IT8022087A1 - EXTRENO (2,3-d) DIIDROISOSSAZOLDIOLI AND PROCESS OF PREPARATION OF THE SAME - Google Patents

Description

Description of the invention entitled:

"" EXTRENO [2, 3-d] DIIDROISOSSAZOLDIOLI AND PROCESS

OF PREPARATION OF THE SAME ""

SUMMARY

where Q represents hydrogen, methyl, ethyl, vinyl or ethinyl and R stands for hydrogen or methyl, which can be used as inhibitors of pituitary gonadotropins or as anabolic agents, are prepared by condensation of the corresponding

methylene-17? -hydroxy-5? - estran -3-oni with hydroxylamine.

DESCRIPTION OF THE INVENTION

The present invention relates to estreno [2,3-d] -dihydroisoxazoldiols and their preparation.

U.S. Patent No. 3. 135? 743 describes steroid [2, 3-d] isoxazoles having metabolic, hormonal and antihormonic characteristics, in particular anabolic characteristics. U.S. Patent No. 3. 145.200 describes steroid [3,2-c] isoxazoles having endocrinological characteristics including myotropic, anabolic, inhibitory on the pituitary or pituitary gland and inhibitory characteristics.

on the synthesis of steroids. U.S. Patent No. 3. 100,171 discloses androstane [3, 2-c] -5'-hydroxy-A-isoxazolin? having metabolic and hormone characteristics, in particular inhibitory characteristics

on the pituitary, and anabolic and myotro characteristics? piche. Kaneko, Nakamura, Yamato and Kurokawa (chem.Pharm.

Bull. , Vol. 17, No. 1, pp. 11-22, 1969) describes the preparation of the compound having the formula of strut

ture

starting from the corresponding 2-hydroxymethylene-3-ketone and hydroxylamine and dehydration of the compound of Formula E with acid to form the corresponding [2, 3-d] isoxazole (ibid., pp. 15-16). Myotropic / androgenic index equal to 0.9 is shown for the compound of Formula E.

The present invention accomplishes

them having the structural formula:

in which

Q represents hydrogen, methyl, ethyl, vinyl or ethynyl; And

R represents hydrogen or methyl.

The compounds of Formula I can be used as inhibitors of pituitary gonadotropins or as anabolic agents. Compounds of Formula I wherein Q represents vinyl or ethynyl are preferred. Compounds of Formula I wherein Q represents ethyls are particularly preferred. Particularly preferred embodiments are represented by the compounds of Formula I in which Q represents vinyl or ethynyl and R represents methyl,

Is it possible to use the compounds of the invention to inhibit the pituitary-gonadal axis in a mammal by administering a quantity to the mammal itself. of a compound of Formula I sufficient to reduce gonadal function.

The compounds can be incorporated into a pharmaceutical composition in a solid or liquid dosage form for oral, parenteral or intravaginal administration with a pharmaceutically acceptable carrier.

In one aspect of the process, the invention consists of the process for preparing a compound of Formula I which comprises condensation of the corresponding

having the structural formula:

with hydroxylamine or a hydroxylamine salt in an aqueous solvent mixture under neutral or slightly acidic conditions at room temperature or with cooling?

The compounds of Formula II are known and are generally prepared by condensation of the corresponding ones

oni with ethyl formate under alkaline conditions. To carry out the process form of the invention yes

can? ' use hydroxylamine or any

salt of the same? Readily available salts, e.g., hydroxylamine hydrochloride and hydroxylamine sulfate, are preferred, while hydroxylamine hydrochloride is particularly preferred.

The non-aqueous part of the solvent mixture can? be made up of any solvent capable of dissolving the compound of Formula II sufficiently to effect the condensation with the hydroxylamine or hydroxylamine salt without completely injuring the condensation itself? water-miscible solvents, in particular alcoholic solvents, are preferred. Have tertiary-butyl alcohol and ethanol been used successfully?

Pu? be used any acid or any base capable of producing the desired neutrality conditions? or of slight acidity? without interfering with condensation. Compounds of Formula I are formed even under more severe conditions. than slightly acidic, for example, in the presence of acetic acid, or p-toluenesulfonic acid? Using aqueous ethanol as a solvent mixture, quantity? approximately equimolar of a compound of Formula II and hydroxylamine hydrochloride, approximately half quantity? molar of sodium carbonate and a quantity? approximately double molar of sodium bicarbonate, and thus obtaining a pH value in the range between 6.5 and 6.8, good yields of the corresponding compound of Formula I were obtained.

