RU2039560C1 - Medicinal preparation possessing antitumor activity - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: object of this invention is extension of range of antitumor drugs used for treating hormone-dependent forms of carcinoma. This object is accomplished by using medicinal preparation already known in the art, that is, diacetate-6-methylpregna -4,6-diene-3-β,17a -diol-20-one (acetomepregenol), for new purpose. EFFECT: novel application of disclosed medicinal preparation as hormone antitumoral drug. 14 tbl

Description

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Известно, что соединение I обладает гестагенным действием. Ацетомепрегенол применяют в качестве контрацептивного и лечебного средства при акушерской и гинекологической патологии (1).The invention relates to medicine and relates to a known compound of a number of oxyprogesterone, namely 6-methylpregn-4,6-diene-3 β-17α-diol-20-one diacetate [acetomepregenol (1), mepregenol diacetate (2), diamol (3) ] of the formula (1):
CH ₃ -

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Compound I is known to have a gestagenic effect. Acetomepregenol is used as a contraceptive and therapeutic agent for obstetric and gynecological pathologies (1).

 For the first time, it is proposed to use compound I in medicine as an antitumor agent.

известный в медицинской практике под названием окси прогестерона капронат, широко применяемый для лечения запущенных форм рака молочной железы и рака матки.Known hormonal antitumor drug-caprotan pregn-4-en-17 α -ol-3,20-dione of the formula A:

Known in medical practice as hydroxy progesterone capronate, widely used to treat advanced forms of breast cancer and uterine cancer.

 This drug is administered only parenterally and in massive doses up to 5 g per week. In addition, with prolonged use of the drug, adaptation to it is observed and, as a consequence, the cessation of action.

Препарат Б известен в медицинской практике под международным названием мегестрола ацетата и применяется для лечения запущенных карциономы груди и рака тела матки.The closest in structure and pharmacological action to the invention is the steroidal antitumor preparation 6-methyl-pregna-4,6-diene-17 α -ol-3,20-dione acetate of the formula B:

The drug B is known in medical practice under the international name megestrol acetate and is used to treat advanced carcinoma of the breast and cancer of the uterus.

 However, the drug can only be administered orally in relatively high doses (up to 320 mg per day), there is no selectivity for hormonal action (antiandrogenic activity is clearly expressed).

 Since the characteristic of most antitumor hormone preparations is the adaptation of the body to the substance used and weakening until their action ceases, the search for new hormonal antitumor drugs is necessary.

 The aim of the invention is a new tool for the treatment of hormone-dependent forms of cancer with improved antitumor activity.

 This is achieved by using the known medicinal substance 6-methyl-pregn-4,6-diene-3 β 17 α-diol-20-ondiacetate of the formula I in a new quality, namely, as a hormonal antitumor agent.

The antitumor activity of the claimed preparation of the formula I (acetomepregenol) was studied in experiments:
in rats with Walker carcinoma-sarcoma (example I, tab. 1-4);
with a tumor of RMK-1 (example 2, tab. 5-10);
on C ₅₇ Bl mice with 755 adenocarcinoma (Example 3, Table 11);
on mice SNR with spontaneous mammary tumors (example 4, table. 12);
in mice TWO ₂ and C ₃ N with tumors of the mammary glands, as well as a comparison of the activity of acetomepregenol (1) and megestrol acetate (B) when administered orally (example 5, table 13);
on F ₁ DB mice with carcinosarcoma 755, as well as the first and second generations of a transplanted breast tumor (OM) and a comparison of the activity of drug I and oxyprogesterone capronate (A) with parenteral administration (Example 6, Table 14).

 PRI me R 1. The effect of acetomepregenol on Walker's carcinosarcoma in rats.

 The study was conducted on 38 female rats weighing 100-110 g. Walker's carcinosarcoma was subcutaneously transplanted. The drug was administered as an aqueous suspension orally in doses of 1.5, 15 and 150 mg / kg daily for 14 days. starting the day after the inoculation of the tumor.

