Abstract
As a therapeutic target for antitumor necrosis factor (TNF)-α interventions, UbcH5c is one of the key ubiquitin-conjugating enzymes catalyzing ubiquitination during TNF-α-triggered nuclear factor kappa B (NF-κB) activation. In the present study, three series of analogues were designed and synthesized from α-santonin, and their UbcH5c inhibitory activities were screened by Western blotting and NF-κB luciferase assay. Further BIAcore, in-gel fluorescence imaging, and immunoprecipitation assays demonstrated that compound 6d exhibited robust and specific inhibition of UbcH5c, exceeding that of the positive compound 1 (IJ-5). Mechanistic investigations revealed that compound 6d preferentially bound to and inactivated UbcH5c by forming a covalent adduct with its active site Cys85. Furthermore, compound 6d exhibited potent anti-inflammatory activity against complete Freund's adjuvant-induced adjuvant arthritis in vivo. These findings suggest that the novel α-santonin-derived UbcH5c inhibitor 6d is a promising lead compound for the development of new antirheumatoid arthritis (RA) agent.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Anti-Inflammatory Agents, Non-Steroidal / chemical synthesis
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Anti-Inflammatory Agents, Non-Steroidal / pharmacokinetics
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Anti-Inflammatory Agents, Non-Steroidal / pharmacology
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Benzofurans / chemical synthesis
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Benzofurans / pharmacokinetics
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Benzofurans / pharmacology*
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Drug Stability
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Fibroblasts / metabolism
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HeLa Cells
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Humans
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I-kappa B Kinase / metabolism
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Male
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Mice
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Microsomes, Liver / metabolism
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NF-kappa B / metabolism
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Rats, Sprague-Dawley
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Santonin / analogs & derivatives*
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Santonin / chemical synthesis
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Santonin / pharmacokinetics
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Santonin / pharmacology*
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Signal Transduction
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Structure-Activity Relationship
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Tumor Necrosis Factor-alpha / metabolism
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Ubiquitin-Conjugating Enzymes / antagonists & inhibitors*
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Ubiquitination
Substances
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8-((2-bromobenzyl)oxy)-6,9-dimethyl-3-methylene-3a,4,5,9b-tetrahydronaphtho(1,2-b)furan-2(3H)-one
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Anti-Inflammatory Agents, Non-Steroidal
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Benzofurans
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IKBKG protein, human
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NF-kappa B
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Tumor Necrosis Factor-alpha
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Santonin
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UBE2D3 protein, human
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Ubiquitin-Conjugating Enzymes
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I-kappa B Kinase