Yang Xiao-Xia, Hu Ze-Ping, Chan Sui Yung, Zhou Shu-Feng
Department of Pharmacy, Faculty of Science, National University of Singapore, Science Drive 4, Singapore 117543, Singapore.
Clin Chim Acta. 2006 Mar;365(1-2):9-29. doi: 10.1016/j.cca.2005.08.021. Epub 2005 Sep 30.
A variety of therapeutic drugs can undergo biotransformation via Phase I and Phase II enzymes to reactive metabolites that have intrinsic chemical reactivity toward proteins and cause potential organ toxicity. A drug-protein adduct is a protein complex that forms when electrophilic drugs or their reactive metabolite(s) covalently bind to a protein molecule. Formation of such drug-protein adducts eliciting cellular damages and immune responses has been a major hypothesis for the mechanism of toxicity caused by numerous drugs. The monitoring of protein-drug adducts is important in the kinetic and mechanistic studies of drug-protein adducts and establishment of dose-toxicity relationships. The determination of drug-protein adducts can also provide supportive evidence for diagnosis of drug-induced diseases associated with protein-drug adduct formation in patients. The plasma is the most commonly used matrix for monitoring drug-protein adducts due to its convenience and safety. Measurement of circulating antibodies against drug-protein adducts may be used as a useful surrogate marker in the monitoring of drug-protein adducts. The determination of plasma protein adducts and/or relevant antibodies following administration of several drugs including acetaminophen, dapsone, diclofenac and halothane has been conducted in clinical settings for characterizing drug toxicity associated with drug-protein adduct formation. The monitoring of drug-protein adducts often involves multi-step laboratory procedure including sample collection and preliminary preparation, separation to isolate or extract the target compound from a mixture, identification and determination. However, the monitoring of drug-protein adducts is often difficult because of short half-lives of the protein adducts, sampling problem and lack of sensitive analytical techniques for the protein adducts. Currently, chromatographic (e.g. high performance liquid chromatography) and immunological methods (e.g. enzyme-linked immunosorbent assay) are two major techniques used to determine protein adducts of drugs in patients. The present review highlights the importance for clinical monitoring of drug-protein adducts, with an emphasis on methodology and with a further discussion of the application of these techniques to individual drugs and their target proteins.
多种治疗药物可通过I相和II相酶进行生物转化，生成对蛋白质具有内在化学反应性并可能导致潜在器官毒性的反应性代谢物。药物-蛋白质加合物是亲电药物或其反应性代谢物与蛋白质分子共价结合时形成的蛋白质复合物。这种引发细胞损伤和免疫反应的药物-蛋白质加合物的形成，一直是众多药物毒性机制的主要假说。监测蛋白质-药物加合物在药物-蛋白质加合物的动力学和机制研究以及剂量-毒性关系的建立中很重要。药物-蛋白质加合物的测定还可为诊断患者中与蛋白质-药物加合物形成相关的药物性疾病提供支持性证据。由于血浆方便且安全，它是监测药物-蛋白质加合物最常用的基质。检测针对药物-蛋白质加合物的循环抗体可作为监测药物-蛋白质加合物的有用替代标志物。在临床环境中，已对包括对乙酰氨基酚、氨苯砜、双氯芬酸和氟烷在内的几种药物给药后血浆蛋白加合物和/或相关抗体进行了测定，以表征与药物-蛋白质加合物形成相关的药物毒性。药物-蛋白质加合物的监测通常涉及多步骤实验室程序，包括样品采集和初步制备、从混合物中分离以分离或提取目标化合物、鉴定和测定。然而，由于蛋白质加合物的半衰期短、采样问题以及缺乏针对蛋白质加合物的灵敏分析技术，药物-蛋白质加合物的监测往往很困难。目前，色谱法（如高效液相色谱法）和免疫法（如酶联免疫吸附测定法）是用于测定患者药物蛋白质加合物的两种主要技术。本综述强调了临床监测药物-蛋白质加合物的重要性，重点是方法学，并进一步讨论了这些技术在个别药物及其靶蛋白中的应用。
