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Protective Effect of a Novel RIPK1 Inhibitor, Compound 4–155, in Systemic Inflammatory Response Syndrome and Sepsis

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Excessive inflammatory response is a critical pathogenic factor for the tissue damage and organ failure caused by systemic inflammatory response syndrome (SIRS) and sepsis. In recent years, drugs targeting RIPK1 have proved to be an effective anti-inflammatory strategy. In this study, we identified a novel anti-inflammatory lead compound 4–155 that selectively targets RIPK1. Compound 4–155 significantly inhibited necroptosis of cells, and its activity is about 10 times higher than the widely studied Nec-1 s. The anti-necroptosis effect of 4–155 was mainly dependent on the inhibition of phosphorylation of RIPK1, RIPK3, and MLKL. In addition, we demonstrated that 4–155 specifically binds RIPK1 by drug affinity responsive target stability (DARTS), immunoprecipitation, kinase assay, and immunofluorescence microscopy. More importantly, compound 4–155 could inhibit excessive inflammation in vivo by blocking RIPK1-mediated necroptosis and not influence the activation of MAPK and NF-κB, which is more potential for the subsequent drug development. Compound 4–155 effectively protected mice from TNF-induced SIRS and sepsis. Using different doses, we found that 6 mg/kg oral administration of compound 4–155 could increase the survival rate of SIRS mice from 0 to 90%, and the anti-inflammatory effect of 4–155 in vivo was significantly stronger than Nec-1 s at the same dose. Consistently, 4–155 significantly reduced serum levels of pro-inflammatory cytokines (TNF-α and IL-6) and protected the liver and kidney from excessive inflammatory damages. Taken together, our results suggested that compound 4–155 could inhibit excessive inflammation in vivo by blocking RIPK1-mediated necroptosis, providing a new lead compound for the treatment of SIRS and sepsis.

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Funding

This work was funded by grants from the National Key R&D Program of China (2021YFA1302200), the National Natural Science Foundation of China (82022065, 81772124), Shanghai “Shuguang” Project (21SG38), and the Science and Technology Commission of Shanghai Municipality (21S11900800).

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  1. Zhong-Yi Ling and Quan-Zhen Lv contributed equally to this work.

Authors and Affiliations

  1. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, China

    Zhong-Yi Ling & Yan Wang

  2. School of Pharmacy, Second Military Medical University, Shanghai, 200433, China

    Zhong-Yi Ling, Quan-Zhen Lv, Jiao Li, Ren-Yi Lu, Lin-Lin Chen, Wei-Heng Xu, Yan Wang & Chun-Lin Zhuang

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  1. Zhong-Yi Ling
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  2. Quan-Zhen Lv
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  7. Yan Wang
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Contributions

All authors contributed to the study conception and design. CLZ and YW conceived and designed this study; QZL, RYL, LLC, and WHX designed experiments and analyzed data; ZYL performed experiments and analyzed data; JL designed and synthesized compound 4–155; QZL and YW wrote and revised the manuscript.

Corresponding authors

Correspondence to Yan Wang or Chun-Lin Zhuang.

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This study was performed in line with the approval granted by the Animal Care Committee of the Second Military Medical University (Shanghai, China).

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Ling, ZY., Lv, QZ., Li, J. et al. Protective Effect of a Novel RIPK1 Inhibitor, Compound 4–155, in Systemic Inflammatory Response Syndrome and Sepsis. Inflammation 46, 1796–1809 (2023). https://doi.org/10.1007/s10753-023-01842-1

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