High Throughput Membrane-Water Partition Coefficients

高通量膜-水分配系数

• 批准号: 6992526
• 负责人: George E Wright
• 金额: $ 25.54万
• 依托单位: GLSYNTHESIS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2007-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6992526
• 关键词: cell membrane; drug screening /evaluation; fluorescence; fluorescence spectrometry; high throughput technology; lipid bilayer membrane; mathematics; pharmacokinetics; technology /technique development; vesicle /vacuole; water

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.

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，通过有机溶剂和水之间的分配来测量，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：水分配系数与药物在细胞膜中的分配更为相关。在这个项目中，我们建议利用荧光体技术来有效地确定药物的真实膜：水分配系数。 “Fluorosomes-intra”是荧光标记的膜脂质双层囊泡，用于测量药物进入膜的速率。通过改变测试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，我们将利用含有细胞膜中存在的脂质的膜建立简单、快速的分配系数测定。该测定将适用于使用标准分光荧光计的多种分子，并且可以适应多孔板格式和机器人技术。具体目标是：建立测定药物膜水分配系数的荧光体内部测定条件和数学数据分析方法；优化 Fluorosome-intra 技术用于膜：水分配系数的常规测量；将 Fluorosome-intra 的结果与已公布的已知药物的膜：水分配系数值进行比较，以验证 Fluorosome-intra 的结果。我们将使用 Fluorosome-intra 获取一组扩展的测试药物的分配系数，以便与通过其他技术获得的已发表结果进行比较和关联。荧光体内部方法能够极大地增强我们确定和理解药物和其他小分子在水和生物膜之间分配的能力。该项目旨在利用该技术的特点，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。实施荧光体技术来获取膜：水分配系数将极大地促进药物发现。