Discovery of potent and selective inhibitors of human NLRP3 with a novel mechanism of action
Kevin Wilhelmsen, Aditi Deshpande, Sarah U. Tronnes, Maitriyee Mahanta, MATTHEW BANICKI, MARY COCHRAN, Samantha Cowdin, Kristen Fortney, George D. Hartman, Robert E. Hughes, Rusty L. Montgomery, CLAUDIA PORTILLO, Paul Rubin, Taiz Salazar, Yan Wang, Shijun Yan, Barry A. Morgan, Assem Duisembekova, Romane Riou, Michael Marleaux, Inga V. Hochheiser, Hannes Buthmann, Dominic Ferber, Jane Torp, Wei Wang, Melanie Cranston, Chloe M. McKee, Thea Mawhinney, Emma McKay, Fehime Kara Eroğlu, Jasmin Kuemmerle‐Deschner, Alexander N.R. Weber, Bénédicte F. Py, Matthias Geyer, Rebecca C. Coll
Abstract
The NLRP3 inflammasome is an intracellular protein complex that causes inflammation via the release of IL-1β and pyroptosis. NLRP3 activation is associated with many age-related inflammatory diseases, and NLRP3 inhibition is a promising therapeutic strategy. We previously performed a DNA-encoded library screen to identify novel NLRP3-binding molecules. Herein we describe the characterization of BAL-0028 as a potent and specific inhibitor of NLRP3 signaling. Notably, BAL-0028 is a poor inhibitor of mouse NLRP3 but inhibits human and primate NLRP3 with nanomolar potency. Using cellular and biochemical analyses, we demonstrate that BAL-0028 binds to the NLRP3 NACHT domain at a site that is distinct from the MCC950-binding pocket. Using humanized NLRP3 mice, we show that a derivative of BAL-0028, BAL-0598, inhibits NLRP3 activation in vivo in a peritonitis model. Finally, we demonstrate that both BAL-0028 and BAL-0598 inhibit select hyperactive NLRP3 mutations associated with autoinflammatory diseases more potently than MCC950. BAL-0028 and BAL-0598 thus represent a new modality for NLRP3 inhibition in inflammatory diseases.