钩吻素子在人与小型猪、SD大鼠、恒河猴、比格犬肝微粒体中的体外代谢比较  

In vitro koumine metabolism of liver microsomes in humans, minipigs, SD rats, Rhesus monkeys and Beagle dogs

作　　者：魏文增[1] 叶丽香[2] 黄慧玲[1] 丘宏强[3] 林菁[1,2] 俞昌喜[1] WEI Wen-zeng YE Li-xiang HUANG Hui-ling QIU Hong-qiang LIN Jing YU Chang-xi(College of Pharmacy, Fujian Medical University, Fuzhou 350108 Fujian Center for Safety Evaluation of New Drug, Fujian Medical University, Fuzhou 350108 Union Hospital, Fujian Medical University, Fuzhou 350001)

机构地区：[1]福建医科大学药学院,福州350108 [2]福建医科大学福建省新药安全性评价中心,福州350108 [3]福建医科大学附属协和医院,福州350001

出　　处：《中南药学》2016年第10期1045-1049,共5页Central South Pharmacy

基　　金：福建省科技计划重点项目(No.2013Y0088);福建省自然科学基金(No.2014J01417)

摘　　要：目的采用超高效液相色谱法(UPLC)研究钩吻素子在肝微粒体的体外代谢,比较其在人与各种实验动物代谢的相似情况,作为体内代谢研究的动物选择和分析的依据。方法建立UPLC法测定条件;通过对孵育时间、肝微粒体蛋白含量和钩吻素子浓度等条件的考察,优化钩吻素子与肝微粒体的反应体系;分析比较钩吻素子在各种属肝微粒体中代谢产物种类、生成量和代谢速率等差异。结果所建立的测定方法灵敏度、分离度高,经方法学考察表明其线性关系、准确度等指标均符合要求。在优化的反应体系下,钩吻素子在人、小型猪、SD大鼠、恒河猴、比格犬肝微粒体的代谢产物分别可检测到2、2、6、6、7种,主要代谢产物(代谢物Ⅴ)相同,占各种属所有代谢物的百分比分别为97.73%、58.09%、89.53%、90.40%和79.06%;其他代谢产物中,除了小型猪的1个(占41.91%)、比格犬的2个(分别占6.69%和7.35%)以外,均在3%以下。钩吻素子在人肝微粒体代谢较慢,且仅代谢22.28%,而在比格犬、恒河猴代谢近50%,代谢速率和代谢率高低顺序为:恒河猴、比格犬、SD大鼠、小型猪、人。结论不同种属肝微粒体对钩吻素子的代谢具有一定的差异,其中小型猪的主要代谢产物构成与人有明显差别,SD大鼠、恒河猴、比格犬的主要代谢产物与人相同,但代谢率和代谢速率以及其他代谢产物的构成方面也有一定的差异。Objective To investigate the inter-species difference of koumine metabolism in human, minipigs, SD rats, Rhesus monkey and Beagle dog liver microsomes by UPLC, as a surrogate model for metabolic studies in vivo. Methods A UPLC method was established to determine koumine and its metabolites. The incubation systems of koumine with liver microsomes in various species were optimized in terms of incubation time, microsomal protein content and koumine concentration. Koumine metabolism in the liver microsomes and the differences in various species were determined and the metabolic rates of koumine compared, as well as the constitution and relative amounts of metabolite. Results The method proved highly sensitive and good resolution for koumine metabolism. The linear calibration curves of koumine were obtained at 0.004-0.1 g·L^（-1） and 0.1-2.5 g·L^（-1）. The accuracy was 103.90%-110.1% and 103.6%-111.0%. The UPLC chromatographic determination of koumine and its metabolites in the liver microsomes were conducted under the optimized reaction system： the concentration of liver microsomes was 0.5 g·L^（-1）, concentration of koumine was 0.25 g·L^（-1）, incubation time was 2 h for Rhesus monkey, 3 h for SD rats and Beagle dog, and 4 h for humans and minipigs. We detected 2, 2, 6, 6 and 7 kinds of koumine metabolites in the liver microsomes of humans, minipigs, SD rats, Rhesus monkeys and Beagle dogs. The major metabolite percentages in all metabolites based on the chromatographic peak area were 97.73%, 58.09%, 89.53%, 90.40% and 79.06%, respectively. Another metabolite in minipig liver microsomes accounted for 41.91%, and 2 in Beagle dogs accounted for 6.69% and 7.35%. The rest metabolites were all below 3%. Koumine was metabolized slowlier and less in human liver microsomes than that in other animal species. The metabolism percentage of koumine was only 22.28% in humans, while nearly 50% in Beagle dogs and Rhesus monkeys. The metabolic ratio and speed in descending order were Rhesus monkeys, Beagle dogs, SD

关 键 词：钩吻素子 肝微粒体 体外代谢 超高效液相色谱法 种属差异 

分 类 号：R969.1[医药卫生—药理学]