Discovery of potent and selective inhibitors of human NLRP3 with a novel mechanism of action

  • J Exp Med. 2025 Nov 3;222(11):e20242403. doi: 10.1084/jem.20242403.
Kevin Wilhelmsen  1 ,  Aditi Deshpande  1 ,  Sarah Tronnes  1 ,  Maitriyee Mahanta  1 ,  Matthew Banicki  1 ,  Mary Cochran  1 ,  Samantha Cowdin  1 ,  Kristen Fortney  1 ,  George Hartman  1 ,  Robert E Hughes  1 ,  Rusty Montgomery  1 ,  Claudia P Portillo  1 ,  Paul Rubin  1 ,  Taiz Salazar  1 ,  Yan Wang  1 ,  Shijun Yan  1 ,  Barry A Morgan  2 ,  Assem Duisembekova  3 ,  Romane Riou  3 ,  Michael Marleaux  4 ,  Inga V Hochheiser  4 ,  Hannes Buthmann  4 ,  Dominic Ferber  4 ,  Jane Torp  4 ,  Wei Wang  5 ,  Melanie Cranston  5 ,  Chloe M McKee  5 ,  Thea J Mawhinney  5 ,  Emma C McKay  5 ,  Fehime K Eroglu  6  7 ,  Jasmin Kümmerle-Deschner  7 ,  Alexander N R Weber  6  8 ,  Bénédicte F Py  3 ,  Matthias Geyer  4 ,  Rebecca C Coll  5
Affiliations
  • 1. BioAge Labs , Emeryville, CA, USA.
  • 2. HitGen Pharmaceuticals Inc. , Houston, TX, USA.
  • 3. CIRI, Centre International de Recherche en Infectiologie, Univ Lyon, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, ENS de Lyon , Lyon, France.
  • 4. Institute of Structural Biology, University of Bonn , Bonn, Germany.
  • 5. Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast , Belfast, UK.
  • 6. Department of Innate Immunity, Institute of Immunology, Tübingen, Germany.
  • 7. Department of Pediatrics I, Pediatric Rheumatology and Autoinflammation Reference Center, University Hospital Tübingen, Tübingen, Germany.
  • 8. Cluster of Excellence 2180 "Image-guided and Functionally Instructed Tumor Therapies" and Cluster of Excellence 2124 "Controlling Microbes to Fight Infection", University of Tübingen , Tübingen, Germany.
PMID: 40892070 DOI: 10.1084/jem.20242403
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.

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