR)-4-diazo-2l-hydroxy-2 methylpregnan-3-one 30 (b) Cholesterol 3 (c) Stearyl alcohol 3 (d) White wax 8 (e) White petrolatum qs 100 * Trade Mark ... .

`` 11 Si4008 WATER SOLUBLE OINTMENT DOSE
. _ ... .
(a) (5a,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one (b) Polyethylene glycol 400 40 (c) Polyethylene glycol O qs 100 PASTE
(a) (5a,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one (b) Starch 25 (c) Zinc oxide 25 (d) White pretrolatum qs 100 AEROSOL FOAM
(a) (5a~20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one (b) Emulsifying wax 3 (c) Stearyl alcohol 2 (d) Diglycol stearate 2 15 (e) Propylene glycol . 92 T.he compounds in treating acne and an oily skin condition may be used in combination with other anti-acne preparations, antiseptics, antiinfective agents, keratolytic agents, for example, resorcinol, comedolytic agents, or agents having a retinoic acid-like action, corticoids or other antiinflammatory agents, thioglycolates, ethyl lactate or benzoyl peroxide. The followjng formulations are illustrative of pharmaceutical preparations for topical application comprising a compound in combination with a keratolytic agent.
AEROSOL FOAM
(a) (5a~2oR)-4-diazo-2l-hydroxy-2o- 1 methylpregnan-3-one (b) Resorcinol monoacetate 30 (c) Emulsifying wax NF 3 (d) Stearyl alcohol 2 (e) Diglycol stearate 2 (f) Propylene glycol 91 J,~>
11~4V08 Dissolve the drug substance in the propylene glycol. Add the emulsifying wax and heat to about 70C. Stir while cooling to room temperature. Charge a suitable aerosol unit with the concentrate and 6.9 g of dichlorodifluoro-5 methane:cryfluorane (20:80).
A GEL
(a) (5~,20R)-4-diazo-21-hydroxy-20- 0.85 9 methylpregnan-3-one (b) Resorcinol 0.85 9 (c) Alcohol 78.9 ml 10 (d) Isopropyl myristate 5.0 g (e) Polyethylene glycol 400 10.0 9 ~f) Carbopol 940 (carboxypolymethylene)0-75 9 (g) Triethylamine qs (g) Purified water qs ad 85 g Disperse the Carbopol 940 in the isopropyl myristate. To 38 ml of alcohol add 7 ml of purified water and the poly-ethylene glycol 400 and mix. Combine the two phases and mix until well dispersed. Add sufficient triethylamine to give a neutral p~. Dissolve the drug substance and the resorcinol in the balance of the alcohol and mix well into the batch. Add and mix sufficient purified water to provide 85 9 of finished product.
For oral administration the compounds can be formu-lated into solid or liquid preparations, such as, capsules, pills, tablets, troches, powders, solutions, suspensions or emulsions. The compounds can be applied in the form of an aerosol containing finely divided particles of the active ingredient or a solution, suspensionJ or emulsion of the active ingredient. The solid unit dosage forms can be a capsule which can be of the ordinary gelatin type containing the active compound and a carrier, for example, lubricants and inert filler such as lactose, sucrose, and corn starch. In another embodiment, an active compound of the invention can be tableted with conventi~nal tablet bases such as lactose, sucr~se and corn starch in combination with binders such as acacia, ~..
~i~ * Trade Mark ..... .

~ - 9J3 11540()8 c~rn starch or gelatin, disintegrating agents such as potato starch or alginic acids and a lubricant such as stearic acid or magnesium stearate.
For parenteral administration the compounds may be administered as injectable dosages of a solution or sus-pension of the compound in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid such as water-in-oi1 with or without the addition of a surfactant and other pharmaceutically accept-able adju~ants. Illustrative of oils which can be em-ployed in these preparations are those of petroIeumJ
animal, vegetable or synthetlc origin/ for example, peanut : oil, soybean oil and mineral oil. In general, water, saline, aqueous dextrose and related sugar solutions, ethanols and glycols, such as, propylene glycol or poly-ethylene glycol are preferred 1iquid carriersJ particularly for injectable solutions.
The compounds can be administered in the form of a depot injection or implant preparation which can be formu-lated in such a manner as to permit a sustained releaseof the active ingredient. The active ingredient can be compressed into pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. Implants may employ inert materials such as biodegradable polymers and synthetic silicones, for example, Silastic, silicone rubber manufactured by the Dow-Corning Corporation.
The following are illustrative pharmaceutical formu-lations suitable for oral or parenteral administration which may be employed in practicing the present invention:
TABLET
(a) (5~J20R)-4-diazo-21-hydroxy-20- 75 g methylpregnan-3-one (b) Lactose 1.216 Kg (c) Corn starch 0.~ Kg * Trade Mark J ~
S4~)0~3 Mix the active ingredient, the lactose and corn starchuniformly. Granulate with 10% starch paste. Dry to a moisture content of about 2.5%. Screen through a No. 12 mesh screen. Add and mix the fo1lowing:
(a) Magnesium Stearate 0.015 Kg (b) Corn starch qs ad 1.725 Kg Compress on a suitable tablet machine to a weight of 0.115 g/tablet.
SOFT GELATIN CAPSULE
(a) (5~,20R)-4-diazo-21-hydroxy-20- 0.25 Kg methylpregnan-~-one (b) Polysorbate 80 0.25 Kg (c) Corn oil qs ad 25.0 Kg Mix and fill into 50,000 soft gelatin capsules.
IM DEPOT INJECTION
Each 1 ml contains the following:
(a) (5~,20R)-4-diazo-21-hydroxy-20- 5.0 mg methylpregnan-3-one (b) Anhydrous chlorobutanol 5.0 mg (c) Aluminum monostearate 50.0 mg (d) Peanut oil qs ad 1.0 ml Dissolve or disperse the ingredients in the peanut oil.
DEPOT-IMPLANT
(a) (5,20R)-4-diazo-21-hydroxy-20- 5 mg methylpregnan-3-one (b) Dimethylsiloxane 240 mg (c) Catalyst qs Disperse the drug substance in the fluid dimethylsiloxane.
Add the catalyst and cast into a suitable monolytic struc-ture.
Alternatively, the drug substance may be enclosed by a precastJ polydimethylsiloxane envelope.
AlternativelyJ the drug substance may be dispersed in a suitable amount of hydroxyethyl acrylate subsequently polymerized and cross-linked by the addition of ethylene-dimethacrylate, and an oxidizing agentJ to yield a 3-dimensional ethylene glycomethacrylate moldable gel (Hydron) TFade Mark - ~ ' '. ' ' - , ~
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~15~V013 . .

