Discovery of novel non-sulfonylurea NLRP3 inflammasome inhibitors for the treatment of multiple inflammatory diseases.

Targeting NLRP3 inflammasome has emerged as a promising therapeutic strategy for various inflammatory diseases. In this work, to discover safe and effective novel NLRP3 inhibitors, we designed and synthesized a series of new non-sulfonylurea NLRP3 inflammasome inhibitors. Among them, the representative compound B6 specifically and potently inhibited the activation of NLRP3 inflammasome with an IC50 of 10.69 nM, exhibiting better potency compared to MCC950 (IC50 = 14.08 nM). Furthermore, B6 showed improved tolerability in both human hepatic cell lines and mouse primary hepatocytes (cell viability >95 %) compared to MCC950 (cell viability <82 %) at 500 μM. Mechanistically, B6 did not inhibit LPS-induced priming of the NLRP3 inflammasome, but significantly blocked NLRP3 inflammasome assembly by directly binding to NLRP3 and disrupting the NEK7-NLRP3 interaction. In vivo studies demonstrated that B6 was more effective than MCC950 in multiple NLRP3-driven diseases model, including systemic inflammation, peritonitis, and colitis. These findings suggest that B6 can be developed as a promising drug candidate for treating NLRP3-driven diseases.