Isoginkgetin as a novel USP8 inhibitor discovered by structure-based virtual screening exerts anticancer effects in ovarian cancer

Ubiquitin-Specific Protease 8 (USP8) gene has been identified as being significantly amplified and overexpressed in Ovarian carcinoma (OC), and thus led to unfavorable clinical outcomes, implying that inhibition of USP8 may be a therapeutic strategy for OC. Herein, a structure-based discovery and bioassays were conducted to identify novel inhibitors of USP8. we employed a novel platform that integrates docking/molecular dynamics (MD) simulation/binding free energy calculations/root-mean-square deviation (RMSD)/USP8 enzyme activity assay to screen a library of bioactive compounds including 30000 compounds. we identified that Isoginkgetin exhibited promising inhibitory activity on USP8 with an IC50 value of 12.68 μM. Moreover, Isoginkgetin could significantly inhibit OC cell growth by suppressing cell proliferation and reducing colony formation, and promoting Fe²⁺ accumulation, ROS production, thereby triggering cell death. These suppressive effects were successfully reversed by Ferrostatin-1 (Fer-1). Mechanistically, Isoginkgetin decreased USP8 levels, leading to ubiquitin degradation and Glutathione Peroxidase 4 (GPX4) instability, and it stimulated Ferroptosis. Ultimately, our results support Isoginkgetin as a potent inhibitor of USP8, the inhibition of which may be a promising therapeutic strategy for OC.

Deubiquitinating enzymes; Isoginkgetin; Ovarian carcinoma; USP8; Virtual screening.