Liu Houfu, Yu Na, Lu Sijie, Ito Sumito, Zhang Xuan, Prasad Bhagwat, He Enuo, Lu Xinyan, Li Yang, Wang Fei, Xu Han, An Gang, Unadkat Jashvant D, Kusuhara Hiroyuki, Sugiyama Yuichi, Sahi Jasminder
Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.)
Drug Metab Dispos. 2015 Jul;43(7):1008-18. doi: 10.1124/dmd.115.064170. Epub 2015 Apr 23.
Organic anion-transporting polypeptide (OATP) 1A2 has the potential to be a target for central nervous system drug delivery due to its luminal localization at the human blood-brain barrier and broad substrate specificity. We found OATP1A2 mRNA expression in the human brain to be comparable to breast cancer resistance protein and OATP2B1 and much higher than P-glycoprotein (P-gp), and confirmed greater expression in the brain relative to other tissues. The goal of this study was to establish a model system to explore OATP1A2-mediated transcellular transport of substrate drugs and the interplay with P-gp. In vitro (human embryonic kidney 293 cells stably expressing Oatp1a4, the closest murine isoform) and in vivo (naïve and Oatp1a4 knock-out mice) studies with OATP1A2 substrate triptan drugs demonstrated that these drugs were not Oatp1a4 substrates. This species difference demonstrates that the rodent is not a good model to investigate the active brain uptake of potential OATP1A2 substrates. Thus, we constructed a novel OATP1A2 expressing Madin-Darby canine kidney (MDCK) II wild type and an MDCKII-multidrug resistance protein 1 (MDR1) system using BacMam virus transduction. The spatial expression pattern of OATP1A2 after transduction in MDCKII-MDR1 cells was superimposed to P-gp, confirming apical membrane localization. OATP1A2-mediated uptake of zolmitriptan, rosuvastatin, and fexofenadine across monolayers increased with increasing OATP1A2 protein expression. OATP1A2 counteracted P-gp efflux for cosubstrates zolmitriptan and fexofenadine. A three-compartment model incorporating OATP1A2-mediated influx was used to quantitatively describe the time- and concentration-dependent apical-to-basolateral transcellular transport of rosuvastatin across OATP1A2 expressing the MDCKII monolayer. This novel, simple and versatile experimental system is useful for understanding the contribution of OATP1A2-mediated transcellular transport across barriers, such as the blood-brain barrier.
有机阴离子转运多肽（OATP）1A2因其在人血脑屏障的腔面定位和广泛的底物特异性，有潜力成为中枢神经系统药物递送的靶点。我们发现人脑中OATP1A2 mRNA表达水平与乳腺癌耐药蛋白和OATP2B1相当，远高于P-糖蛋白（P-gp），并证实其在脑中的表达高于其他组织。本研究的目的是建立一个模型系统，以探索OATP1A2介导的底物药物跨细胞转运以及与P-gp的相互作用。使用OATP1A2底物曲坦类药物进行的体外研究（稳定表达最接近的小鼠同工型Oatp1a4的人胚肾293细胞）和体内研究（未处理的和Oatp1a4基因敲除小鼠）表明，这些药物不是Oatp1a4的底物。这种种属差异表明，啮齿动物不是研究潜在OATP1A2底物脑内主动摄取的良好模型。因此，我们使用杆状病毒转导构建了一种新型的表达OATP1A2的Madin-Darby犬肾（MDCK）II野生型细胞系和MDCKII-多药耐药蛋白1（MDR1）系统。转导后MDCKII-MDR1细胞中OATP1A2的空间表达模式与P-gp重叠，证实其定位于顶膜。随着OATP1A2蛋白表达增加，OATP1A2介导的佐米曲坦、瑞舒伐他汀和非索非那定跨单层细胞的摄取增加。OATP1A2可抵消P-gp对共底物佐米曲坦和非索非那定的外排作用。一个包含OATP1A2介导的内流的三室模型用于定量描述瑞舒伐他汀在表达OATP1A2 的MDCKII单层细胞中从顶膜到基底外侧的跨细胞转运的时间和浓度依赖性。这个新颖、简单且通用的实验系统有助于理解OATP1A2介导的跨细胞转运对血脑屏障等屏障的作用。