 The results of measuring the volume of tumors at different periods of observation are presented in table. 1.

 The data table. 1 allow us to conclude that the ability of the drug of formula (1), applied at a dose of 150 mg / kg daily orally, statistically significantly inhibit the growth of Walker carcinosarcoma in rats by 37.5-48.9% When using the drug in doses of 1.5 and 15 mg / kg, a stimulating effect on tumor growth was not observed, and a tendency to its inhibition was observed.

 The dynamics of body weight in rats with Walker carcinoma treated with drug I are presented in table. 2.

 When considering the data of the dynamics of body mass, presented in table. 2, attention is drawn to the absence of differences in body weight of rats of the control group and rats treated with drug I at a dose of 150 mg / kg during tumor growth. Given that the drug in the specified dose inhibits tumor growth, these data indicate the absence of toxic effects of the drug at a dose of 150 mg / kg.

 In addition to the use of acetaminophenol as a substance, studies were conducted on the effect of the tablet form of acetaminophenol on tumor growth in rats with Walker carcinosarcoma (see Table 3). The experiments were performed on 56 female rats weighing 140-150 g. The next day after inoculation of the tumor, acetomepregenol (tablet form, 1 tablet contains 25 mg of acetomepregenol) was administered daily through a probe in a volume of 1 ml of distilled water in doses of 15, 150, 300 mg / kg . The introduction of the drug continued until the death of the animals.

The moment of the appearance of the tumor after transplantation and the day of the death of the animal were recorded. The body weight of the animals and the tumor volume in cm ^{3 was} determined every 2-3 days.

 The results of experimental studies (Table 3) showed that metomepregenol in doses of 150-300 mg / kg causes significantly significant inhibition of Walker carcinosarcoma growth. When the drug was administered at a dose of 150 mg / kg, the tumor growth was inhibited by 43.5% (P <0.01), and when a double dose of 300 mg / kg was introduced, the tumor growth was inhibited by 49.7% (P <0.01). The drug at a dose of 15 mg / kg caused moderate inhibition of tumor growth of 19.9% (P>> 0.05).

 A study of the effect of the drug on rat body weight (Table 4) showed that the drug in the doses applied did not cause a noticeable decrease in animal body weight. Acetomepregenol, reducing the tumor volume of Walker's carcinosarcoma, did not have a toxic effect on the animal organism.

 As can be seen from the table. 4, acetomepregenol does not affect the rate of occurrence of Walker carcinosarcoma tumor.

 PRI me R 2. The effect of acetomepregenol on a tumor of RMK-1 in rats.

 The study was conducted on 37 female rats weighing 100-110 g. The tumor RMK-1 was subcutaneously transplanted. Acetomepregenol (substance) was administered orally at a dose of 150 mg / kg daily from the first day after inoculation during the entire time of observation of the animals.

 The experimental results are presented in table. 5 show that the drug at a dose of 150 mg / kg had an inhibitory effect on the growth of this tumor.

 When comparing tumor volume indices in rats of the control group and rats treated with drug I at a dose of 150 mg / kg, carried out at a later date of PMK-1 inoculation (30-41 days), even more significant differences were revealed (Table 6).

 The inhibitory effect of drug I on tumor growth of PMK-1 for 30-41 days is presented in table. 6.

 As can be seen from the data table. 5 and 6, drug I, applied at a dose of 150 mg / kg daily, inhibits tumor growth of RMK-1 by 2-4 times.

 Presented in the table. 7 data on the dynamics of body weight in rats with RMK-1 treated with the drug indicate the absence of significant differences with the indices of the control group.

 Taking into account the existing tendency to inhibit the growth of the tumor of RMK-1 from 12-14 days after vaccination under the influence of drug I at a dose of 150 mg / kg, the obtained data can be considered as the absence of toxic effects of the drug used in the specified dose.

 In addition, studies have been conducted on the effect of the tablet form of acetomepregenol on tumor growth in rats with RMK-1. The experiments were performed on 64 female rats weighing 140-150 g body weight (table. 8).

