SE457535B - STEROID DERIVATIVES IN THE ANDROSTAN SERIES CONTAINING ATM MINSTONE AND CHLORPHENACYL ESTER GROUP - Google Patents

Description

457 535 10 15 20 25 30 35 2 sarcoma W-256 in rats and mammary gland adenocarcinoma Ca-755 in mice), and in mammary gland cancer of certain types in mice and rats (e.g. HK, R-13762), which is practically insensitive to alkylating agents.

These compounds are less toxic than those containing chlorphenacyl. If optimal therapeutic doses of chlorphenacyl for mice are 1-1.5 mg / kg body weight when chlorphenacyl is introduced over a period of 5-10 days, these doses of dehydroepiandrosterone derivatives (DEA mustard) are equal to 15-20 mg / kg at the same cure time for introduction of this derivative. However, these known compounds have a toxic (toxic) effect on the organism, namely that they cause neurotoxicity, lymphopenia and necrosis, in the liver and kidneys (compare, for example, Sofjina ZP, Lagova ND, Valoueva IM, Kouzjmina ZV, Shkodinskaja EN, Kholetsky AM, “ collections from the Soviet Union's first conference on chemotherapy for malignant tumors ", Riga, 1968, pp. 441-4Å3; M. Wall, G. Abernethy, F. Carroll, D. Taylor," J. Med. Chem., 1969, vol 12, volume 5, pp. 810-818; U. Schaeppi, J. Heyman, R. Fleischman, H. Rosenkrantz, V. Glievski, R. Phelan, D. Cooney, R. Davis, “Canc. Chem . Rep. ", Year 1975, part 3, vol 4, volume 1, pp. 85-95; Lagova ND, the book" Current problems in experimental chemotherapy for tumors ", volume 1, pp. 222-225).

BACKGROUND OF THE INVENTION The main object of the present invention is to provide new derivatives of steroids in the androstane series, Chernogolovka in 1980, which steroid derivatives exhibit antitumor and hormone activity, in particular a targeted effect on tissue shot tablets and in these emerging tumors.

The steroid derivatives proposed according to the present invention in the androstane series are a new type of derivative not previously described in the literature.

This object is achieved according to the present invention in that the steroid derivatives in the androstane series consist of compounds of the general formula: wherein R 1 represents -OH, = O or -OCO-CH 2 -C 6 H 4 -N (CH 2 CH 2 Cl) 2, R 2 -OCO-CH-CH -CHN (CH 264 'N fl cocu represents -OH, CH 2 Cl) 2 or 2 3 -OCO- (CH 2) n -C 6 H 4 -N (CH 2 CH 2 X) 2, where n is 1 or 3 and X is CL, Br or J; R 3 represents -CH 3 or -H, provided that at least one of R 1 and R 2 is always a chlorophenacyl ester group.

The steroid derivatives of the androstane series (hormone cytostatic) proposed according to the present invention contain, as steroids, testosterone metabolites or synthetic analogues thereof and consist of esters of Sa-androstane substituted at 2-, 3- and 17-positions.

The compounds of the present invention consist of amorphous or fine crystalline powdery substances which are white in color (or colored in white with a yellow tint), are water-insoluble and soluble in organic solvents. The compounds are stable for a long time when stored in the dark at a temperature of between 5 ° C and 0 ° C. When stored in light, they turn yellow.

The compounds proposed according to the present invention are 178- [O-di (2-chloroethyl) aminophenyl acetate] -Sa-androstan-178-ol-3-one (II: 17B ~ [D-di (2-chloroethyl) ami nophenyl butyrate] -5u-androstan-17B-ol-3-one (II); 17B- {o-di (2-chloroethyl) aminophenyl-N-octylalanyl] -5d-androstan-17B-ol--3-one ( III); 178- [p-di (2-chlorotyl) aminophenyl-N-acetylalanyl] -2a-methyl-5α-androstan-17B-ol-3-one (IV); 3B- [D-di (2) - -coretyl) aminophenyl acetate] -Sa-androstanediol-36, 173 (V): 176- [p-di (2-chloroethyl) aminophenyl acetate] -Su-androstanediol-36, 178 (VI); 178- [p-di ( 2-bromomethyl) aminophenyl acetate] -5α-androstan-176-01-3-one (VII): 176-1D-di (2-di (2-di (2-iodoethyl] aminophenylacetate] -5n-androstan-17B-ol- 3-one (VIII). 457 535 '10 15 20 25 30 35 4 The following steroid derivatives are suitably used in the androstane series: f / \ "ococH2c6H4N (cH2cn2cL) 2 / '(I) H (c1cu2cH2) 2Nc6H4cu2oco __í ::: ¿\ / g 1H (V) OCOCH 2 C 6 H 4 N (CH 2 CH 2 Cl) 2 HO 1 H (VI) The compounds proposed according to the present invention show considerable antitumor activity and other gene and anabolic hormone activity, the nature of which is dependent on the structure of a hormone component (hormonal part of the molecule of these compounds and the structure and site of association of the alkylation group with the steroid.

