Orthopedics Department, Affiliated Zhongshan Hospital of Dalian University, Dalian, China.
J Pharm Pharmacol. 2013 Apr;65(4):521-7. doi: 10.1111/jphp.12014. Epub 2012 Dec 24.
The aim of this work was to identify the uridine glucuronosyltransferase (UGT) isoforms involved in the metabolism of the broad-spectrum antiviral drug arbidol.
A human liver microsome (HLM) incubation system was employed to catalyse the formation of arbidol glucuronide. The glucuronidation activity of commercially recombinant UGT isoforms towards arbidol was screened. A combination of kinetic analysis and chemical inhibition study was used to determine the UGT isoforms involved in arbidol's glucuronidation.
The arbidol glucuronide was detected when arbidol was incubated with HLMs in the presence of UDP-glucuronic acid. The Eadie-Hofstee plot showed that glucuronidation of arbidol was best fit to the Michaelis-Menten kinetic model, and K(m) and apparent V(max) were calculated to be 8.0 ± 0.7 μm and 2.03 ± 0.05 nmol/min/mg protein, respectively. Assessment of a panel of recombinant UGT isoforms revealed that UGT1A1, UGT1A3 and UGT1A9 could catalyse the glucuronidation of arbidol. Kinetic analysis and chemical inhibition study demonstrated that UGT1A9 was the predominant UGT isoform involved in arbidol glucuronidation in HLMs.
The major contribution of UGT1A9 towards arbidol glucuronidation was demonstrated in this study.
本研究旨在鉴定参与广谱抗病毒药物利巴韦林代谢的尿苷二磷酸葡萄糖醛酸转移酶（UGT）同工酶。
采用人肝微粒体（HLM）孵育体系催化利巴韦林生成葡萄糖醛酸缀合物。筛选商业重组 UGT 同工酶对利巴韦林的葡萄糖醛酸化活性。采用动力学分析和化学抑制研究相结合的方法，确定参与利巴韦林葡萄糖醛酸化的 UGT 同工酶。
当利巴韦林在 UDP-葡萄糖醛酸存在下与 HLMs 孵育时，检测到利巴韦林葡萄糖醛酸缀合物。Eadie-Hofstee 图表明，利巴韦林的葡萄糖醛酸化最符合米氏-门坦动力学模型，计算得出 K(m)和表观 V(max)分别为 8.0±0.7μm 和 2.03±0.05nmol/min/mg 蛋白。对一系列重组 UGT 同工酶的评估表明，UGT1A1、UGT1A3 和 UGT1A9 可以催化利巴韦林的葡萄糖醛酸化。动力学分析和化学抑制研究表明，UGT1A9 是 HLMs 中参与利巴韦林葡萄糖醛酸化的主要 UGT 同工酶。
本研究证明 UGT1A9 对利巴韦林葡萄糖醛酸化的贡献较大。
