RU2102083C1 - Method of antitumor agent preparing - Google Patents

Abstract

FIELD: medicine, oncology. SUBSTANCE: medicinal lignin in the form of an aqueous-alkaline suspension is subjected for ultrasonic oscillation and oxidized with oxygen-containing gas. After reaction mass cooling and separation of solid phase the end product is isolated from an acidified solution. Ultrasonic oscillation is carried out at irradiation power 40.0 ± 0.5 Wt, oscillation frequency 22.0 ± 0.5 kHz for 20-25 min. Oxidation with oxygen-containing gas is carried out at 190-200 C, under pressure 2.0-2.5 MPa for 0.5-3.0 h. EFFECT: decreased tumor proliferation. 2 cl, 5 tbl

Description

 The present invention relates to the field of production of drugs, in particular to the production of antitumor drugs, and may find application in practical oncology.

 A known method of obtaining an antitumor agent (Ed. St. USSR N 1701323, class A 61K 45/05, publ. 1991), in which an aqueous solution containing DNA and sodium chloride is mixed with an aqueous solution of potassium tetrachloride platinum chloride. Before mixing, ammonium chloride, potassium chloride and potassium acetate are added to the latter, with each solution being heated to the melting temperature of DNA. Sodium citrate is added to the solution containing DNA and sodium chloride. The method allows to obtain a tool that combines high antitumor activity with a pronounced immunomodulating effect.

 Of particular interest are herbal preparations. Such preparations are, in particular, taxanes obtained from yews of the genus Taxus. So, politaxel is a herbal preparation obtained from the bark of the Pacific yew (Voprosy Onkologii, 1994, v.40, NN 9-12, p. 259).

 The objective of the invention is to develop a method that allows to obtain effective preparations having antitumor activity from raw materials of plant origin.

 The problem is solved in that a method for producing an antitumor agent from raw materials of plant origin is proposed, in which a medical lignin in the form of a water-alkaline suspension is oxidized with an oxygen-containing gas after ultrasonic treatment and after cooling the reaction mass and separating the solid phase from the acidified solution, the target product is isolated.

 Ultrasonic exposure is carried out at a radiation power of 40 ± 0.5 W for 20-25 minutes at an oscillation frequency of 22 ± 0.5 kHz.

The oxidation with oxygen-containing gas is carried out at a temperature of 190-200 ^o C, a pressure of 2.0-2.5 MPa for 0.5-3 hours

 As medical lignin, a product of alkaline hydrolysis of lignin, enterosorbents based on lignin, such as Polyphepan, Lignosorb, can be used.

 "Polyphepan" nonspecific enterosorbent (reg. N 80/1211/3) is a brown powder, odorless and tasteless, with a moisture content of 65% consists mainly of lignin and contains no more than 20% residual polysaccharides (hydrocellulose).

 Lignosorb (polyphepan paste) VFS 42-2203-93.

 All substances used as feedstock are approved for oral administration.

 Special studies have shown that during the oxidative-hydrolytic destruction of medical lignin under the conditions of the proposed method, the formation of substances harmful to the body, such as polyaromatic hydrocarbons, nitrosoamines and polychlorinated dicyclo-p-dioxins, does not occur.

The study of the toxicity of the product obtained by the proposed method on healthy SHR mice of both sexes weighing 18-20 g showed that a dose of 300 mg / kg of the mass causes necrosis at the injection site, and the LD ₅₀ is 640 mg / kg of mass.

 The antitumor activity of the obtained product was evaluated by its effect on tumor growth in various kinds of mice (Ehrlich solid tumor, Ca 755 carcinosarcoma, spontaneous breast tumor, Walker carcinosarcoma). As shown by experimental data, the administration of the drug to mice reduces tumor growth by 60-70% (69.4-74.8% for Ehrlich tumor, 50-62% for Ca 755 carcinosarcoma, from 50% until the tumor completely disappears for the breast tumor, up to 88% for walker carcinosarcoma).

 The following examples illustrate the proposed method and the properties of the product obtained by the proposed method.

Example 1. In the initial aqueous-alkaline suspension of medical lignin (Polyphepan brand, reg. N 80/1211/3) of a non-specific enteric sorbent, a process of developed acoustic cavitation is created at a radiation power of 40 ± 0.5 W / cm ² , vibration frequency 22 ± 0.5 kHz for 23 ± 2 min followed by oxidation with atmospheric oxygen.

The composition of the suspension g
Polyphepan 500
Alkali (sodium hydroxide) 10
The density of the suspension is 1:10.