IM INJECTIONS
A. Oil type:
(a) (5~20R)-4-diazo-21-hydroxy-20- 25 mg methylpregnan-3-one (b) BHA, BHT aa 0.01~ w/v 5 (c) Peanut oil or sesame oil qs 1.0 ml ~. Suspension type:
(a) (5a,20R)-4-diazo-21-hydroxy-20- 25 mg methylpregnan-3-one (b) Sodium carboxymethylcellulose 0.5~ w/v (c) Sodium bisulfite 0.02% w/v 10 (d) Water for injection, qs 1.0 ml BUCCAL OR SUBLINGUAL TABLET
(a) (5~,20R)-4-diazo-21-hydroxy-20- 1 methylpregnan-3-one (b) Calcium stearate 1%
(c) Calcium saccharin 0.02 15 (d) Granular mannitol qs Mix and compress on a suitable tablet machine to a weight of 0.115 g/tablet.
The compounds herein described can be prepared from the ~4-~ keto precursor~ R

~
o wherein R is as already described, but may be blocked, e.g., forming a siloxy ether or a ketal as appropriate and desirable during the reaction sequence. For instance the reaction sequence set out below, and followed in Example 1 employs a di-methyl tertiary butyl siloxy ether blocking group on the 17-substituent to protect the 17-substituent during the reaction sequence used to add a 4-benzoyl group. Then R may be unblocked and transformed to the ultimate substituent, e.g., oxidized to aldehyde or to an '~1 , .

~ , 11540l~E~ M-903 CIP

acid oxidation level then esterified, etc., before the diazo transfer reaction is carried out.
When R contains an alcoho', the starting material will be the alcohol itself. If R contains an aldehyde, acid or ester the corresponding alcohol is utilized as the starting material, and later the alcohol group is trans-formed into the aldehyde, etc.
The alcohol function is protected through forming the dimethyl-t-butyl silyl ether by the technique described by E. J. Corey et al., J. Am. Chem. Soc. 94, 6190 (lg72).
When RzCOCH3 (i.e., the starting material is proges-terone) the side chain is protected by first forming the ketal C~H3 The (protected) ~-keto ~4 steroid, e.g., Compound A
of the reaction se~quence flow sheet which follows, is subjected to dissolving metal reduction using Li in NH3 in aniline or tert-butyl alcohol for the proton donor at -78 to -33C for 1 to 60 minutes as generally described by G. Stork et al., J. Am. Chem. Soc. 96, 7114 (1974).
Then the enolate ion is trapped with trimethylsllyl chloride and the resulting enol ether, e.g., Compound B, isolated.

,, , 1~ 59~1~()8 M ~ 93 C I P

O~ ~ A

OS i~

B
~S iO H
~ OS i~

OH ~ C

O D O C~H5 O \C~3Hs R ~!

O C~H, ~2 OH

0~

11~40~8 The enolate anion is then regenerated from the enol ether using alkyl lithiums, e.g., methyl or butyl lithium in ethers, such as, tetrahydrofuran or diethyl ether at 0 - 25C for 1 to 60 minutes, and is reacted with benzoyl acid chloride, or a lower alkyl Cl-4 acid chloride alternatively, for 1-20 minutes at -100 to -70C in~
e.g., diethyl ether or tetrahydrofuran (Compound C).
Thereafter the alcohol function is unblocked with F ~ or acid as described by E. J. Corey et al., J. A~.
Chem. Soc. 9~, 6190 (1972) or else by reaction with a tetrafluoroborate salt such as the lithium, sodium, zinc, tin, magnesium, silver, potassium, triphenylcarbenium, trialkyloxonium (e.g., methyl, ethyl, propyl, butyl) tetra-fluoroborate in aprotic solvents, acetonitrile, dimethyl formamide, dimethylacetamide, ethers, methylene chloride, chloroform or combinations thereof at temperatures of from about 0 to 100C for 1 to 72 hours. The procedure will effect cleavage of the tert-butyldimethylsilyl ether to the corresponding alcohols, both primary and secondary.
(Compound D) When R=OH, CH20H or CH-CH20H or other alcohols as defined by R the diazo transfer reaction (to compound E) may follow directly, by treatment of equivalent amounts of Compound D, and trialkylamines, such as, triethylamine, with sodium hydride to generate the enolate, then p-toluenesulfonyl azide is added to introduce the diazo function as descrlbed by J. B. Hendrlckson et al., J. Org.
Chem. 33, 3610 (1968), usual1y performed in ether or tetrahydrofuran at 0 to 25C, 1 to 24 hours.

