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地阐明肝、肾、肠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。关于肾脏中的转运蛋白，我们构建了表达OAT 1或OAT 3的LLC-PK 1细胞，并观察了该细胞系中几种化合物的跨细胞转运，表明顶端外排转运蛋白的参与。我们还通过Mrp 4基因敲除小鼠和基因表达系统阐明了Mrp 4参与阿德福韦酯和头孢唑肟在肾脏的顶端外排。对于肝脏，我们建立了通过诱导代谢酶预测人体清除率变化的方法，并显示了当CYP 3A 4被联合给药药物诱导时，CYP 3A 4底物的药代动力学的良好预测。此外，我们首次阐明了在基底膜中表达的Mrp 3参与非索非那定和甲氨蝶呤的药代动力学（联合给药）。 ...更多信息 改变形式）。在小肠中，我们发现BCRP和SULT都在下部表达比上部多，并且在小鼠小肠中，4-MUS和硫酸米诺地尔向管腔侧的肠分泌在下部比在上部高，表明SUIT和BCRP协同工作以进行有效的解毒。我们阐明了BCRP也可作为外排转运蛋白，限制几种药物（丹曲林、哌唑嗪）和致癌化合物（MelQx、PhIP）在脑和睾丸中的分布。在这些组织中，P-gp和BCRP对总体外排的相对贡献取决于化合物的理化性质。建立了定量预测小肠内CYP 3A 4介导的药物相互作用的方法学，并证明表观肠容积法能较好地预测咪达唑仑和酮康唑在小肠内的药物相互作用。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