The advantage of the compounds proposed according to the present invention is that they act directly on tissues functioning as target plates and tumors occurring in these tissues, which contributes to the spectrum of antitumor activity of these compounds having special properties compared with known hormone cytostatics. The proposed parenteral (subcutaneous) method of administering hormone cytostatics proposed in accordance with the present invention ensures that the compounds become less toxic and their biological action becomes more targeted compared to oral administration (in the oral mode of administration of hormonal cytostatics), since the parenteral administration is not leads to the molecule being cleaved into a hormonal and a highly toxic alkylation fragment.

The antitumor activity (anti-tumor activity) of the compounds proposed according to the present invention was tested on a number of strains of unvaccinated solid tumors (swellings) and leukoses in mice: mammary adenocarcinoma ca-755, Louis-Lung carcinoma LL, cervical cancer (cervical cancer) LHK-5, Waker's carcinosarcoma W-256, and lymphoid leukosis L-1210.

For comparison with the compounds of the ovarian tumor ÄST proposed according to the present invention, dihydrotestosterone (Su-androstan-178-ol-3-one), dehydroepiandrosterone and chlorphenacyl or the preparation sarcolysin (B- {0- -di (2- chloroethyl) aminophenyl] -o-alanine, chlorohydrate), which are included in the molecule of most of the compounds of the present invention, as well as such known hormonal cytostatics as ester of dehydroepiandrosterone with chlorphenacyl (DEA-mustard) and preparations estracyt ( estra-1,3,5 (10) -trien-3,176-diol, 3N- [bis- (2-chloroethyl)] carbamate-17-sodium phosphate).

The compounds were introduced into the animal subcutaneous in oil at doses calculated in mg per kg of the animal's body weight, for a period of 5-10 days after time intervals, which are approved for examination on corresponding tumors (24 or 48 hours after the re-inoculation of the tumor). ): The estracyte was similarly introduced into a 0.9% solution of sodium chloride.

To investigate the targeted action (targeting) of the compounds, spontaneous and / or cancer-induced tumors arising in tissue target plates were used. In these cases, treatment was started after the tumors had reached a certain size, at which spontaneous regression of the tumors could not occur.

One criterion for the antitumor activity was an increase in the lifespan of the animals and an inhibition of the growth of the solid tumors, the lifespan being expressed in% being equal to T ~ C _ C T is the average lifespan of the treated animals; 100, where C is the average lifespan of the reference animals, while the% expressing inhibition rate of the solid tumors is equal to ----- - 100, where 0 denotes the growth of the tumor. average volume (weight) in the group of treated animals; K denotes the mean volume (weight) of the tumor in the reference group.

One criterion for the antitumor effect when influencing spontaneous and induced tumors was a cure rate expressed in% for the animals and a change in the tumor size compared with the original tumor size.

The studies were performed on so-called linear mice and their hybrids from the previous generation.

Results obtained by examining the antitumor activity of the compounds of the present invention on some of the unvaccinated tumors are set forth in Table I.

