RU2308285C2 - Agent based on polysaccharides from calamus root enhancing of anti-tumor and anti-metastasis activity of cytostatic preparations - Google Patents

Abstract

FIELD: medicine, oncology.

SUBSTANCE: claimed agent represents polysaccharides from calamus root (Acorus calamus Z.) rootstock. Said polysaccharides enhance anti-tumor and anti-metastasis activity of known cytostatic preparations.

EFFECT: new agent enhancing of anti-tumor and anti-metastasis activity of cytostatic preparations.

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Description

The invention relates to medicine, specifically to oncology, and can be used to increase the effectiveness of the cytostatic treatment of patients with malignant tumors.

Closest to the proposed invention is a tool with antitumor and antimetastatic activity that increases the effectiveness of cytostatic drugs - zymosan [1].

Cytostatic drugs have a number of toxic phenomena (impaired blood formation, immunosuppression, etc.), which limit their continued use in highly effective doses for the treatment of patients with malignant tumors. Thus, increasing the antitumor and antimetastatic activity of cytostatic drugs while reducing their toxicity is an urgent task.

The invention is aimed at solving a problem urgent for clinical oncology: increasing the effectiveness of drug therapy for malignant neoplasms with decreasing the course dose of cytostatic drugs, and as a result, reducing the negative consequences of their use.

To solve this problem, an agent is used that increases the antitumor and antimetastatic activity of cytostatics, containing the sum of water-soluble polysaccharides from the roots of calamus marsh.

Natural rhizomes of calamus (finely ground powder) are part of the preparations Vicair and Vicalin, used to treat gastritis and gastric ulcer. Calamus rhizomes are also used as a decoction, as a means of improving digestion [2]. There are experimental data proving the effectiveness of a 40% extract from rhizomes of calamus swamp on a model of ulcerative pathology [3].

New is that as a means of increasing the antitumor and antimetastatic activity of cytotoxic drugs, water-soluble polysaccharides from rhizomes of calamus swamp are used.

These significant features are not identified from the medical, scientific and patent literature. They do not explicitly follow from the prior art for specialists. This tool has passed an experimental study in the oncopharmacology laboratory of the Research Institute of Pharmacology of the Tomsk Scientific Center of the Siberian Branch of the Russian Academy of Medical Sciences in Tomsk and at the Department of Chemistry of Siberian State Medical University.

RESEARCH RESULTS OF PHARMACOLOGICAL ACTIVITY

New properties of polysaccharides from rhizomes of calamus marsh were discovered experimentally. An aqueous solution of polysaccharides obtained by the standard procedure for isolating water-soluble polysaccharides from plant materials [7] was injected into laboratory animals. The experiments were performed on female mice of the C57BL / 6 line weighing 19-20 g at the age of 3 months. As a model of experimental malignant growth, hematogenously metastatic Lewis lung carcinoma (3LL) was used, transplanted intramuscularly at 5 million tumor cells per mouse. To conduct the experiment, all mice with 3LL inoculated were divided into groups:

1. “Control” - mice receiving daily 0.1 ml of ampoule water for injection intraperitoneally for 12 days.

2. “Cyclophosphamide” - mice that received cyclophosphamide once intraperitoneally at a dose of 125 mg / kg on the 11th day of tumor development.

3. “Calamus polysaccharides” - mice that received an aqueous solution of polysaccharides from calamus rhizomes of bogs intraperitoneally daily at a dose of 10 mg / kg (diluted with ampoule water for injection) for 12 days, starting from the 7th day of tumor development.

4. “Cyclophosphamide + calamus polysaccharides” - mice treated with cyclophosphamide once intraperitoneally at a dose of 125 mg / kg on the 11th day of tumor development and a solution of polysaccharides from calamus rhizomes of bog marsh intraperitoneally (on the day of CF administration 4 hours later) daily at a dose of 10 mg / kg within 12 days, starting from the 7th day of tumor development.

