Tunable Cysteine-Targeting Electrophilic Heteroaromatic Warheads Induce Ferroptosis

Once considered potential liabilities, the modern era witnesses a renaissance of interest in covalent inhibitors in drug discovery. The available toolbox of electrophilic warheads is limited by constraints on tuning reactivity and selectivity. Following our work on a class of ferroptotic agents termed CETZOLEs, we discovered new tunable heterocyclic electrophiles which are capable of inducing ferroptosis. The biological evaluation demonstrated that thiazoles with an alkyne electrophile at the 2-position selectively induce ferroptosis with high potency. Density functional theory calculations and NMR kinetic studies demonstrated the ability of our heterocycles to undergo thiol addition, an apparent prerequisite for cytotoxicity. Chemoproteomic analysis indicated several potential targets, the most prominent among them being GPX4 protein. These results were further validated by western blot analysis and the cellular thermal shift assay. Incorporation of these heterocycles into appropriate pharmacophores generated highly cytotoxic agents such as the analogue BCP-T.A, with low nM IC50 values in ferroptosis-sensitive cell lines.

Automated summary

This article explores the renewed interest in covalent inhibitors in drug discovery and the discovery of a new class of tunable heterocyclic electrophiles capable of inducing ferroptosis. The results show that these heterocycles selectively induce ferroptosis with high potency. Further analysis suggests that these compounds can undergo thiol addition and their potential target is the GPX4 protein. Incorporation of these heterocycles into appropriate pharmacophores generates highly cytotoxic agents.