Press Barry
MaRS Centre, Ontario Institute for Cancer Research, Toronto, ON, Canada.
Methods Mol Biol. 2011;763:139-54. doi: 10.1007/978-1-61779-191-8_9.
In vitro permeability assays are a valuable tool for scientists during lead compound optimization. As a majority of discovery projects are focused on the development of orally bioavailable drugs, correlation of in vitro permeability data to in vivo absorption results is critical for understanding the structural-physicochemical relationship (SPR) of drugs exhibiting low levels of absorption. For more than a decade, the Caco-2 screening assay has remained a popular, in vitro system to test compounds for both intestinal permeability and efflux liability. Despite advances in artificial membrane technology and in silico modeling systems, drug compounds still benefit from testing in cell-based epithelial monolayer assays for lead optimization. This chapter provides technical information for performing and optimizing the Caco-2 assay. In addition, techniques are discussed for dealing with some of the most pressing issues surrounding in vitro permeability assays (i.e., low aqueous solubility of test compounds and low postassay recovery). Insights are offered to help researchers avoid common pitfalls in the interpretation of in vitro permeability data, which can often lead to the perception of misleading results for correlation to in vivo data.
体外通透性测定对于先导化合物优化过程中的科学家而言是一种有价值的工具。由于大多数发现项目都聚焦于口服生物利用度药物的开发，因此将体外通透性数据与体内吸收结果相关联，对于理解吸收水平较低的药物的结构 - 物理化学关系（SPR）至关重要。十多年来，Caco - 2筛选测定一直是一种常用的体外系统，用于测试化合物的肠道通透性和外排倾向。尽管人工膜技术和计算机模拟系统取得了进展，但药物化合物在基于细胞的上皮单层测定中进行先导优化测试仍有益处。本章提供了进行和优化Caco - 2测定的技术信息。此外，还讨论了处理体外通透性测定中一些最紧迫问题（即测试化合物的低水溶性和测定后回收率低）的技术。文中提供了一些见解，以帮助研究人员避免在解释体外通透性数据时常见的陷阱，这些陷阱常常会导致与体内数据相关性产生误导性结果的认知。
