Abstract
Necroptosis is an inflammatory form of programmed cell death that has been implicated in various human diseases. Compound 2 is a more potent analogue of the published compound 1 and inhibits necroptosis in human and murine cells at nanomolar concentrations. Several target engagement strategies were employed, including cellular thermal shift assays (CETSA) and diazirine-mediated photoaffinity labeling via a bifunctional photoaffinity probe derived from compound 2. These target engagement studies demonstrate that compound 2 binds to all three necroptotic effector proteins (mixed lineage kinase domain-like protein (MLKL), receptor-interacting serine/threonine protein kinase 1 (RIPK1) and receptor-interacting serine/threonine protein kinase 3 (RIPK3)) at different levels in vitro and in cells. Compound 2 also shows efficacy in vivo in a murine model of systemic inflammatory response syndrome (SIRS).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line, Tumor
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Female
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Humans
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Mice, Inbred C57BL
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Necroptosis / drug effects*
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Phenylurea Compounds / metabolism
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Phenylurea Compounds / pharmacokinetics
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Phenylurea Compounds / therapeutic use*
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Protein Binding
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Protein Kinase Inhibitors / metabolism
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Protein Kinase Inhibitors / pharmacokinetics
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Protein Kinase Inhibitors / therapeutic use*
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Protein Kinases / metabolism
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Receptor-Interacting Protein Serine-Threonine Kinases / antagonists & inhibitors
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Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
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Signal Transduction / drug effects*
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Sulfonamides / metabolism
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Sulfonamides / pharmacokinetics
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Sulfonamides / therapeutic use*
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Systemic Inflammatory Response Syndrome / drug therapy
Substances
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Phenylurea Compounds
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Protein Kinase Inhibitors
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Sulfonamides
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MLKL protein, human
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MLKL protein, mouse
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Protein Kinases
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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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Ripk1 protein, mouse
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Ripk3 protein, mouse