Table I Antitumor activity of the steroid derivatives proposed according to the present invention in the androstane series No. Compound Dose, Treatment days mg / kg after re-inoculation per day of the tumor 1 2 3 4 1 lvß-tp-a-z-chloroethine-aminophenyl acetate] -5a-androstane- 17B-o1-3- -one (I) 25-50 2-6 2 17B ~ [p-di (2-chloroethyl) aminophenyl butyrate] -5a- -androstan-17ß-ol-3- -one (II) 30- 2-6 3 176- [p-di (2-chloroethyl) -aminophenyl-N-acetyl-alanyl] -5a-androstan-176-ol-3-one (III) 10-100 2-6 4 178- [p-di (2-chloroethyl) amino-phenyl-N-acetylalanil] -2a-methyl-5a-androstan-17B--ol-3-one (IV) 10-100 10 15 20 25 30 35 457 535 7 Table 1 (continued) 1 2 3 4 5. 36 - [p-di (2-chloroethyl) aminophenyl-10-100 2-6 nyl acetate] -5a-androstandiol-35, 17β (V) 5-30 2-6 6. 178- [p-di (2-chloroethyl) amino-phenylacetate] -Sa -androstanediol-33, 173 (VI) 5-50 2-6 7. 178- [p-di (2-bronethyl) amino - phenyl acetate] -5α-androstane-17B-01-3-One (VII) 10-50 2-6 8. 178- [p-di (2-iodoethyl) amino-phenyl acetate] -5α-androstane-17B-o1 -3-one (VIII) 25-100 2-6 9. Dihydrotestosterone 1 0-15 2-6 10. Chlorophenacyl 1.5-2 2-6 ll. Sarcolysis 2-3 2-6 7 2-6 12. DBA mustard 10-15 2-11 50 2-6 13. Estracyte 100 2-6 No. Maximum inhibition of tumor Lifespan increase, growth,% Ca-755 LL W-256 Ca-755 L-1210 W-256 1 5 6 7 8 9 10 1 98 66 99 59 41 160 2 71 30% -ig healing 47 3 98 71 122 82 4 96 58 79 82 5 95 66 31 28 6 95 60 32 25 7 97 54 8 65 47 9 16 +77 10 33 97 11 83 58 32 90 12 10 57 13 +6 29 4 457 535 10 15 20 25 30 35 8 The sign "+" denotes stimulation of tumor growth.

Table 1 shows that the compounds proposed according to the present invention show a strong antitumor effect with respect to a number of unvaccinated tumors, which is expressed in a considerable inhibition (inhibition) of the growth of the tumors (by 58-99%) and / or an increase in the treated the lifespan of the animals by 25-160%. The advantages of the compounds of the present invention are particularly marked with respect to the treatment of Ca-755. The compounds not only inhibit the growth of Ca-775 by 65-98% but the lifespan of the animals by 31-122%. The hormone cytostatics known under the name DEA mustard in the androstane series and estracyt did not show any antitumor activity with respect to this tumor.

It can also be seen from Table 1 that the compounds proposed according to the present invention differ from alkylating agents, chlorphenacyl and sacrolysine, which are included in the molecule of these compounds, in terms of the intensity and spectrum of antitumor action and the toxicity. Therapeutic doses of the compounds of the present invention vary between 5 and 150 mg / kg when administered daily for a period of 5 days, while maximum tolerable doses of chlorophenacyl and sarcolysin under the same treatment conditions are 1.5 and 3 mg / kg, respectively. In compounds 178- [p-di (2-chloroethyl) aminophenylacetate]. androstanediol-3-B, 178 (V) and 178- - [p-di (2-chloroethyl) aminophenyl acetate] -Su-androstanediol-36, 178 (VI) a more marked direct effect and differences in the effect on tumors were observed, which arise in hormone-dependent tissues, such as unvaccinated cervical cancer LHK-5, ovarian tumors and on induced and spontaneous ovarian and mammary gland tumors in mice and rats.

At the same time, all of these compounds were less active in the treatment of animals with leukosis L-1210 (Table 1), indicating that these compounds exhibit a less intense alkylating effect. The data reported in Table 2 show that compound I has little effect on the growth of the ovarian tumors, while compound VI and especially compound V cause an intense depression of the growth of this tumor. It is also shown in Table 2 that cervical cancer LHK-5 and ovarian tumors are insensitive or not at all sensitive to dihydrotestosterone and chlorphenacyl, while the estrogen cytotoxic drug astracyte even stimulates the growth of cervical cancer LHK-5. The data presented in Table 3 show that compounds I, V and VI show advantages over the known androgen testosterone propiones in terms of their antitumor activity with respect to induced and spontaneous ovarian and mammary gland tumors in mice and rats.

Examination of the overall toxic effect of the compounds of the present invention has shown that these compounds are relatively low in toxicity.

Mice tolerate five-fold introduction of these compounds at a dose of from 5 to 10 mg / kg. Most compounds of the invention exhibit a considerable therapeutic dose range (Table 4) and cause a high antitumor effect at doses which differ from 3 to 10 times the absolute value.

