人常见肝UGTs活性与抑制体外评价体系的建立与验证  

Establishment and validation of an In Vitro evaluation system for human liver UDP-glucuronosyltransferase activity and drug inhibition study

作　　者：张静[1] 李航 张雪侠 刘帅兵[1] ZHANG Jing;LI Hang;ZHANG Xue-xia;LIU Shuai-bing(Dept of Pharmacy,the First Affiliated Hospital of Zhengzhou University;Henan Integrative Medicine Hospital,Institute of Chinese Medicine,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院药学部 [2]河南省中西医结合医院中药研究所,河南郑州450052

出　　处：《中国药理学通报》2024年第10期1990-1996,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年基金(No 81603204)。

摘　　要：目的 本研究旨在建立一种可靠的体外评价体系,用于评估人肝UDP-葡萄糖醛酸转移酶活性及药物对人肝UGTs的抑制作用,并验证该体系的准确性和稳定性。方法 选择了人肝微粒体UGTs酶6种主要亚型的6个探针底物,在优化的温孵条件下通过测定其代谢产物生成反映人肝微粒体UGTs酶的活性,代谢产物浓度测定利用经过确证的液相色谱-质谱法(LC-MS/MS)进行。同时应用已知抑制剂对所建立的温孵体系进行验证。结果 建立的LC-MS/MS分析方法符合生物分析要求;在本实验温孵体系下,已知的UGT抑制剂均表现出明显的抑制作用。结论 该研究成功建立了一种准确可靠的体外评价体系,该方法可应用于评估药物对UGTs酶的体外抑制作用,该研究可为药物研发和临床合理用药提供重要指导。Aim To establish a reliable in vitro evaluation system to assess human liver UGTs activities and the inhibitory effect of drugs on human liver UGTs and validate its accuracy and stability.Methods Six probe substrate of the six major subtypes of human liver microsomal UGT enzymes were selected.The activity of human liver microsomal UGT enzymes was determined under optimized incubation conditions using a metabolite generation method.The concentrations of metabolites were determined using a validated liquid chromatography-tandem mass spectrometry(LC-MS/MS)method.Known inhibitors were used to verify the established incubation system.Results The established LC-MS/MS analysis method complied with the requirements of bioanalysis.In the incubation system of this experiment,known UGT inhibitors exhibited significant inhibitory effects.Conclusions This study successfully establishes an accurate and reliable in vitro evaluation system,which can be used to assess the in vitro inhibition of drugs on UGT enzymes,providing important guidance for drug development and rational clinical medication.

关 键 词：UDP-葡萄糖醛酸转移酶 人肝微粒体 探针底物法 抑制作用 液相色谱-串联质谱 药物相互作用 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R345[医药卫生—基础医学] R977.3