IN VIVO INVESTIGATIONS USING MICRODIALYSIS SAMPLING

描述：(逐字摘自申请者的摘要)这个项目的目标是 开发一系列技术来研究药物在生物体内的传输 障碍。从体外模型系统到体内模型系统的连续技术 将开发动物系统。这些技术将基于一种组合 微渗析采样和毛细管电泳法。这些技术将 提供在早期药物开发中进行高通量筛选的方法 将与体内研究相关。 一种基于囊泡和细胞修饰毛细管的高通量方法 将发展电泳法(CE)。在这项技术中，水泡或者 人工的或生物衍生的，否则细胞将被用作迁移 CE中的修饰符。将测试化合物分配到囊泡或细胞中 将根据它们的渗透性修改测试化合物的迁移 进入囊泡或细胞。迁移相对于正常CE的变化应该是 与膜透性相关。这种方法将提供高吞吐量 通过允许同时测定混合物中化合物的渗透率。 我们还将开发使用细胞培养模型的体内微透析技术 生物屏障。利用细胞培养的扩散池方法 确定渗透率的模型将通过在细胞上生长来修改 微透析探头。这将提高细胞培养的产量。 技术通过改善系统中的质量传输。此外， 微透析的连续采样能力将提供动态 测定以及平衡测量。细胞包衣微渗析 探测器也将可重复使用。 最后，将继续使用微透析取样进行活体研究 研究体内跨越生物屏障的运输。重点将放在 跨胃肠道粘膜、跨胎盘和跨 血脑屏障。相同的一组测试化合物将用于所有 体外和体内研究提供了体外和体内的相关性。 这些方法的发展也将提供一系列的技术 增加了复杂性，但降低了吞吐量。

DESCRIPTION: (verbatim from applicant's abstract) The goal of this project is to develop a range of techniques to study drug transport across biological barriers. A continuum of techniques from in vitro model systems through in vivo animal systems will be developed. The techniques will be based on a combination of microdialysis sampling and capillary electrophoresis. These techniques will provide methods for high throughput screening in early drug development that will be correlated to in vivo studies. A high throughput method based on vesicular and cell-modified capillary electrophoresis (CE) will be developed. In this technique, vesicles, either artificial or biologically derived, or cells will be used as migration modifiers in CE. Partitioning of the test compounds into the vesicles or cells will modify the migration of the test compounds based on their permeability into the vesicle or cell. The change in migration relative to normal CE should correlate to membrane permeability. This approach will provide high throughput by allowing simultaneous permeability determinations of compounds in a mixture. We will also develop in vivo microdialysis techniques using cell culture models of biological barriers. The diffusion cell approach to using cell culture models to determine permeability will be modified by growing the cells on microdialysis probes. This will improve the throughput of cell culture techniques by improving the mass transport in the system. In addition, the continuous sampling capabilities of microdialysis will provide for kinetic determinations as well as equilibrium measurements. Cell-coated microdialysis probes will also be reusable. Finally, work will continue on the in vivo studies using microdialysis sampling to study transport across biological barriers in vivo. The focus will be on transport across the gastro-intestinal mucosa, across the placenta, and across the blood-brain-barrier. The same set of test compounds will be used in all of the in vitro and in vivo studies to provide an in vitro to in vivo correlation. The development of these methods will also provide a continuum of techniques of increasing complexity but decreasing throughput.