When R is other than o ~ or one of the alcohol containing groups conversion at the 17-position to the desired 17-substituent is carried out immediately prior to the diazo transfer reaction (as in Compound D to F
then to Compound G).

11$4~ M-903 CIP

-18a-The alcohol group may be ozidized to the aldehyde or ketone by standard procedures, for example, using pyridinium chlorochromate as described by E. J. Corey et al., Tet. Letts., 2647 (1975), or using chromium trioxide/pyridine (R. W. Ratcliffe, Org. Syn., 1973) or to the acid using Jones Reagent (R. Bowden et al., J.
Chem. Soc., 39 (1966). The acid may be converted to alkyl esters, using diazomethane for the methyl ester, and the alcohol saturated with HCl gas for higher ethyl es~ers, or via the acid chloride and the alcohol.

~.S4008 When R is OCO-alkyl these are obtained from the alco-hol by reaction with a C1~6 alkyl acid chloride or anhy-dride in pyridine as solvent (0-25C, 1 to 24 hours).
The diazo transfer reaction can then be conducted on the aldehydes, ketones, acids tUsing 2 equivalents of NaH), and esters.
In the instance wherein R = -COCH3 the actual start-ing material for the reaction sequence is the 20-ketal, the ketal protecting group being removed prior to the diazo transfer reaction by exchange in acetone or propa-none using ~-toluenesulfonic acid as the catalyst at 25-60C for 1 to 24 hours.

<o The furan (R = ~ ) may be utilized directly without need for any blocking groups and the reaction se~uence proceeds directly from the starting compound to the ultimate diazo compound.
The following examples describe preparation of the 4-diazo compounds of this invention.

Testosterone dimethyl t-butylsilylether (Compound A) A mixture of testosterone (1.0 g, 3.5 mmole), t-butyldimethylsilyl chloride (627 mg, 4.2 mmole) and imi-dazole (287 mg, 4.2 mmole) in dimethylformamide (4 ml) is stirred overnight at 40C. The mixt~lre is then poured into ice water and the resulting precipitate filtered off and recrystallized from methanol to afford 1.35 g.

3-Trimethylsiloxy-17~-(dimethyl-t-butylsiloxy)-5~-an-drost-3-ene (Compound B) Testosterone dimethyl-t-butylsilyl ether (12.0 g, 29.8 mmole) in tetrahydrofuran (70 ml) is added to ammonia containing aniline (2.7 g, 29.8 mmole) and lithium (625 mg, 89 mmole). After 1 hour the blue solution is treated dropwise with isoprene until the blue color is dissipated. The ammonia is allowed ~lS~o~

to evaporate and the residue dried under vacuum (O.S mm).
Tetrahydrofuran (50 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride (12 ml) and triethylamine (12 ml) which had previously been centrifuged. After 15 minutes the mixture is diluted with pentane and washed with chilled 0.5 M HCl, then chilled a~ueous sodium bicarbonate, then dried (MgSO4) and concentrated. The residue is crystallized from ethyl acetate to afford 6.6 g. (The mother liquors are chromatographed on silica gel. Elution with 10~
ether-pentane affords a fraction which is recrystallized from ethyl acetate (2.2 g).) M.P. 126C.

4-Benzoyl-17~~(dimethyl-t-butylsiloxy)-5~-androstan-3-one (Compound C) To the enol ether (4.89 g, 10.27 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M solution, 11.3 mmole). After 1 hour at 25C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.45 g, 10.3 mmole) in ether (30 ml) at -70C.
After 5 minutes aqueous ammonium chloride is added and the products isolated by ether extraction. The residue, after evaporation of the ether, is recrystallized from carbon tetrachloride to afford 2.0 g.
4-Benzoyl-17~-hydroxy-5~-androstan-3-one (Compound D) The silyl ether (1.64 g, 3.2 mmole) in dichlorome-thane (50 ml) is treated with tritylfluoroborate (1.27 g, 3.84 mmole) for 1 hour at 25C. This solution is then washed with aqueous ammonium chloride, dried and evapor-ated. The residue is chromatographed on silica gel. Elu-tion with 1~ methanol-chloroform afforded a fraction which is recrystallized from ethyl acetate-pentane (1.0 g).
4-Diazo-17~-hydroxy-5~-androstan-3-one (Compound E) The diketone (3.5 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50~
dispersion) in tetrahydrofuran (5 ml). After 30 minu-tes, tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether ' . . ~

1~ S41)0~3 is then added, the mixture filtered, then washed with water, dried and evapora~ed. The residue is chromato-graphed, the fraction eluted with 70% ether-petrol being collected. Recrystallization from chloroform-hexane afforded yellow crystals (26 mg), m.p. 171C.

