Abstract
Necroptosis is a regulated form of cell death involved in several disease models including in particular liver diseases. Receptor-interacting protein kinases, RIPK1 and RIPK3, are the main serine/threonine kinases driving this cell death pathway. We screened a noncommercial, kinase-focused chemical library which allowed us to identify Sibiriline as a new inhibitor of necroptosis induced by tumor necrosis factor (TNF) in Fas-associated protein with death domain (FADD)-deficient Jurkat cells. Moreover, Sib inhibits necroptotic cell death induced by various death ligands in human or mouse cells while not protecting from caspase-dependent apoptosis. By using competition binding assay and recombinant kinase assays, we demonstrated that Sib is a rather specific competitive RIPK1 inhibitor. Molecular docking analysis shows that Sib is trapped closed to human RIPK1 adenosine triphosphate-binding site in a relatively hydrophobic pocket locking RIPK1 in an inactive conformation. In agreement with its RIPK1 inhibitory property, Sib inhibits both TNF-induced RIPK1-dependent necroptosis and RIPK1-dependent apoptosis. Finally, Sib protects mice from concanavalin A-induced hepatitis. These results reveal the small-molecule Sib as a new RIPK1 inhibitor potentially of interest for the treatment of immune-dependent hepatitis.
Keywords:
Concanavalin A -induced hepatitis; RIPK1 inhibitor; kinase inhibitor; molecular docking; necroptosis.
© 2017 Federation of European Biochemical Societies.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alkaloids / chemistry
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Alkaloids / pharmacology*
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Animals
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Apoptosis / drug effects
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Apoptosis / genetics
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Caspase 3 / genetics
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Caspase 3 / immunology
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Cell Line, Transformed
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Concanavalin A
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Cycloheximide / pharmacology
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Fibroblasts / cytology
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Fibroblasts / drug effects
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Fibroblasts / metabolism
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Gene Expression Regulation
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HT29 Cells
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Hepatitis, Animal / chemically induced
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Hepatitis, Animal / genetics
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Hepatitis, Animal / immunology
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Hepatitis, Animal / prevention & control*
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Humans
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Imidazoles / pharmacology
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Immunologic Factors / chemistry
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Immunologic Factors / pharmacology*
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Indoles / pharmacology
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Jurkat Cells
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Male
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Mice
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Molecular Docking Simulation
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Necrosis / chemically induced
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Necrosis / genetics
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Necrosis / immunology
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Protein Kinase Inhibitors / chemistry
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Protein Kinase Inhibitors / pharmacology*
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Receptor-Interacting Protein Serine-Threonine Kinases / antagonists & inhibitors
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Receptor-Interacting Protein Serine-Threonine Kinases / genetics*
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Receptor-Interacting Protein Serine-Threonine Kinases / immunology
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Signal Transduction
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Spiro Compounds / chemistry
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Spiro Compounds / pharmacology*
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TNF-Related Apoptosis-Inducing Ligand / pharmacology
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Tumor Necrosis Factor-alpha / pharmacology
Substances
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Alkaloids
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Imidazoles
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Immunologic Factors
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Indoles
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Protein Kinase Inhibitors
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Spiro Compounds
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TNF-Related Apoptosis-Inducing Ligand
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TNFSF10 protein, human
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Tumor Necrosis Factor-alpha
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necrostatin-1
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sibirine
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Concanavalin A
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Cycloheximide
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RIPK1 protein, human
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RIPK3 protein, human
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Receptor-Interacting Protein Serine-Threonine Kinases
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CASP3 protein, human
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Caspase 3