Structure-activity relationship study of RSL3-based GPX4 degraders and its potential noncovalent optimization

Ferroptosis is an iron-dependent, non-apoptotic form of cell death involving in various disease processes. Mechanistically, Glutathione Peroxidase 4 (GPX4) which belongs to the redox Enzyme can convert lipid hydroperoxides into innocuous lipid alcohol to protect cells from Ferroptosis. Therefore, targeting manipulation of GPX4 may represent a promising strategy for regulating cell redox homeostasis and Ferroptosis. In this work, we designed, synthesized and evaluated a series of RSL3-based GPX4 degraders using PROTAC strategy. The structure-activity relationship of these compounds with different E3 Ligase ligands, linker lengths and chemical compositions was systematically studied. Compound R17 with carbon chain linker and lenalidomide E3 ligand was selected as the most potent GPX4 degrader for degrading GPX4 protein in nanomolar level either in wild tumor cells or in drug-resistant tumor cells. We also optimized the POI ligand of R17 with chloracetylamine replaced to propionamide to construct noncovalent GPX4 degrader NC-R17. Such noncovalent modification led to a moderate GPX4 degradation activity and represents a promising strategy for the development of noncovalent GPX4 PROTACs. In general, we screened a set of GPX4 degraders to give the compound R17 with excellent protein degradation activity, and further optimization gave the noncovalent degrader NC-R17 with moderate efficacy. These results lay a firm foundation for the discovery of novel anti-tumor drugs targeting GPX4 and offer the proof of concept for the design of noncovalent GPX4 PROTACs.