RU2792613C1 - Antitumor agents based on macro- and microelement-containing polygalacturonates (options) - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to a water-soluble agent based on polygalacturonate, which has selective cytotoxic activity against A549 lung carcinoma, MCF-7 breast adenocarcinoma and M-HeLa cervical carcinoma, which is potassium cobalt polygalacturonate (PCCo) or its composition with sodium-cobalt-polygalacturonate (PNaCo) in a mass ratio of 1-2:1-2, and also refers to a water-soluble polygalacturonate-based agent with selective cytotoxic activity against A549 lung carcinoma, MCF-7 breast adenocarcinoma and cervical carcinoma M-HeLa, which is a composition containing sodium-cobalt-polygalacturonate (PNaCo) in a mass ratio of 3:1 and a metal complex of pectin biopolymer sodium-metal-polygalacturonate, respectively, where the biogenic metal is zinc, manganese, chromium or magnesium (PGNaZn, PGNaMn, PGNaCr, PGNaMg).

EFFECT: obtaining antitumor agents based on macro- and microelement-containing polygalacturonates.

2 cl, 1 dwg, 3 tbl

Description

The claimed group of inventions relates to the chemical and pharmaceutical industry, in particular, to metal complexes of pectin biopolymers and compositions based on them, which have antitumor activity against A549 lung carcinoma, MCF-7 breast adenocarcinoma, M-HeLa cervical carcinoma and can find application in medicine and pharmacology.

Cancer is currently the second leading cause of death after cardiovascular disease. There are more than 200 types of human malignant tumors, and between tumors of even one organ there are differences in the histological structure, in growth characteristics, in interaction with the body, etc. The main disadvantage of anticancer drugs available in the pharmacy network is high toxicity, low bioavailability and lack of selectivity for normal cells of living organisms.

Therefore, reducing the toxicity of known anticancer drugs while maintaining their effectiveness remains an urgent task. It is known from the literature that in order to increase the effectiveness of the treatment of tumor diseases, it is necessary to create new drugs that more selectively act on tumor cells, optimize the methods of using antitumor agents, improve chemotherapy as a link or stage in the complex and combined treatment of patients with malignant tumors, and use pharmacological agents to reduce side effects of anticancer drugs, etc. [Pankratov Andrey Aleksandrovich. Abstract of the dissertation "Antineoplastic efficacy and toxicity of binary catalytic systems based on cobalt metal complexes: Experimental study" for the degree of Ph.D. specialty 14.00.14. - 31 p.].

The use of biopolymers, including pectin polysaccharides, is a promising approach to solving this problem. Pectins are attracting more and more attention of researchers as potential drugs and biologically active additives in the treatment of malignant tumors, especially in the inhibition of metastasis and rehabilitation after chemotherapy. Pectin polysaccharides are plant biopolymers of a polyuronide nature with functional properties (complexing and gelling ability), which makes them promising as radioprotectors for removing radionuclides from the body, as well as in the food industry (E440). Pectins are part of the cell walls of plants [Ovodov Yu.S./ Modern ideas about pectin substances // Bioorganic chemistry, 2009. V. 35. No. 3. pp.293-310 (review); Graham B. Seymour, J. Paul Knox / Pectins and their Manipulation // U.S.A. Canada CRC Press. 2002. 250 rubles; Pectin: Chemical Properties, Uses and Health Benefits (Food Science and Technology) / Ed. Ph.L. Bush. New York: Nova science publishers, 2014. 268 rubles], they are rich in vegetables and fruits eaten. The main industrial types of commercial pectin produced in the world are apple and citrus pectins.

Pectin polysaccharides and products of their modification have a wide range of biological activity: immunomodulatory, anti-inflammatory, antibacterial, antioxidant and antitumor activity [S.T. Minzanova, V.F. Mironov, D.M. Arkhipova, A.V. Khabibullina, L.G. Mironova, Y.M. Zakirova, V.A. Milyukov. Biological Activity and Pharmacological Application of Pectic Polysaccharides: A Review // Polymers. 2018, 10(12), 1407. DOI: 10.3390/polym10121407].

It is known that native pectins extracted from plant materials, and to a greater extent pectins modified in various ways, reduce the proliferation, migration and adhesion of cancer cells and induce apoptosis in various tumor lines [Maxwell, E.G.; Colquhoun, I. J.; Chau, H.K.; Hotchkiss, A.T.; Waldron, K. W.; Morris, V.J.; Belshaw, N.J. Rhamnogalacturonan I containing homogalacturonan inhibits colon cancer cell proliferation by decreasing ICAM1 expression // Carbohydr. Polym. 2015. 132, 546-553]. Maxwell E.G. et al evaluated the effectiveness of pectins from different sources (potato, sugar beet, larch and citrus fruits) for the treatment of colon cancer. Colon and rectal cancer is the third most common diet-related cancer in the world. Studies have shown that increased consumption of fruits and vegetables helps reduce the risk of developing this disease. The authors found that by varying the dose of rhamnogalacturonan-I extracts of potato pectin, it is possible to reduce the proliferation of colon cancer cells.

The patent document [US5895784 (A), - 1999-04-20] discloses a method for treating cancer in mammals for inhibiting metastasis of primary tumors by oral administration of a therapeutic amount of modified pectin, namely, pH-modified pectin having a molecular weight of ~ 1-15 kDa (mainly 10 kDa), or the claimed composition, which contains a mixture of dry citrus pectin with a modified pH and a pharmaceutically acceptable digestible carrier. In the Dunning (R3327) model of prostate adenocarcinoma, developed by Dunning from a spontaneously occurring adenocarcinoma that, when injected with 1 x 10 ⁶ MLL cells into the rat thigh, results in disease. During approximately 25 days of widespread infection, an in vivo study of the average number of metastases (%) in the lungs versus the concentration of modified citrus pectin (0.01%, 0.1%, 1.0%) is illustrated. It was found that the concentration of modified citrus pectin 1.0% provides the maximum reduction in metastases. The technical result - the reduction of metastases - is provided by pectin with a modified pH, which binds to proteins that bind carbohydrates on the surface of tumor cells, for example, demethoxylated polygalacturonic acid.

