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A sinomenine derivative protects life-threatening inflammatory injuries via covalently binding to a novel allosteric inhibition site of IRF3

Acta Pharmacologica Sinica (2026)Cite this article

Abstract

The severe inflammation associated with infectious or inflammatory diseases significantly contributes to mortality. Interferon regulatory factor 3 (IRF3) represents a potential anti-inflammatory target, but the development of IRF3 inhibitors has not yielded satisfactory results to date. In this study, we established a phenotype-based high-throughput screening system to conduct activity-guided hierarchical screening of clinical frequently used anti-inflammatory and anti-rheumatic herbal extracts and compounds. Employing a Gaussia-luciferase reporter system driven by the IFNB1 promoter, we identified sinomenine as a potent type I interferon (IFN) inhibitor from a set of 28 anti-inflammatory herbal products. Furthermore, among 24 synthesized sinomenine derivatives modified by various electrophilic groups, Sim-9 (2.5–10 μM) dose-dependently inhibited IFN responses triggered by TLRs, RLRs, and STING activation in mouse RAW264.7 cells and in human THP-1 cells, HT-29 cells and A549 cells. We demonstrated that Sim-9, by covalently binding to Cys222, induced a conformational change in the pLxIS motif-binding surface of IRF3, thus blocking its interaction with upstream adapters, including TRIF, MAVS and STING, and subsequent homodimerization of IRF3 itself, which were all essential for activation of type I IFN responses. In in vivo experiments, we showed that injection of Sim-9 (30, 60 mg/kg, i.p.) effectively protected against devastating inflammation in cecal ligation and puncture (CLP)-induced sepsis in mice, and improved cerulein-induced pancreatitis by inhibiting IRF3. Our study discovers Sim-9 as a novel covalent allosteric inhibitor of IRF3 and reveals that the pLxIS motif binding surface represents a previously uncharacterized druggable target for IRF3 activation, providing a promising therapeutic strategy for the treatment of severe inflammatory injuries.

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Fig. 1: The discovery of small molecule compound with anti-type I IFN activities.
Fig. 2: Sim-9 inhibits the signal transduction of type I IFN.
Fig. 3: Sim-9 inhibits the dimerization and nuclear translocation of IRF3 in response to different stimulus.
Fig. 4: Sim-9 directly binds to IRF3.
Fig. 5: Sim-9 allosterically inhibits IRF3 by binding to Cys222.
Fig. 6: Sim-9 induces conformational changes of IRF3 by binding to Cys222.
Fig. 7: Sim-9 protects mice from CLP-induced sepsis.
Fig. 8: Sim-9 facilitates the recovery of acute pancreatitis.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (82322075 to RPL; General project 82104365 to BX), the National Key Research and Development Program (2022YFC3502100), the Beijing “High-grade, Precision and Advanced” Project, and Beijing Key Laboratory for Basic and Development Research on Chinese Medicine.

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  1. These authors contributed equally: Shuo Li, Bing Xu, Ming-jun Lu

Authors and Affiliations

  1. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China

    Shuo Li, Bing Xu, Ming-jun Lu, Qian-wen Wu, Wen-qing Qin, Zi-qi Dai, Jin-zhao Bai, Hai-min Lei & Run-ping Liu

  2. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 100029, China

    Xiao-jiao-yang Li

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Contributions

SL, investigation, data curation, formal analysis, methodology, visualization, and writing-original manuscript; MJL and QWW, investigation, data curation, methodology. WQQ, methodology and visualization; ZQD, methodology, data curation, and visualization; XJYL, methodology, writing-review and editing. JZB, methodology. HML, conceptualization, funding acquisition, project administration, and resources. BX and RPL, conceptualization, funding acquisition, project administration, and resources, supervise, writing-review and editing. All authors agree to be accountable for all aspects of work, ensuring integrity and accuracy.

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Correspondence to Bing Xu, Hai-min Lei or Run-ping Liu.

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Li, S., Xu, B., Lu, Mj. et al. A sinomenine derivative protects life-threatening inflammatory injuries via covalently binding to a novel allosteric inhibition site of IRF3. Acta Pharmacol Sin (2026). https://doi.org/10.1038/s41401-025-01723-3

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