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6E11, a highly selective inhibitor of Receptor-Interacting Protein Kinase 1, protects cells against cold
C Delehouzé1, S Leverrier-Penna2,3, F Le Cann2,3,4,5
1Sorbonne Universités, UPMC Univ Paris 06, CNRS USR3151, Protein Phosphorylation and Human Disease Laboratory, Station Biologique, F-29688, Roscoff, France.
Abstract:
Necroptosis is a programmed cell death pathway that has been shown to be of central pathophysiological relevance in multiple disorders (hepatitis, brain and cardiac ischemia, pancreatitis, viral infection and inflammatory diseases). Necroptosis is driven by two serine threonine kinases, RIPK1 (Receptor Interacting Protein Kinase 1) and RIPK3, and a pseudo-kinase MLKL (Mixed Lineage Kinase domain-Like) associated in a multi-protein complex called necrosome. In order to find new inhibitors for use in human therapy, a chemical library containing highly diverse chemical structures was screened using a cell-based assay. The compound 6E11, a natural product derivative, was characterized as a positive hit. Interestingly, this flavanone compound: inhibits necroptosis induced by death receptors ligands TNF-α (Tumor Necrosis Factor) or TRAIL (TNF-Related Apoptosis-Inducing Ligand); is an extremely selective inhibitor, among kinases, of human RIPK1 enzymatic activity with a nM Kd; has a non-ATP competitive mode of action and a novel putative binding site; is weakly cytotoxic towards human primary blood leukocytes or retinal pigment epithelial cells at effective concentrations; protects human aortic endothelial cells (HAEC) from cold hypoxia/reoxygenation injury more effectively than necrostatin-1 (Nec-1) and Nec-1s. Altogether, these data demonstrate that 6E11 is a novel potent small molecular inhibitor of RIPK1-driven necroptosis.
Insights
A new compound, 6E11, effectively inhibits RIPK1-driven necroptosis, a programmed cell death pathway implicated in various diseases. This potent flavanone derivative shows high selectivity and protects cells from injury, offering therapeutic potential.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Necroptosis is a programmed cell death pathway crucial in diseases like hepatitis and ischemia.
- It involves RIPK1, RIPK3, and MLKL, forming the necrosome complex.
- Targeting necroptosis offers therapeutic potential for various inflammatory and ischemic conditions.
Purpose of the Study:
- To identify novel small molecule inhibitors of RIPK1-driven necroptosis.
- To characterize the inhibitory potential and mechanism of action of identified compounds.
- To evaluate the therapeutic efficacy of promising inhibitors in cellular models.
Main Methods:
- Screening of a diverse chemical library using a cell-based assay.
- Biochemical assays to determine inhibitor selectivity and binding kinetics (Kd).
- Assessment of cytotoxicity and protective effects in human cell lines (HAEC, leukocytes, RPE).
Main Results:
- The natural product derivative 6E11 was identified as a potent necroptosis inhibitor.
- 6E11 selectively inhibits human RIPK1 kinase activity with nanomolar affinity (nM Kd).
- It acts via a non-ATP competitive mechanism and demonstrates superior protection against cold hypoxia/reoxygenation injury compared to Nec-1/Nec-1s.
Conclusions:
- 6E11 is a novel, potent, and selective small molecule inhibitor of RIPK1.
- Its unique non-ATP competitive mechanism and efficacy present a promising therapeutic candidate.
- 6E11 warrants further investigation for treating RIPK1-mediated diseases.
