Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy

作者：Ke‐Gang Linghu, Tian Zhang, Guangtao Zhang, Guang-Tao Zhang, Wenjun Zhang, Peng Lv, Wen-Jun Zhang, Wen-Jun Zhang, Mingming Zhao, Shi-Hang Xiong, Qiu-Shuo Ma, Ming-Ming Zhao, Meiwan Chen, Hu Y, Changsheng Zhang, Hua Yu · 发表于：Journal of Pharmaceutical Analysis · 年份：2023 · DOI：10.1016/j.jpha.2023.07.009 · 被引用次数：24 · 研究领域：Genomics, phytochemicals, and oxidative stress、Glutathione Transferases and Polymorphisms、Free Radicals and Antioxidants

Activation of nuclear factor erythroid 2-related factor 2 (Nrf2) by Kelch-like ECH-associated protein 1 (Keap1) alkylation plays a central role in anti-inflammatory therapy. However, activators of Nrf2 through alkylation of Keap1-Kelch domain have not been identified. Deoxynyboquinone (DNQ) is a natural small molecule discovered from marine actinomycetes . The current study was designed to investigate the anti-inflammatory effects and molecular mechanisms of DNQ via alkylation of Keap1. DNQ exhibited significant anti-inflammatory properties both in vitro and in vivo . The pharmacophore responsible for the anti-inflammatory properties of DNQ was determined to be the α, β-unsaturated amides moieties by a chemical reaction between DNQ and N -acetylcysteine. DNQ exerted anti-inflammatory effects through activation of Nrf2/ARE pathway. Keap1 was demonstrated to be the direct target of DNQ and bound with DNQ through conjugate addition reaction involving alkylation. The specific alkylation site of DNQ on Keap1 for Nrf2 activation was elucidated with a synthesized probe in conjunction with liquid chromatography-tandem mass spectrometry. DNQ triggered the ubiquitination and subsequent degradation of Keap1 by alkylation of the cysteine residue 489 (Cys489) on Keap1-Kelch domain, ultimately enabling the activation of Nrf2. Our findings revealed that DNQ exhibited potent anti-inflammatory capacity through α, β-unsaturated amides moieties active group which specifically activated Nrf2 sig...