5. "Echinacea purpurea polysaccharides" (used as a comparison drug) - mice treated with echinacea purpurea polysaccharides intraperitoneally daily at a dose of 10 mg / kg (diluted with ampoule water for injection) for 12 days starting from the 7th day of tumor development.

6. “Cyclophosphamide + echinacea polysaccharides” - mice treated with cyclophosphamide once intraperitoneally at a dose of 125 mg / kg on the 11th day of tumor development and a solution of echinacea purpurea polysaccharides intraperitoneally purple (on the day of CF administration after 4 hours) daily at a dose of 10 mg / kg for 12 days, starting from 7 days of tumor development.

At the end of the experiment, animals were killed, observing the "Rules for the work using experimental animals", approved by the Ministry of Health of the Russian Federation. Mice were opened, internal organs revised, tumors isolated, metastases counted. The effectiveness of the treatment courses was evaluated by the antitumor and antimetastatic effect of the drugs [4, 5].

The results were processed using nonparametric Wilcoxon-Mann-Whitney criteria (u) and the Fisher angular transformation (φ). The differences were considered significant at P <0.05 [6].

The results of a study of the combined use of cyclophosphamide and polysaccharides from rhizomes of calamus marsh showed that in mice with combined treatment there was a more pronounced inhibition of growth of the primary tumor node (inhibition of tumor growth was 31% compared with 8% in the group of animals treated only with cytostatic, P <0 01), while there was also a significant inhibition of the development of the metastasis process (the average number of metastases per mouse decreased 3.1 times, and their area - 2.8 times compared with these displays mice that received only CF, P <0.05). The inhibition index of metastasis in the described group was the highest and amounted to 90% versus 61% in the “CF” group. Attention should be paid to the independent antitumor effect of calamus polysaccharides - the inhibition of tumor growth in the group of mice treated with this drug, isolated, was 15% (P <0.01). The deaths of animals in the groups “Cyclophosphamide + calamus polysaccharides” and “calamus polysaccharides” were not detected (Table 1).

At the same time, with the combination of cyclophosphamide with Echinacea purpurea polysaccharides, the increase in the antitumor and antimetastatic effect of the cytostatic was less pronounced than in the case of combining CF with calamus polysaccharides. Thus, inhibition of tumor growth in mice treated with CF and Echinacea polysaccharides was 22% versus 8% in the CF group and 31% in the CF + calamus polysaccharides group; the number of metastases decreased by 1.4 times, and their area - by 2.8 times compared with these data in mice treated only with CF. The metastasis inhibition index in this group of mice was less than in the Cyclophosphamide + calamus polysaccharides group and amounted to 74% versus 90%. In addition, no independent antitumor activity was found in echinacea polysaccharides, unlike calamus polysaccharides. The death of 30% of mice in the combined use of cytostatic and echinacea polysaccharides, 10% of mice in the CF group and 10% of mice in the group treated with echinacea polysaccharides only was also observed (Table 1).

Thus, experiments on female C57BL / 6 female mice with transplanted Lewis lung carcinoma showed the ability of polysaccharides from calamus rhizomes to inhibit the growth of the primary tumor, increase the antitumor and antimetastatic activity of cyclophosphamide, which exceeds that of the reference agent, echinacea polysaccharides, toxin purity and , judging by the absence of dead animals in the groups “Calamus Polysaccharides” and “Cyclophosphamide + Calamus Polysaccharides”.

The effect of polysaccharides of calamus swamp and zymosan on the development of Lewis lung carcinoma (3LL) in 1/6 C57B mice under the conditions of 5-fluorouracil (5-FU) therapy.

The experiment was carried out on female mice of the C57B line 1/6 weighing 18-20 g. Lewis lung carcinoma was transplanted with 5-6 million tumor cells per mouse, intramuscularly (in the hind paw). The calamus and zymosan polysaccharides were started to be administered intraperitoneally at a dose of 10 mg / kg from the 7th day after tumor inoculation and continued for 12 days. 5-Fluorouracil was administered once intramuscularly at a dose of 80 mg / kg on the 14th day of tumor development. Evaluation of the effectiveness of therapeutic effects was performed on 21 days after transplantation of Lewis lung carcinoma: the mass of the primary tumor node, the percentage of inhibition of tumor growth, the incidence rate, the number and area of metastatic nodes in the lungs were determined.

