Evaluation of animal models for intestinal first-pass metabolism of drug candidates to be metabolized by CYP3A enzymes via in vivo and in vitro oxidation of midazolam and triazolam
Evaluation of animal models for intestinal first-pass metabolism of drug candidates to be metabolized by CYP3A enzymes via in vivo and in vitro oxidation of midazolam and triazolam
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DOI：
10.3109/00498254.2012.751517
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发表时间：
2013-07-01
期刊：
影响因子：
1.8
通讯作者：
Narimatsu, S.
中科院分区：
文献类型：
--
作者：
Kuze, J.;Mutoh, T.;Narimatsu, S.
1.寻找一种合适的评价方法，用于评价新药候选物、葡萄柚汁（GFJ）和溶媒的肠道首过代谢（自来水）预处理的小鼠或大鼠口服给予咪达唑仑（MDZ）或三唑仑（TRZ），并通过液相色谱/MS/MS/MS测定母体化合物及其代谢物的血液水平。在小鼠中，仅观察到GFJ对TRZ的血药浓度升高的显著作用。使用小鼠、大鼠和人肠和肝微粒体组分的体外实验表明，GFJ抑制MDZ（尤其是TRZ）的肠微粒体氧化。MDZ的底物抑制导致肠和肝微粒体组分中1 '-羟基化减少，但不导致4-羟基化减少。小鼠肠和肝微粒体组分中MDZ氧化和底物抑制的动力学特征与人微粒体中的动力学特征非常相似，但与大鼠微粒体中的动力学特征不同。此外，MDZ在小鼠、大鼠和人肠微粒体中引起细胞色素P450 3A依赖性TRZ 1 '-羟基化的机制失活，其效力相似.这些结果是在小鼠和大鼠中获得的数据分析中的有用信息，用于评价待被CYP 3A酶代谢的候选药物的首过效应。
1. To search an appropriate evaluation methodology for the intestinal first-pass metabolism of new drug candidates, grapefruit juice (GFJ)- and vehicle (tap water)-pretreated mice or rats were orally administered midazolam (MDZ) or triazolam (TRZ), and blood levels of the parent compounds and their metabolites were measured by liquid chromatography/MS/MS. A significant effect of GFJ to elevate the blood levels was observed only for TRZ in mice.2. In vitro experiments using mouse, rat and human intestinal and hepatic microsomal fractions demonstrated that GFJ suppressed the intestinal microsomal oxidation of MDZ and especially TRZ. Substrate inhibition by MDZ caused reduction in 1'-hydroxylation but not 4-hydroxylation in both intestinal and hepatic microsomal fractions. The kinetic profiles of MDZ oxidation and the substrate inhibition in mouse intestinal and hepatic microsomal fractions were very similar to those in human microsomes but were different from those in rat microsomes. Furthermore, MDZ caused mechanism-based inactivation of cytochrome P450 3A-dependent TRZ 1'-hydroxylation in mouse, rat and human intestinal microsomes with similar potencies.3. These results are useful information in the analysis of data obtained in mouse and rat for the evaluation of first-pass effects of drug candidates to be metabolized by CYP3A enzymes.