Antitumor activity of the steroid derivatives in the androstane series with respect to from so-called Tissue-shoot tablets derived from tumors Compound Maximum inhibition of tu- Percentage of tender growth (5) Lifespan increase Increase Cervical- Ovarian- Ovarian- cancer LHK-5 tumor tumor l 2 3 4 lveíp-aixz- chlorethyl1) amino- phenyl1acetate] -5u- 17B-ol--3-one (I) 82 37 0 457 535 10 15 20 25 30 35 10 Table 2 (cont.) zβ-ip-di (z-chloroethyl) aminophenyl acetate] -5anadrostanediol-33, 175 (V ) 80 17ß- [p-di (2-chloroethyl) -aminophenyl acetate] -5α-androstanediol-36, 178 (VI) 79 17S- [p-di (2-chloroethyl) -aminophenyl-N-acetyla-1anil] -Sa -androstan- -178-ol-3-one (III) 87 17B- [p-di (2-chloroethyl) -aminophenyl-N-acetyl-1anil] -2u-methyl-5u-androstan-17-ol-3- -one (IV) 99 Dihydrotestesterone 13 Chlorphenacyl 35 Estracyte +322 Antitumor effect of sterio on inducing and spotting-derived tumors 3 4 94 25 73 33 97 57 92 37 54 0 25 0 29 0 Table 3 ddeivatives in the androstane series, from tissues-shot tablets no Tumors l 2 l According to biskindam ovarian tumors induced in nñss 2 According to Biskindam induced ovarian tumors in r 3 Spontaneous mammary gland tuners in mice 4 DMA-induced mammary glands in rats Decrease in size or stimulation (+) of tumor growth compared to tumor size at baseline, 176- [p-di (2-chloroethyl) aminophenyl acetate] -5u-androstan-17B-ol-3-one (I) 3 tumor growth, cessation 27 tunurtil growth ceases 60 10 15 20 25 30 35 457 535 ll Table 3 (continued) no Reduction in size or stimulation (+) of tumor growth compared to tumor size at baseline,% 17ß- [p-ai (z-chloroethyl) amino- Sterone- fen 38- [p-di (2-chloroethyl) -Testo- Refe- aminophenyl acetate] -5a- androstane-178 -ol-3-one (V) phenylacetate] - propio--5a-androstanate-176-ol-3-one (VI) 1 4 5 6 7 1 67 +256 +21 2 64 +32 3 35 24 + 33 +86 4 growth +280 ten ceases Table 4 Maximum tolerable doses (MTD) and range of therapeutic doses of the steroid derivatives in the androstane series (mice; five-fold introduction) Compound mg / kg MTD Therapeutic doses 1 2 3 176- [p-di (2-chloroctyl) aminophenyl acetate] -5α-androstane-17β-o1-3-one (I) 20-25 457 535 10 15 20 25 30 35 12 Table 4 (continued) 1 2 3, * 176- (p-di (2-chloroethyl) aminophenyl butyrate] -Sa-androstan-176-ol--3-one (II) 50 30 178 - [p-di (2-chloroethyl) aminophenyl-N-acetylalanil] -5a-androstane-178-01-3-one (III) 100 10-100 17s- [p-di <2-1 <1ore1; y ' 1) aminophenyl-N-acetylalanil] -2a-methyl-50-androstan-176-ol-3-one (IV) 100 10-100 3Bfp-di (2-chloroethyl) aminophenyl acetate] -5a-androstanediol--35,175 (V) 5-30 176- (p-di (2-chloroethyl) aminophenyl acetate] -Su-androstanediol-33,17ß (VI) 50 10-30 176- [p-di (2-bromomethyl) aminophenyl] acetate] -5-α-androstan-17B-ol-3-one (VII) 50 10-50 176- [p-di (2-iodoethyl) aminophenyl acetate] -5a-androstan-178-01-3-one ( VIII) 100 25-100 Chlorphenacyl 3 1,5-2 Sarcolysin * 5 2-3 For the study of the direct action of the compounds of the present invention on tissue bulkheads, their hormonal properties were determined by biological (androgenic and anabolic effects) and biochemical (specific binding at dihydrotestosterone and estradiol 176 receptors) tests. The androgenic and anabolic activity was determined depending on the weight change of the prostate and levator ani in castrated rats and was expressed as an index (the ratio between the so-called relative weight of each organ after exposure and the relative weight of said organ of the reference rats, which was assumed to be equal with 1). The specific binding at the dihydrotestosterone and estradiol receptors (the free binding sites) was examined in the ventral prostate of normal and pre-castrated (two weeks before the study) rats receiving therapeutic drug regimens.

