RU2454424C1 - 13-ETHYL-GONA-1,3,5( 10), 8 (9)-TETRAENE-3,17β-DIOL DIACETATE, HAVING ANTI-IMPLANTATION AND ANTIOXIDANT ACTIVITY - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a diacetate of racemic 18-ethyl-gona-1,3,5(10),8(9)-tetraene-3,17β-diol, having anti-implantation and antioxidant activity with low uterotropic action. Presence of antioxidant activity in said steroid is essential since compounds with such action can be agents for preventing oestrogen-dependent breast cancer.

EFFECT: compounds exhibit anti-implantation and antioxidant activity with low uterotropic action, which is an advantage over agents used in practice.

1 cl, 1 ex, 4 tbl

Description

The invention relates to medicine and can be used in the synthesis of biologically active analogues of steroid estrogens, in particular, in the preparation of racemic 18-ethylgon-1,3,5 (10), 8 (9) -tetraen-3,17β-diol diacetate, possessing anti-implantation and antioxidant activity with reduced uterotropic effect, which suggests its use as a contraceptive.

A patent is known in which a method for the synthesis of 13-Alkylgona-1,3,5 (10), 9 (11) -tetraenes [1] is proposed, and it is also reported that it has estrogenic and lipid-lowering activity, which is closest to the claimed invention and selected as a prototype. Later, this steroid in optically active form was used as an intermediate to obtain other compounds, but its biological properties were not studied [2].

The technical result of the claimed invention is to obtain a substance with anti-implantation and antioxidant effects with reduced uterotropic activity.

The indicated technical result is achieved by the synthesis of 13-ethylgon-1,3,5 (10), 8 (9) -tetraen-3,17β-diol diacetate.

The synthesis scheme of the steroid is shown in example 1.

Example 1. To a solution of 1 g of the known compound (3-hydroxy-13-ethylgon-1,3,5 (10), 8 (9) -tetraen-17-one) [3] in 40 ml of THF, 10 ml are added with stirring 1M solution of lithium aluminum hydride in THF, the mixture is boiled for 2 hours. Then the reaction mixture is cooled to room temperature, the excess solvent is decomposed with ethyl acetate and poured into 1 l of a 3% hydrochloric acid solution. The reduction products are extracted with three 100 ml portions of ethyl acetate, the combined extracts are washed with water until neutral, dried with anhydrous sodium sulfate. The desiccant is filtered off, the residue is dissolved in 40 ml of pyridine, 25 ml of acetic anhydride are added, the reaction mixture is left for a day at room temperature. After the usual treatment [1], the target compound is isolated by crystallization from methanol. Obtain 1.03 g (78%) of the steroid, so pl. 145-146.5 ° C (according to literature data, mp 141-143 ° C [1]).

Mass spectrum, m / z (I _Rel ,%): 368 (35, M +), 326 (51), 266 (19), 237 (100), 209 (27), 181 (14), 160 (37 ), 133 (12), 107 (16).

¹³ C NMR Spectrum, δ, ppm: 9.81; 18.41; 21.00; 21.15; 21.91; 23.66; 24.44; 27.95; 28.16; 29.92; 43.88; 48.28; 82.56; 118.77; 120.21; 122.55; 125.97; 133.53; 133.76; 136.62; 148.39; 169.67; 171.10.

Found,%: C 74.99; H 7.70. C ₂₃ H ₂₈ O ₄ . Calculated,%: C 74.97; H 7.66.

Mass spectra were obtained by chromatography-mass spectrometry on an Aligent 6850 instrument with an Aligent 5973 mass-selective detector. Chromatographic analysis was performed on an HP-5MS capillary column with a length of 30 m, an inner diameter of 0.25 mm, and a film thickness of the stationary phase of 0.25. The velocity of the carrier gas (helium) through the column is 1.3 ml / min. We used linear programming of the column thermostat temperature from 220 to 230 ° C at a speed of 5 C / min. Evaporator temperature 330 ° С, flow division 20: 1. The mass selective detector provided scanning of mass spectra obtained by electron impact ionization (70 eV) in the range from 50 to 800 m / z at a speed of 2 scans per second. The obtained information was processed using the ADMIS program.

Spectra were obtained at 295 ° K on a Bruker DPX-300 spectrometer with an operating frequency of 75.468 MHz. For registration, a solution of 50 mg of the substance in 0.6 ml of CDCl _{3 was used} . Chemical shifts were measured with respect to TMS by assigning a solvent value (CDCl ₃ / CHCl ₃ = 99.9 / 0.1) of a standard value of 76.90 ppm.

The anti-implantation activity of the 3.17β-dihydroxy-18-ethylgon-1,3,5 (10), 8 (9) -tetraene racemic diacetate was investigated as follows. Males were planted in female rats weighing 160-180 g with a steady estrous cycle, daily for 3 days, from the date of detection of sperm in vaginal smears (first day of pregnancy), the drug was injected subcutaneously in an oil solution. After 20 days, the number of fruits was recorded. The results of experimental studies are presented in table 1.

