药物对人肝CYP450酶诱导和抑制作用体外评价体系的建立与验证

发布时间：2018-06-25 16:10

本文选题：人肝微粒体 + 人原代肝细胞　；参考：《药学学报》2017年12期

【摘要】：建立药物对CYP450酶诱导与抑制作用的体外评价方法并进行验证。LC-MS/MS法测定人肝微粒体CYP450酶7种主要亚型的8个底物代谢产物;在优化的温孵条件下,底物法测定人肝微粒体CYP450酶的活性;应用已知抑制剂对所建立的温孵体系进行验证。LC-MS/MS法测定人肝细胞CYP450酶3种主要亚型的底物代谢产物;底物法测定人原代肝细胞CYP450酶的活性,同时收集温孵后的人肝细胞,应用Taqman荧光探针法测定主要亚型m RNA的表达量;应用已知诱导剂对所建立的温孵体系进行验证。建立的LC-MS/MS分析方法符合生物分析要求;已知抑制剂和诱导剂在本实验温孵体系下均表现出明显的抑制作用或诱导作用。建立的实验方法准确、可靠,可用于体外评价药物对CYP450酶的诱导或抑制作用。
[Abstract]:An in vitro evaluation method of drug induced and inhibitory effects on CYP450 enzyme was established and 8 substrate metabolites of 7 main subtypes of human liver microsomal CYP450 enzyme were tested by.LC-MS/MS method. The activity of human liver microsomal CYP450 enzyme was determined by substrate method under the optimized incubation condition; the incubated system was tested by the known inhibitor. .LC-MS/MS method was used to determine the substrate metabolites of 3 main subtypes of human hepatocyte CYP450 enzyme; the activity of CYP450 enzyme in human primary hepatocytes was determined by substrate method, and the cultured human hepatocytes were collected at the same time. The expression of the main subtype m RNA was measured by Taqman fluorescence probe. The established incubating system was established by using the known inducer. The LC-MS/MS analysis method conforms to the biological analysis requirements; the known inhibitors and inducers have obvious inhibition or induction effect under the incubation system of this experiment. The experimental method established is accurate and reliable, and can be used to evaluate the effect of drugs on the induction or inhibition of CYP450 enzyme in vitro.
【作者单位】：南京医科大学第一附属医院;中国药科大学药物质量与安全预警教育部重点实验室;中国药科大学药物分析系;南京科利泰医药科技有限公司;
【基金】：国家自然科学基金资助项目(81503160,81673515) 江苏省自然科学基金资助项目(BK20161591) 江苏省“六大人才高峰”项目(2014-YY-001) 江苏省医学重点人才基金项目(H201108) 药物质量与安全预警教育部重点实验室资助项目(DQCP2015MS07)
【分类号】：R96

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