Pu? A range of reaction temperatures between about -40 ° C and about 40 ° C be employed. The compounds of Formula I in which Q represents ethyl or vinyl are also prepared by catalytic hydrogenation or partial catalytic hydrogenation of the corresponding compounds of Formula I in

which 0 represents ethynyl. Effective catalyst-solvent combinations for the preparation of

places of Formula I, where Q represents ethyl, are palladium on carbon and ethanol, ethyl acetate or tetrahydrofuran. An effective combination catalyst-

solvent for the preparation of the compounds of Formula I

where Q represents vinyl? consisting of ball-

god on strontium carbonate and pyridine.

The following examples illustrate the invention

tion. Product structures are deduced from known structures of starting materials and reaction courses

preparations foreseen and are confirmed by melting temperature ranges (i.f.), infrared spectroscopic analysis, spectroscopic analysis

of nuclear magnetic resonance, spectroscopic analysis

mass pica and / or thin layer chromatography, which are also used to evaluate the purity of starting materials and products.

EXAMPLE 1

TO? A solution of hydroxylamine hydrochloride

(0.84 g) in water (10 ml) was added under stirring.

tion to a solution of

in tertiary-butyl alcohol (40 ml). The stirring was continued at room temperature for 1 hour. A solution of sodium acetate trihydrate (1.63) in water

(10 ml) was then added and stirring continued for 30 minutes. Water (250 ml) was added

and the mixture was extracted with two portions (50 ml

each) of chloroform-methanol (9: 1). The extracts

they were dried, filtered and stripped

solvents. Following crystallization of the

acetonitrile residue (30 ml) at room temperature

it was obtained

4 ', 5'-dihydroisoxazole -

the compound of Formula I wherein Q represents ethynyl and R stands for methyl.

Thin layer chromatography showed that the

crude product of the above reaction consisted mainly of the compound of Formula I wherein Q

represents ethinyl and R stands for methyl, and secondly in the corresponding androstane-isoxazoline of US patent N? 3,100,171 previously mentioned.

The relationship between the two products was not changed

following the addition of acetic acid or ptoluene sulfonic acid to the reaction mixture.

B. A solution of sodium bicarbonate (50 g)

in water (500 ml) was added under stirring a

room temperature to a solution of 1

one (100 g) in ethanol (2 1) A part of the steroid precipitates. The agitation continued

for 10 minutes. A solution of hydroxylamine hydrochloride (24.6 g) in water (50 ml) was then added for 25 minutes, and stirring was continued for

3 hours. Acetic acid (20 ml) was then added

drop by drop, and the stirring was continued

for 1 hour. The mixture was cooled in a bath

of ice (at 5? C). The resulting solid was collected, washed first with water and then with methanol, and dried (under vacuum at 60 ° C), obtaining

17 id

A solution of the combined products (166 g) of several of such drugs of 17a? Ethinyl-10pme l

hot dimethylformamide diol was filtered to remove turbidity. Moderately warm water was added to the filtrate until the first crystallization was observed and the mixture was cooled in ice-water. The solid was collected, washed

with cold methanol (5? C), and dried (under vacuum a

i.f. 212-214? C). In order to eliminate a light yellow color, the product was suspended with acetone, collected and dried again (136 g, i.f. 212-214? C).

EXAMPLE 2

A quantity? enough of a solution of sodium carbonate (3.35 g) in water (about 30 ml)

was added under stirring to a suspension of

17a-methyl-10p-methyl-2-hydroxymethylene-17? -Hydroxy-5aestranr-3-one (25 g) in ethanol (300 ml) to obtain a pH of 6.5-6.8. The solution was cooled to (5.7 g), sodium bicarbonate (13.7 g)

and water (about 125 ml) and a solution of hydroxylamine hydrochloride (5.7 g), sodium bicarbonate (13.7 g) and

water (about 125 ml) was added dropwise

under agitation. The temperature was then left

rise to room temperature overnight

three the agitation was continued. The mixture / cooled in water, obtaining a coarse gel, which was collected, washed with dimethylformamide-water

(50%) and dried (17.3 g). Water was added

to a filtered solution of the gel in dimethylformams

de (200 ml) until the solution became cloudy?