4-Benzoyl-5~-androstane-3,17-dione 4-Benzoyl-17~-hydroxy-5-androstan-3-one made as in Example 1 (788 mg, 2 mmole) in 2 ml of CH2C12 is added to pyridinium chlorochromate (650 mg, 3 mmole) suspended in CH2C12 (2 ml) at 25C. After 2 hours, ether is added and the solvent decanted. This is then filtered through*Flo-risil, the eluate evaporated and the residue recrystal-lized from chloroform-heptane.
4-Diazo-5 -androstane-3,17-dione The triketone (350 mg, O.B mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70% ether-petrol being collected. Re-crystallization from chloroform-hexane afforded yellow crystals (35 mg).

The reaction sequence employed in this example is illustrated in the following formula sequence:

iB~ Trade Mark .

~1~545~ M-90~ C I P

il CH20H

0~ ' ~ 0~

~CH20S i ~ CH205 i~

-sio~ 0~

~CH.05 + ~CHzOH

O C~Hs O C~H5 ~CH20H
~P
0~

- . . ..

(20R)-21-Hydroxy-20-methyl~ n-4-en-3-one To the aldehyde starting material (16.4 g, 50 mmole) in ethanol (250 ml) and THF (50 ml) at 0C is added a solution of NaBH4 (675 mg, 12.5 mmole) in ethanol (125 ml) dropwise. The mixture is stirred at 25C overnight, then neutralized by the addition of glacial acetic acid, then concentrated on the rotary evaporator. The residue is taken up in chloroform and washed with saturated aqueous NaHCO3, then brine, then dried and concentrated. The residue is recrystallized from chloroform-pentane to af-ford a colorless solid (12.6 g, 77~). M.P. 132.5C.
(20R)-21-tDimethyl-t-butylsiloxy)-20-methylpregn-4-en-3-one A mixture of (20R)-21-hydroxy-20-methylpregn-4-en-3-one, t-butyldimethylsilyl chloride (10 g, 30.3 mmole) and imidazole in dimethylformamide (50 ml) is stirred overnight at 40C. The mixture is then poured into ice water and the resulting precipitate filtered off and re-crystallized from methanol to afford 8.6 g.
(5~,20R)-3-Trimethylsiloxy-21-(dimethyl-t-butylsiloxy)-20-methYlpregn-3-ene (20R)-21-(Dimethyl-t-but~lsiloxy)-20-methylpregn-4-en-3-one (2.0 g, 4.5 mmole) in tetrahydrofuran (20 ml) is added to ammonia containing aniline (420 mg, 4.5 mmole) and lithium (100 mg, 15 mmole). After 1 hour the blue solution is treated dropwise with isoprene until the blue color is dissipated. The ammonia is allowed to evaporate and the residue dried under vacuum (0.5 mm). Tetrahydro-furan ~20 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride (4 ml) and triethylamine (4 ml) which had previously been centrifuged. After 15 minutes the mixture is diluted with pentane and washed with chilled 0.5 M HCl, then chilled aqueous sodium bicarbonate, then dried (MgSO4) and con-centrated. The residue is crystallized from ethyl acetate to afford 1.6 g. M.P. 113C.
(5~,20R)-4-Benzoyl-21-(dimethyl-t-butylsiloxy)-20-methyl-~regnan-3-one To the enol ether (5~,20R)-3-trimethylsiloxy-21-(dimethyl-t-butylsiloxy)-20-methylpregn-3-ene (5.08 g, 9.8 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M solution, 11.3 mmole). After 1 hour at .~..... . .
.

1 15~008 25C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.54 g, 1.27 mmole) in ether (30 ml) at -70C. After 5 minutes aqueous ammonium chloride is added and the products isolated by ether extraction. The residue, after evaporation of the ether, is recrystallized from chloroform-heptane to afford 450 mg~ M.P. 169C.
(5~,20R)-4-Benzoyl-21-hydroxy-20-methylpre~nan 3-one The silyl ether (5~,20R~-4-benzoyl-21-(dimethyl-t-butylsiloxy)-20-methylpregnan-3-one (1.06 g, 2.0 mmole) in dichloromethane (50 ml) is treated with tritylfluoro-borate (660 mg, 2 mmole) for 1 hour at 25C. This solu-tion is then washed with aqueous ammonium chloride, dried and evaporated. The residue is chromatographed on silica gel. Elution with 1% methanol-chloroform affords a frac-tion which is recrystallized from ethyl acetate-pentane (500 mg). M.P. 236-238C.
(5~,20R?-4-Diazo-21-hydroxy-20-methylpregnan-3-one The (5~,20R)-4-benzoyl-21-hydroxy-20-methylpregnan-3-one (436 mg, 1.0 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dispersion) in tetrahydrofuran (S ml). After 30 minutes, tosylazide (196 mg, 1.0 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mix-ture filtered, then washed with water, dried and evapora-ted. The residue is chromatographed on silica gel, the fraction eluted with 70% ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (30 mg), M.P. (210 decomp.), of (5~,20R)-4-diazo-21-hydroxy-20-methylpregnan-3-one.

