Fluorosome Technique for Drug Permeability Studies

描述（由申请人提供）：我们已经验证了一种新的方法，“荧光体-反式技术”，用于测量药物的膜透性。47种上市药物的结果显示与人体口服吸收高度相关。该方法采用特定的荧光团囊泡制剂，并检测与化合物通过脂质双分子层转移速率成正比的荧光变化。使用合适的读板分光光度仪，该方法已适用于多孔板，并正在开发用于高通量筛选。该方法快速且廉价，只需要微摩尔浓度，不需要动物或活细胞，不需要无菌条件，不使用放射性标记材料或昂贵的检测方法。它适用于机器人技术，并补充了用于早期筛选候选药物的其他工具。在第二阶段，我们将开发一种预测肠细胞膜通透性和药物口服吸收的产品。我们将标准化现有和新形式的荧光体的生产和分析条件，增加与肠细胞膜的相关性，增加化合物覆盖范围，并扩大荧光体测定的pH范围。我们将制备一种用于血脑屏障渗透的荧光体。在多孔板中筛选化合物文库的高通量方法、数据采集程序以及数据计算和显示的软件修改将得到实施，后者将通过BMG Labtech公司的分包合同进行。现代药物发现的一个主要瓶颈是预测大型文库中哪些新的化学实体在动物中具有最佳的处置特性。良好的口服吸收是其关键特性，目前用于测量透性的细胞培养和人工脂质模型存在显著的缺点，包括低通量。实施检测所需的荧光体试剂、缓冲液和软件将根据许可出售给制药和生物技术用户和cro。氟体反式技术有望成为一种高效、低成本的替代技术，可应用于制药工业的高通量需求。

DESCRIPTION (provided by applicant): We have validated a novel method, the "Fluorosome-trans Technique", to measure membrane permeabilities of drugs. The results for 47-marketed drugs show high correlation with human oral absorption. This method employs specific fluorophore-vesicle preparations, and detects fluorescence changes that are proportional to the rate of transfer of compounds through the lipid bilayer. With a suitable plate reader spectrofluorimeter, the method has been adapted to multi-well plates and is being developed for high throughput screening. The method is rapid and inexpensive, requires only micromolar concentrations, employs no animals or living cells, does not require sterile conditions, and uses no radiolabeled materials or expensive detection methods. It is adaptable to robotics and complementary to other tools used in early screening of drug candidates. In phase II we will develop a product that will be predictive of intestinal cell membrane permeability and of the oral absorption of drugs. We will standardize manufacturing and assay conditions for current and new forms of Fluorosomes, increase relevance to intestinal cell membranes, increase compound coverage, and extend the pH range of the Fluorosome assay. We will prepare a Fluorosome for blood-brain barrier permeability applications. High throughput methods for screening compound libraries in multi-well plates, data acquisition procedures, and software modification for data calculation and display will be implemented, the latter via a subcontract with BMG Labtech Inc. A major bottleneck in modern drug discovery is the prediction of which new chemical entities from large libraries possess optimal disposition properties in animals. Good oral absorption is a key property, and the cell culture and artificial lipid models currently used for measuring permeabilities have significant disadvantages, including low throughput. The Fluorosome reagents, buffers and software needed to implement the assays will be sold under license to pharma and biotech users and CROs. The Fluorosome-trans Technique promises to become an efficient, low cost alternative and one which can be applied to the high throughput needs of the pharmaceutical industry.