The oxidation is carried out in a reactor with mechanical stirring for 3 hours. Process conditions:
Temperature 195 ± 5 ^o C
Pressure 2.0-2.5 MPa
Air consumption 5 l / min
The reaction mass is cooled to room temperature and the precipitate is separated from the solution by filtration. The filtrate is acidified with sulfuric acid to pH 2-3. The precipitate was separated by filtration, washed with a water-alcohol mixture, then with distilled water until a pH of 7 ± 0.5 was established and dried at 105 ^° C. The product was a dark brown powder, odorless and tasteless, insoluble in water.

The elemental composition of the selected product in terms of organic matter, wt. C 61.36, H 3.56, N -0.41, (S + 0) 32.66 (calculated by difference). The substance decomposes without melting, starting from 170 ^o C.

The results of infrared Fourier spectra show that the resulting product contains bands of 1700 and 1600 cm ^-1 close in intensity, characteristic of carboxyl groups, an intense band in the region of 1200 cm ^-1 , a broad band in structure with local maxima of 3400, 3200, 3100 and 2600 cm ^-1 , as well as bands 2926, 2815, 1460 cm ^-1 and bands 2955 and 2870 cm ^-1 characteristic of methylene groups.

The resulting product has paramagnetism. Symmetrical EPR signals are observed in the product with a q factor of 2,000 ± 0.001 close to the value for a free electron (q = 2.00) and a width of H = 6.1 ± 0.1 Oersted. The concentration of PMC is at the level of (2.5 ± 0.4) • 10 ¹⁸ per gram.

 The product contains 3.0-3.6 mg • equiv of COOH groups and 6.0-6.5 mg • equiv of phenolic hydroxyls and can be classified as polyoxycarboxylic acids in structure.

 Example 2. 200 mg of the product was dissolved in 2 ml of 0.2 M tetrasubstituted sodium pyrophosphate, heated in a water bath to complete dissolution, the volume was adjusted to 10 ml with physiological saline, and sterilized in a water bath for 10 minutes. For administration to animals, the solution was diluted with sterile saline to the desired concentration.

The solid Ehrlich tumor was transplanted into outbred female mice weighing 18-20 g subcutaneously by the introduction of 10 ⁶ tumor cells. With daily intramuscular administration at a dose of 25 mg / kg for 5 days of the product obtained by the proposed method, an antitumor effect is observed (see table. 1). By day 19, the tumors disappeared in two mice. A single intramuscular injection of the product at a dose of 100 mg / kg to mice with a solid Ehrlich tumor causes significant inhibition of tumor growth (see table. 2).

Example 3. To mice C ₅₇ BI, subcutaneous suspension of 10 ⁶ tumor cells was inoculated with carcinosarcoma 755. Within 5 days, starting from the third after transplantation, intramuscularly or intraperitoneally or once intramuscularly on the third day after transplantation of tumors, the product obtained by the proposed method was administered at a dose 100 mg / kg. Five-time administration of the product causes a moderate inhibition of tumor growth of 62% (see table. 3).

 With intraperitoneal administration of the product, the maximum antitumor effect is detected on day 14, and with intramuscular administration on day 7 after tumor transplantation. The effectiveness of a single, intramuscular injection of a product is close to its effectiveness with repeated administration.

 Example 4. To mice with a spontaneous tumor of the mammary gland, a product obtained by the claimed method was once administered at a dose of 25 and 50 mg / kg. The effect of the product on the growth of spontaneous mammary tumor in mice is presented in table. 4.

 A single intramuscular injection of the product at a dose of 25 mg / kg of mouse N 1 led to a twofold decrease in the tumor on day 5 after administration. A single administration of the product intramuscularly at a dose of 50 mg / kg of mouse N 2 led to a decrease in tumor volume and the tumor disappeared by day 12.

 Example 5. Male rats weighing 100-150 g with a tumor of a carcinosarcoma Walker were injected intramuscularly daily for 5 days at a dose of 50 mg / kg. The most effective inhibition of the growth of Walker carcinosarcoma tumors was observed on the 7th day after their transplantation (see Table 5).

Claims (2)

 1. A method of obtaining an antitumor agent by treating plant materials, characterized in that the medical lignin in the form of a water-alkaline suspension is oxidized with an oxygen-containing gas after ultrasonic treatment and after cooling the reaction mass and separating the solid phase from the acidified solution, the target product is isolated. 2. The method according to p. 1, characterized in that the ultrasonic treatment is carried out at a radiation power of 40.0 ± 0.5 W, a frequency of sound vibrations of 22.0 ± 0.5 kHz for 20 25 minutes, and oxidation with an oxygen-containing gas at a temperature of 190,200 ^o C, a pressure of 2.0 to 2.5 MPa for 0.5 to 3.0 hours

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