Xanthatin directly targets ZDHHC7 to inhibit NLRP3 palmitoylation and inflammasome activation, alleviating sepsis and acute lung injury

Aberrant activation of the NLRP3 inflammasome drives the pathogenesis of diverse inflammatory diseases, including acute respiratory distress syndrome (ARDS) and sepsis. Among the post-translational modifications regulating NLRP3, S-palmitoylation contributes to NLRP3 localization and inflammasome assembly, but pharmacological modulators of this process remain limited. Here, we identified that Xanthatin, a bioactive sesquiterpene lactone, as a potential and specific inhibitory activity against NLRP3 inflammasome activation. Xanthatin dose-dependently suppressed IL-1β maturation and pyroptosis in both murine and human macrophages, with an IC 50 of approximately 2.09 µM. Mechanistically, Xanthatin specifically blocked the assembly of the NLRP3 inflammasome complex. Using a chemoproteomic strategy combining cellular thermal shift assay with mass spectrometry, we identified the palmitoyltransferase ZDHHC7 as a molecular target engaged by Xanthatin. We demonstrated that Xanthatin binds to ZDHHC7, thereby abrogating the ZDHHC7-mediated palmitoylation of NLRP3 and its subsequent interaction with apoptosis-associated speck-like protein containing a CARD (ASC). In vivo, Xanthatin treatment significantly attenuated ARDS, peritonitis, and sepsis in mouse models. However, it conferred no additional therapeutic benefit in Nlrp3 ⁻ /⁻ mice. Collectively, this study presents Xanthatin as the first-in-class lead compound for developing palmitoyltransferase ZDHHC7 inhibition and elucidates the ZDHHC7 as a promising pharmacological target for the treatment of NLRP3-driven inflammatory diseases.