5-((7-Chloro-6-fluoro-1h-indol-3-yl) methyl)-3-methylimidazolidine-2,4-dione as a RIP1 inhibitor protects LPS/D-galactosamine-induced liver failure

Life Sci. 2021 May 15:273:119304. doi: 10.1016/j.lfs.2021.119304. Epub 2021 Mar 1.

Abstract

Aims: Necroptosis, an inflammatory form of regulated necrosis mediated by receptor-interacting kinase 1 (RIP1), RIP3, and pseudokinase mixed lineage kinase domain-like protein (MLKL) is extensively implicated in liver inflammatory disease. Thus identification small-molecule inhibitor of necroptosis has emerged as a potential therapeutic strategy to prevent liver damage. In this study, we identified 5-((7-chloro-6-fluoro-1 h-indol-3-yl) methyl)-3-methylimidazolidine-2,4-dione (F-nec) as a novel potent necroptosis inhibitor.

Main methods: To find out the potent chemical inhibitors of necroptosis, human monocytic U937 cells were treated with a combination of tumor necrosis factor alpha (TNFα) and a pan-caspase inhibitor z-VAD-fmk. LPS and D-galactosamine (LPS/GalN) were further employed to simulate acute liver failure to explore therapeutic potency of F-nec in vivo. In addition, a specific inhibitor of c-Jun NH (2)-terminal kinases (JNK) SP600125 and its activator anisomycin are used to elucidate its mechanisms in acute liver failure therapy. Necroptosis pathway related proteins were tested by western blot.

Key findings: In this study, we identified F-nec as a novel potent RIP1 inhibitor which efficiently blocked TNFα-induced necroptosis in human and mice cells. Furthermore, pre-treatment of F-nec could prevent hepatic necrosis by reducing RIP1-mediated necroptosis also effectively ameliorated LPS/GalN induced acute liver failure by attenuating cell death signaling-stimulated JNK pathway activation and then suppressing JNK-triggered inflammation.

Significance: Altogether, this study demonstrates that F-nec is a potent inhibitor of RIP1 and highlights its great potential for use in the treatment of RIP1-driven inflammatory liver diseases.

Keywords: Acute liver failure; Inflammation; JNK pathway; Necroptosis; RIP1 inhibitor.

MeSH terms

  • Animals
  • GTPase-Activating Proteins / antagonists & inhibitors*
  • Galactosamine / toxicity*
  • Humans
  • Indoles / chemistry*
  • Lipopolysaccharides / toxicity*
  • Liver Failure, Acute / chemically induced
  • Liver Failure, Acute / drug therapy*
  • Liver Failure, Acute / metabolism
  • Liver Failure, Acute / pathology
  • Mice
  • Mice, Inbred C57BL
  • Necroptosis*
  • Protective Agents / pharmacology*
  • Reactive Oxygen Species / metabolism
  • Signal Transduction
  • U937 Cells

Substances

  • GTPase-Activating Proteins
  • Indoles
  • Lipopolysaccharides
  • Protective Agents
  • Ralbp1 protein, mouse
  • Reactive Oxygen Species
  • Galactosamine