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

Abstract

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.

Keywords: RIPK1; compound 4–155; necroptosis; sepsis.; systemic inflammatory response syndrome.