Kietic analysis and evaluation of drug absorption and excretion using cultured cells.

使用培养细胞进行药物吸收和排泄的动力学分析和评估。

基本信息

批准号：
07557145
负责人：
INUI Ken-ichi
金额：
$ 4.42万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

The characterization of absorption, distribution, metabolism, excretion of xenobiotics is a important process to evaluate the drug efficacy and safety in preclinical stages. We developed the in vitro experimental systems, the intestinal/renal membrane vesicles and the cultured epithelial cells, to save the use of experimental animals. In this study, we tried to analyze the drug transport across monolayrs of renal (LLC-PK_1, OK,MDCK) and intestinal (Caco-2) cells, and to evaluate the rate limiting step in drug absorption and excretion, as follows :1.Experimental system using cultured cell monolayrs : The intestinal and renal epithelial cells were grown on the microporous membrane filters. We examined the apical-to-basolateral and basolateral-to apical transport of drugs in relation to the cellular accumulation. This experimental system was shown to be useful for evaluate the drug transport in intestinal/renal epithelial cells.2.Toxicity of drugs in cultured cell : The toxicity of drug in renal cells was evaluated by measuring enzyme activities of apical membrane and lysosome. We established the optimal conditions to estimate the nephrotixicity of drugs.3.Kinetic analysis of drug transport in cell monolayrs : We developed a simple model to estimate the clearance from apical medium to cells, cells to apical medium, cells to basolateral medium, and basolateral medium to cells. The kinetic analysis of drug transport in cell monolayrs gave the useful information about the rate limiting step of drug transport and the activity of drug transporter in intestinal and renal epithelium.

外源物质的吸收、分布、代谢、排泄表征是临床前阶段评价药物疗效和安全性的重要过程。我们开发了体外实验系统，肠/肾膜囊泡和培养上皮细胞，以节省实验动物的使用。在本研究中，我们尝试分析肾细胞（LLC-PK_1、OK、MDCK）和肠细胞（Caco-2）单层的药物转运，并评估药物吸收和排泄的限速步骤，如下：1使用培养细胞单层的实验系统：肠和肾上皮细胞在微孔膜过滤器上生长。我们检查了药物从顶端到基底外侧和基底外侧到顶端的转运与细胞积累的关系。该实验系统可用于评估肠/肾上皮细胞中的药物转运。2.药物在培养细胞中的毒性：通过测量顶膜和溶酶体的酶活性来评估药物在肾细胞中的毒性。我们建立了估计药物肾毒性的最佳条件。3.单层细胞中药物转运的动力学分析：我们开发了一个简单的模型来估计从顶端介质到细胞、细胞到顶端介质、细胞到基底外侧介质和基底外侧的清除率培养基至细胞。单层细胞中药物转运的动力学分析提供了有关药物转运限速步骤以及肠和肾上皮中药物转运蛋白活性的有用信息。