绑定属性的描述人类22:发展[3 h]维拉帕米radioligand-binding化验。
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Doppenschmitt年代,哥P Regardh CG Andersson结核病,Hilgendorf C, Spahn-Langguth H
绑定属性的描述人类22:发展[3 h]维拉帕米radioligand-binding化验。
J Exp其他杂志》1999年1月,288 (1):348 - 57。
- PubMed ID
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9862789 (在PubMed]
- 文摘
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交互exsorptive转运蛋白22 (P-gp)是一个可能的来源的药物药物动力学特性,包括吸收剂量依赖性,药物之间的相互作用,肠分泌,和有限的血脑屏障通透性。在建立体外与P-gp药物相互作用的分析方法,没有直接量化与P-gp配体的亲和力。相反,他们测量膜渗透的结果和一个受体结合过程;这可能会导致困难的解释结果。量化的分析输送药物的亲和力是radioligand-binding分析原理的基础上提出的。这样做的优势直接量化药品和P-gp之间的交互。由于可逆和竞争互动与P-gp众多基质,一个radioligand-binding试验是由[3 h]维拉帕米和[3 h]长春花碱放射性配体和人类肠道Caco-2细胞,过表达与P-gp培养在长春花碱的存在或使转染与多药耐药性基因MDR-1受体制备。试验于96年执行——孔板和有潜力用作高通量的方法。清晰的感应P-gp的表达是Caco-2证明细胞生长在长春花碱的存在,以及在转染细胞,尽管规模较小。放射性配体都是绑定到P-gp显示。 Verapamil was the radioligand of choice for further investigations due to its lower nonspecific binding to the transporter preparation. Kinetics as well as specificity of the binding of verapamil to the P-gp preparation were demonstrated. A two-affinity model was found to adequately describe the data derived from saturation as well as from competition experiments, in accordance with previous findings on two exsorption sites for P-gp. The binding properties of [3H]verapamil and [3H]vinblastine to a P-gp preparation derived from induced Caco-2 cells are described. The concentration-dependent displacement of the radioligand by nonlabeled substrates for P-gp should be a suitable principle for the determination of drug affinity to the respective binding sites at the human intestinal multidrug transporter P-gp.