High Throughput Membrane-Water Partition Coefficients

描述（由申请人提供）：用于药物筛选和设计的关键分子特性是分配系数，通过在有机溶剂和水之间分配来测量，但更常见的是使用基于辛醇：水衍生的疏水性常数的算法来计算。然而，膜：水分配系数与药物分配到细胞膜中更相关。在本计画中，我们提出利用萤光体技术来有效地测定药物的真实膜：水分配系数。“Fluorosomes-intra”是荧光标记的膜脂质双层囊泡，其测量药物进入膜的速率。通过改变试验药物和脂质双层的浓度，并通过平衡或动力学算法分析数据，我们将建立一种简单，快速的分配系数测定法，膜含有存在于细胞膜中的脂质。该测定将适用于使用标准荧光分光光度计的宽范围的分子，并且可以适用于多孔板格式和机器人。具体目标是：建立药物膜-水分配系数测定的Fluorosome-intra分析条件和数学数据分析方法，优化Fluorosome-intra技术用于药物膜-水分配系数的常规测定，并将Fluorosome-intra测定结果与已知药物的膜-水分配系数测定值进行比较验证。我们将获得一组扩展的测试药物与荧光体内的分配系数与其他技术获得的已发表的结果进行比较和相关性。荧光体内的方法有能力大大提高我们的能力，以确定和了解水和生物膜之间的药物和其他小分子的分配。该项目旨在利用该技术的功能，将扩展药物化学家和药物发现科学家可用的工具，并利用该技术的高通量能力。荧光体技术的实施，以获取膜：水分配系数将大大促进药物发现。

DESCRIPTION (provided by applicant): A key molecular property used for drug screening and design is the partition coefficient, measured by partitioning between an organic solvent and water, but more commonly calculated with algorithms based on octanol:water-derived hydrophobicity constants. However, it is the membrane:water partition coefficient that is more relevant to partitioning of drugs into cell membranes. In this project we propose to exploit Fluorosome Technology to efficiently determine true membrane:water partition coefficients of drugs. "Fluorosomes-intra" are fluorescently labeled membrane lipid bilayer vesicles that measure drug entry rates into the membrane. By varying the concentrations of both test drug and lipid bilayer, and analysis of data by equilibrium or kinetic algorithms, we will establish a simple, rapid partition coefficient assay with membranes containing lipids that exist in cell membranes. The assay will .be applicable to a wide range of molecules using standard spectrofluorimeters, and can be adapted to multiwell plate formats and robotics. The specific aims are: to develop the Fluorosome-intra assay conditions and mathematical data analysis methods for the measurement of membrane:water partition coefficients of drugs; to optimize the Fluorosome-intra technique for the routine measurement of membrane:water partition coefficients, and; to validate the results of Fluorosome-intra in comparison with published values of membrane:water partition coefficients of known drugs. We will acquire partition coefficients of an expanded set of test drugs with Fluorosome-intra for comparison and correlation with published results obtained by other techniques. Fluorosome-intra methodology has the capacity to greatly enhance our ability to determine and understand the partitioning of drugs and other small molecules between water and biomembranes. This project seeks to exploit features of the technique that will extend the tools available to the medicinal chemist and drug discovery scientist, and to take advantage of the high-throughput capacity of the technique. The implementation of the Fluorosome Technique to the acquisition of membrane:water partition coefficients will thus greatly facilitate drug discovery.