IL44903A - 17beta-hydroxy-16,16-dimethylestr-4-en-3-one its preparation and pharmaceutical compositions containing it - Google Patents

Abstract

Preparation of 17 beta -hydroxy-16,16-dimethylestr-4-en-3-one and its unexpected antiandrogenic and antisebum activities are disclosed. [US3853926A]

Description

44903/2 -|»-4— τβοκ *ηΒ»ι 6,,16-»ο ιπ»π- βΐ7 ο»7·>3θη ηιπρη '-I'som ina^n , † ηκ-3 urns 17 β-Hydroxy-l 6,16-dimethylestr÷4-en-5-one , preparation and pharmaceutical compositions containing them G.D. SEARLE & CO.

C:- 42941 t 173-HYDROXY-16,16-DIMETHYLESTR- -EN-3-ONE This invention relates to a new, useful, and unobvious steroid having the chemical formula and further characterized by antiandrogenic activity and the capacity to suppress the secretion of sebum.

The antlandrogenic activity of the steroid of this invention is remarkable because the corresponding l6-monomethyl steroid, a compound prepared by the instant inventor a number of years ago as disclosed in Example 7 of U.S. 3,049,555, has the opposite biological effect: it is androgenic. The monomethyl steroid also promotes anabolism, a possibly complicating side-effect which the dimethyl steroid of this invention does not share. Likewise androgenic and anabolic is another steroid which, were it not for unexpected difference in activity, might be considered closely-related, namely, 171 $-hydroxyspiro[cyclopropane- a compound patented by the instant inventor in U.S. 3,280,156 and described in Example 9 thereof.

Tabulated immediately below are the results of standardized tests for anabolic and androgenic activity showing the unexpected absence of such activity which dis tinguishes 178-hydroxy-l6,l6-dimethylestr-4-en-3-one. The procedure was essentially that described by Saunders and Drill, Proc. Soc . Exp. Biol. Med.. , 6 6 (1957). Male Badger strain rats were castrated at 22-24 days of age; and to each of a group of 5 or more such animals, beginning 19-21 days later, the compound to be assayed, dissolved or suspended in corn oil, was administered intramuscularly in equally divided doses on each of 7 successive days. Commonly, the initial intramuscular total dose is 5 mg. of compound in 0.7 ml. of corn oil. A second group of 5 or more animals likewise and concurrently administered corn oil alone served as controls. On the day after treatment was concluded, the animals were sacrificed; and (1) the seminal vesicle and ventral prostate glands and (2) the levator ani muscles were excised and dissected free of extraneous tissue. Fluid was expressed from the vesicles (but not the prostates or muscles), whereupon the glands and muscles were blotted and weighed. A compound is considered androgenic if the mean weight of the vesicles in the group of animals treated therewith is significantly (P ≤ 0.01) greater than the corresponding weight in the control group and there is a proportionate increase in the mean prostate weight for treats vis-a-vis controls. A compound is anabolic if the mean weight of the levator ani muscles in the group of animals treated is significantly (P < 0.01) greater than the corresponding weight in the control group. Compounds found active are commonly reassayed at progressively lower doses to determine potency. In the table, 176-hydroxy-l6 ,16-dimethylestr-iJ-en-S-one is designated "dimethyl", "mono-methyl" refers to 176-hydroxy-l68-methylestr-il-en-3-one, and "cyclopropyl" refers to 17 ' 3-hydroxyspiro[cyclopropane- 1 jie'-estr-j-enl-S'-one. Each steroid was administered intramuscularly.

Total Mean Wt. (mg.) Dose Seminal Ventral Levator Compound (mg.) Vesicles Prostate Ani cyclopropyl 1.0 10.9* 11.2* 08.2* control 0 7 - 5 9.2 52. cyclopropyl II 0.5 9.3* 10.3 00.7* 0.2 9.1* 11.4 69*2* II 0.1 7.8 10.0 60.4 control 0 7.6 9.3 56.2 *statistically greater than concurrent controls at the 1$ level of probability (P S 0.01) **statisticaily greater than concurrent controls at the 5# level of probability (P s? 0.05) The antiandrogenic utility of 17P-hydroxy-l6, 16- dimethylestr-4-en-3-one is evident from the results of a test for its capacity to inhibit the testosterone-stimulated growth of peminal vesicle and prostate glands in castrated immature male rats. The procedure is as follows: 20 male Charles River rats are castrated at 22 days of age and separated into 2 equal groups such that for each animal in each group there is a littermate in the other. When the mixture of 8 mg. of testosterone, 8 mg. of Carbowax (polyethylene glycol), and 16 mg. of Flexo Wax C Light (noncrystalline hydrocarbon wax melting at 60-64 °C.) is inserted under the skin of each rat dorsally in the neck region.