[WO2009096789 (A1), 2009-08-06] proposes a composition comprising galactooligosaccharide, fructooligosaccharide and pectin hydrolyzate in the ratio (5-9):1:(2-10), respectively, designed to stimulate the activity of natural killer cells (NK -cells, large granular lymphocytes) of people whose activity of these cells is reduced by 10-40% compared to healthy people, including cancer patients, to prevent the progression of the disease. The authors have shown that the use of this composition increases the activity of NK cells, determined using the commercially available test kit NKTEST®, manufactured by ORPEGEN Pharma, by 30-50%, compared with the activity of NK cells before the claimed composition enters the body. Predominantly acid oligosaccharides (derived from high methoxyl pectin) have a methylation degree of 70%, less than 50%, and even less than 20%. The pectin hydrolyzate was administered in an amount of 10 mg to 100 g per day, or 100 mg to 50 g per day, or 500 mg to 20 g per day. In addition, it is proposed to use a composition with the additional inclusion of N-acetylcysteine, and additionally, the composition may include proteins, fats, easily digestible carbohydrates, vitamins and minerals - at least 10% milk protein and 10-50% fat of the total calorie content of the composition.

The invention [CN113025674 (A), 2021-06-25] discloses a method for obtaining an enzymolysis product of lemon pectin (modified citrus pectin, MCP) with antitumor activity. The method includes the following steps: triple hydrolysis with MCP pectinase in buffer (at pH NaAc/HAc buffer 4.2) at a temperature of 50°C, enzymolysis of MCP with pectinase at room temperature overnight, heating in a boiling water bath for 10 min, centrifugation at 4000 rpm for 15 min after cooling to remove inactivated pectinase, dialysis in a dialysis bag with separation of a fraction with a molecular weight of 1000 Da to obtain a mixed product after enzymolysis of MCP, and then sequential separation by column chromatography and purification of the enzymolysis product using DEAE-cellulose and Sephadex to obtain five purified pectin enzymatic hydrolysis products with different monosaccharide composition. The invention discloses the possibility of using the lemon pectin enzymolysis product against MCF-7 human breast cancer cells and A549 non-small cell lung cancer cells. The antitumor activity of each component of MCP enzymatic hydrolysis was assessed by inhibition of proliferation of human breast cancer cells MCF-7 with high expression of galectin-3 and non-small cell lung cancer A549. Under the condition of introducing a dose of 63-2000 μg / ml, after treatment of MCF-7 cells with five components, the proliferative activity of MCF-7 cells decreases to 53.4%, 55.4%, 61.0%, 36.7% and 45.9%, respectively; the proliferative activity of A549 cells decreases to 89.3%, 64.8%, 53.4%, 57.4%, and 82.8%, respectively.

A review of the authors is known [Zhang, W., Xu, P., & Zhang, H. Pectin in cancer therapy: A review // Trends in Food Science & Technology, 2015. 44. 258-271. DOI: 10.1016/j.tifs.2015.04.001], which summarizes data on the potential of commercial pectins and pectins from non-traditional plant sources in cancer therapy. The necessity of additional study of fundamental issues for the promotion of pectin in cancer therapy is substantiated: a detailed description of the role of galectin-3 (Gal-3), modification of pectin as such, as well as their pharmacokinetics and pharmacodynamics.

Known work [Lionel Leclere, Maude Fransolet, Francois Cote, Pierre Cambier, Thierry Arnould, Pierre Van Cutsem, Carine Michiels. Heat-Modified Citrus Pectin Induces Apoptosis-Like Cell Death and Autophagy in HepG2 and A549 Cancer Cells // PLOS ONE. 2015, 1-24 DOI: 10.1371/journal.pone.0115831], in which the authors evaluated the cytotoxicity of thermally fragmented citrus pectin and native citrus pectin against hepatocellular carcinoma (HepG2) and lung carcinoma (A549) cell lines after 24 hours and 48 hours of incubation. HepG2 cells and A549 cells were incubated in comparison to control medium, 50 μM etoposide, 3 mg/ml thermally fragmented citrus pectin, or 3 mg/ml native citrus pectin. The thermally fragmented citrus pectin fraction showed antitumor activity against HepG2 cancer cells: it gives a cell death rate of 95% at a concentration of 5 mg/ml and has an IC _{50 of} 3 mg/ml. The antitumor activity of native citrus pectin is very low.

Known work of the authors [Delphi L., Sepehri H. Apple pectin: A natural source for cancer suppression in 4T1 breast cancer cells in vitro and express p53 in mouse bearing 4T1 cancer tumors, in vivo. Biomed. Pharmacother. 2016. 84. 637-644. DOI: 10.1016/j.biopha.2016.09.080], which describes the ability of apple pectin to induce cancer cell death and suppress tumor growth in vivo in a mouse breast cancer model. The results of in vitro studies showed that 0.1% concentration of pectin acid can induce apoptosis, inhibit the growth of tumor cells (p<0.001) and reduce cell adhesion. In addition, in vivo studies in the 4T1 breast cancer animal model have shown that pectic acid can inhibit tumor growth by overexpressing p53 and increasing apoptotic cells.

The work [Lin, L.; Wang, P.; Du, Z.; Wang, W.; Cong, Q.; Zheng, C.; Jin, C.; Ding, K.; Shao, C. Structural elucidation of a pectin from flowers of Lonicera japonica and its antipancreatic cancer activity, Int. J Biol. macromol. 2016. 88. 130-137. DOI: 10.1016/j.ijbiomac.2016.03.025], the authors of which showed that the pectin polysaccharide from the flowers of Lonicera japonica is promising for the creation of a powerful drug for the treatment of pancreatic cancer on its basis. Pectin polysaccharide can inhibit the growth of PANC-1 pancreatic cancer cells at a concentration of 1 mg/ml with an inhibition ratio of 52.1%.

Authors' results [Hadjira Hamai-Amara, Khalida Abdoun-Ouallouche, Assia Nacer-Khodja, Karima Abdelhafid, Abdelouafi Benmouloud & Assia Djefal-Kerrar. Optimization of the extraction of orange peel pectin and evaluation of its antiproliferative activity towards HEp2 cancer cells // Euro-Mediterranean Journal for Environmental Integration. 2020. 5, article number: 43] demonstrate the pro-apoptotic effect of chemically modified and irradiated pectin isolated from orange peels on the laryngeal cancer cell line (HEp2). The strongest proapoptotic activity against HEp2 cells was observed after 72 hours of contact with chemically modified and irradiated orange peel pectin. With this modified pectin, cancer cell mortality of 95% (IC ₉₅ 5 mg/mL) was achieved with an IC _{50 of} 3.18 mg/mL.

There is a review by M. Khotimchenko, which discusses various forms of drug delivery systems based on pectin (hydrogels, tablets, films, microspheres, nanoparticles, etc.) and presents an extensive literature on pectin biopolymers as a means of drug delivery to the colon. It has been shown that pectins can suppress tumor growth by inhibiting cell growth, reducing cell adhesion [M. Khotimchenko. Pectin polymers for colon-targeted antitumor drug delivery // International Journal of Biological Macromolecules. 2020. 158. 1110-1124. DOI: 10.1016/j.ijbiomac.2020.05.002].

