Discovery and validation of indole nitroolefins as novel covalent GPX4 inhibitors for inducing ferroptosis in urological cancers.

Objective: Ferroptosis represents a form of cell death characterized by the accumulation of iron dependent lipid peroxidation. This process culminates in membrane damage and cell lysis. One pivotal surveillance mechanism is induced by glutathione peroxidase 4 (GPX4). Furthermore, inhibition of GPX4 has been reported to hold a promise effect in cancer therapeutics. Methods: Computer-aided docking and small molecule probe were used for designed compounds. Flow cytometry was used to evaluate the ferroptosis. Animal experiments were taken to evaluate the Results: Based on our prior research, a series of twenty compounds with covalent binding potential was designed and synthesized. Under systematic evaluation, our team identified two small molecules 14 and 16, which significantly stabilized GPX4 thermal denaturation. Further investigations revealed that treatment with compounds 14 and 16 led to an increase in lipid peroxidation, oxidative stress, and other markers (C11, Fe Conclusions: Compounds 14 and 16 deserve further works as lead compounds of novel docking models for finally discovering effective anti-tumor drug. Future research is needed to dissect their mechanism and exploits this scaffold for GPX4 inhibitor development.