Using (5~,20R)-4-benzoyl-3-oxopregnane-20-carboxalde-hyde; (5~,20R)-4-benzoyl-3-oxopregnane-20-carboxylic acid esters; 4-benzoyl-5~-androstane-3,17-dione; 4-benzoyl-5~-pregnane-3,20-dione etc., the corresponding 4-diazo-steroids may be prepared by the diazo transfer reaction as in Example 2.
~ i;

llS~008 EXAMPLE: _4A
(5~, 20R) -4-Benzoyl-3-oxopregnane-20-carboxaldehYde (5~,20R)-4-Benzoyl-21-hydroxy-20-methylpregnan-3-one of Example 3 (872 mg, 2 mmole) in dichloromethane (2 ml)

5 is added to pyridinium chlorochromate (650 mg, 3 mmole) suspended in dichloromethane (2 ml) at 25C. After 2 hours ether (20 ml) is added and the solvent decanted.
This is then filtered through Florisil, the eluate evaporated and the residue recrystalliæed from chloroform-heptane.(5~,20R)-4-Diazo-3-oxopreg ~
The diketone (350 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70% ether-petrol being collected. Recry-stallization from chloroform-hexane affords yellow cry-stals (15 mg).

(5~,20R)-4-BenzoYl-3-oxopregnane-20-carboxylic acid The alcohol of Example 3 (872 mg, 2 mmole) in acetone (5 ml) is treated with excess Jones reagent at 25C for 4 hours. Saturated sodium chloride is then added and the mixture extracted with chloroform. The organic phase is washed wellllwith aqueous sodium chloride, then dried and evaporated. The residue is recrystallized from methanol.
Treatment with ethereal diazomethane in excess over-night, followed by evaporation of the solvent leads to the methyl ester.
Treatment of 300 mg of acid with a saturated solu-tion of isobutylene in methylene chloride (30 ml) over-night at 25C containing H2SO4 (2 drops) as catalyst Y ~n~
* Trade Mark .

o~

affords the t-butyl ester. Conduct of the diazo transfer reaction on the 20~-carboxylate is below exemplified by transformation of the t-butyl ester.
t-Butyl (5~,2QR)-4-diazo-3-oxopregnane-20-carbox~late The t-butyl ester diketone (400 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50~ dispersion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mmole) in tetrahydro-furan is added and the mixture stirred overnight at 25C.
Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chro-matographed, the reaction eluted with 70~ ether-petrol being collected. Recrystallization from chloroform-hexane affords yellow crystals (40 mg).

The reaction sequence employed in this example is illustrated in the following formula sequence.

1154001!~ M-903 C I P

CH2 OH CH2 OS i+
O ~ 0~

~-SiO~
~H, 0~ )0~
o' CoH ~5 N2 ~iS4~

17~-(Dimethyl-t-butylsiloxymethyl)androst-4-en-3-one A mixture of 17~-hydroxymethylandrost-4-en-3-one (1.0 g, 3.5 mmole), t-butyldimethylsilyl chloride (627 mg, 4.2 mmole) and imidazole (287 mg, 4.2 mmole) in dimethyl-formamide (4 ml) is stirred overnight at 40C. The mix-ture is then poured into ice water and the resulting pre-cipitate filtered off and recrystallized from methanol to afford 1.35 g.
3-Trimethylsiloxy-17~-(dimethyl-t-butylsiloxymethyl)-5a-androst-3-ene 17~-(Dimethyl-t-butylsiloxymethyl)androst-4-en-3-one (12.0 g, 29.8 mmole) in tetrahydrofuran (70 ml) is added to ammonia containing aniline (2.7 g, 29.8 mmole) and lithium (625 m, 89 mmole). After 1 hour the blue solution is treated dropwise with isoprene until the blue color dissipates. The ammonia is allowed to evaporate and the residue dried under vacuum (0.5 mm). Tetrahydrofuran (50 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride (12 ml) and triethylamine (12 ml) which had previously been centri-fuged. After 15 minutes the mixture is diluted with pen-tane and washed with chilled 0.5 M HCl, then chilled aque-ous sodium bicarbonate, then dried (MgSO4) and concen-trated. The residue is crystallized from ethyl acetate to afford 6.6 g. The mother liquors are chromatographed on silica gel. Elution with 10~ ether-pentane affords a fraction which is recrystallized from ethyl acetate (2.4 g)-4-Benzoyl-17~-(dimethyl-t-butylsilox~methyl)-5~-andros-tan-3-one To the enol ether (4.89 g, 10.27 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M
solution, 11.3 mmole). After 1 hour at 25C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.45 g, 10.3 mmole) in ether (30 ml) at 70C. After 5 minutes aqueous ammonium chloride is added and the products isolated by ether extraction.

~.lS~

The residue, after evaporation of the ether, is recrys-tallized from carbon tetrachloride to afford 2.2 9.
4-Benzoyl-17~-hydroxymethyl-5a-androstan-3-one The silyl ether (1.64 g, 3.2 mmole) in dichloro-methane (50 ml) is treated with tritylfluoroborate (1.27g, 3.84 mmole) for 1 hour at 25C. This solution is then washed with aqueous ammonium chloride, dried and eva-porated. The residue is chromatographed on silica gel.
Elution with 1~ methanol-chloroform affords a fraction which is recrystallized from ethyl acetate-pentane (800 mg).
_-Diazo-17g-hydroxymethyl-5a-androstan-3-one The diketone (350 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes,tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70~ ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (30 mg).