Beginning on the day of implant, the compound to be tested — dissolved or suspended in sesame oil, corn oil, or other physiologically inert vehicle — is administered subcutaneously to each animal in. one group at a dose of 10 mg. per kg. daily for 24 successive days. The quantity of vehicle is such that a l80-gm. rat receives approximately 0.1 ml. at the beginning of the test and —■ with increasing body weight — approximately 0.2 ml. at the end. Each animal in the second group receives concurrent subcutaneous daily injections of vehicle likewise adjusted to body weight, and these animals serve as controls. On the day after treatment is concluded, the animals are sacrificed; and' the seminal vesicle and ventral prostate glands arc excioed and dincected free of extraneous tissue. Fluid is expressed from the vesicles (but not the prostates), whereupon the glands are blotted 1 and weighed. A compound is considered antiandrogenic if the mean weight of the vesicles in the group of animals treated therewith is significantly (P ≤ 0.01) less than the corresponding weight in the control group and there is a proportionate decrease in the mean prostate weight for treats vis-a-vis controls. 17β- Hydroxy-l6,l6-dimethylestr-4-en-3-one was active in this test.

The antisebum utility of 178-hydroxy-l6,l6- dimethylestr-4-en-3-one is evident from the results of a standardized test for its capacity to decrease the amount of hair fat in rats, carried out essentially as described by P. J. Ebling in Proc . Roy . Soc . Med. , 62, 890 (1969). 17S-Hydroxy-l6,l6-dimethylestr-4-en- 3-one was active subcutaneously at the 5 mg. per kg. dose level in this test.

The compound of the instant invention is prepared by the acid hydrolysis of a 16,16-dimethyl- estran-173-ol wherein the A-ring. contains a covered 3- keto-A^ structure. A covered structure refers to a steroid structure which can be hydrolyzed to yield h the 3-keto-A system. Examples of such covered structures are : (alkyl) enol ether) (a ketal) wherein in the alkyl group has 1 to 5 carbon atoms .

Examples of acids which are suitable for conducting the hydrolysis are strong mineral acids (e.g., hydrochloric acid, sulfuric acid), sulfonic acids and strong organic acids (e.g., trichloroacetic acid). Solvents in which the steroid will dissolve such as tetrahydrofuran , acetone or lower alkanols (e . . , methanol) are used. The reaction is usually run at room temperature and pressure.

The ' invention will appear more fully from the examples which follow. Those examples are given by way of illustration only and are not to be construed as limiting the invention either in spirit or in scope as many modifications both in materials and methods will be apparent to those skilled in the art. Temperatures are given in degrees Centigrade (°C.) and quantities of materials in parts by weight unless parts by volume is specified. The relationship between parts by weight and ..parts by volume is the same as that existing between grams and milliliters.

PREPARATION OF STARTING MATERIALS EXAMPLE A A solution of 377 parts of 3-methoxy-l6 ,16-dimethylestra-l, 3,5 (10 )-trien-17-one and 30 parts of sodium tetrahydroborate (1-) in 12,000 parts of 2-propanol was heated at the boiling point under reflux overnight. The resultant solution was cooled, acidified with a slight excess of acetic acid, concentrated to approximately 1/10 volume by vacuum distillation, and thereupon diluted with sufficient water to effect precipitation. After chilling, the precipitate was filtered off, washed with water, dried in air, and then recry-stallized from a mixture of dichloromethane and ethyl acetate. The product thus isolated is 3-methoxy-l6 ,16-d.imethylestra-l,3,5(10)-trien-173-ol, melting at 130-133°C. and further characterized by a specific rotation — at 27°C. in 1.09 chloroform solution — of +72.5° (referred to the D line of sodium).

EXAMPLE B A solution of 31 ^ parts of 3-methoxy-l6,l6- dimethylestra-l,3,5(10)-trien-17S-ol in i)500 parts of tetrahydrofuran was added to a stirred solution of 4000 parts of tert-butyl alcohol in 105,000 parts of redistilled liquid ammonia. Sufficient sodium metal was introduced portionwise with continued stirring to maintain a deep blue color for 2 hours, whereupon the ^ammonia was distilled off and the residual mixture steam distilled. Prom the chilled distillate, the granular solid was filtered out, washed with water, and dried to 60°C. in vacuo. The product thus isolated is 3-methoxy-l6,l6-dimethylestra-2,5(10)-dien-173-ol melting in the range 140-147° C.

• EXAMPLE C A solution of 350 parts of 3,3-dimethoxy- I6,l6-dimethylestr-5(10)-en-17-one in tetrahydrofuran was mixed with 500 parts of lithium aluminum hydride at -10°C. for 5 minutes. 900 Parts of ethyl acetate was added followed by 50 parts of water, 50 parts by volume of ^ sodium hydroxide and 150 parts of water. The reaction mixture was then filtered, the solvent distilled and the residue recrystallized from pentane to yield 3, 3-dimethoxy-l6 ,l6-dlmethylestr- (10 )-en-17S -ol melting at about 99-101°C.