The specific binding of dihydrotestosterone and estradiol 176 was determined in cytosolic tissue fractions by precipitation of egg whites (proteins) by protamine sulfate after complete incubation of cytosols with saturating concentrations of [3 H] -dihydrotestosterone and [3 H] -estradiol 176 ° C. The nuclear receptors were determined from a fraction of purified nuclei, which were previously extracted with a 0.4 solution of KCL, by precipitating a hormone-receptor complex by means of protamine sulphate.

The table shows this regarding the androgenic and anabolic action of the compounds proposed according to the invention.

It can be seen from Table 5 that most of the proposed steroid derivatives in the androstane series show weak anabolic activity, while their androgenic activity is highly dependent on the structure of the hormonal part of the compound. Like androgens, compounds I and III are stronger than the others. Compounds I and III also show the longest duration of androgenic action.

However, the androgenic activity of the compound I is considerably weaker than the androgenic activity of such a known androgen as testosterone propionate.

It has been found that the steroid derivatives proposed in the androgen series according to the present invention bind specifically to the receptors for the respective hormone receptors. For example, compound I binds to dihydrotestosterone cytoplasmic receptors with subsequent reaction with respective acceptor zones for chromations both in normal tissues (ventral prostate and uterus in rats and in tumors in animals and humans, which tumors are derived from tissues-shot plates for genes (mammary gland).

The hormonal properties of the compounds I, V and VI proposed according to the invention were also manifested in the effect on the reception of estradiol 17β in the tissue of the ventral prostate, observing differences between the compounds. In intact rats, for example, the specific binding of [3 H] -estradiol 178 in cytosol was increased fivefold after introduction of compound V. After introduction of compound I, said binding increased tenfold.

After introduction of compound VI, no specific binding of [3 H] -estradiol 176 i was found (the binding was completely suppressed). cytosol The compounds proposed according to the present invention thus significantly altered the animals' sensitivity to estrogens.

Compounds I, V and VI also differ from each other with respect to the metabolism in tissue and tablet tablets for androgens. These differences were already evident at one level for the major androgen metabolism enzyme, namely 5α-reductase, and at one level for 36-, Ba-, Su- and 7α-hydroxylases.

The steroid derivatives proposed according to the present invention in the androstane series thus differ from each other with respect to the spectrum of antitumor action, total toxic action, nader-shot tablets. Hormonal action and metabolism in tissue- Each of the compounds is characterized by the typical features of biological action for the said compound only, so it is of independent interest.

Table 5 Androgenic and anabolic properties of hormocytostatic, namely the steroid derivatives in the androstane series No. Compound Dose (mg / kg) time interval (hours) number of injections 1 2 _ 3 1. 176- [p-di (2-chloroethyl) aminophenyl) acetate] -Sa-androstan-17B-ol- X -3-one (I) 25/24 10 2. 35- [p-di (2-chloroethyl) aminophenyl acetate] -Sß-androstanediol-38, 17ß (V ) 20/24 x 10 10 15 20 25 30 35 15 457 555 Table 5 (cont.) 1 2 3 3. 176-fp-di (2-chloroethyl) aminophenyl acetate] -5α-androstandio1- -38,176 (VI) 20 / 24 x 10 4. 176- [p-di (2-chloroethyl) aminophenyl- N-acetylalanil] -5a-androstane- 25/24 x 10 17ß-01-3-One (III) 50/24 X 10 5. 178 - [p-di (2-chloroethyl) aminophenyl-N-acetylalanil] -2a-25/24 x 10 methyl-5a-androstan-176-01- 50/24 x 10 -3-one (IV) 6. DEA mustard 5/24 x 10 10/24 x 10 7. Dihydrotestosterone 15/24 x 10 8. Dihydroepiandrosterone 0.5 / 24 x 10 1/24 x 10 9. Testosterone propionate 15/24 x 10 10. Sarcolysin 2/24 x 10 11. Chlorphenacyl 1/24 x 10 Table 5 (cont.) No. 24 hours after treatment '' Anabolic Androgenic ling cut index. index 1 4 5 1 2.4 1: o 1, 2 10, 1 1.2, 6 3 10 1.4 1 3 1 1.7 2.0 10 1.2 1.1 4 1 2.1 6, 2 10 1.3 4.5 457 535 16 Table 5 (continued) l 4 5 6 1 1.7 1 6 10 1.2 0: 8 5 1 2.3 1.5 5 10 1.6 1.8 1 1.3 1.4 6 l 1.2 2.2 1 1.6 1.1 7 10 1.1 0.9 10 1 1.5 1.2 8 l 1.1 1.4 1 2.7 23.9 9 10 2.1 8.2 15 10 1 0.8 1.6 11 1 1.0 0.9 20 25 30 35 When examining the hormonal kinetics of 17ê [p-di (2-chloroethyl) aminophenyl acetate] -5u-androstane -17B- -ol-3-one (I) an interesting law was discovered, namely that the stimulation period for organ-shot tablets (prostate, testicle, seminal vesicles) was alternated - after a time interval of 1.5 months - with a period for profound depression of said organs, while at the depths which received only dihydrotestosterone, the normal weight of all organs was restored at this time. In addition, a long-term inhibitory effect of compound I was observed on the thyroid gland of male and female rats (analogous to the action of dihydrotestosterone) and on the uterus and ovaries, the normal weight of these organs not being restored 3 months after the introduction of the preparation. . In female rats with estrogen-active follicular cysts on the ovary, the temporary "chemical castration effect" of the compound I was observed. After a therapeutic course of treatment with the compound I, the rats ceased to have a continuous, for them typical, heat.