It was shown that the treatment of mice with 3LL polysaccharides of calamus swamp led to a decrease in the mass of the tumor node by 23% (P <0.01), but did not cause a significant change in metastasis compared to those in the control group of animals (Table 2). In contrast to polysaccharides, calamus zymosan, when used alone in mice with carcinoma, did not affect the growth of the primary tumor formation, but the area of metastatic lesion in the lungs of animals with this treatment tended to decrease compared to the control.

During mototherapy with 5-fluorouracil, the tumor mass in mice with 3LL decreased (P <0.01) by only 14% compared with that in the control group of animals. The addition of marsh calf polysaccharides to the cytostatic treatment regimen significantly increased the tumor inhibition percentage to 29% (P <0.05) relative to this indicator in the group of treated 5-FU mice only. Zimosan in combination with an antitumor drug inhibited the growth of the primary tumor node by 25% (P <0.05) compared to that in the group of animals treated with cytostatic in isolation.

The most pronounced effect on the dissemination of malignant neoplasms was exerted by polysaccharides of calamus swamp in combination with 5-fluorouracil. The metastasis inhibition index in this group of 3LL mice was 49% versus 42% in animals that received only cytostatic, and 40% in the group of combined use of 5-FU and zymosan (Table 2).

Thus, in this experiment, the most optimal of all the proposed therapeutic regimens was a combination of polysaccharides of calamus swamp and 5-fluorouracil. In the group of animals treated with this treatment, the highest percent inhibition of growth of the main tumor node and the inhibition index of metastasis were observed. The zymosan polysaccharide isolated from the yeast membrane, with isolated use and in combination with cytostatic (5-FU), showed less pronounced antitumor and antimetastatic activity.

Thus, polysaccharides from rhizomes of calamus marsh in an experiment on laboratory animals increased the antitumor and antimetastatic activity of widely used cytostatic preparations (cyclophosphamide and 5-fluorouracil) more efficiently than the known drugs taken as reference preparations (zymosan and polysaccharides of Echinacea purpurea), in addition , polysaccharides from rhizomes of calamus swamp showed independent antitumor activity, which is described for them for the first time. In this regard, a promising direction is the development of drugs for the complex treatment of malignant neoplasms based on polysaccharides of calamus swamp.

Bibliography

1. Balitsky K.P., Veksler I.G., Struk V.I., Smelkova M.I., Vinnitskaya V.K., Legun A.D. The use of certain polysaccharides in combination chemotherapy for malignant neoplasms. // Actual problems of oncology and medical radiology. - 1970, - v. 2, - p. 360-363.

2. Mashkovsky M.D. Medicines Ed. 13th, v. 1. - Kharkov, 1997 .-- 314 p.

3. Guriev AM, Krylova SG, Razina T.G., Zueva EP, Kalinkina GI, Amosova EN, Yusubov MS Experimental study of antiulcer properties of marsh calf (Acorus calamus L .). // Russian pharmacies. 2003. No. 10.

4. Arkhipov S.A., Junker V.M. The change in the intensity of metastasis to the lungs of transplanted tumors of mice depending on the size of the transplant dose of tumor cells. // Research on the induction and metastasis of tumors in experimental animals. - Novosibirsk, 1984. - S.14-32.

5. Sof'ina Z.P. et al. Experimental evaluation of antitumor drugs in the USSR and the USA. - M.: Medicine, 1980 .-- 296 p.

6. Gubler EV Computational methods of analysis and recognition of pathological processes. - L .: Medicine, 1978.- 193 p.

7. Methods for the study of carbohydrates (translated from English by V. A. Nesmeyanov. / Under the editorship of prof. Harlin) - M .: Mir, 1975. - 445 p.

