Potent Benzothiazole-Triazole RIPK1/RIPK3 Dual-Targeting Inhibitors for the Treatment of Systemic Inflammatory...

Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and RIPK3 play a central role in necroptosis for systemic inflammatory response syndrome (SIRS). Single-target RIPK1 inhibitors have shown limited clinical progress, and no RIPK3 inhibitors have reached clinical trials, supporting the development of dual RIPK1/RIPK3 inhibitors. In this study, novel benzothiazole-triazole derivatives were designed by hybridizing benzothiazole inhibitor SZM-594 and RIPK1 inhibitor GSK'772. Most compounds exhibited potent nanomolar antinecroptotic activity and low cytotoxicity. Selected compounds were confirmed to effectively inhibit RIPK1 (K_d < 10 nM) and RIPK3 (K_d = ∼50 nM) in the nanomolar range. Compounds 10, 38, and 43 alleviated TNF-α-induced SIRS in mice by restoring body temperature, improving survival, and reducing IL-1β/IL-6 levels in the serum and multiple organs. Mechanistically, they blocked RIPK1, RIPK3, and MLKL phosphorylation, specifically inhibiting necroptosis rather than apoptosis. This study provides novel lead compounds for RIPK1/RIPK3 dual-targeting intervention and promising candidates for necroptosis-related inflammatory diseases.