Table 1 Anti-implantation effect of 3,17β-dihydroxy-13-ethylgon-1,3,5 (10), 8 (9) -tetraene diacetate (1) Injected drug Dose, mg / kg body weight per day The number of animals in the experience Contraceptive effect,% The control - 10 0 one 0.1 fifteen 87 one 0.5 fifteen one hundred

In addition, in the course of research, no pathology was revealed in the development of fetuses and deformities in fetuses under the influence of the drug in animals that retained their pregnancy. The size and weight of the fetus and placenta in this case did not differ from those in animals of the control group.

The uterotropic effect of the drug in effective doses is noticeably less than that of 17α-ethinyl estradiol used in medical practice (table 2).

table 2 Uterotropic effect of racemic diacetate 3,17β-dihydroxy-13-ethylgon-1,3,5 (10), 8 (9) -tetraene (1) Injected drug Dose, mg / kg body weight per day The number of animals in the experience Uterus Weight, mg The control - 6 13.4 ± 0.8 Ethinyl estradiol 0.05 6 98.5 ± 11.5 one 0.1 5 16.6 ± 1.4 one 0.5 6 34.6 ± 2.8 one 1.5 6 71.0 ± 8.4 one 5.0 5 88.5 ± 10.5

The antioxidant properties of 3,17β-dihydroxy-18-methyl-estra-1,3,5 (10), 8 (9) -tetraene diacetate were studied in accordance with [5–9]. The research results presented in tables 3 and 4 were processed according to the student - reliably significant differences are indicated in comparison with the control experiments.

Table 3 Antioxidant properties of 3,17β-dihydroxy-13-ethylgon-1,3,5 (10), 8 (9) -tetraen diacetate in rat brain (dose - 1.5 mg / kg body weight per day) Group of animals Schiff bases, cu / mg lipids Diene conjugates, nmol / mg lipids Triene conjugates cu / mg lipids Klein coefficient Malondialdehyde, nmol / mg, protein Intact rats 133 ± 9 3.7 ± 0.4 0.067 ± 0.005 0.186 ± 0.017 1.35 ± 0.08 Intact rats, 91 ± 5 2.6 ± 0.1 0.050 ± 0.005 0.148 ± 0.10 1.14 ± 0.06 received P <0.01 P <0.01 P <0.05 P <0.05 P <0.05 steroid 1

Table 4 Antioxidant properties of 3,17β-dihydroxy-13-ethylgon-1,3,5 (10), 8 (9) -tetraene diacetate (1) in rat liver (dose - 1.5 mg / kg body weight per day) Group of animals Schiff bases, cu / mg lipids Diene conjugates, nmol / mg lipids Triene conjugates, cu / mg lipids Klein coefficient Malonic dialdehyde, nmol / mg protein Intact rats 173 ± 11 5.8 ± 0.4 0.123 ± 0.008 0.248 ± 0.010 1.43 ± 0.10 Intact 134 ± 6 4.6 ± 0.3 0.163 ± 0.009 0.214 ± 0.14 0.98 ± 0.03 steroid 1 rats P <0.01 P <0.05 P <0.05 P <0.05 P <0.01

Thus, as shown by numerous experimental studies conducted at the laboratory facilities of St. Petersburg State University and the Research Institute of Obstetrics and Gynecology, the steroid [1] exhibits anti-implantation and antioxidant activity with reduced uterotropic activity, which is an advantage compared to those used in practice drugs. The presence of antioxidant activity in the claimed steroid is very important, since compounds with such an action can be used as prophylactic agents for estrogen-dependent breast cancer [4], which is especially relevant today to maintain health, given the tendency of this disease to significant growth observed in recent years.

Literature

1.13-Alkylgona-1,3,5 (10), 9 (11) -tetraenes // U.S. Pat. 3391170 (1968) - prototype

2.8 (9) -Dehydroestradiol derivatives // PCT Int. Appl. WO 98/16544.

3. Sorokina IB, Barkova TI, Zakharychev A.V., Chigir R.N., Ananchenko S.N., Torgov I.V. // Estrogen and antitumor activity in a series of transformed analogues of estrone and estradiol. Izv. USSR Academy of Sciences, ser. Chem. 1973, No. 5. S.664-670.

4. Felty Q., Singh K., Roy D. Estrogen-induced G1 / S transition to go-arrested estrogen-dependent breast cancer cells is regulated by mitochondrial oxidant signaling // Oncogene. 2005. V.24. N 31. P. 483-4893.

5. Folch J., Less M., Sloane-Stanly G. M. A simple method for the isolation and purification total lipids from animal tissues // J. Biol. Chem. 1957. V.226. P.497-509.

6. Bartlet G. Phosphorus assay in colomn chromathography // J. Biol. Chem. 1959, V.23. P.466-473.

7. Shvedova A.A., Polensky N.B. // Method for determination of lipid hydroperoxides conjugates in tissue extracts. The study of synthetic and natural antioxidants (edited by E.B. Burlakova): M. - 1992. P.74-75.

8. Bidlack W.R., Tappel A.L. Fluorescent products of phospholipids during lipid peroxidation // Lipids. 1973. P.8.

9. Andreeva L.I., Kozhemyakin L.A., Kashkun A.A. Modification of the method for determining lipid peroxides in the test with TBA // Labor. a business. 1988. No. 11. S.41-43.

Claims (1)

Anti-implantation and antioxidant agent, which is

diacetate 13-ethylgon-1,3,5 (10), 8 (9) -tetraen-3,17β-diol.