Charactering the metabolism of cryptotanshinone by human P450 enzymes and uridine diphosphate glucuronosyltransferases in vitro  

作　　者：Jin ZENG Yu-juan FAN Bo TAN Hui-zong SU Yue LI Lin-lin ZHANG Jian JIANG Fu-rong QIU 

机构地区：[1]Department of Clinical Pharmacology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203,China

出　　处：《Acta Pharmacologica Sinica》2018年第8期1393-1404,共12页中国药理学报（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China （No 81173118） and Shanghai Key Lab of Traditional Clinical Medicine （Grant C14dZ2273200）.

摘　　要：Cryptotanshinone （CT） is the main active component in the root of Salvia miltiorrhiza Bunge （SMB） that displays antibacterial, anti-inflammatory and anticancer activities. In this study, we characterized phase I and phase II metabolism of CT in human liver microsomes in vitro and identified the metabolic enzymes （CYPs and UGTs） involved. The metabolites of CT generated by CYPs were detected using LC-MS/MS and the CYP subtypes involved in the metabolic reactions were identified using chemical inhibitors of CYP enzymes and recombinant human CYP enzymes （CYPIA2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4）. Glucuronidation of CT was also examined, and the UGT subtypes involved in the metabolic reactions were identified using recombinant human UGT enzymes （1A1, 1A3, 1A4, 1A5, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15 and 2B17）. After adding NADPH to the human liver microsomes incubation system, CT was transformed into 6 main dehydrogenation and hydroxylation metabolites. CYP2A6, CYP3A4 and CYP2C19 were the major contributors to the transformation of its hydroxylation metabolites. CYP2C19, CYPIA2 and CYP3A4 were the major contributors to the transformation of its hydrogenation metabolites in human liver microsomes. This study showed that the metabolites at m/z of 473 were mediated by UGTIA9 and that the metabolites at m/z of 489 were mediated by UGT2B7 and UGT2B4. CT was extensively metabolized by UGTs following metabolism by CYPs in the liver.Cryptotanshinone （CT） is the main active component in the root of Salvia miltiorrhiza Bunge （SMB） that displays antibacterial, anti-inflammatory and anticancer activities. In this study, we characterized phase I and phase II metabolism of CT in human liver microsomes in vitro and identified the metabolic enzymes （CYPs and UGTs） involved. The metabolites of CT generated by CYPs were detected using LC-MS/MS and the CYP subtypes involved in the metabolic reactions were identified using chemical inhibitors of CYP enzymes and recombinant human CYP enzymes （CYPIA2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4）. Glucuronidation of CT was also examined, and the UGT subtypes involved in the metabolic reactions were identified using recombinant human UGT enzymes （1A1, 1A3, 1A4, 1A5, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15 and 2B17）. After adding NADPH to the human liver microsomes incubation system, CT was transformed into 6 main dehydrogenation and hydroxylation metabolites. CYP2A6, CYP3A4 and CYP2C19 were the major contributors to the transformation of its hydroxylation metabolites. CYP2C19, CYPIA2 and CYP3A4 were the major contributors to the transformation of its hydrogenation metabolites in human liver microsomes. This study showed that the metabolites at m/z of 473 were mediated by UGTIA9 and that the metabolites at m/z of 489 were mediated by UGT2B7 and UGT2B4. CT was extensively metabolized by UGTs following metabolism by CYPs in the liver.

关 键 词：CRYPTOTANSHINONE P450 enzymes uridine diphosphate glucuronosyltransferases metabolic characteristics human liver microsomes 

分 类 号：Q343.1[生物学—遗传学] TS974.1[轻工技术与工程]