Authors [Kamonrak Cheewatanakornkool, Sathit Niratisai, Somkamol Manchun, Crispin R. Dass, Pornsak Sriamornsak. Characterization and in vitro release studies of oral microbeads containing thiolated pectin - doxorubicin conjugates for colorectal cancer treatment // Asian journal of pharmaceutical sciences. 2017. 12 509-520; Cheewatanakornkool K, Niratisai S, Manchun S, Dass CR, Sriamornsak P. Thiolated pectin-doxorubicin conjugates: Synthesis, characterization and anticancer activity studies // Carbohydr. Polym. 2017. V. 174. P. 493-506. DOI: 10.1016/j.carbpol.2017.06.115] thiolated pectin-doxorubicin (DOX) conjugates were obtained, characteristics, physicochemical properties and morphology of thiolated pectins and thiolated pectin-DOX conjugates were determined. The in vitro anticancer activity of thiolated pectin-DOX conjugates was significantly higher than that of free DOX in human colon carcinoma and bone osteosarcoma cells, but was not significantly different from that of native DOX in human prostate cancer cells. This makes thiolated pectin-DOX conjugates promising for DOX delivery aimed at the treatment of colorectal cancer.

Authors [Indu Hira, Reena Kumari, Adesh Kumar Saini, Henok Gullilat, Vipin Saini, Anil Kumar Sharma, Reena V. Saini. Apoptotic Cell Death Induction Through Pectin, Guar Gum and Zinc Oxide Nanocomposite in A549 Lung Adenocarcinomas // Biointerface Research in Applied Chemistry. 2022. 12. 2. 1856-1869. DOI: 10.33263/BRIAC122.18561869] revealed in vivo anticancer potential of the Pec-gg-ZnO nanocomposite against A549 lung carcinoma, cervical carcinoma (HeLa), and prostate adenocarcinoma (PC-3) cells. The IC ₅₀ values were 83.67 (0.10 µg/mL, 87.25 (0.03 µg/mL) and 85.95 (0.03 µg/mL for A549, HeLa, and PC-3 cells, respectively. than pectin (IC ₅₀ 781.70 (0.09 µg/mL) and guar gum (IC ₅₀ 374.20 (0.08 µg/mL).

In [Antineoplastic activity of pectins of various molecular weights / E.A. Leontiev, V.V. Vikharev, A.A. Kalitnik, M.Yu. Khotimchenko // Pacific Medical Journal, 2020. No. 4 (82). 68-72. DOI: 10.34215/1609-1175-2020-4-68-72] an analysis of the dependence of the antitumor properties of various pectins on structural features and molecular weight is given. To study the antitumor effect of polysaccharides, cell lines of mouse breast adenocarcinoma C127, cervical carcinoma HeLa, human colon carcinoma SW620, human breast adenocarcinoma HTB-30, and normal retinal cells RPE were used. The evaluation of the antitumor and cytotoxic effects of pectins was carried out using the test for metabolic activity by the MTT assay and by in vivo staining of living and dead cells with fluorescent DNA-binding dyes Hoechst and propidium iodide. It was shown that samples of citrus pectins in the entire selected range of concentrations had a significant antitumor effect on the C127 adenocarcinoma cell line, reducing metabolic activity and cell proliferation, and also exhibiting a pronounced cytotoxic effect.

Anticancer drugs containing metal ions play an important role in tumor chemotherapy.

High cytotoxic activity of biogenic metal coordination compounds is known in the literature [D.A. Guk, O.O. Krasnovskaya, E.K. Beloglazkina. Russ. Chem. Rev., 2021. 90(12). 1566 - 1623 (review). https://doi.org/10.1070/RCR5016?locatt=label:RUSSIAN] and catalytic systems (transition metal complexes (Co, Fe, Mn, etc.), as well as their prospects as potential antitumor agents. The review shows how varying the structure of the ligand can significantly change not only the cytotoxicity of the coordination compound, but also the mechanism of binding to DNA, and in some cases make another mechanism of cytotoxic action the main one.

In [Pankratov Andrei Aleksandrovich. Abstract of the dissertation "Antineoplastic efficacy and toxicity of binary catalytic systems based on cobalt metal complexes: Experimental study" for the degree of Ph.D. specialty 14.00.14. - 31 pp.] the author studied the antitumor and toxic properties of binary catalytic systems based on complexes of cobalt and ascorbic acid on experimental animals in vivo.

The applicant in the prior art has not identified pectin metal complexes that exhibit cytotoxic activity against the tumor cell line A549 (human lung carcinoma), MCF-7 (breast adenocarcinoma). Known primary data on the inhibition of the viability of cervical carcinoma cells M-HeLa when exposed to sodium-cobalt-polygalacturonate (PGNaCo), the value of IC ₅₀ was 2.00% ± 0.20, obtained by the author's team of the applicant [Minzanova S.T., Voloshina A.D., Arkhipova D.M., Krayushkina A.V., Mironova L.G., Sapunova A.S., Kufelkina A.A., Kulik N.V., Mironov V.F., Milyukov V.A. Synthesis and antitumor activity of cobalt complexes with sodium polygalacturonate // Proceedings of the XIII International Conference "New and non-traditional plants and prospects for their use", Sochi, June 4-8, 2018 - Moscow: RUDN University, 2018. - P. 634-637].

The analysis of literature data demonstrates that the response of tumor cells to the action of pectins depends both on the type of compound used (pectins from various plant sources, pectins with different properties, such as molecular weight, viscosity, degree of esterification), and on the sensitivity of the cells themselves to pectins. In addition, some pectin preparations are poorly soluble in water, due to which, in order to achieve a therapeutic effect, it becomes necessary to increase the dose of the drug, which in turn causes severe side effects. It is known that with an increase in the solubility of drugs for oral administration, their effectiveness increases, so the creation of bioavailable (water-soluble or fat-soluble) drugs based on pectin biopolymers remains an urgent task.

Lung cancer and breast cancer are serious medical and social problems. In developed countries, lung cancer is the most common malignant tumor and the most common cause of death from cancer. According to the International Agency for Research on Cancer, about one million new cases of lung cancer are registered annually in the world, and 60% of cancer patients die as a result of this disease. In Russia, lung cancer ranks first among oncological diseases, its share in this pathology is twelve percent, and in fifteen percent of cases in deceased cancer patients, lung cancer was diagnosed. According to WHO data in 2000, men suffer from lung cancer much more often: lung cancer caused the death of 32.0% of men, and 7.2% of women who had any malignant neoplasms [V.A. Nidyulin B.V. Erdniev. On the epidemiology of lung cancer // Medical Bulletin of Bashkortostan: review article // Bashkir State Medical University, 2009. 4 (1). 66-71. ISSN 1999-6209].

