A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases

Abstract

The NOD-like receptor (NLR) family, pyrin domain–containing protein 3 (NLRP3) inflammasome is a component of the inflammatory process, and its aberrant activation is pathogenic in inherited disorders such as cryopyrin-associated periodic syndrome (CAPS) and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease and atherosclerosis. We describe the development of MCC950, a potent, selective, small-molecule inhibitor of NLRP3. MCC950 blocked canonical and noncanonical NLRP3 activation at nanomolar concentrations. MCC950 specifically inhibited activation of NLRP3 but not the AIM2, NLRC4 or NLRP1 inflammasomes. MCC950 reduced interleukin-1β (IL-1β) production in vivo and attenuated the severity of experimental autoimmune encephalomyelitis (EAE), a disease model of multiple sclerosis. Furthermore, MCC950 treatment rescued neonatal lethality in a mouse model of CAPS and was active in ex vivo samples from individuals with Muckle–Wells syndrome. MCC950 is thus a potential therapeutic for NLRP3-associated syndromes, including autoinflammatory and autoimmune diseases, and a tool for further study of the NLRP3 inflammasome in human health and disease.

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Figure 1: MCC950 inhibits NLRP3 inflammasome activation in response to canonical and noncanonical NLRP3 stimuli.
Figure 2: MCC950 does not inhibit NLRC4, AIM2, TLR signaling or priming of NLRP3.
Figure 3: MCC950 blocks NLRP3-dependent ASC oligomerization.
Figure 4: MCC950 does not block K+ efflux, Ca2+ flux or direct NLRP3 and ASC interactions.
Figure 5: MCC950 is active in vivo, and treatment of mice with MCC950 attenuates EAE.
Figure 6: MCC950 inhibits NLRP3 activation in a mouse model of MWS and in PBMCs from human subjects with MWS ex vivo.

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Acknowledgements

We thank A. Kitanovic (German Center for Neurodegenerative Diseases) for image analysis (Fig. 3c), S. Corr (Trinity College Dublin) for providing S. typhimurium UK-1 strain and H.M. Hoffman (University of California, San Diego) for providing Nlrp3 (A350VneoR) mice. This work was supported by the following funding bodies, grants and fellowships. L.A.J.O'N. and R.C.C.: Science Foundation Ireland (G20598). D.L.K. and J.J.C.: Intramural Research Program of the National Human Genome Research Institute, US National Institutes of Health. S.C.H., C.E.S. and K.H.G.M.: Science Foundation Ireland (11/PI/1036) and (07/SRC/B1144). I.V.: Australian Research Council (FT130101215). G.N.: US National Institutes of Health (DK091191) and (DK095782). E.L.: German Research Foundation (SFB645, SFB670, SFB704, TRR57), European Research Council (ERC, InflammAct) and Excellence Cluster ImmunoSensation. S.L.M.: VESKI innovation fellowship and National Health and Medical Research Council of Australia (1032065) and (1057815). K.S.: Queensland Smart Futures Fund and Australian Research Council (FT130100361). L.A.J.O'N.: ERC Advanced Grant (E12435).

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R.C.C. performed and analyzed the experiments described in Figures 1b–j, 2, 3a,b and 4f–h and Supplementary Figure 6b–g; helped analyze the experiments described in Figure 5a–c and Supplementary Figure 6h,i; and wrote the manuscript. A.A.B.R. synthesized MCC950, conducted formulation for in vivo studies, determined compound pharmacokinetics, helped write the manuscript and provided advice. J.J.C. performed the experiments described in Figure 6f and Supplementary Figure 8. S.C.H. performed and analyzed the experiment described in Figure 5d–g. R.M.-P. performed the experiments described in Figure 4a,b. M.C.I. and I.V. performed the experiments described in Figure 4c–e. L.S.D. performed the experiment described in Figure 5a–c. B.G.M. and A.S. performed the experiments described in Figure 3c and Supplementary Figure 7. D.E.C. performed the experiments described in Supplementary Figure 6a. M.S.B. performed the NMR analysis described in Supplementary Figures 1, 2, 3, 4, 5 and Supplementary Table 1. M.H. performed the experiments described in Supplementary Figure 6h,i. C.E.S. helped analyze data from the experiment described in Figure 5d–g. G.N., E.L. and D.L.K. oversaw a portion of the work. K.H.G.M. conceived ideas and oversaw a portion of the work. S.L.M. performed and analyzed the experiments described in Figure 6a–e. K.S., S.L.M. and M.A.C. conceived ideas, oversaw a portion of the work, reviewed the manuscript and provided advice. L.A.J.O'N. conceived ideas, oversaw the research program and wrote the manuscript.

Corresponding authors

Correspondence to Matthew A Cooper or Luke A J O'Neill.

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The authors declare no competing financial interests.

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Supplementary Figures 1–9 and Supplementary Tables 1–5. (PDF 12971 kb)

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Coll, R., Robertson, A., Chae, J. et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med 21, 248–255 (2015). https://doi.org/10.1038/nm.3806

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