The resulting fine crystals were collected, suspended with ether, washed with ether and acetone and dried (7.37 g). A second crop was obtained from the ether-acetone wash liquids

(1.47 g) The two crops and the corresponding product (1, 97 g) of a preliminary preparation on a smaller scale were combined, suspended with ether, collected and dried, obtaining 17a-methyl-10? -

diol (i.f. 19 7-198? C), the compound of Formula I in which Q and R both represent methyl?

EXAMPLE 3

A. A solution of 17a? Ethinyl-10? -methyl-5? -estr-2-eno [2, 3-d] -4 ', 5'-dihydroisoxazole ^ (14.08 g) in pyridine (300 ml) was hydrogenated over palladium hydroxide on strontium carbonate (2%, 2.0 g ) in an apparatus of Parr. The colloidal mixture was subjected to pyridine stripping. Ethyl acetate was added and the mixture was stripped again. A mixture of the residue and hot ethyl acetate was filtered through infusor earth (Super-Cel) and the filter cake was washed with hot ethyl acetate. The filtrate was stripped of ethyl acetate and the residue

(12.50 g, i.f. 160? 165? C) was recrystallized from acetonitrile (7.20 g, i.f. 175.5-176? C). The product

was combined with the product (3.20 g) of a preliminary preparation on a smaller scale and the combined products were recrystallized from acetonitrile to obtain

the compound of Formula I, where Q represents vinyl and R stands for methyl.

B. Following condensation of 17a-vinyl-10? -Methyl-2-hydroxymethylene 17? -Hydroxy-5a-estran-3-one with hydroxylamine or a hydroxylamine salt, under neutral or weakly acidic conditions at room temperature or with cooling is achieved as well

EXAMPLE 4

Similarly to Example 2 and maintaining the pH at 6.0-6.5, 10p-methyl -2-hydroxymethylene-17? -Hydroxy-5a-estran-3-one (25 g) was condensed with hydroxylamino hydrochloride ( 5.7 g). The crude product (15.82 g, i.f. 181, 5-18 3 ° C) was combined with the product

(4.85 g, i.f 178-1 79? C) of a previous preparation on a smaller scale and the combined products were recrystallized from dimethylformamide-water, obtaining 10? Methyl-5? -extr-2- eno [2 3-d] -4 ', 5'-dihydroisoxazol-3? , 17 ?? diol (14.95 g, i.f. 179-181? C), the compound of Formula I in which Q represents hydrogen and R is

for methyl?

EXAMPLE 5

Similarly to that of Examples 2 and 4,

17a-ethinyl-2-hydroxymethylene-17? -Hydroxy-5a-estranm-3-one (20 g) was condensed with hydroxylamino hydrochloride (4.9 g)? Was the raw, resinous Product extracted with ether and methylene dichloride? The extracts were washed with diluted aqueous sodium hydroxide, dried over potassium carbonate, treated with activated carbon, filtered and stripped of the solvents, obtaining a solid in three fractions

(12.73g, 1.48g, 2.33g) The first two fractions were combined and recrystallized from dimethylformamide water and the product was dried under vacuum and heating (first at 50-60 ° C, and subsequently

at 85-100 ° c), obtaining 17? -ethinyl-5a-extr-2-eno [2, 3-d] -4 ', 5'-dihydroisoxazol-3? , 17? -Diol (8.40 g, i.f. 182-18 5? C) the compound of Formula I in which Q represents ethynyl and R stands for hydrogen.

EXAMPLE 6 A. Following condensation of

with hydroxylamine or a hydroxylamine salt under neutral or slightly acidic conditions at room temperature or with cooling, 17a-ethyl-

the compound of Formula I wherein Q represents ethyl and R stands for methyl.

B. Following hydrogenation of 1

-dihydroisoxazol-

under pressure on palladium on carbon in ethanol is obtained as well

BIOLOGICAL CHARACTERISTICS OF THE COMPOUNDS

As previously established, the compounds of Formula I can be used as inhibitors of pituitary gonadotropins or as anabolic agents. These uses have been demonstrated in rat and monkey tests, which indicate similar uses in the treatment of humans. Ownership suitable for reducing the gonadal function of compounds of Formula I were shown by the following tests carried out on the rat.

Evidence relating to impaired endocrine balance

Mature male rats, each weighing about 200 g, are treated orally or subcutaneously with an aqueous solution / suspension of tragacanth (1%) or with a solution / suspension in ethanol (10%) - oil cotton (90%) of the test compound twice a day for 14 consecutive days. Control rats receive vehicle only. The day following the last treatment the rats are weighed and then killed. The testicles, ventral prostate, seminal vesicles, thymus, adrenal glands, and pituitary gland are cut, dried with paper towels and weighed. The activity? of pituitary gonadotropins is evaluated by the relative weights of the testes, ventral prostates and seminal vesicles of treated and control rats.