Every tenth woman faces breast cancer in Russia. The most common type of oncological pathology of the mammary glands is adenocarcinoma, it can be of two types: as a neoplasm that develops in the lobules of the mammary gland (10%); or as a tumor that occurs in the thoracic ducts (90%). Diagnosis of breast adenocarcinoma necessarily includes standard clinical blood and urine tests, and hormonal levels are also assessed. In addition, blood is donated for oncological markers (the index of the specific antigen CA15-3 is of the greatest importance). According to indications, genetic testing is carried out for certain genes [Surgical diseases: textbook / edited by A.F. Chernousov. M.: GEOTAR-MEDIA, 2010. 664 p. - ISBN 978-5-9704-1278-7].

Cervical cancer is a malignant neoplasm that occurs in the area of the cervix. Histologically, its two main varieties are distinguished: carcinoma and squamous cell carcinoma. The incidence of cervical cancer significantly exceeds the incidence of other tumors of the female reproductive system. At the beginning of the 21st century, there is an increase in patients with advanced cervical cancer: the proportion of stage IV cancer, according to various sources, is 37.1% - 47.3% [Paliychuk O.V., Polishchuk L.Z. The role of infectious factors in the etiology and pathogenesis of intraepithelial neoplasia and cervical cancer // Reproductive health of women. 2007. 32(3). 10-16; Modern approaches to the surgical treatment of the cervix. Report of Morkhov K.Yu. at the oncological congress "White Nights". 2015; https://www.youtube.com/watch?v=sX0Jkgu2-o0].

The technical problem to be solved by the invention consists in expanding the arsenal of water-soluble agents based on pectin biopolymer, which at the same time have selective cytotoxic activity against the tumor cell line of human lung carcinoma A549, breast adenocarcinoma MCF-7 and cervical carcinoma M-HeLa, practically no toxic effect on normal human cells.

The technical result provided by the invention is the realization of the specified purpose by the invention.

The technical problem is solved, and the technical result is achieved by the proposed group of inventions related to objects of the same type of the same purpose and providing the same technical result (options) - agents that have a selective antitumor effect in lung carcinoma A549, breast adenocarcinoma MCF-7 and cervical carcinoma M-HeLa, while not affecting normal human cells by means - water-soluble metal complexes of pectin polysaccharides with biogenic macro- and microelements and pharmacological compositions based on them.

In one embodiment of the invention, the claimed agents are water-soluble sodium cobalt polygalacturonate (PNaCo), or potassium cobalt polygalacturonate (PGKCo), or mixtures thereof in a mass ratio of 1-2:1-2.

In another embodiment of the invention, the claimed agents are new, previously not described, water-soluble pharmacological compositions containing sodium cobalt polygalacturonate (PNaCo) and a metal complex of a pectin biopolymer selected from the group of sodium zinc polygalacturonate, or sodium manganese polygalacturonate, or sodium chromium-polygalacturonate, or sodium-magnesium-polygalacturonate (PGNaZn, PGNaMn, PGNaCr, PGNaMg), in a mass ratio of 3:1, respectively.

At the same time, selective cytotoxic activity is declared:

water-soluble sodium cobalt polygalacturonate (PGNaCo) against A549 lung carcinoma and MCF-7 breast adenocarcinoma (PGNaCo activity against M-HeLa cervical carcinoma is known from the prior art);

water-soluble potassium-cobalt-polygalacturonate (PGKCo), water-soluble pharmacological compositions, which are a mixture of potassium-cobalt-polygalacturonate (PGKCo) and sodium-cobalt-polygalacturonate (PGNaCo) in a mass ratio of 1-2:1-2, and

water-soluble pharmacological compositions containing sodium-cobalt-polygalacturonate (PGNaCo) and a pectin biopolymer metal complex selected from the group of sodium-zinc-polygalacturonate, or sodium-manganese-polygalacturonate, or sodium-chromium-polygalacturonate, or sodium-magnesium-polygalacturonate (PGNaZn, PGNaMn, PGNaCr, PGNaMg), in a mass ratio of 3:1, respectively, -

against A549 lung carcinoma, MCF-7 breast adenocarcinoma, and M-HeLa cervical carcinoma.

According to the hazard classification of substances according to the degree of impact on the body [GOST 12.1.007.76 and Izmerov N.F., Sanotsky I.V., Sidorov K.K., Parameters of toxicometry of industrial poisons with a single injection (reference book), M .: Medicine, 1977 pp.196-197] the claimed antitumor agents do not exhibit toxic effects on experimental animals. When administered orally to mice in doses of 5000-15000 mg/kg, they do not cause the death of animals. This allows us to classify the claimed funds to the class of low-toxic (low-hazard) compounds.

Metal complexes based on sodium polygalacturonate with a degree of salt formation of 100% - PGNaCo, PGNaZn, PGNaMn, PGNaCr and PGNaMg - are known from the prior art, namely, information about the structure, characteristics and method of obtaining [RU2219187 C1, 20.12.2003; S.T. Minzanova, V.F. Mironov, A.B. Vyshtakalyuk, O.V. Tsepaeva, L.G. Mironova, A.Z. Mindubaev, I.R. Nizameev, K.V. Kholin, V.A. Milyukov. Complexation of pectin with macro- and microelements. Antianemic activity of Na, Fe and Na, Ca, Fe complexes // Carbohydrate Polymers, 2015. 134. 524-533 (DOI: 10.1016/j.carbpol.2015.07.034); V.F. Mironov, A.N. Karaseva, O.V. Tsepaeva, A.B. Vyshtakalyuk, S.T. Minzanova, V.I. Morozov, V.V. Karlin, E.R. Yunusov, A.Z. Mindubaev. Some new aspects of complex formation of pectin polysaccharides with d-metal cations. // Chemistry and computer modeling. Butler messages. 2003. 4(1-3). S.45-50; Vasilii A. Milyukov, Anna V. Khabibullina, Daria M. Arkhipova, Vladimir F. Mironov, Ayrat R. Khamatgalimov, Irina S. Ryzhkina, Leysan I. Murtazina, Lubov G. Mironova, Alexandra B. Vyshtakalyuk, Andrey V. Nemtarev, Nail G. Nazarov, Kirill V. Kholin, Irek R. Nizameev, and Salima T. Minzanova. Synthesis, Physicochemical Properties and Anti-Fatigue Effect of Magnesium, Zinc and Chromium Polygalacturonate Based Composition // ChemistrySelect. 2019. 4 (14). 4331-4338. DOI:10.1002/slct.201803812; RU2564949 C1, 10.10.2015].

