Nolte Whitney M, Tessman Robert T, Goldman Jennifer L
Children's Mercy Hospital, University of Missouri-Kansas City, Division of Clinical Pharmacology, Toxicology, and Therapeutic Innovation, 2401 Gillham Rd., Kansas City, MO 64108.
Med Chem Res. 2020 Jul;29(7):1238-1246. doi: 10.1007/s00044-020-02570-z. Epub 2020 Jun 4.
Modification of endogenous proteins by drugs and drug metabolites are thought to be a cause of idiosyncratic adverse drug reactions (IADRs). Trimethoprim (TMP) is a commonly prescribed antibiotic that has been implicated in IADRs; however, there is no known mechanism by which this drug or its metabolites modify proteins. This study describes the results of screening trimethoprim and its primary metabolites for the ability to covalently modify human serum albumin (HSA). The first step of the screen was reactions of the compounds with HSA followed by western blotting with antisera specific to drug-modified proteins. Compounds with positive signal in the western blot were then screened using an untargeted peptide profiling method to discover modified peptides. This strategy identified two sites in HSA that are modified by incubation with a TMP metabolite, α-hydroxy trimethoprim (Cα-OH-TMP).
药物和药物代谢产物对内源性蛋白质的修饰被认为是导致特异质性药物不良反应（IADR）的一个原因。甲氧苄啶（TMP）是一种常用的抗生素，与IADR有关；然而，目前尚不清楚该药物或其代谢产物修饰蛋白质的机制。本研究描述了筛选甲氧苄啶及其主要代谢产物共价修饰人血清白蛋白（HSA）能力的结果。筛选的第一步是化合物与HSA的反应，然后用针对药物修饰蛋白质的抗血清进行蛋白质印迹分析。然后使用非靶向肽谱分析方法对蛋白质印迹中信号呈阳性的化合物进行筛选，以发现修饰的肽段。该策略确定了HSA中两个因与TMP代谢产物α-羟基甲氧苄啶（Cα-OH-TMP）孵育而被修饰的位点。