The 4-benzoyl-l7~-hydroxymethyl-5a-androstan-3-one can be oxidized to the corresponding 17~ aldehyde or 17 acid steroids, then converted to the 4-diazo derivative.
(A) Methyl 4-benzoyl-3-oxo-5a-androstane-17~-carboxylate The alcohol (872 mg, 2 mmole) in acetone (5 ml) is treated with excess Jones reagent at 25C for 4 hours.
Saturated sodium chloride is then added and the mixture extracted with chloroform. The organic phase is washed well with aqueous sodium chloride, then dried and evaporated. For recovery of the 17~-carboxylic acid the residue is recrystallized from methanolO For production of 17~-carboxylate esters, e.g., the methyl ester, the residue is treated with an excess of etheral diazo-~.~

~1541301~

methane. After 10 minutes the solvents are evaporated and the residue recrystallized from methanol.
Methyl 4-diazo-3-oxo-S~-androstane-17~-carboxylate The diketone (350 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mm91e) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70% ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (30 mg).
4-Diazo-3-oxo-5~-androstane-17~-carboxylic acid The 17~-carboxylic acid (350 mg, 0.8 mmole) in tetra-hydrofuran (2.0 ml) is added to sodium hydride (96 mg, 2 mmole of a 50% dispersion) in tetrahydrofuran t5 ml).
After 30 minutes, tosylazide (157 mg, 0.8 mmole) in te-trahydrofuran is added and the mixture stirred overnight at 25C. Acetic acid (60 mg) is then added followed by ether, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the reac-tion eluted with 70% ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (20 mg).
(B) 4-Benzoyl-3-oxo-5~-androstane 17~-carboxaldehyde The alcohol (872 mg, 2 mmole) in dichloromethane (2 ml) is added to pyridinium chlorochromate (65 mg, 3 mmole) suspended in dichloromethane (2 ml) at 25C. After 2 hours ether (20 ml) is added and the solvent decanted.
This is then filtered through Florisil, the eluate eva-porated and the residue recrystallized from chloroform-heptane.
4-Diazo-3-oxo-5~-androstane-17~-carboxaldehYde . ~
The diketone (350 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50 ~! * Trade Mark .

~ ~ ~j4~ 8 dispersion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mmole~ in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the reac-tion eluted with 70% ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (26 mg).

One g of 4-benzoyl-17~-hydroxy-5-androstan-3-one prepared as in Example 1 in pyridine (5 ml) and acetic anhydride (5 ml) is maintained at 25C for 10 hours, then diluted with ether. The ether solution is washed with 1 N
HCl, saturated NaHCO3, then dried, evaporated and the residue crystallized from ethyl acetate to afford the 17-acetate.
4-Diazo-17~-acetoxy-5~-androstan-3-one The triketone (350 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50~ dis-persion) in tetrahydrofuran ~5 ml). After 30 minutes,tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70% ether-petrol being collected. Re-crystallization from chloroform-hexane affords yellow crystals (30 mg).

Conversion of progesterone is illustrated in this example.
3-Trimethyl5l10xy-20-ethylenedioxy-5c~-pregn-3-ene Progesterone 20-ethylenedioxy ketal (10.7 g, 30 mmole) in tetrahydrofuran (70 ml) is added to ammonia containing aniline (2.7 g, 30 mmole) and lithium (625 mg, 89 mmole).
After 1 hour the blue solution is treated dropwise with isoprene until the blue color dissipates. The ammonia ..-~i.l. ~
. ~-11~41~138 is allowed to evaporate and the residue dried under vacuum (0.5 mm). Tetrahydrofuran (50 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride ~12 ml) and triethylamine (12 ml) which had previously been centrifuged. After 15 minutes the mixture is diluted with pentane and washed with chilled 0.5 M HCl, then chilled aqueous sodium bicar-bonate, then dried (MgSO4) and concentrated. The residue is crystallized from ethyl acetate to afford 6.0 g.
4-Benzoyl-20-ethylenedioxy-5~-pregnan-3-one To the enol ether (4.4 g, 10.27 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M solution, 11.3 mmole). After 1 hour at 25C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.45 g, 10.3 mmole) in ether (30 ml) at -70C.
After 5 minutes aqueous ammonium chloride is added and the products isolated by ether extraction. The residue, after evaporation of the ether, is recrystallized from carbon tetrachloride to afford 2.1 g.
The acetal (1.1 g) in acetone (100 ml) containing ~-toluenesulfonic acid (100 mg) is stirred overnight at 25C, then the solvent evaporated. The residue is dis-solved in ether and washed with aqueous NaHCO3, then dried and evaporated to afford the triketone (800 mg).
4-Diazo-5~-pregnane-3,20-dione -The triketone (355 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70% ether-petrol being collected. Recry-stallization from chloroform-hexane affords yellow cry-stals (40 mg).
~., '~j , 11~4008 Conversion of 4',5' dihydrospiro landrost-4-ene-17,2' ~3'H) furan]-3-one is illustrated in the following formula sequence:

0~ ~ iO~

0--~ ' O ~
~\ N2 4',5'-Dihydrospiro[3-trimethylsiloxy-5~-androst-3-ene-17,2'(3'H)furan]
Cyclic ether (9.8 g, 30 mmole) in tetrahydrofuran (70 ml) is added to ammonia containing aniline ~2.7 g, 30 mmole) and lithium (625 mg, 89 mmole). After 1 hour the blue solution is treated dropwise with isoprene until the blue color dissipates. The ammonia i5 allowed to eva-porate and the residue dried under vacuum (0.5 mm).
Tetrahydrofuran (50 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride (12 ml) and triethylamine (12 ml) which had previously been centrifuged. After 15 minutes the mixture is diluted with pentane and washed with chilled aqueous sodium bicarbonate, then dried (MgS04) and concentrated.
The res due is crystallized from ethyl acetate to afford 7.0 g.

