GSK461364 Inhibits NLRP3 Inflammasome by Targeting NEK7 Phosphorylation.

NLRP3 inflammasome is a multiple protein complex sensing exogenous or endogenous stimuli, and aberrant activation of the NLRP3 inflammasome is implicated in various inflammatory disorders. While numerous small-molecule compounds targeting NLRP3 inflammasome activity have been developed, most have encountered limited success in clinical translation. Through screening of a kinase compound library, GSK461364 is identified as a potent and selective NLRP3 inflammasome inhibitor. Notably, GSK461364 confers significant protective effects in murine models of LPS-induced endotoxemia and DSS-induced colitis. Mechanistic study reveals that GSK461364 exerts its inhibitory effects via targeting Polo-like Kinase 1(PLK1). Specifically, that PLK1-mediated phosphorylation of NEK7, likely occurring at evolutionarily conserved serine residues (Ser221 and Ser260), is shown to enhance NEK7-NLRP3 binding, a critical step for NLRP3 inflammasome assembly. These findings not only establish GSK461364 as a novel therapeutic candidate for NLRP3-driven inflammatory diseases but also provide new insights into the regulatory mechanisms governing inflammasome activation through post-translational modification.