Comparative studies on in vitro methods for evaluating in vivo function of MDR1 P-glycoprotein
Comparative studies on in vitro methods for evaluating in vivo function of MDR1 P-glycoprotein
复制标题
DOI：
10.1023/a:1013358126640
复制
发表时间：
2001-12-01
影响因子：
3.7
通讯作者：
Sugiyama, Y
中科院分区：
文献类型：
--
作者：
Adachi, Y;Suzuki, H;Sugiyama, Y
目的. MDR 1 P-糖蛋白（P-gp）在决定药物处置中起重要作用。本研究的目的是建立体外模型来预测P-gp在体内的功能。作为体外方法，检查了12种化合物穿过单层Caco-2和MDR 1转染细胞的跨细胞转运。这些化合物刺激ATP水解的能力也使用表达P-gp的分离的膜级分测定。作为描述体内P-gp功能的参数，我们计算了mdr 1a/1b基因敲除小鼠中化合物的脑-血浆浓度比除以野生型小鼠中相同的比例。观察到12种化合物的体外单层通量比与体内P-gp功能之间存在良好的相关性。虽然所有刺激ATP水解的化合物都能被P-gp显著转运，但有些化合物能被P-gp转运而不影响ATP水解。总之，跨P-gp表达细胞单层的体外通量比可用于预测体内P-gp功能。体外ATP水解的程度也可能是体内预测的有用参数，特别是在高通量筛选程序中消除P-gp底物。
Purpose. MDR1 P-glycoprotein (P-gp) plays an important role in determining drug disposition. The purpose of the present study was to establish in vitro models to predict the in vivo function of P-gp.Methods. As an in vitro method, the transcellular transport of 12 compounds across the monolayer of Caco-2- and MDR1-transfected cells was examined. The ability of these compounds to stimulate the ATP hydrolysis was also determined using the isolated membrane fraction expressing P-gp. As a parameter to describe the in vivo P-gp function, we calculated the brain-to-plasma concentration ratio of compounds in mdr1a/1b knockout mice divided by the same ratio in wild type mice.Results. A good correlation was observed between the in vitro flux ratio across the monolayer and in vivo P-gp function for 12 compounds. Although all compounds that stimulated ATP hydrolysis were significantly transported by P-gp, some compounds were transported by P-gp without significantly affecting ATP hydrolysis.Conclusion. Collectively, the in vitro flux ratio across monolayers of P-gp-expressing cells may be used to predict in vivo P-gp function. The extent of ATP-hydrolysis in vitro may also be a useful parameter for in vivo prediction, particularly for eliminating P-gp substrates in high-throughput screening procedures.