. ,, ~1~41;~(3~3 4',5'-Dihydrospiro[4-benzoyl-5~-androstane-17,2'~3'-H)-_ furan]-3-one .
To the enol ether (4.5 g, 10.27 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M solution, 11.3 mmole). After 1 hour at 25C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.45 g. 10.3 mmole) in ether (30 ml) at -70C.
After 5 minutes aqueous ammonium chloride is added and ~he products isolated by ether extraction. The residue, after evaporation of the ether, is recrystallized from carbon tetrachloride to afford 2.2 g.
4',5'-Dihydro[~-diazo-5~-androstane-1_,2'(3'-H)furan]-3-one The diketone (320 mg, 0.8 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dis-persion) in tetrahydrofuran (5 ml). After 30 minutes,tosylazide (157 mg, 0.8 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mixture filtered, then washed with water, dried and evaporated. The residue is chromatographed, the frac-tion eluted with 70~ ether-petrol being collected. Recry-stallization from chloroform-hexane affords yellow cry-stals (35 mg).
Alternatively, the protection-deprotection sequence may be achieved by beginning with an aldehyde, for exam-ple, as illustrated in the following sequence by compoundH protected as an acetal by treatment of 1 equivalent of aldehyde with 1 equivalent of ethyleneglycol using ~-toluenesulfonic acid as a catalyst in benzene or toluene solution at a temperature of about 80 to 120C and re-moving excess water, for example, by using a Dean Starktrap. The aldehyde, protected as the acetal, is then sub-jected to dissolving metal reduction using lithium in ammonia in aniline or tert-butanol for the proton at -78 to 33C for 1 to 60 minutes. The enolate ion is trapped with trimethylsilyl chloride as described above. The enolate anion is regenerated using alkyl lithiums, , ., ~ . ~

. .

' .

' e.g., methyl or butyl lithium in ethers, such as, tetra-hydrofuran or diethyl ether at 0 to 25C for 1 to 60 minutes and is reacted with benzoyl chloride or a lower alkyl acid chloride alternatively for 1 to 20 minutes at -100 to -70C in, e.g., diethyl ether or tetrahydrofuran to give Compound L.
The aldehyde function is regenerated by treatment with acetone and butanone in the presence of a catalytic amount of p-toluenesulfonic acid or mineral acid for 1 to 24 hours at 25 to 50C. The diazo group may then be introduced at the 4-position of the aldehyde derivative directly, or the aldehyde derivative first may be con-verted to the corresponding acid by procedures generally known in the art, for example, by oxidation with silver oxide or by Jones reagent or reduction to the corres-ponding alcohol by procedures generally known in the art, by treatment with borohydride in lower alcohols, e.g., ethanol or tetrahydrofuran at 0 to 25C for 1 to 12 hours with subsequent introduction of the diazo group. The diazo transfer is achieved using ~-toluenesulfonyl azide as described hereinabove.

(5a,20R)-4-Diazo-3-oxopregnan-20-carboxaldehyde (20R)-21-Ethylenedioxy-20-methylpregn-4-en-3-one (2.0 g, 4.5 mmole) in tetrahydrofuran (20 ml) is added to ammonia containing aniline (420 mg, 4.5 mmole) and lithium (100 mg, 15 mmole). After 1 hour the blue solution is treated dropwise with isoprene until the blue color dissipates. The ammonia is allowed to evaporate and the residue dried under vacuum (0.5 mm). Tetrahydro-furan (20 ml) is then added, the solution cooled to 0C, and treated with a solution of trimethylsilyl chloride (4 ml) and triethylamine (4 ml) which had previously been centrifuged. After 15 minutes the mix-ture is diluted with pentane and washed with chilled 0.5M HCl, then chilled aqueous sodium bicarbonate, then dried (MgSO4) and concentrated. The residue is crystallized from ethyl acetate to afford 1.6 g.
To the enol ether (5~,20R)-3-trimethylsiloxy-21-ethylenedioxy-20-methylpregn-3-ene (5.08 g, 9.8 mmole) in ether (20 ml) is added methyl lithium (5.5 ml of a 2.05 M
solution, 11.3 mmole). After 1 hour at 25~C the solution is taken up in a syringe and added slowly to a solution of benzoyl chloride (1.54 g, 1.27 mmole) in ether (30 ml) at -70C. After 5 minutes aqueous ammonium chloride is added and the products isolated by ether extraction. The resi-due, after evaporation of the ether, is recrystallized from chloroform-heptane to afford 450 mg.
The acetal (1 g) in acetone (200 ml) containing ~-toluenesulfonic acid (50 mg) is stirred overnight at 25C, then concentrated. The residue is crystallized from di-chloromethane-heptane to afford the aldehyde.
The (5~,20R)-4-benzoyl-3-oxopregnane-20-carboxalde-hyde (436 mg, 1.0 mmole) in tetrahydrofuran (2.0 ml) is added to sodium hydride (48 mg, of a 50% dispersion) in tetrahydrofuran (5 ml). After 30 minutes, tosylazide (196 mg, 1.0 mmole) in tetrahydrofuran is added and the mixture stirred overnight at 25C. Ether is then added, the mix-ture filtered, then washed with water, dried and eva-porated. The residue is chromatographed on silica gel, the fraction eluted with 70% ether-petrol being collected.
Recrystallization from chloroform-hexane affords yellow crystals (30 mg), of (5~,20R)-4-diazo-3-oxopregnane-20-carboxaldehyde.