The characteristics of metal complexes based on sodium and potassium polygalacturonates, including potassium-cobalt-polygalacturonate (PCCo), previously unknown from the prior art, are presented in Table 1. The metal content in the claimed products was determined by atomic emission spectroscopy at the NPP iCAP 6300 DUO ICP. The specific optical rotation of pectin metal complexes was measured on a Perkin-Elmer 341 polarimeter (concentration c is given in g/100 ml, wavelength 589 nm, temperature 20°C). The elemental composition of the samples was determined on a Euro EA - CHNSO Elemental Analyzer instrument using a streptocide standard (C ₆ H ₈ O ₂ N ₂ S).

The method for obtaining potassium polygalacturonate (PGK) with a 100% degree of salt formation is similar to that described in [RU2281957 C1, 20.08.2006] method for obtaining sodium polygalacturonate (PGNa) by treating pectin with alkali at 50-60°C, but differs in that bringing the pectin solution to pH 9 is carried out by adding 0.1 n. KOH solution instead of NaOH. Potassium polygalacturonate (PGK) - beige powder, IR spectrum (KBr, v , cm- ¹ ): 1610 (C=O).

The method for producing potassium-cobalt-polygalacturonate (PGKCo) is similar to the method for producing sodium-cobalt-polygalacturonate (PNaCo) [RU 2281957 C1, 20.08.2006], but differs in that potassium polygalacturonate is used as the starting compound instead of sodium polygalacturonate. Potassium-cobalt-polygalacturonate (PGKCo) - amorphous powder of lilac color, IR spectrum (KBr, v , cm- ¹ ): 1609 (C=O).

Sodium and potassium are alkali metals of the first group of the main subgroup, found in all known biological systems. Water-soluble pharmacological compositions are obtained, which are a mixture of potassium-cobalt-polygalacturonate (PGKCo) and sodium-cobalt-polygalacturonate (PGNaCo) by simply mixing in a mass ratio of 1-2: 1-2 water-soluble PGNaCo and PGACo with thorough mixing, for example, in a ball mill within 3-5 minutes.

As already mentioned, the antitumor properties of sodium cobalt polygalacturonate (PGNaCo) against cervical carcinoma M-HeLa were previously described by us [Minzanova S.T., Voloshina A.D., Arkhipova D.M., Krayushkina A.V., Mironova L.G., Sapunova A.S., Kufelkina A.A., Kulik N.V., Mironov V.F., Milyukov V.A. Synthesis and antitumor activity of cobalt complexes with sodium polygalacturonate // Proceedings of the XIII International Conference "New and non-traditional plants and prospects for their use", Sochi, June 4-8, 2018 - Moscow: RUDN University, 2018. P. 634-637], however the ability of PGNaCo to inhibit the growth of A549 lung carcinoma and MCF-7 breast adenocarcinoma cells, as well as cytotoxic activity against A549 lung carcinoma, MCF-7 breast adenocarcinoma, and M-HeLa cervical carcinoma potassium cobalt polygalacturonate (PCCo) and mixtures potassium-cobalt-polygalacturonate (PGKCo) and sodium-cobalt-polygalacturonate (PGNaCo) in a mass ratio of 1-2:1-2, are not known to the applicant from the prior art.

The proposed pharmacological compositions according to the second variant of the invention, containing sodium-cobalt-polygalacturonate (PNaCo) with one of the pectin biopolymer metal complexes (PNaZn, PNaMn, PNaCr, PNaMg) are also new, not previously described. These pharmacological compositions are obtained by simple mixing in a mass ratio of 3:1 PGNaCo with a metal complex of a pectin biopolymer with macro- and microelements with thorough mixing, for example, in a ball mill for 3-5 minutes.

The ability to inhibit the growth of cancer cells of A549 lung carcinoma, MCF-7 breast adenocarcinoma and M-HeLa cervical carcinoma of pharmacological compositions based on PGNaCo with one of PGNaZn, PGNaMn, PGNaCr, PGNaMg at a mass ratio of 3:1 is also not known to the applicant from the prior art.

Table 2 shows the content of metals in the claimed compositions.

Maintaining normal homeostasis is associated with the decomposition and synthesis of proteins, carbohydrates and fats in the body, the possibility of passing through and regulation of which is determined by metal ions. In the case of two- and multicomponent compositions, the types of interaction between metal ions are of particular importance. The end result of the physiological effect may be due to synergy or antagonism. The mass ratio of PGNaCo with the metal complex of pectin biopolymer (PGNaZn, PGNaMn, PGNaCr, PGNaMg) in the claimed pharmacological compositions is due to a decrease in cytotoxic activity with a decrease in the proportion of PGNaCo in the composition.

Below is a brief description and the biological role of biogenic metals in the claimed products (pharmacological compositions): microelements (cobalt, zinc, chromium and manganese), as well as a macroelement - magnesium.

Cobalt (Co) is an important redox trace element widely distributed in mammalian cells. This metal is involved in a number of redox processes, alkylation and the generation (destruction) of free radicals that occur in a living cell. The most common mechanism of cytotoxicity found for cobalt(II) coordination compounds is DNA binding and subsequent triggering of apoptosis [D.A. Guk, O.O. Krasnovskaya, E.K. Beloglazkina // Russ. Chem. Rev., 2021, 90 (12) 1566 - 1623. DOI: 10.1070/RCR5016?locatt=label:RUSSIAN].

Malignant tumors develop more easily against the background of a reduced level of zinc (Zn). Zinc is involved in cell metabolism and is contained in the active center of more than 200 metalloenzymes. Zinc supplementation stimulates the production of white blood cells, one of the antitumor components of the immune system, and supports the overall activity of neutrophils and T-lymphocytes. Taking into account physiological needs, a daily prophylactic dose of 15 - 25 mg of zinc is recommended [D.A. Guk, O.O. Krasnovskaya, E.K. Beloglazkina. Russ. Chem. Rev., 2021, 90(12). 1566-1623, DOI: 10.1070/RCR5016?locatt=label:RUSSIAN; Gray, L. Zinc. Marshall Cavendish / L. Gray. 2006. - R. 9].

Magnesium (Mg), like zinc, has an immunomodulatory effect. Magnesium is found in all tissues of the body and is necessary for the normal functioning of cells. Participates in most metabolic reactions, in the regulation of the transmission of nerve impulses and in muscle contraction, has an antispasmodic and antiplatelet effect. The most general effect of Mg ²⁺ on any tissue is that Mg ²⁺ ions stabilize the structure of the transfer RNA that controls the overall rate of protein resynthesis. With magnesium deficiency, destabilization of transport - non-coding RNA occurs (the number of dysfunctional RNA molecules increases), which is accompanied by a decrease and slowdown in the rate of synthesis of protein structures of cells with a relative predominance of apoptosis processes (one of the mechanisms of aging). The daily requirement for magnesium is 350-500 mg [Russian Medical Journal: https://www.rmj.ru/articles/kardiologiya/Roly_deficita_magniya_v_ patogeneze_metabolicheskogo_sindroma/#ixzz7EwdOkfBz].