In laparotomies and histological examination of ovarian logs, it was found that the burning stopped due to follicular atresia and atrophy of the interstitial tissue of the ovaries, which in estrogen is estrogenative. The chemical castration effect was also discovered in lactating rats, which after a therapeutic treatment course with compound I, i.e. 17β- [p-di (2-chloroethyl) aminophenyl acetate] -Su-androstan-176-ol-3-one lost during a time of 5-7 months fertility. These data show that compound I exhibits contraceptive properties.

It has been found that compound I also exhibits alkylating properties, interfering with the transcription and transport of RNA and causing structural defects in DNA, cells (spleen) and in tumor cells (sarcoma S-37). However, those which recover more rapidly in the normal latter case show that the antitumor effect is selective.

Thus, with reference to compound I as an example, it has been found that hormone cytostatic, namely the steroid derivatives in the androstane series, exhibit the double mechanism of biological action, these compounds exhibiting alkylating and hormonal effects.

The process for the preparation of the steroid derivatives in the androstane series is simple to carry out and does not require any complicated process equipment.

The compounds of the present invention are synthesized according to known process techniques by reacting substituted 36, 176-2a-androstane with chlorohydride ac cytotoxic phenylalkanoic acid or n-amino-β-phenylpropionic acid in an aprotonic solvent in the presence of an organic base at boiling point of the solvent for a period of 8-24 hours and that the final reaction product is isolated by preparative column chromatography or by subsequent substitution of halogens in the reaction product against Br or J or by subsequent reduction of a keto group in the reaction product.

The synthesis of the compounds by this method can be represented by the following reaction scheme: 457 555 18 5 s R R Rocë, Q2 RÅ R * R I H 5 | % “3 A; §å> / ñ: § >>” / H 5 5 P2 R Ri 10 å C A. Rl = CH; = o; R2 = -ca; R3 = -CH3, -H; R1 = R2 = -CH.

B. nl = -on; = o; -ococn2c6H4N (cH2ca2c1) 2; R2 = -cs, -oco (cH) c H -N (cH CH cl), where n = 1.3 2 n 6 4 2 2 2 20 -ococHcH2c6H4-N (cH2cH2c1) 2; R3 = -CH3, -H.

NHcocn3 c. R1 = -on, = o, -ococa2c6H4N (cH2cn2c1) 2; H2 = -on, -oco r 2 n 6 4 2 2 2 25 n = 1.3; x = cl, er, J; -ococHcn2c6H4N (cH2cn2c1) 2: Nncocn3 R3 = -CH3, -H.

The compounds proposed according to the present invention are synthesized by reacting chlorohydride of p-di (2-chloroethyl) aminophenylacetic acid with 5a-androstane substituted at 38- or 178- or 2a-positions with subsequent reduction of the esterification products by sodium boro-boron or or bromine 35 or iodine.

The preparation of the compounds of the present invention is further illustrated below by the following examples.

Example 1 Preparation of 176- [p-di (2-chloroethyl) aminophenyl acetate] -Sa-androstan-176-ol-3-one (I).

