Biochemical and Comparative Proteomic Analyses Delineate the Anti‐Ovarian Carcinogenic Roles of Modified Calycosin

Ovarian cancer, the eighth leading cause of cancer-related deaths globally, is projected to result in approximately 307,000 deaths by 2040. So, identifying novel therapeutic compounds is critical to improving the survival rate of patients with ovarian cancer. Calycosin, derived from Astragalus root, has demonstrated anti-cancer properties, suggesting its possible use for treating ovarian cancer. In the present study, we synthesized and evaluated a series of calycosin derivatives (H1-H10) to enhance its therapeutic efficacy against ovarian cancer. Among these, calycosin derivative H10 exhibited the most potent anti-cancer activity, effectively inhibiting cell proliferation, migration, and colony formation abilities in SKOV3 and A2780 ovarian cancer cell lines. In addition, H10 induced G0/G1 cell cycle arrest and dose-dependent apoptosis in these cells. Further, comparative proteomic analysis coupled with Ingenuity Pathway Analysis was used to delineate the molecular mechanisms underlying the anti-ovarian cancer effect. Our results demonstrated that H10 modulated key biological processes related to DNA damage response, chromatin and kinase activities, ferroptosis, FoxO signaling, and p53 signaling in ovarian carcinoma. Specifically, H10 regulated a protein cluster comprising RAD51AP1, USP1, USP22, DDX11, ACSL4, GPX4, NCOA4, CCNB1, and CDK1, which are critical to ovarian tumorigenicity. Functional assays confirmed H10's ability to induce cell cycle arrest, senescence, and apoptosis, while proteomic analysis further highlighted its regulatory role in cell cycle regulation and ferroptosis. These findings identify calycosin H10 as a promising therapeutic candidate for ovarian cancer, offering novel insights into its molecular mechanisms of action.