Whitby Landon R, Obach R Scott, Simon Gabriel M, Hayward Matthew M, Cravatt Benjamin F
The Skaggs Institute for Chemical Biology and Department of Chemical Physiology, The Scripps Research Institute , 10550 N. Torrey Pines Rd., La Jolla, California 92307, United States.
Pfizer Worldwide Research and Development , Eastern Point Road, Groton, Connecticut 06340, United States.
ACS Chem Biol. 2017 Aug 18;12(8):2040-2050. doi: 10.1021/acschembio.7b00346. Epub 2017 Jun 21.
Idiosyncratic liver toxicity represents an important problem in drug research and pharmacotherapy. Reactive drug metabolites that modify proteins are thought to be a principal factor in drug-induced liver injury. Here, we describe a quantitative chemical proteomic method to identify the targets of reactive drug metabolites in vivo. Treating mice with clickable analogues of four representative hepatotoxic drugs, we demonstrate extensive covalent binding that is confined primarily to the liver. Each drug exhibited a distinct target profile that, in certain cases, showed strong enrichment for specific metabolic pathways (e.g., lipid/sterol pathways for troglitazone). Site-specific proteomics revealed that acetaminophen reacts with high stoichiometry with several conserved, functional (seleno)cysteine residues throughout the liver proteome. Our findings thus provide an advanced experimental framework to characterize the proteomic reactivity of drug metabolites in vivo, revealing target profiles that may help to explain mechanisms and identify risk factors for drug-induced liver injury.
特异质性肝毒性是药物研究和药物治疗中的一个重要问题。被认为修饰蛋白质的反应性药物代谢产物是药物性肝损伤的主要因素。在此，我们描述了一种定量化学蛋白质组学方法，用于在体内鉴定反应性药物代谢产物的靶点。用四种代表性肝毒性药物的可点击类似物处理小鼠，我们证明了广泛的共价结合，主要局限于肝脏。每种药物都表现出独特的靶点谱，在某些情况下，显示出特定代谢途径的强烈富集（例如，曲格列酮的脂质/甾醇途径）。位点特异性蛋白质组学表明，对乙酰氨基酚与整个肝脏蛋白质组中的几个保守的、功能性（硒代）半胱氨酸残基以高化学计量比反应。因此，我们的研究结果提供了一个先进的实验框架，以表征体内药物代谢产物的蛋白质组反应性，揭示可能有助于解释机制和识别药物性肝损伤风险因素的靶点谱。