A suspension of 30 g (0.1 mmol) of 5α-androstan-17β-ol-3-one in 500 ml of dry benzene is added with a calculated amount of triethylamine and 38 g (0.1 mmol) of chlorohydride of p-di (2-chloroethyl) aminophenylacetic acid. The resulting mixture is stirred at a temperature of 80 ° C for a period of 24 hours and triethylamine chloride is separated. The benzene solution is washed with an aqueous solution of KHCO3 and water and evaporated. The residue is dissolved in benzene, introduced into a chromatography column with alumina and eluted with benzene. The benzene eluate is evaporated and the residue in the form of an oil is crystallized from ether. The product yield is 27.89 (49.2% calculated as starting hormone), while the melting point of the product is 99-101 ° C (etef).

Analysis for C 31 H 43 NO 3 Cl 2: Found: C 67.85; H 7.89; Cl 12.96; Calculated: C 67.89; H 7.84; Cl 12.95; Example 2 Preparation of 17β- [p-di (2-chloroethyl) aminophenyl butyrate] -Sa-androstan-178-01-3-one (II).

This compound is prepared analogously to the synthesis of the compound of Example 1.

In the esterification reaction, 5α-androstan-17β-ol-3-one and chlorohydride of p-di (2-chloroethyl) aminophenylbutyric acid are introduced. The reaction mixture is stirred at a temperature of 80 ° C for a period of 17 hours. Subsequent process steps are carried out in the same manner as in Example 1.

The product yield is 45.5%, calculated as the starting hormone, while the melting point of the product is 100-102 ° C.

Analysis for C 33 H 47 Cl 2 NO 3: C 68.65; H 8.15; Cl 11.98; C 68.77; H 8.16; Cl 12.32 ;.

Found: Calculated: 457 555 Example 15 Preparation of 176- [p-di (2-chloroethyl) aminophenyl- -N-acetylalanil] -5a-androstan-17B-ol-3-one (III) .

This compound is prepared analogously to the synthesis of the compound of Example 1. In the esterification reaction, Sa-androstan-176-ol-3-one and chlorohydride of Q-N-acetylamino-B1-di (2-chloroethyl) amino] phenylalanine are introduced into methylene chloride.

The reaction mixture is stirred at a temperature of 40 ° C for a period of 9 hours. Subsequent process steps are carried out in the same manner as in Example 1.

The esterification product is chromatographed on a silica gel column, the eluent being a mixture of benzene and ethyl acetate in a mutual ratio of 1: 1. The product yield is l4.5%, calculated as the output hormone.

Analysis for C 34 H 18 Cl 2 N 2 O 4: Found: C 65.87; H 7.72; Cl ll, ll; Calculated: C 65.94; H 7.75; Cl ll, 47 ;.

Example Gel 4 Preparation of 178- [p-di (2-chloroethyl) aminophenyl- -N-acetylalanil] -2a-methyl-5a-androstan-176-ol-3-one (IV).

This compound is prepared analogously to the synthesis of the compound of Example 1.

In the esterification reaction, 2u-methyl-5a-androstan-176-ol-3-one and chlorohydride of α-N-acetylamino-S [p-di (2-chloroethyl) amino] phenylalanine are introduced into methylene chloride.

The reaction mixture is stirred at a temperature of 40 ° C for a period of 9 hours. Subsequent process steps are carried out in the same way as in Example 1.

The esterification product is chromatographed on a silica gel column, the eluent being a mixture of benzene and ethyl acetate in a 1: 1 mixing ratio. The product yield is 30% calculated as the starting hormone. Analysis for C 35 H 55 OCl 2 N 2 O 4: Found: C 66.41; H 7.96; Cl 10.9l; Calculated: C 66.34; H 7.96; Cl ll, 2 ;.

Example 5 Preparation of 36- [p-di (2-chloroethyl) aminophenyl acetate] -Sa-androstanediol-38,17B (V). 4.2 g of 38-Lp-di (2-chloroethyl) aminophenylacetate] -5a-androstan-176-one are mixed with 90 ml of dry methanol, then 0.4 g of sodium borohydride are added at a temperature of 5 ° C. The reaction mixture is stirred for a period of 1 hour and acidified by adding acetic acid to a pH of 6, after which the solution is evaporated, the residue is washed with water and dried over sodium sulphate.

From the benzene-containing solution 4 g of amorite powder are obtained. The product yield is 95%, calculated as the original keto compound.