In a similar test performed on a mature female rat, the uterus and ovaries are severed in place of the testes, ventral prostate and seminal vesicles.

Evidence related to activity? androgenic / anabolic Neutered immature male rats, each weighing approximately 70 g, are treated orally or subcutaneously with an aqueous solution / suspension of tragacanth gum (1%) or with a solution / suspension in ethanol (10%) -cotton oil (90%) of the test compound once a day for 10 consecutive days? Control rats receive vehicle only? Is testosterone propionate used as a gold standard and is it administered subcutaneously? Are the rats weighed and then sacrificed the day after the last treatment? Are the levator ani, ventral prostate and seminal vesicles of each rat severed, blotting paper and weighed? Activities androgenic and anti-androgenic are evaluated by the weights of the ventral prostate and seminal vesicles of treated rats, control rats and reference standard rats. Activities Are myotropic and anti-myotropic evaluated by levator ani weights of rats? treated, control rats and reference standard rats? The activity? myotropic is considered as an index of activity? anabolic?

Proof related to the activity? uterotropic / anti-estrogen Immature female rats, each weighing approximately 40 g, are treated orally or subcutaneously with an aqueous solution / suspension of tragacanth gum (1%) or a solution / suspension in ethanol (10%) cotton oil (90%) of the test compound, once a day for three consecutive days. Control rats receive vehicle only. Estradiol is used as a reference standard and is administered subcutaneously. The day after the last treatment the rats are weighed and then sacrificed. The uterus of each rat is cut, dried with paper towels and weighed. The activity? uterotropic and antiestrogenic is evaluated by the weights of the uterus of treated rats, control rats and standard reference rats.

Results of the previous tests carried out on the compounds of Examples 1 to 5 are reported in Table I.

the

,

i i i

100.

Monkey Tests - Compound of Example 1

Female rhesus monkeys whose last three menstrual cycles have been regular are treated orally with the test compound for 90 consecutive days, and effects on menstrual cycles are observed. In this test, the compound of Example 1, which is the compound of Formula I in

which 0 represents ethinyl and R stands for methyl and which according to what previously established, represents a preferred species of the present invention, caused amenorrhea at a dosage of 6.25 mg-per monkey, per day.

Comparison Tests - Compound of Example 1

The compound of Example 1 was compared in the test relating to the alteration of the endocrine balance in mature male rats and in the test relating to activity. andr? gen ^ myotropic in immature female rats, with the corresponding compound of the prior art dii & r? danazol of Example 27 of U.S. Pat. N? 3,135,743 previously mentioned. The results of the two comparison tests are shown in Table II and Table III.

TABLE II Impaired endocrine balance in mature male rats

TABLE III? Activities androgenic / myotropic in immature male rats

TABLE II Impaired endocrine balance in mature male rats

The table above shows how both compounds caused dose-related increases in the weights of the ventral prostate and seminal vesicles. The compound of Example 1, but not the compound of formula B ', caused significant decreases in body weight gains at all dosages. The compound of formula B1 caused a significant increase in the weight of the levator ani muscle at the higher dosage. high. Although the compound of Example 1 showed a relative potency equal to 0.8 (95% confidence limits between 04 and 1.6), referring to the comparison of the weights of the ventral prostate, the two compounds were found to be equipotent within limits of experimental error.

Although the compound of Example 1 and the compound of formula B 'were found to be approximately equipotent in tests carried out on the rat, the compound of Example was found to be at least 15 times higher. potent of the compound of formula B 'in causing amenorrhea or oligomenorrhea in the monkey. Therefore, while the minimum oral dose of the Example 1 compound to induce amenorrhea in regular-cycle female rhesus monkeys was 6.25 mg per monkey per day. androgenic / myotropic in immature male rats

TABLE III

The above Table demonstrates that both compounds caused dose-related decreases in the weights of the ventral prostate and seminal vesicles and significant decreases in body weight gains at all dosages. The compound of Formula B, but not the compound of Example 1, caused a significant decrease in the weight of the testes at the higher dose. high. Although the compound of Example 1 showed a relative potency equal to 2.1 (95% confidence limits between 0.8 and 3.6), referring to the comparison of the weights of the ventral prostate, the two compounds are, within the limits of the experimental error, equipotent.