Discovery of potent and selective inhibitors of human NLRP3 with a novel mechanism of action
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.
Authors
- Romane Riou (ORCID: https://orcid.org/0000-0002-2345-8503)
- Sarah U. Tronnes (ORCID: https://orcid.org/0000-0001-5357-046X)
- Rusty L. Montgomery (ORCID: https://orcid.org/0000-0002-7193-2468)
- Fehime Kara Eroğlu (ORCID: https://orcid.org/0000-0003-2364-4282)
- Rebecca C. Coll (ORCID: https://orcid.org/0000-0002-7359-6023)
- Bénédicte F. Py (ORCID: https://orcid.org/0000-0001-6716-7807)
- Alexander N.R. Weber (ORCID: https://orcid.org/0000-0002-8627-7056)
- MARY COCHRAN
- George D. Hartman (ORCID: https://orcid.org/0000-0002-8016-8369)
- Jane Torp (ORCID: https://orcid.org/0000-0003-3257-8483)
- Shijun Yan (ORCID: https://orcid.org/0000-0001-5046-8427)
- Emma McKay (ORCID: https://orcid.org/0009-0003-6632-2010)
- Michael Marleaux (ORCID: https://orcid.org/0000-0002-1994-8227)
- Jasmin Beate Kuemmerle-Deschner (ORCID: https://orcid.org/0000-0002-6365-6598)
- Kevin Wilhelmsen (ORCID: https://orcid.org/0000-0001-6518-6369)
- CLAUDIA PORTILLO
- Matthias Geyer (ORCID: https://orcid.org/0000-0002-7718-5002)
- Inga V. Hochheiser (ORCID: https://orcid.org/0000-0001-6649-2624)
- Paul Rubin
- Chloe M. McKee (ORCID: https://orcid.org/0009-0004-2882-165X)
- Dominic Ferber (ORCID: https://orcid.org/0000-0003-2633-2858)
- Maitriyee Mahanta
- Samantha Cowdin
- MATTHEW BANICKI
- Aditi Deshpande (ORCID: https://orcid.org/0000-0002-5184-5658)
- Thea Mawhinney (ORCID: https://orcid.org/0009-0003-8657-2455)
- Robert E. Hughes (ORCID: https://orcid.org/0009-0006-5650-2802)
- Yan Wang
- Barry A. Morgan
- Wei Wang (ORCID: https://orcid.org/0000-0002-8156-9754)
- Hannes Buthmann
- Assem Duisembekova (ORCID: https://orcid.org/0009-0000-9997-1517)
- Melanie Cranston (ORCID: https://orcid.org/0009-0003-9068-2593)
- Taiz Salazar
- Kristen Fortney
Institutions
- Université Claude Bernard Lyon 1 (FR)
- École Normale Supérieure de Lyon (FR)
- Queen's University Belfast (GB)
- Centre National de la Recherche Scientifique (FR)
- University of Bonn (DE)
- Inserm (FR)
- Ligand Pharmaceuticals (United Kingdom) (GB)
- Innovation Cluster (Canada) (CA)
- University Children's Hospital Tübingen (DE)
- Institute of Immunology (HR)
- Bellicum Pharmaceuticals (United States) (US)
- Centre International de Recherche en Infectiologie (FR)
- Joint BioEnergy Institute (US)
- University of Tübingen (DE)
Publication Details
- Journal
- The Journal of Experimental Medicine
- Published
- 2025-09-02
- DOI
- https://doi.org/10.1084/jem.20242403
- Citations
- 8
- Primary Topic
- Inflammasome and immune disorders
- Type
- article
- Field-Weighted Citation Impact
- 6.45
Funders
- Academy of Medical Sciences
- Queen's University Belfast
- Deutsche Forschungsgemeinschaft
- Agence Nationale de la Recherche
- Institut National de la Santé et de la Recherche Médicale
- Eberhard Karls Universität Tübingen
- Queen's University
- Studienstiftung des Deutschen Volkes
- Centre National de la Recherche Scientifique
- Université Claude Bernard Lyon 1
- Rheinische Friedrich-Wilhelms-Universität Bonn
- Association Neurofibromatoses et Recklinghause
- École Normale Supérieure de Lyon
- Directorate for Biological Sciences
- Biotechnology and Biological Sciences Research Council
- H2020 European Research Council