Analysis for C 31 H 45 Cl 2 NO 3: Found: C 67.59; H 8.25; Cl 12.88; Calculated: C 67.90; H 8.16; Cl 12.95 ;.

Example 6 Preparation of 178- [p-di (2-chloroethyl) aminophenyl acetate] -Sa-androstanediol-36,178 (VI).

This compound is prepared in the same manner as the compound of Example 5.

176- [p-di (2-chloroethyl) aminophenyl acetate] -5u-androstan-3-one is introduced into the reduction reaction. Subsequent process steps are performed in the same manner as in Example 5.

The reduction product is chromatographed on a silica gel column, the eluent being a mixture of benzene and ethyl acetate in a 4: 1 mixing ratio. Fractions are collected which contain a 3a-OH isomer (with an Rf value of 0.41) and a 36- -OH isomer (with an Rf of 0.33). The yield of the final β-isomer is 67%, calculated as the original 3-keto compound. 457 535 10 15 20 25 30 35 22 Analysis for C 31 H 45 Cl 2 NO 3: Found: C 67.54; H 8.26; Cl 13.03; Calculated: C 67.59; H 8.25; Cl 12.88 ;.

Example 7 Preparation of 17β- [p-di (2-bromomethyl) aminophenyl acetate] -Sa-androstan-176-ol-3-one (VII).

To a solution of 3 g of 176- [p-di (2-chloroethyl) aminophenylacetate] -Sa-androstan-1776-ol-3-one in 40 ml of methyl ethyl ketone is added 5.7 g of lithium bromide (in about seven times excess), after which the reaction mixture is heated at a temperature of 80 ° C for a period of 35 hours. The solvent is evaporated and the residue is dissolved in benzene. The benzene solution is chromatographed on a column of alumina and the final product is isolated in the form of an oil which is easily crystallized from ether. The melting point of the product is between 106 and 108 ° C.

The product yield is 54% calculated as the starting compound.

Analysis for C 31 H 43 Br 2 NO 3: Found: C 58.28; H 6.79; Br 25.09; Calculated: C 58.38; H 6.80; Br 25.08 ;.

Example Gel 8 Preparation of 176- [p-di (2-iodoethyl) aminophenyl acetate 1-5u-androstan-17B-ol-3-one (VIII).

To a solution of 5 g of 17β- [p-di (2-chloroethyl) aminophenylacetate] -α-androstan-1776-ol-3-one in 50 ml of methyl ethyl ketone is added 7.5 g of sodium iodide (in an approximately six-fold excess), after which the reaction mixture is heated at the boiling point of the solvent of 79.5-80 ° C for a period of 72 hours. After the solvent has been evaporated, the residue is extracted with benzene and chromatographed on a column of alumina, the eluent being benzene. From the benzeneuylate a solid phase substance is isolated, which is crystallized from ether. is 3.29 g (48.5% Product yield calculated as the starting hormone), the melting point of the product being between 110 and 112 ° C. Analysis for C 31 H 43 J 2 NO 3: Found: C 50.70; H 5.87; J 34.07; Calculated: C 50.90; H 5.88; J 34.74 ;.

Industrial Applicability The steroid derivatives of the androstane series proposed according to the present invention can be used in various fields of medicine, for example gynecology and oncology such as antitumor preparations and contraceptives.

Claims (4)

1. Steroid derivatives in the androstane series, characterized in that they consist of compounds of the general formula - wherein R 1 represents -OH, = O or -OCO- CH 2 -CGHA-N (CH 2 CH 2 Cl) 2, R 2 represents -one, -oco-CH-cuz-c 6 H 4 -N (cH 2cH 2 Cl) 2 or NHCOCH 3 -OCO- (CH 2) n -CGH 4 -N (CH 2 CH 2 X) 2, where n is 1 or 3 and X 20 is Cl, Br, or J; R 3 represents -CH 3 or -H, provided that at least one of R 1 and R 2 is always a chlorophenacyl ester group. 2. A derivative according to claim 1, characterized in that it is a compound of the formula: ococH2c6H4u 30 <3% H 10 15 20 3. A derivative according to claim 1, 457 535 characterized in that it is a compound of the formula: fl (C1cH2CH2) 2Nc6H4OCO 4. t HO Derivatives according to claim 1, OH characterized in that it is a compound of the formula: OCOCH zcs fl q N (cH2cH2c1) 2 ~, VI