PREPARATION OF FINAL PRODUCTS EXAMPLE 1 methoxy-l6,l6-dlmethylestra-2,5 (10)-dien-17(3 -ol in 80 parts of methanol at room temperature under a nitrogen atmosphere was added approximately parts of 18% hydrochloric acid. Approximately 1 1/2 hours later, the resultant solution was diluted with 200 parts of water, and the mixture thus obtained was chilled.

Insoluble solids were filtered out, washed with water, dried in air, and recrystallized from a mixture of methanol and ethyl acetate to give 173-hydroxy-l6 ,16-dimethylestr-J-en-S-one melting at about 172-175°C.

EXAMPLE 2 The entire quantity of 3 , 3-dimethoxy-l6 , 16-dimethylestr-5(10 )-en-17S-ol prepared in Example C was dissolved in 4,000 parts of methanol. 500 Parts by volume. of 6N hydrochloric acid was added to the solution at room temperature and stirred. After 3 hours the acid was neutralized with sodium hydroxide, the solution was concentrated, and diluted portionwise with water. The solid which formed was separated by filtration, washed with water and air dried. The solid was then recrystallized twice from ethyl acetate to give 173-hydroxy-16 ,l6-dimethylestr-4-en-3-one , melting at about I67-175°C EXAMPLE 3 The steroid of this invention can be administered in dosage unit form including, but not necessarily limited to, sterile solutions or suspensions for intramuscular injection, intravaginal or rectal compositions such as suppositories, lozenges for sublingual adminis- tration, and salves or lotions (including sprayable solutions or mixtures) for topical application.

As is well-known in the pharmacological art, the appropriate dose in any given instance depends upon the nature of the condition treated and its severity, the route of administration, the species of mammal · involved and its size and individual idiosyncrasies, etc. In general, and insofar as consistent with such factors, daily parenteral dosages of from 1 to about 5 mg. per kg. of body weight are suggested.

EXAMPLE Preparation of sterile solution. A sterile solution containing 10 mg. per ml. of 173 -hydroxy-16 ,16-dimethylestr-^-en-S-one was prepared by dissolving 10 gm. thereof in 50 ml. of benzyl alcohol, diluting to 1 liter with polyethylene glycol 400, and then was filtered to remove microorganisms.

EXAMPLE 5 Preparation of sterile suspension. A sterile suspension was prepared by mixing 100 gm. of sorbitol with 30 mg. of polyethylene glycol 400 and mg. of benzyl alcohol, whereupon sterilization was effected by filtration and to the filtrate was added 250 mg. of sterile micronized 178-hydroxy-l6,l6-dimethylestr- -en-3-one .

The resultant mixture was pulverized and smoothed in a sterile mill.

EXAMPLE 6 Preparation of suppositories. Suppositories were prepared to contain, individually, 259.:mg. of 178- hydroxy-16 by molding a mixture of .250 gm. of the micronized steroid, 800 gm. of polyethylene glycol 6000, 600 gm. of polyethylene glycol 15*10, and 250.gm. of water into 1000 suppositories.

' EXAMPLE 7 ^ Preparation of lozenges . Lozenges were prepared to contain, individually, 60 mg. of 173-hydroxy-16 ,l6-dimethylestr- -en-3-one by compressing a mixture of 60 mg. of the micronized steroid, 75 gm. of powdered polyethylene glycol ^000, 75 gm. of powdered polyethylene glycol 6000, and 150 gm. of mannitol into 1000 lozenges.

EXAMPLE 8 Preparation of a salve. A mixture of 15 parts of stearyl alcohol, Ί parts of cetyl alcohol., and 20 parts of polyethylene glycol 400 was melted, whereupon 10 parts of 170-hydroxy-l6 ,l6-dimethylestr-4-en-3-6ne was added with stirring. The resultant mixture was heated to 70-75°C. arid then added to 50 parts of water containing 1 part of sodium lauryl sulfate at the same temperature. The mixture thus obtained was stirred until it begins to cool and solidify.

Other vehicles which can be combined with 173-hydroxy-l6 ,l6-dimethylestr-4-en-3-one to prepare a composition adapted to topical application comprise fatty alcohols, fatty acids, liquid petrolatum, glycerol monostearate, glycerol, propylene glycol and other polyalcohols , ethylene glycol polymers, surfactants vegetable oils, fats, esters of fatty acids and fatty alcohols, water and hydrous or anhydrous emulsion bases

Claims (1)

1. WHAT WE CLAIM A process for the preparation of which comprises the acid hydrolysis of wherein the contains a covered hereinbefore The process for the preparation of which comprises the hydrolysis of with hydrochloric The process for the preparation of which comprises the hydrolysis of with hydrochloric Pharmaceutical compositions as an active together with pharmaceutical carriers For insufficientOCRQuality