Abstract: Objective To screen and identify  novel  anti⁃ovarian cancer active compounds from the traditional Chinese medicine sophora tonkinensis radix, and investigate their effects on apoptosis and autophagy of ovarian cancer cells. Methods Flavonoids components from sophora tonkinensis radix were systematically isolated by silica gel column chromatography and vacuum rotary evaporation techniques. The inhibitory activity of separated fractions on ovarian cancer cell growth was evaluated. High performance liquid chromatography⁃mass spectrometry (HPLC⁃MS) was employed to characterize and identify anti⁃cancer active monomers, while nuclear magnetic resonance (1H NMR) and electrospray ionization (ESI) mass spectrometry were used for structural confirmation. The inhibitory effects of the sophoranochromene and sophoradin on six ovarian cancer cell lines（A2780， NIH: OVCAR3, SKOV3, Caov⁃3, Caov⁃4, ES⁃2） and normal ovarian epithelial cells IOSE80 were assessed using real⁃time cell analysis (RTCA) system. Network pharmacology and molecular docking were employed to predict interaction targets between the active monomers and ovarian cancer cells. Apoptosis was measured via flow cytometry, and Western blotting was performed to analyze the cleavage of apoptosis⁃related proteins (PARP, C⁃caspase⁃3) and the expression of autophagy markers (p62, LC3B). Results Sophoranochromene and sophoradin were identified as anti⁃ovarian cancer active monomers from the effective fraction SdgfF of sophora tonkinensis radix. RTCA system confirmed that the IC50 values of both monomers on A2780, NIH: OVCAR3, SKOV3, Caov⁃3, Caov⁃4 and ES⁃2 cells were lower than those for IOSE80 cells (all P<0.0001). Flow cytometry revealed significantly increased apoptosis rates in A2780 cells treated with either compound (all P<0.05). Ten core targets for sophoranochromene⁃ovarian cancer and 16 core targets for sophoradin⁃ovarian cancer were identified. Western blotting demonstrated elevated C⁃PARP expression in A2780 cells treated with sophoranochromene (P<0.05), while sophoradin treatment significantly reduced p62 and increased LC3B⁃I/II expression (P<0.05). Conclusions Sophoranochromene and sophoradin exert anti⁃ovarian cancer effects by inducing apoptosis and autophagy in cancer cells, demonstrating potential as novel therapeutic agents for ovarian cancer.

Key words: Ovarian cancer, Medicinal plants, Sophora tonkinensis Radix, Sophoranochromene, Sophoradin