Chromium (Cr) is one of the biogenic elements that is part of the tissues of plants and animals. In animals, chromium is involved in the metabolism of lipids, proteins (part of the trypsin enzyme), and carbohydrates. A decrease in the content of chromium in food and blood leads to a decrease in the growth rate, an increase in cholesterol in the blood. The biological activity of chromium is explained by the ability of Cr ³⁺ ions to form complex compounds. For example, Cr ³⁺ ions are involved in stabilizing the structure of nucleic acids. Chromium has an effect on hematopoiesis, has the ability to activate trypsin, as it is part of crystalline trypsin in the form of a labile compound capable of splitting off chromium ions. Chromium salts inhibit alcoholic fermentation, accelerate the work of insulin; affect carbohydrate metabolism and energy processes. The body of an adult contains 6 mg of chromium [AP Avtsyn, AA Zhavoronkov, MA Rish. Mikroelementozy cheloveka // Human Micronutrient Balance Disorders. Moscow. 1991. 496 p.].

An important trace element is manganese, the biogenic function of Mn ²⁺ ions is to regulate the activity of enzymes. Therefore, Mn ²⁺ ions have a wide range of biological effects: they affect hematopoiesis, mineral metabolism, growth, reproduction, etc. In addition, Mn ²⁺ ions stabilize the structure of nucleic acids. Manganese protects the walls of arteries, making them resistant to the formation of atherosclerotic plaques. Manganese is vital for brain function, for the formation of skin pigment, is part of proteins and enzymes, and has a pronounced antioxidant activity. The daily requirement for manganese is 2.5 - 5.0 mg [AP Avtsyn, AA Zhavoronkov, MA Rish. Mikroelementozy cheloveka // Human Micronutrient Balance Disorders. Moscow. 1991. 496 p.].

Study of the antitumor activity of the claimed agents

Evaluation of the cytotoxic effect of the claimed agents on cells of tumor lines was carried out in vitro on cell cultures of human lung carcinoma A549, breast adenocarcinoma MCF-7 (pleural fluid) and cervical carcinoma M-HeLa from the collections of the Institute of Cytology of the Russian Academy of Sciences (St. Petersburg); liver cells (Chang liver from the collection of the Research Institute of Virology of the Russian Academy of Medical Sciences (Moscow) were used as normal cell lines).

The cytotoxic effect of the claimed agents is determined by counting viable cells of A549, MCF-7 and M-HeLa tumor cultures, as well as cultures of normal liver cells (Chang liver) using the multifunctional Cytell Cell Imaging system (GE Helthcare Life Science, Sweden) using the Cell application Viability BioApp, which allows you to accurately count the number of cells, assess their viability based on fluorescence intensity [A.D. Voloshina, A.S. Sapunova, N.V. Kulik, M.G. Belenok, I.Yu. Strobykina, A.P. Lyubina, S.K. Gumerova, V.E. Kataev, Antimicrobial and cytotoxic effects of ammonium derivatives of diterpenoids steviol and isosteviol, Bioorg. Med. Chem. 32 (2021) 115974. https://doi.org/10.1016/j.bmc.2020.115974].

For cultivation of cells using a standard nutrient medium "Needle" produced by the Moscow Institute of Poliomyelitis and Viral Encephalitis. M.P. Chumakov company "PanEco" with the addition of 10% fetal calf serum and 1% non-essential amino acids.

Cell sieving is carried out on a 96-well Eppendorf panel at a concentration of 100,000 cells/ml in each well in a volume of 150 μl of medium and cultivated in a CO ₂ incubator at 37°C. medium, and 150 μl of solutions of the studied drug in given dilutions are added to the wells. Compound dilutions are prepared directly in the growth medium.

The cytotoxic effect of the claimed agents is determined in the following concentrations, wt. %: 0.047%, 0.093%, 0.1875%, 0.375%, 0.75%.

Calculation of the inhibitory concentration at which 50% of the cells die - IC ₅₀ - is done using the online calculator: MLA - "Quest Graph Calculator (IC50") AAT Bioquest, Inc, June 4, 2021, https://www.aatbio .com/tools/ic50-calculator [V. Kaur, M. Kumar, A. Kumar, K. Kaur, V. Singh Dhillon, S. Kaur, Pharmacotherapeutic potential of phytochemicals: Implications in cancer chemoprevention and future perspectives // Biomed. Pharmacother 2018 97 564-586 DOI: 10.1016/j.biopha.2017.10.124].

O, Villarreal M,

L, Meneses-Acosta A,

V. Cytotoxicity, post-treatment recovery, and selectivity analysis of naturally occurring podophyllotoxins from bursera fagaroides var. fagaroides on breast cancer cell lines // Molecules. 2016; 21 (8). 1013. DOI: 10.3390/molecules21081013].The selectivity index (SI) is calculated as the ratio of the IC ₅₀ value for normal cells to the IC ₅₀ value for cancer cells; compounds with SI≥10 are considered highly selective [

Oh Villarreal M

L, Meneses-Acosta A,

V. Cytotoxicity, post-treatment recovery, and selectivity analysis of naturally occurring podophyllotoxins from bursera fagaroides var. fagaroides on breast cancer cell lines // Molecules. 2016; 21(8). 1013. DOI: 10.3390/molecules21081013].

The results of the claimed antitumor drugs in comparison with antitumor drugs of plant origin are presented in table 3. As reference drugs, the original citrus pectin, the methanol extract of Thymus vulgaris L. (common thyme) are used [Balakina A.A., Kuzmina E.A., Drewova A. .N., Mubarak M.M., Karsunkina N.P., Kalashnikova E.A., Cherednichenko M.Yu. Evaluation of the cytotoxic effect of extracts from medicinal plants on the M-HeLa cell line. Electronic scientific and production journal "AgroEcoInfo"] and the drug Arglabin, which is a lyophilisate of biologically active sesquiterpene lactone isolated from wormwood Artemisia glabella (Karaganda Pharmaceutical Complex, LLP) [N.A. Neverova, A.N. Zhabaeva, A.A. Levchuk, V.A. Babkin, A.R. Beisenbaev, L.I. Larina, A.N. Sapozhnikov, S.M. Adekenov. Study of the physicochemical properties of mechanically processed arglabin and its mechanocomposites based on arabinogalactan. Chemistry of plant raw materials. 2019. 4. 105-112. DOI: 10.14258/jcprm.2019046259].