 The next day after transplantation of tumors, acetomepregenol (tablet form, 1 tablet contains 25 mg of acetomepregenol) was administered daily through a probe in a volume of 1 ml of distilled water at doses of 15, 150, 300 mg / kg. The introduction of the drug continued until the death of the animals. Animals of the control group through one probe received distilled water in the same volume.

The moment of the appearance of the tumor after transplantation and the day of death of the animals were recorded. The body weight of the animals and the tumor volume (in cm ³ ) were determined every 2-3 days.

 The effect of acetomepregenol (tablet form) on the rate of onset of rat tumor PMK-1 is presented in Table. 8.

 The results of the studies showed that acetomepregenol did not significantly affect the rate of onset of PMC-1 tumor after inoculation.

 There were also no significant changes in the average life expectancy of rats with a PMK-1 tumor treated with acetomepregenol in various doses, compared with control animals (Table 9).

 The effect of acetomepregenol (tablets) on the growth of PMC-1 tumor in rats was also studied (see Table 10).

As can be seen from the table. 10, acetomepregenol inhibits the growth of PMK-1 tumor in rats with the administration of the drug at a dose of 15 mg / kg at 38.7% 150 mg / kg at 50.6% 300 mg / kg at 68.8% The disappearance of PMK-1 tumors in the control group of animals was 15%
Thus, studies have shown that the drug, reducing the volume of tumors PMK-1, does not have a toxic effect on the animal organism, does not cause a noticeable decrease in animal body weight.

 The obtained experimental data allow us to conclude that acetomepregenol has pronounced carcinolytic properties, which are manifested in a significant inhibition of the growth of Walker and PMK-1 carcinosarcoma tumor strains.

 PRI me R 3. The effect of acetomepregenol on adenocarcinoma 755 in mice.

The study was conducted on 27 female mice of the C ₅₇ Bl line. Acenocarzionoma 755 is subcutaneously interrupted. The animals received acetomepregenol orally in the form of an aqueous suspension in doses of 150 and 300 mg / kg from the 3rd day of inoculation to the 7th.

 Measurement of the volume of tumors produced on the 11th day after inoculation showed that acetomepregenol at a dose of 150 mg / kg inhibits the growth of adenocarcinoma 755 by 50% with an increase in dose to 300 mg / kg, no effect was observed (Table 11).

The effect of acetomepregenol on the growth of adenocarcinoma 755 in C ₅₇ Bl mice is presented in Table. eleven.

 PRI me R 4. The effect of acetomepregenol on spontaneous breast tumors in SHR mice.

 The study was conducted on 10 SHR female mice over the age of 1 year with spontaneous mammary tumors. The mice of the experimental groups received acetomepregenol daily at a dose of 150 mg / kg orally. Acetomepregenol administration continued throughout the observation period. According to the evaluation of the results, the animals were distributed in pairs in such a way that each mouse receiving acetomepregenol was compared with a control mouse with similar localization and with similar initial tumor sizes.

 Data on the measurement of the volume of tumor nodes and the survival of mice are presented in table. 12.

 The data table. 12 show that acetomepregenol has a therapeutic effect in spontaneous mammary tumors in SHR mice. Along with the delayed tumor growth in two of the five cases, a partial regression of the tumor nodes was also observed (experiments 2 and 3), and in both cases the tumor volume decreased by almost 90% compared with the initial tumor volume.

 Monitoring the survival of mice showed that mice with spontaneous tumors treated with acetomepregenol lived longer than the control ones (experiment 1, 2, and 4).

The activity of acetomepregenol against spontaneous mammary tumors in mice favorably distinguishes acetomepregenol from its analogue of hydroxyprogesterone capronate (A), which did not affect the growth rate of the first generation on one of the six mammary tumors studied in C ₃ H and DD mice.

