Development of the quantitative prediction method of pharmacokinetics with considering the function of metabolic enzymes and transporters

本研究的目的是定量阐明肝脏、肾脏和肠道中表达的代谢酶和转运蛋白在药物吸收和排泄中的协同作用。关于肾脏的转运蛋白，我们构建了表达OAT1或OAT3的LLC-PK1细胞，并观察到该细胞系中几种化合物的跨细胞转运，提示顶端外排转运蛋白的参与。我们还利用mrp4基因敲除小鼠和基因表达系统阐明了mrp4参与阿德福韦和头孢唑酮在肾脏顶端外排的作用。在肝脏方面，我们建立了预测代谢酶在人体内清除量变化的方法，并对联合用药诱导细胞色素P3A4的底物的药代动力学进行了较好的预测。此外，我们首次明确了基底膜表达的mRP3参与了非索非那定和甲氨蝶呤(un…)的药代动力学。更多的改变形式)通过使用基因敲除小鼠。在小鼠的小肠中，BCRP和SULT在下半部的表达均高于上半部，而4-MUS和米诺地尔在小鼠小肠下部向肠腔侧的分泌量高于上半部，表明Suit和BCRP协同作用达到了有效的解毒效果。我们阐明了BCRP也是几种药物(丹曲林、哌唑嗪)和致癌化合物(MelQx、PhIP)在脑和睾丸中分布的限制的外流转运体。在这些组织中，P-gp和BCRP对总流出的相对贡献由化合物的物理化学性质决定。此外，我们还建立了定量预测CyP3A4介导的小肠药物相互作用的方法学，并用表观肠道体积证明了咪达唑仑和酮康唑在小肠内药物相互作用的良好预测。较少

The purpose of this study is to clarify the cooperative roles of metabolic enzymes and transporters expressed in liver, kidney and intestine in the drug absorption and excretion quantitatively. Regarding the transporters in kidney, we constructed the OAT1- or OAT3-expressing LLC-PK1 cells and observed the transcellular transport of several compounds in this cell line, suggesting the involvement of apical efflux transporters. We also clarified the involvement of Mrp4 in the apical efflux of adefovir and ceftizoxime in kidney by using Mrp4 knockout mice and gene expression systems. As for the liver, we established the methodology for the prediction of the change in the clearance by the induction of metabolic enzymes in humans and showed the good prediction of the pharmacokinetics of substrates of CYP3A4 when CYP3A4 was induced by coadministered drugs Moreover, we first clarified that Mrp3 expressed in basal membrane is involved in the pharmacokinetics of fexofenadine and methotrexate (un … More changed form) by using knockout mice. In the small intestine, we found that both BCRP and SULT are expressed more at the lower part compared to the upper part, and that intestinal secretion of 4-MUS and minoxidil sulfate to luminal side at the lower part was higher than that at the upper part in small intestine in mice, indicating that SUIT and BCRP cooperatively work for the efficient detoxification. We clarified that BCRP also works as a efflux transporter for the limit of the distribution of several drugs (dantrolene, prazosin) and carcinogenic compounds (MelQx, PhIP in brain and testis. In these tissues, the relative contribution of P-gp and BCRP to the overall efflux is determined by the physicochemical properties of compounds. Also, we established the methodology for the quantitative prediction of drug-drug interaction mediated by CYP3A4 in the small intestine and demonstrated the good prediction of drug-drug interaction between midazolam and ketoconazole in the small intestine by using apparent intestinal volume. Less