Table 1
The effect of polysaccharides from rhizomes of calamus marsh and echinacea purpurea during the intraperitoneal route of administration on the effectiveness of cyclophosphamide treatment in mice with Lewis lung carcinoma The observation group, the dose of the drug (number of animals) Tumor mass (X ± m), g Inhibition of tumor growth,% The frequency of metastasis,% The number of metastases per 1 mouse (X ± m) Area of metastases per 1 mouse (X ± m), mm ² Lung mass (X ± m), mg IMI,% 1. Control (10) 6.58 ± 0.17 - one hundred 19.20 ± 2.35 20.42 ± 4.19 187.10 ± 11.40 - 2.PC 125 mg / kg × 1 (9) 6.06 ± 0.24 8 89 8.33 ± 1.78 1-2P _u <0.01 4.40 ± 1.00 1-2P _u <0.01 186.89 ± 5.77 61 3. Calamus polysaccharides 10 mg / kg × 12 (10) 5.57 ± 0.25 1-3P _u <0.01 fifteen one hundred 21.90 ± 3.18 33.48 ± 8.50 197.40 ± 13.12 0 4. CF + polysaccharides calamus 10 mg / kg × 12 (10) 4.54 ± 0.32 1-4P _u <0.01 2-4P _u <0.01 31 70 1-4P _φ <0.001 2.70 ± 1.25 1-4P _u <0.01 2-4P _u <0.05 1.56 ± 0.95 1-4P _u <0.01 2-4P _u <0.05 169.80 ± 5.83 90 5. Echinacea polysaccharides 10 mg / kg × 12 (9) 5.80 ± 0.27 12 one hundred 20.22 ± 2.80 32.30 ± 8.43 201.22 ± 10.57 0 6.PC + Echinacea polysaccharides 10 mg / kg × 12 (7) 5.14 ± 0.32 1-6P _u <0.01 2-6P _u <0.01 22 86 5.86 ± 1.44 1-6P _u <0.01 1.56 ± 0.54 1-6P _u <0.01 2-6P _u <0.05 171.00 ± 3.74 74

table 2
The effect of polysaccharides from calamus rhizomes (PS calamus) of marsh and zymosan on the development of Lewis lung carcinoma in female mice of the C57B line 1/6 and the effectiveness of treatment with 5-fluorouracil (5-FU) The observation group, the dose of the drug (number of animals) Tumor mass (X ± m), g Inhibition or stimulation (+) of tumor growth,% The frequency of metastasis,% The number of metastases per 1 mouse (X ± m) Area of metastases per 1 mouse (X ± m), mm ² Lung mass (X ± m), mg IMI,% 1. Control (10) 5.61 ± 0.17 - one hundred 20.50 ± 2.06 31.64 ± 13.80 222.70 ± 25.25 - 2.5 FU 80 mg / kg × 1 (10) 4.83 ± 0.17 1-2P <0.01 fourteen one hundred 11.90 ± 3.14 1-2P <0.05 5.13 ± 1.33 1-2P <0.01 173.20 ± 10.45 42 3. PS calamus 10 mg / kg × 12 (10) 4.30 ± 0.27 1-3P <0.01 23 one hundred 21.70 ± 3.11 24.13 ± 9.73 213.90 ± 22.34 +6 4. Aira PS 10 mg / kg × 12 + 5-FU 80 mg / kg × 1 (9) 3.97 ± 0.34 2-4P <0.05 29th one hundred 10.44 ± 1.96 6.56 ± 2.10 179.89 ± 8.03 49 5. Zymosan 10 mg / kg × 12 (9) 5.71 ± 0.26 +2 one hundred 17.00 ± 3.09 8.99 ± 3.06 198.11 ± 20.68 17 6. Zimozan 10 mg / kg × 12 + 5-FU 80 mg / kg × 1 (8) 4.19 ± 0.252-6P <0.05 25 88 13.88 ± 4.11 6.65 ± 3.08 183.13 ± 6.39 40

Claims (1)

The use of polysaccharides from rhizomes of calamus marsh (Acorus calamus L.) as a means of increasing the antitumor and antimetastatic activity of cytostatic drugs.