PRI me R 5. The effect of acetomepregenol (I) on the growth of the first generation of transplantable mammary tumors in mice TWO ₂ . Comparison of the activity of I and megestrol acetate (B).

A study of the effect of acyloxyprogesterone derivatives (the claimed drug and the known drug (B)) on the growth of the first generation of a breast tumor in TWO ₂ mice was carried out on female mice of this line. The source of the tumor was a spontaneous breast tumor found in a nulliparous female TWO ₂ . Under aseptic conditions, the tumor was removed, crushed, and a 10% suspension in physiological saline was prepared. Inoculation was performed intramuscularly with 10 ⁶ cells contained in 0.2 ml of suspension. Transplantation was positive in 100% of the animals. Tumors began to be distinctly determined by the 12th day after inoculation. The drugs were administered orally from 3 days after inoculation at a dose of 150 mg / kg daily five times a week for two weeks.

The study was conducted on 33 female mice of the TWO ₂ line.

 The first group (10 mice) served as a control; mice from 3 days after inoculation were ingested with a 0.4 ml starch paste probe, which was used to prepare a suspension of insoluble preparations administered to animals of the other three groups. Mice of the control group died by day 28. The tumor mass at the time of the death of the animals exceeded 3000 mg.

 Mice of the second group received megestrol acetate (B), the third group of the claimed drug (I) is acetomepregenol.

 Tumor volumes were measured 16, 18, and 20 days after inoculation. Megestrol acetate (B) inhibited tumor growth by 28-30%. Acetomepregenol (I) was the most effective drug, because under its influence, tumor growth was inhibited by 45-67%. Life expectancy of mice treated with acetomepregenol (I) was higher than the duration life of mice of other groups (table. 13).

b) The effect of acetomepregenol (I) on the growth of a spontaneous breast tumor in C ₃ N. mice.

In a study in mice C ₃ H female mice was intramuscularly intertwined mammary tumor spontaneously arose-line female mouse C ₃ N. transplantation in all mice was positive, although tumor growth rate was lower compared to tumors in mice TWO _2. The mice of the first group served as a control, the mice of the second group were injected with a preparation of I at a dose of 150 mg / kg for 2 weeks, starting from the 3rd day after inoculation.

The measurement of the volume of the tumors, made on the 48th day after inoculation, showed that under the influence of acetomepregenol, the growth of tumors was inhibited by 81%
PRI me R 6. The study of the antitumor activity of acetaminophenol (I) with parenteral administration. Comparison of the activity of I and oxyprogesterone capronate (A). The antitumor activity of acetomepregenol was studied with the introduction of an intramuscularly 3% and 1.5% suspension in stone oil and an intraperitoneal 1.5% suspension in a 10% solution of polyvinylpyrrolidone.

 Evaluation of antitumor activity was carried out in comparison with the activity of oxyprogesterone capronate (A).

The experiments were performed on 88 female DB 1 mice with an average initial weight of 23-27 g with 755 carcinoma, as well as the first and second generations of a transplanted mammary tumor (OM) obtained from spontaneous OMF in TWO ₂ mice. The drug was administered daily for 7-9 days. Administration was started 24 hours later (carcinoma 755) or 9 (1st generation of prostate cancer) and 5 (2nd generation) days after tumor transplantation. Animals were sacrificed 48 hours later (carcinoma 755) or 96 hours after the last administration of the drug, then its tolerance (according to the general condition and change in body weight during treatment compared to the control, Cr) and the percentage of inhibition of tumor growth in such mice were evaluated compared with the control (J _t ,).

 Acetomepregenol (I) and oxyprogesterone capronate (A) were administered at previously determined optimal doses indicated in the table. 14 (α> 0.95).

 As can be seen from the table. 14, acetomepregenol with intraperitoneal and intramuscular administration has a clear inhibitory effect on tumor growth.

