Li Yang, Xu Chunxia, Xu Jinjin, Qin Zifei, Li Shishi, Hu Liufang, Yao Zhihong, Gonzalez Frank J, Yao Xinsheng
College of Pharmacy, Jinan University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, Jinan University, Guangzhou, China.
J Pharm Pharmacol. 2020 Dec;72(12):1771-1786. doi: 10.1111/jphp.13324. Epub 2020 Jul 9.
Bavachin is a bioactive natural flavonoid with oestrogen-like activity. Here, we aimed to investigate its metabolic and disposal fates involving in CYPs, UGTs and efflux transporters.
Phase I metabolism and glucuronidation were performed by human liver microsomes (HLM). Reaction phenotyping and activity correlation analysis were performed to identify the main CYP and UGT isozymes. Chemical inhibition and gene knock-down approaches were employed to explore the function of BCRP and MRPs.
Five phase I metabolites (M1-M5) and three glucuronides (G1-G3) were identified. The CL values for M4 and G1 by HLM were 127.99 and 1159.07 μl/min per mg, respectively. Reaction phenotyping results suggested CYP1A1 (208.85 μl/min per mg) and CYP2C9 (107.51 μl/min per mg), and UGT1A1 (697.19 μl/min per mg), UGT1A7 (535.78 μl/min per mg), UGT1A8 (247.72 μl/min per mg) and UGT1A9 (783.68 μl/min per mg) all participated in the metabolism of bavachin. In addition, activity correlation analysis also supported the results above. Furthermore, the metabolism exhibited marked species differences, and rabbits were the appropriate model animals. Moreover, MRP4 was identified as the main contributor based on chemical inhibition and gene silencing approaches.
CYP1A1 and CYP2C9, UGT1A1, UGT1A7, UGT1A8 and UGT1A9, and MRP4 all played important roles in the metabolism and disposition of bavachin.
补骨脂二氢黄酮是一种具有雌激素样活性的生物活性天然黄酮类化合物。在此，我们旨在研究其涉及细胞色素P450酶（CYPs）、尿苷二磷酸葡萄糖醛酸基转移酶（UGTs）和外排转运蛋白的代谢及处置命运。
采用人肝微粒体（HLM）进行Ⅰ相代谢和葡萄糖醛酸化反应。通过反应表型分析和活性相关性分析来鉴定主要的CYP和UGT同工酶。采用化学抑制和基因敲低方法来探究乳腺癌耐药蛋白（BCRP）和多药耐药相关蛋白（MRPs）的功能。
鉴定出了5种Ⅰ相代谢产物（M1 - M5）和3种葡萄糖醛酸苷（G1 - G3）。HLM对M4和G1的清除率（CL）值分别为127.99和1159.07 μl/min per mg。反应表型分析结果表明，细胞色素P450 1A1（CYP1A1，208.85 μl/min per mg）、细胞色素P450 2C9（CYP2C9，107.51 μl/min per mg）以及尿苷二磷酸葡萄糖醛酸基转移酶1A1（UGT1A1，697.19 μl/min per mg）、尿苷二磷酸葡萄糖醛酸基转移酶1A7（UGT1A7，535.78 μl/min per mg）、尿苷二磷酸葡萄糖醛酸基转移酶1A8（UGT1A8，247.72 μl/min per mg）和尿苷二磷酸葡萄糖醛酸基转移酶1A9（UGT1A9，783.68 μl/min per mg）均参与了补骨脂二氢黄酮的代谢。此外，活性相关性分析也支持上述结果。而且，该代谢表现出显著的种属差异，兔子是合适的模型动物。此外，基于化学抑制和基因沉默方法确定多药耐药相关蛋白4（MRP4）是主要贡献者。
CYP1A1、CYP2C9、UGT1A1、UGT1A7、UGT1A8、UGT1A9以及MRP4在补骨脂二氢黄酮的代谢和处置过程中均发挥着重要作用。