,, .

M-903CI'' 0~ 0 ~ TM50 ~J
O ~H5 or alkyl C1_~

~ ~r,ll ~ OOH

a~ ~ N
O~ \C~Hs or C" ~C~Hs or alkyl Cl_4 alkyl C1_4 ~ ~r 8 ~ ~ CH20H

0~ t~-~ W
alkyl C1_g In the above reaction sequence TMSO means trimethylsilyl-oxy .

1154~08 -38~

CH2o$ i ~ CH20H

~ LiBF, ) ~

To the silyl ether (5.5 9, 10 mMole) in methylene chloride (200 ml) and acetonitrile (125 ml) is added lithium tetrafluoroborate (2.8 9, 30 mMole) and the mix-ture is stirred at room temperature for oO hours. Themixture is then washed with water, aq. NaHC03 and aq.
NaCl then dried (MgS04) and concentrated. The residue is recrystallized from methylene-chloride-heptane to afford the alcohol (3.8 9~ 86~).

,~

Claims (8)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for the preparation of a compound of the formula wherein R is: =O, -OH, -OCO-alkylC1-5, -COOH, -CH2OH, -CHO, -COOalkylC1-6, -COCH3, , , and the 20R isomers of , , or which comprises reacting a compound having the formula wherein R is as defined above being blocked when appro-priate forming a siloxy ether or a ketal, subjecting said compound to dissolving metal reduction and reaction with trimethylsilyl chloride forming thereby a compound of the formula then reacting with an alkyl lithium and with benzoyl chloride or a lower alkyl C1-4 acid chloride forming thereby a compound of the formula then removing any siloxy ether or ketal blocking groups in R, said siloxy ether being removed by treatment with fluoride ion, acid, or a tetrafluoroborate salt and said ketal being removed by treatment with acetone or butanone in the presence of a catalytic amount of p-toluenesulfonic acid, thereafter reacting with sodium hydride or a lower trialkylamine and p-toluenesulfonyl azide forming thereby a compound of the formula

2. A process according to Claim 1 for the pre-paration of a compound of the formula wherein R is the 20R-isomer of , or , which comprises converting a compound of the formula to the 21-acetal, subjecting said compound to dissolving metal reduction and reaction with trimethylsilyl chlo-ride forming a compound of the formula then reacting with an alkyl lithium and with benzoyl chloride or a lower alkyl acid chloride forming a compound of the formula then regenerating the aldehyde function by treatment with acetone or butanone in the presence of a catalytic amount of p-toluenesulfonic acid followed by treatment with sodium hydride or a lower trialkylamine and p-toluenesulfonyl azide with conversion of the aldehyde to the appropriate acid or alcohol as desired prior to treatment with p-toluenesulfonyl azide.

3. A process according to Claim 1 for the pre-paration of (5a,20R)-4-diazo-21-hydroxy-20-methylpreg-nan-3-one which comprises reacting (20R)-21-hydroxy-20-methylpregn-4-en-3-one with t-butyldimethylsilyl chloride to give the corresponding 21-(dimethyl-t-butylsiloxy) ether, subjecting said compound to dis-solving metal reduction and reaction with trimethyl-silyl chloride to give (5a,20R)-3-trimethylsiloxy-21-(dimethyl-t-butylsiloxy)-20-methylpregn-3-ene, then reacting with an alkyl lithium and with benzoyl chloride or a lower alkyl C1-4 acid chloride to give a compound of the formula removing the siloxy group by treatment with fluoride ion, acid, or a tetrafluoroborate salt followed by reaction with sodium hydride or a lower trialkylamine and p-toluenesulfonyl azide to give the desired compound.

4. A process according to Claim 1 for the pre-paration of (5a,20R)-4-diazo-21-hydroxy-20-methylpreg-nan-3-one which comprises reacting (20R)-21-hydroxy-20-methylpregn-4-en-3-one with t-butyldimethylsilyl chloride to give the corresponding 21-(dimethyl-t-butylsiloxy) ether, subjecting said compound to re-duction with lithium and ammonia containing aniline and reaction with trimethylsilyl chloride to give (5a,20R)-3-trimethylsiloxy-21-(dimethyl- -butylsiloxy)-20-methylpregn-3-ene, then reacting with methyl lithium and with benzoyl chloride to give (5a,20R)-4-benzoyl-21-(dimethyl-_-butylsiloxy)-20-methylpregnan-3-one, removing the siloxy group by treatment with trityl fluoroborate, followed by reaction with sodium hydride and p-toluenesulfonyl azide to give the desired compound.

5. A compound of the formula wherein R is: =O, -OH, -OCO-alkylC1-5, -COOH, -CH2OH, -CHO, -COOalkylC1-6, -COCH3, , , and the 20R isomers of , , ~;

or whenever prepared by the process of Claim 1.

6. A compound of the formula wherein R is the 20R-isomer of , or whenever prepared by the process of Claim 2.

7. (5a,20R)-4-Diazo-21-hydroxy-20-methylpregnan-3-one whenever prepared by the process of Claim 3.

8. (5a,20R)-4-Diazo-21-hydroxy-20-methylpregnan-3-one whenever prepared by the process of Claim 4.