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Britannin as a novel NLRP3 inhibitor, suppresses inflammasome activation in macrophages and alleviates NLRP3-related diseases in mice

Acta Pharmacologica Sinica volume 45, pages 803–814 (2024)Cite this article

Abstract

Overactivation of the NLRP3 inflammasomes induces production of pro-inflammatory cytokines and drives pathological processes. Pharmacological inhibition of NLRP3 is an explicit strategy for the treatment of inflammatory diseases. Thus far no drug specifically targeting NLRP3 has been approved by the FDA for clinical use. This study was aimed to discover novel NLRP3 inhibitors that could suppress NLRP3-mediated pyroptosis. We screened 95 natural products from our in-house library for their inhibitory activity on IL-1β secretion in LPS + ATP-challenged BMDMs, found that Britannin exerted the most potent inhibitory effect with an IC50 value of 3.630 µM. We showed that Britannin (1, 5, 10 µM) dose-dependently inhibited secretion of the cleaved Caspase-1 (p20) and the mature IL-1β, and suppressed NLRP3-mediated pyroptosis in both murine and human macrophages. We demonstrated that Britannin specifically inhibited the activation step of NLRP3 inflammasome in BMDMs via interrupting the assembly step, especially the interaction between NLRP3 and NEK7. We revealed that Britannin directly bound to NLRP3 NACHT domain at Arg335 and Gly271. Moreover, Britannin suppressed NLRP3 activation in an ATPase-independent way, suggesting it as a lead compound for design and development of novel NLRP3 inhibitors. In mouse models of MSU-induced gouty arthritis and LPS-induced acute lung injury (ALI), administration of Britannin (20 mg/kg, i.p.) significantly alleviated NLRP3-mediated inflammation; the therapeutic effects of Britannin were dismissed by NLRP3 knockout. In conclusion, Britannin is an effective natural NLRP3 inhibitor and a potential lead compound for the development of drugs targeting NLRP3.

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Fig. 1: Britannin suppressed NLRP3-mediated pyroptosis.
Fig. 2: Britannin specifically inhibited activation of the NLRP3 inflammasome.
Fig. 3: Britannin inhibited the inflammasome assembly via the interaction between NLRP3 and NEK7.
Fig. 4: Britannin directly binds to the NACHT domain of NLRP3.
Fig. 5: Britannin prevented gouty arthritis by suppressing NLRP3 inflammasome activation.
Fig. 6: Britannin inhibited NLRP3 activation to protect against ALI.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (82360805 to JS), Zhejiang Province Traditional Chinese Medicine Science and Technology Project (2024ZF056 to JS), and Zhejiang Provincial Key Scientific Project (2021C03041 to GL).

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Author notes

  1. These authors contributed equally: Jing-jing Shao, Wei-feng Li

Authors and Affiliations

  1. Department of Cardiology and Medical Research Center, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325035, China

    Jing-jing Shao, Ju-lian Min, Gao-jun Wu & Guang Liang

  2. Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Jing-jing Shao, Wei-feng Li, Xiao-hong Long & Hao-wen Xu

  3. School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, 311399, China

    Jin-feng Sun, Hao-wen Xu & Guang Liang

  4. Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, School of Pharmaceutical Sciences, Yanbian University, Yanji, 133002, China

    Jin-feng Sun

  5. Affiliated Cangnan Hospital, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Zai-shou Zhuang

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  1. Jing-jing Shao
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Contributions

GL and HWX contributed to the literature search and study design. HWX and JJS participated in the drafting of the article. JJS, WFL, JFS, ZSZ, JLAM, and XHL carried out the experiments. GL and HWX revised the manuscript. JFS and GJW contributed to data collection and analysis.

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Correspondence to Hao-wen Xu or Guang Liang.

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Shao, Jj., Li, Wf., Sun, Jf. et al. Britannin as a novel NLRP3 inhibitor, suppresses inflammasome activation in macrophages and alleviates NLRP3-related diseases in mice. Acta Pharmacol Sin 45, 803–814 (2024). https://doi.org/10.1038/s41401-023-01212-5

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