USP30-IN-20 is an orally active USP30 inhibitor with a Kd of 1.61 μM and an IC50 of 12.8 μM. It induces ferroptosis by promoting the ubiquitination-mediated degradation of GPX4. This compound inhibits the proliferation, migration, and invasion of prostate cancer cells and induces G0/G1 phase arrest while increasing ROS levels. In a mouse xenograft model with PC3 cells, USP30-IN-20 demonstrates significant antitumor activity. It is applicable for advanced prostate cancer research.

USP30:1.61 μM (Kd)

USP30-IN-20 (Compound 8m) exhibits significant antiproliferative activity against prostate cancer cells PC3, DU145, and CD24 - CD44 + cancer stem-like cells with IC50 values of 4.87 μM, 5.44 μM, and 5.94 μM, respectively. At concentrations of 2.5-5 μM over 14 days, it notably reduces the formation of malignant colonies in PC3 and DU145 cells. USP30-IN-20 also induces G0/G1 phase arrest, inhibits cell migration, and decreases invasion capabilities in PC3 and DU145 cells at 2.5-5 μM for 24-48 hours. Furthermore, when applied at 2.5-5 μM for 72 hours, it disrupts tumor sphere formation by significantly reducing the sphere count in these cells. The compound also elevates intracellular and lipid ROS levels, increases malondialdehyde (MDA) content, and decreases glutathione (GSH) levels after 24 hours of treatment at 2.5-5 μM. Additionally, treatment with 5 μM for 24 hours causes mitochondrial shrinkage, loss of cristae and membrane structure, and disruption of mitochondrial membrane potential in PC3 cells, without affecting apoptosis. Lastly, it downregulates GPX4 protein expression in a dose-dependent manner in PC3 and DU145 cells after 24 hours.

USP30-IN-20 (Compound 8m), administered orally at a dosage of 25-50 mg/kg once daily for 18 consecutive days, significantly inhibits the proliferation of PC3 prostate cancer xenografts in nude mice.