The data in Table 3 show that almost all of the claimed agents are superior to the selected reference drugs - methanol extract of Thymus vulgaris L. (common thyme) and citrus pectin in terms of cytotoxicity against cancer cells. At the same time, all of them have a higher selectivity compared to the indicated compared agents: the selectivity index (SI) of the methanol extract of Thymus vulgaris L. (common thyme) in relation to MCF-7 is 2.6, A-549 - 1.1, in relation to M-HeLa - 1.4 ; Citrus pectin does not have selectivity, its cytotoxicity (IC ₅₀ ) against all types of cancer cells and normal liver cells (Chang liver) exceeds 1.5%. The drug Arglabin is an effective broad-spectrum antitumor agent, active, which is illustrated by the data in Table 3, against tumor cells of human lung carcinoma A549 (IC ₅₀ 0.0008), breast adenocarcinoma MCF-7 (IC ₅₀ 0.0008) and cervical carcinoma M-HeLa (IC ₅₀ 0.001). The main disadvantage of the comparison drug Arglabin is the complete lack of selectivity, its cytotoxicity index (IC ₅₀ ) against normal liver cells (Chang liver) is 0.0006. The index of the selectivity index of the drug Arglabin in relation to tumor cells is very low: in relation to A549 and MCF-7 it is 0.8, in relation to M-HeLa - 0.6.

The data in Table 3 show that water-soluble PGNaCo showed high cytotoxicity against tumor cells of human lung carcinoma A549 - IC ₅₀ 0.023% (IC ₅₀ of the methanol extract of Thymus vulgaris L. (common thyme) is 0.34%) with a high selectivity index of 39 (IC ₅₀ 0.9% for Chang liver). At the same time, in relation to breast adenocarcinoma MCF-7, the cytotoxicity of PGNaCo is lower than the cytotoxicity of the methanol extract of Thymus vulgaris L. (common thyme) - IC ₅₀ 2.2% versus IC ₅₀ 0.14%, respectively.

Similarly, potassium-cobalt-polygalacturonate and compositions of sodium-cobalt-polygalacturonate and potassium-cobalt-polygalacturonate in a mass ratio of 1-2:-1-2 show high cytotoxicity against tumor cells of human lung carcinoma A549 and adenocarcinoma of the cervix M-HeLa, lower in relation to tumor cells of breast adenocarcinoma MCF-7: IC ₅₀ PGCCo in relation to A549 is 0.06%, M-HeLa - 0.05%, MCF-7 - 0.8%; for PGNaCo:PGKCo compositions, these indicators are respectively:

at a mass ratio of 1:1 - 0.08%, 0.22% and 0.51%,

at a mass ratio of 1:2 - 0.14%, 0.31% and 0.55%,

at a mass ratio of 2:1 - 0.26%, 0.34% and 0.62%.

At the same time, their cytotoxicity with respect to oncocells is selective, since the IC ₅₀ of PGACo and PGNaCo:PGKCo in relation to Chang liver is greater than 1.0.

The selectivity index of the PGKCo compound in relation to A549 is 17, in relation to M-HeLa - 20, which indicates a high selectivity of this compound in relation to these tumor lines, since compounds with SI (10 are considered highly selective. At the same time, SI PGKCo in relation to MCF-7 is 1.5. The composition of PGNaCo and PGKCo in a mass ratio of 1: 1 is highly selective in relation to human lung carcinoma A549 (SI> 12.5), however, compositions in other ratios selective for all presented cancer cells (table 3).

Compositions containing PGNaCo and a metal complex of a pectin biopolymer with a macro- or microelement in a mass ratio of 3: 1 also exhibit high cytotoxicity against human lung carcinoma tumor cells A549 and less cytotoxicity against tumor cells of breast adenocarcinoma MCF-7, IC ₅₀ against A549 for PGNaCo:PGNaZn - 0.07%, PGNaCo:PGNaCr - 0.1%, PGNaCo:PGNaMn - 0.2%, PGNaCo: PGNaMg - 0.1%; in relation to MCF-7 - for PGNaCo:PGNaZn - 0.12%, PGNaCo:PGNaCr - 0.8%, PGNaCo:PGNaMg - 0.7%, for PGNaCo:PGNaMn> 1.0%. For adenocarcinoma of the cervix, M-HeLa IC ₅₀ is 0.9% for PGNaCo:PGNaZn, 0.2% for PGNaCo:PGNaCr, 0.2% for PGNaCo: PGNaMg.

At the same time, high selectivity is shown by: PGNaCo:PGNaZn - in relation to two types of cancer cells (selectivity indices for A549 - 20, for MCF-7 - 12), PGNaCo:PGNaCr and PGNaCo:PGNaMg as for A549 (selectivity indices 13 and 9, respectively) and for M-HeLa (selectivity indices 7 and 4.5, respectively).

Thus, the claimed agents have high cytotoxicity against A549 human lung carcinoma tumor cells and exhibit high selectivity for normal Chang liver liver cells, which makes them suitable for use as antitumor drugs.

Regarding tumor cells of breast adenocarcinoma MCF-7 effective is the pharmacological composition PGNaCo:PGNaZn (IC₅₀0.12; SI 12). Among the proposed pharmacological compositions based on PGNaCo, the composition PGNaCo:PGNaCr (IC₅₀0.2; SI 7).

Induction of apoptotic effects by the claimed means

To establish the relationship between the cytotoxic effect of the tested claimed antitumor agents and the induction of apoptosis, the apoptotic effect of the claimed agents on cancer cells was determined by flow cytometry, the results are presented for the pharmacological compositions PGNaCo:PGNaCr (3:1) and PGNaCo:PGKCo (1:1) on cancer cells M -HeLa. Apoptosis is a regulated process of programmed cell death, as a result of which the cell breaks up into separate apoptotic bodies bounded by the plasma membrane. Fragments of a dead cell are usually very quickly (on average in 90 minutes) phagocytosed by macrophages or neighboring cells, bypassing the development of an inflammatory reaction [Serbin M. E., Shcherbak E. V. Apoptosis and its molecular effectors // Actual problems of biology, medicine and ecology: Collection / edited by prof., d.m.s. N.N. Ilyinsky. - Tomsk: Siberian State Medical University, 2004. Issue 11].

Apoptotic effects were measured by flow cytometry using the annexin V-Alexa Fluor 647 staining protocol. Values are presented as mean (standard deviation (n=3). L - live cells; D - dead cells; Ea - cells in early apoptosis; La - cells in late apoptosis [Voloshina AD, Sapunova AS, Kulik NV, Belenok MG, Strobykina IYu, Lyubina A, et al. Antimicrobial and cytotoxic effects of ammonium derivatives of diterpenoids steviol and isosteviol // Bioorg. Med. Chem. 2021. 32. 115974 DOI: 10.1016/j.bmc.2020.115974].

