Roles of transporter adaptors as regulatory mechanism for drug absorption and disposition

转运蛋白适配器作为药物吸收和处置调节机制的作用

• 批准号: 18590137
• 负责人: KATO Yukio
• 金额: $ 2.53万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590137/
• 关键词: Transporter; Pharmacokinetics; Adaptor; Protein-protein interaction; Membrane permeation; Gastrointestinal absorption; Small intestine; Finical membrane; タンパク質問相互作用; 薬効・毒性; 機能制御

Many types of xenobiotic transporters have been identified. They generally exhibit multispecific recognition of various types of substrates, and mediate membrane permeation of therapeutic agents, thereby playing important roles in drug absorption and disposition. We have recently proposed that protein-protein interactions involving the xenobiotic transporters may affect their function, localization and expression on plasma membranes based on in vitro experimental data. So-called adaptor proteins that directly interact with the transporters include PDZ (PSD95, Dig and ZOO domain-containing proteins. This project was performed with an aim to clarify pharmacokinetic roles of such adaptor proteins in vivo. Using pdzk1 gene knockout (pdzk1^+) mice, it was found that interaction with a PDZ adaptor PDZK1 is essential for the cell-surface localization of three solute carriers, Slc15a1 (oligopeptide transporter PEPTD, S1c22a5 (carnitine/organic cation transporter OCTN2) and Slco 1a (organic ani … More on transporting polypeptide, OATP1A) in mouse small intestine. Electron microscopy revealed localization of PEPT1 in intracellular vesicular structures in pdzk1^+ mice. In pdzk1^+ mice, gastrointestinal absorption of cephalexin, a substrate of PEPT1 and carnitine, a substrate of OCTN2 was delayed, compared with wild mice. In addition, uptake of estrone sulfate, a substrate of OATP1A from apical membrane of small intestine was also decreased in pdzk1^+ mice. Thus, PDZK1 plays pivotal roles as a regulator of transporters, thereby affecting the absorption of substrates of those interacting transporters. Since PDZ adaptors have multiple PDZ domains in their structure, and each PDZ domain can interact with the cytosolic region of the transporters, it can be speculated that transporters are localized within networks consisting of several transporters and adaptors. We have also found that apical localization of PEPT1 and Slc5a1 (sodium/glucose cotransporter, SGLT1) was almost completely reduced in gene knockout mice for small GTP-binding protein rab8 (rab8^+). Gastrointestinal uptake across the apical membranes of glycylsarcosine, a substrate of PEPT1 and α-methylglucose, a substrate of SGLT1 was concomitantly reduced in rab8^+. Thus, our results demonstrate that rab8 is necessary for the proper localization of the two transporters and digestion of various nutrients which are the substrate of those transporters. Less

已鉴定出许多类型的外源转运蛋白。它们通常表现出对各种类型底物的多特异性识别，并介导治疗剂的膜渗透，从而在药物吸收和处置中发挥重要作用。我们最近根据体外实验数据提出，涉及外源转运蛋白的蛋白质-蛋白质相互作用可能会影响它们的功能、定位和质膜上的表达。所谓的直接与转运蛋白相互作用的接头蛋白包括含有PDZ（PSD95、Dig和ZOO结构域的蛋白）。该项目的目的是阐明此类接头蛋白在体内的药代动力学作用。使用pdzk1基因敲除（pdzk1^+）小鼠，发现与PDZ接头PDZK1的相互作用对于转运蛋白的转运至关重要。 小鼠小肠中三种溶质载体 Slc15a1（寡肽转运蛋白 PEPTD、S1c22a5（肉碱/有机阳离子转运蛋白 OCTN2）和 Slco 1a（有机阴离子转运多肽，OATP1A）的细胞表面定位。电子显微镜揭示了 PEPT1 在细胞内囊泡结构中的定位 pdzk1^+ 小鼠。与野生小鼠相比，在 pdzk1^+ 小鼠中，头孢氨苄（PEPT1 的底物）和肉碱（OCTN2 的底物）的胃肠道吸收延迟。此外，pdzk1^+ 小鼠对硫酸雌酮（小肠顶膜 OATP1A 的底物）的摄取也减少。 因此，PDZK1 作为转运蛋白的调节剂发挥着关键作用，从而影响那些相互作用的转运蛋白的底物吸收。由于PDZ适配器在其结构中具有多个PDZ结构域，并且每个PDZ结构域可以与转运蛋白的胞质区域相互作用，因此可以推测转运蛋白位于由多个转运蛋白和适配器组成的网络内。我们还发现 在小 GTP 结合蛋白 rab8 (rab8^+) 基因敲除小鼠中，PEPT1 和 Slc5a1（钠/葡萄糖协同转运蛋白，SGLT1）的顶端定位几乎完全减少。胃肠道通过顶膜摄取甘氨酰肌氨酸（PEPT1 的底物）和 α-甲基葡萄糖（PEPT1 的底物） SGLT1 在 rab8^+ 中同时减少。因此，我们的结果表明，rab8 对于两种转运蛋白的正确定位以及作为这些转运蛋白底物的各种营养物质的消化是必需的。少

项目成果

マウス小腸における有機アニオン性薬物estrone-3-sulfateのOatp介在吸収

Oatp介导的有机阴离子药物雌酮-3-硫酸酯在小鼠小肠中的吸收

• 发表时间: 2007
• 作者: 大竹 亨、加藤将夫、辻 彰;ほか
• 通讯作者: ほか

Role of SLC xenobiotic transporters and their regulatory mechanisms PDZ proteins in drug delivery and disposition.

• DOI: 10.1016/j.jconrel.2006.06.009
• 发表时间: 2006-11
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: T. Sugiura;Y. Kato;A. Tsuji

Transporter-mediated hepatic uptake of ulifloxacin, an active meabolite of a prodrug-type new quinolone antibiotic prulifloxacin in rats.

转运介导的肝脏对乌利沙星的摄取，乌利沙星是大鼠体内前药型新型喹诺酮抗生素普利沙星的活性代谢物。

• 发表时间: 2007
• 期刊: Drung Metab Pharmacokinet 22
• 作者: Yagi Y;Aoki M;Kato Y;Tsuji A;ほか
• 通讯作者: ほか

Transporter-mediated hepatic uptake of ulifloxacin,an active metabolite of a prodrug-type new quinolone antibiotic prulifloxacin in rats

转运蛋白介导的大鼠肝脏对乌利沙星的摄取，乌利沙星是一种前药型新型喹诺酮抗生素普利沙星的活性代谢物

• 发表时间: 2007
• 期刊: Drug Metab Pharmacokinet 22
• 作者: Yagi Y;Aoki M;Iguchi M;Shibasaki S;Kurosawa T;Kato Y;Tsuji A
• 通讯作者: Tsuji A

トランスポータータンパク質相互作用と活性調節創薬動態-医薬品創製のための考え方と最新情報-

转运蛋白相互作用和活性调节药代动力学 - 药物发现的概念和最新信息 -

• 发表时间: 2006
• 作者: Yukio;Kato;Yoshiyuki;Kubo;Akira;Tsuji;加藤将夫
• 通讯作者: 加藤将夫