Biologically Active Copmounds in the Exrtacts of Speranskia Tuberculata (Bunge) Baill and Their Effect on the Viability of Cancer Cells of Five Different Lines

Abstract

The aim of the work. Identifying the most considerable bioactive compounds in Speranskia tuberculata extracts and studying their effect on the viability of cancer cells of five lines. Materials and Methods. The herb Speranskia tuberculata extract was prepared based on ethyl acetate and petroleum ether in the 12.5-75 μg/ml concentration range. The presence of compounds in the extracts was investigated by the tandem high-performance liquid chromatography-mass spectroscopy (HPLC/MS) method. The MTT cytotoxicity test was used to evaluate the effectiveness of extracts on five commercially available cell lines: A549 (human lung adenocarcinoma), HEPG2 (human hepatocellular carcinoma of the liver), A375 (human malignant melanoma), HELA (human pancreatic carcinoma), RAW264.7 (macrophage-like cell line from mice transformed with Abelson's leukaemia virus). Results and Discussion. The peaks of about 300 compounds were identified in the mass spectra. Fifty-six compounds with the largest peaks were investigated from the point of view of manifestations of anticancer, antitumour, antioxidant and anti-inflammatory activity known from the literature. Only 35 of the 56 peaks have a documented effect on the listed diseases, another 15 compounds are active on other diseases, and 6 showed no activity. Of the 35 compounds belonging to 10 chemical classes, alkaloids, lipids, and polyphenols dominate (20 out of 35). Chemical reactions for phytochemical screening of chemical classes found in extracts are presented. In MTT tests, the viability during treatment of 4 out of 5 cell lines significantly decreases with increasing concentration of extracts. Only RAW264.7 cells were stable - their viability did not fall below 75-85%. Ethyl acetate extracts were the most effective. They maximally reduce viability to 18%, and the IC50 of this extract for all cell lines varied between 49-53 μg/ml. For petroleum ether, the IC50 of three lines was 65-74 μg/ml (for A375, the approximate value reached 114 μg/ml). Conclusions. The peaks of about 300 compounds found in extracts of the herb Speranskia tuberculata were identified by the HPLC/MS. Among the 56 most intense peaks, 35 compounds, according to the literature, have anticancer, antitumour, antioxidant and anti-inflammatory effects. MTT tests showed that the studied extracts significantly reduced the viability of cancer cells in 4 out of 5 lines. IC50 for ethyl acetate extracts is 49-53 μg/ml, and for petroleum ether - 65-74 μg/ml.