M-HeLa cells were treated for 24 hours with the proposed agents PGNaCo:PGNaCr (3:1) and PGNaCo:PGKCo (1:1) at concentrations of 0.1% and 0.2%. The results are shown in the figure. It can be seen that, after 24 hours of treatment for both pharmacological compositions, apoptotic effects were observed in M-HeLa cells. Analysis of apoptotic effects characterizes the dose-dependent nature, especially in the case of late apoptosis. At a concentration of 0.1%, not exceeding IC ₅₀ , the pharmacological composition PGNaCo:PGNaCr (3:1) causes early and late apoptosis in 12.83% and 13.82% of M-HeLa cells, respectively, and the pharmacological composition PGNaCo:PGKCo (1:1) - early and late apoptosis at the same concentration of 5.61%. and 6.64% of M-HeLa cells, respectively. It can be seen that the composition of PGNaCo:PGNaCr (3:1) at a concentration of 0.1% induces apoptosis more actively than PGNaCo:PGKCo (1:1). With an increase in the concentration of compounds to 0.2%, the number of cells in late apoptosis increases: for the PGNaCo:PGNaCr (3:1) composition, more than 2 times (from 13.82% to 28.30%); for the composition PGNaCo:PGKCo (1:1) - more than three times (from 6.64% to 21.34%). The results suggest that the cytotoxicity of the studied pharmacological compositions in M-HeLa cancer cells is due to the apoptotic pathway.

Thus, agents are claimed that expand the arsenal of antitumor water-soluble agents based on pectin biopolymer with bioavailability, low toxicity (LD ₅₀ above 15000 mg/kg) and selective cytotoxic activity against the human lung carcinoma tumor cell line A549, breast adenocarcinoma MCF-7 and M-HeLa cervical carcinomas, which have practically no toxic effect on normal human cells. It has been reliably established that macro- and microelements in the composition of pectin metal complexes, when taken orally, are absorbed into the blood and absorbed by the body. This allows us to expect the prospects of the proposed drugs for the treatment of oncological diseases, and also expands the range of known drugs in this field of medicine. Antitumor drugs based on macro- and trace-containing polygalacturonates (options)

Table 1 Characteristics of the used metal complexes Samples Metal content, % Elemental composition, % [α] _D ²⁰ WITH H N PGNaCo 4.68 (Na);
3.28(Co) 31.91 4.70-4.51 0.40 +201.7
(C 0.5; H ₂ O) PGKCo 5.79(K);
3.50 (Co) 31.00-31.51 4.99-5.12 (0.50 +189.5
(From 0.5,
_H2O ) PGNaZn 3.44 (Na);
3.13 (Zn) 30.63-30.55 5.75-5.87 (0.50 +195.4
(C 0.5; H ₂ O) PGNaCr 4.60 (Na);
2.87(Cr) 31.65-31.67; 5.70-5.63 (0.50 +176.5
(C 0.5; H ₂ O) PGNaMn 4.92 (Na);
2.68 (Mn) 30.45-30.52 5.65-5.71- (0.50 +210.2
(C 0.42; H ₂ O) PGNaMg 4.60 (Na);
2.75 (Mg) 30.62-30.13 4.7-4.66 (0.50 +191.0
(C 0.5; H ₂ O)

Antitumor agents based on macro- and microelement-containing polygalacturonates (options)

Table 2 The content of metals in the claimed compositions Composition Metal content, % PGNaCo: PGKCo (1:1) 3.42 (Co) Na (2.23) K (3.71) PGNaCo: PGKCo (2:1) 3.36(Co) Na (3.12) K (1.93) PGNaCo: PGKCo (1:2) 3.41(Co) Na (1.56) K (3.86) PGNaCo: PGNaZn (3:1) 2.67 (Сo) Na (4.70) Zn (0.96) PGNaCo: PGNaCr (3:1) 2.54 (Co) Na (4.88) Cr (0.36) PGNaCo: PGNaMg (3:1) 2.80 (Сo) Na (4.70) Mg (0.53) PGNaCo: PGNaMn (3:1) 2.45 (Co) Na (4.47) Mn (1.43)

Antitumor agents based on macro- and microelement-containing polygalacturonates (options)

Table 3 Cytotoxicity of the claimed antitumor agents based on macro- and microelement-containing polygalacturonates Sample name Tumor lines normal lines MCF-7 A549 M-HeLa Changliver _IC50 % SI _IC50 % SI _IC50 % SI _IC50 % PGNaCo 2.2±0.2 - 0.023±0.002 39 known 0.9±0.07 PGKCo 0.8 1.3 0.06 >17 0.05 >20 >1.0 PGNaCo: PGKCo (1:1) 0.51±0.03 >2 0.08±0.07 >12.5 0.22±0.01 >4.5 >1.0 PGNaCo: PGKCo (1:2) 0.55±0.04 >1.8 0.14±0.01 >7 0.31±0.01 >3.2 >1.0 PGNaCo: PGKCo (2:1) 0.62±0.04 >1.6 0.26±0.02 >3.8 0.34±0.01 >2.9 >1.0 PGNaCo: PGNaZn 0.12±0.01 12 0.07±0.006 20 0.9±0.07 1.5 1.4±0.1 PGNaCo: PGNaCr 0.8±0.06 1.6 0.1±0.008 13 0.2±0.01 7 1.3±0.1 PGNaCo: PGNaMn >1 0.4 0.2±0.02 2 - - 0.4±0.03 PGNaCo: PGNaMg 0.7±0.06 1.3 0.1±0.009 9 0.2±0.01 4.5 0.9±0.07 Pectin citrus >1.5 1 >1.5 1 >1.5 1 >1.5 Methanol extract of Thymus vulgaris L. (common thyme) 0.14±0.01 2.6 0.34±0.02 1.1 0.25±0.02 1.4 0.36±0.01 Arglabin 0.0008 0.8 0.0008 0.8 0.001 0.6 0.0006

Claims (2)

1. A water-soluble agent based on polygalacturonate with selective cytotoxic activity against lung carcinoma A549, breast adenocarcinoma MCF-7 and cervical carcinoma M-HeLa, which is potassium-cobalt-polygalacturonate (PCCo) or its composition with sodium-cobalt- polygalacturonate (PGNaCo) in a mass ratio of 1-2:1-2. 2. A water-soluble agent based on polygalacturonate with selective cytotoxic activity against lung carcinoma A549, breast adenocarcinoma MCF-7 and cervical carcinoma M-HeLa, which is a composition containing sodium-cobalt-polygalacturonate (PNaCo) in a mass ratio of 3: 1 ) and a metal complex of pectin biopolymer sodium-metal-polygalacturonate, respectively, where the biogenic metal is zinc, manganese, chromium or magnesium (PGNaZn, PGNaMn, PGNaCr, PGNaMg).

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