When the drug was administered intramuscularly to mice with carcinoma 755 (9 injections of acetomepregenol at a dose of 150 mg / day as a 3% suspension in bone oil), the tumor growth was inhibited by 68% (oxyprogesterone capronate slowed the tumor growth in the same experiment at an optimal dose of 1500 mg / day only 49%);
with a decrease in the number of injections of the drug 1 to 7, the inhibition coefficient of tumor growth was 62%
After intraperitoneal administration (seven administrations) of acetomepregenol (I) in the form of a 1.5% suspension in polyvinylpyrrolidone, inhibition of tumor growth reaches 58%. The toxic effect of acetomepregenol (I) on the body was not detected.

 In mice with the first generation of OMF, acetomepregenol inhibited tumor growth by only 38%. Subsequent inoculation, accompanied by a decrease in the hormone sensitivity of the tumor, the antitumor effect of drug I decreased and turned out to be borderline (25% α> 0.95).

 Thus, with parenteral administration, acetomepregenol (I) exhibits a distinct antitumor activity at a dose 10 times lower than oxyprogesterone capronate (A).

 The study of the toxicity of the claimed drug.

 Toxicological studies were carried out on four species of animals, including dogs, it was found that the claimed preparation of formula I does not adversely affect the vital functions of the body (morphological picture of the blood, its coagulation system; liver, kidney, cardiovascular system; respiratory system; central nervous system, endocrine system, etc.).

 A study of the spherical toxicity of the claimed drug showed its lack of teratogenic, allergenic, mutagenic and carcinogenic activities.

 When studying the hormonal spectrum of the physiological activity of compound (I), it was found that the claimed preparation is a pure geogen that does not exhibit extrogenic, anabolic, anti-inflammatory mineralocorticoid, thymolytic, anti-estrogenic, antiandrogenic activity, i.e., it is a multi-hormonal drug devoid of side hormonal effects.

 The inventive preparation I exhibits a high antitumor activity with any route of administration: with parenteral administration (i / m and / b) it is more active than compound A, and when administered orally, it is more active than the closest chemical and pharmacological analogue of drug B.

 Thus, from the presented experimental data it can be seen that the proposed drug of formula I is a new effective antitumor agent.

Acetomepregenol (I) is an odorless white or white to yellowish crystalline powder, readily soluble in chloroform, acetone, soluble in alcohol, vegetable oils, practically insoluble in water. Melting point 191 ± 6 ^о С, specific rotation 77.5 ± 2.5 ^о (1% solution in chloroform), optical density 1% / 1 cm 585 ± 15 at 242 nm (0.001% solution in methanol).

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Пример получения 6-метилпрегна-4,6-диен-3 β17 α -диол-20-она (ацетомепрегенола).Scheme for the preparation of compound I

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An example of obtaining 6-methylpregn-4,6-dien-3 β17 α-diol-20-one (acetomepregenol).

To a stirred suspension of 5 g of 6-methylpregn-4,6-diene-17 α -ol-3,20-dione acetate in 100 ml of methanol was added 0.5 g of sodium borohydride. After further stirring for 15 minutes, the reaction mixture was diluted with water. To complete crystallization, the suspension is stirred for 30 minutes, the precipitate is filtered off, washed with water and dried. Obtain 4.72 g of 17-acetate-6-methylpregn-4,6-diene-3 β17 α-diol-20-one with a melting point of 203-205 ^о С, / α / _D ²⁰ 56.3 ^о (2% - solution in chloroform).

 Acetylate 5 g of 17-acetate-6-methylpregn-4,6-diene-3 β17 α-diol-20-one with a mixture of 10 ml of acetic anhydride and 10 ml of pyridine with stirring at room temperature for 16 hours. The resulting solution was poured into the mixture hydrochloric acid, water and ice and stirred for 1 hour. The precipitate formed is filtered off, washed with a solution of hydrochloric acid, then with water until neutral, dried, and after purification 4.36 g of acetomepregenol are obtained.

Claims (1)

ANTI-TUMOR ACTIVITY SUBSTANCE, characterized in that it is 6-methyl-pregna 4,6-diene-3β, 17α diol-20-one diacetate of the formula

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