NLRP3 inflammasome inhibitor INF39 attenuated NLRP3 assembly in macrophages

INF39 is a nontoxic, irreversible, acrylate-based NLRP3 inhibitor and a further optimization of ethyl 2-((2-chlorophenyl) hydroxyl) methyl) acrylate (INF4E). However, the detail mechanism and the direct target of its anti-inflammatory activity is not clear. Here, we show that INF39 is a specific inhibitor for NLRP3 inflammasome activation. INF39 specifically suppresses NLRP3 activation but not the NLRC4 or AIM2 inflammasomes. INF39 has no effect on K⁺ efflux, ROS generation or mitochondrial membrane potential, which are the upstream events of NLRP3 inflammasome activation. In addition, INF39 has no direct inhibitory effect on GSDMD, which is the downstream event of inflammasomes. More importantly, INF39 inhibits the interaction of NEK7-NLRP3, and subsequently inhibits interaction of NLRP3-NLRP3, NLRP3-ASC, ASC oligomerization and speckle formation. Altogether, our study unveils a deeper anti-inflammatory mechanism for INF39 and suggests it could serve as a lead for developing novel therapeutics combating NLRP3-driven diseases.