GLS1 Inhibitor-4 (compound 41e) is a potent GLS1 inhibitor with an IC50 of 11.86 nM, exhibiting antiproliferative effects, substantial metabolic stability, and strong GLS1 binding affinity. It disrupts glutamine metabolism, induces ROS production, triggers apoptosis, and demonstrates notable antitumor activity [1].

GLS1 Inhibitor-4 (compound 41e) exhibits potent antiproliferative effects with IC50 values of 0.051 µM for HCT116, 0.37 µM for MDA-MB-436, 0.32 µM for CT26, and 1.34 µM for H22 cells, demonstrating high stability in human plasma (96%) and liver microsomes, and binding strongly to the GLS1 protein with a dissociation constant (Kd) of 52 nM. It inhibits colony formation in HCT116 cells dose-dependently at 0.1, 0.5, and 1 µM, reduces key metabolite levels at 100 and 300 nM within 12 hours, increases ROS levels at 30, 50, and 200 nM in 6 hours, induces apoptosis over 24 hours, significantly reduces ATP production, and suppresses aerobic glycolysis by lowering basal and maximal OCRs in HCT116 cells at 1 mmol/L after 12 hours [1].

GLS1 Inhibitor-4, administered intraperitoneally at doses of 50 and 100 mg/kg twice daily for 21 consecutive days, demonstrates dose-dependent antitumor efficacy [1].