NOVEL FLUORESCENT ESTROGEN RECEPTOR LIGAND BINDING ASSAY

最近越来越多的关注雌激素受体（ER）作为治疗 目标和日益关注的影响，环境雌激素 对人类健康的影响产生了对改进的测定方法的重要需求 为了鉴定雌激素受体配体，特别是适用于 涉及高通量筛选形式。 为了满足这一需求，PanVera将研究开发同质 两种人雌激素的体外荧光偏振分析 受体，ER α和ER β。 作为实现这一目标的第一步， I研究将通过1）开发雌激素受体的荧光配体 合成雌激素-荧光团缀合物并表征其 结构和化学和物理性质，2）评价 结合和荧光特性的新的缀合物， 纯化ER α和ER β，优化荧光偏振， 具有最佳性质的那些缀合物的测定条件，3） 通过以下步骤验证用于高通量筛选应用的测定： 筛选一组已知的ER配体和测试化合物， 药物化学品收集，并比较相对结合 与使用标准测定方法获得的那些的亲和力。 二期 研究将把这项技术扩展到其他类固醇激素 受体。 在第三阶段，PanVera将把这项技术商业化 通过其国内和国际分销网络， 制药联盟 拟定商业应用： 第一阶段SBIR研究将导致开发一种专有的， 用于高通量筛选均相荧光偏振分析 雌激素受体配体。 PanVera将把这些检测方法商业化 立即通过其国内和国际研究产品 分销网络 II期研究将产生类似的测定 其他类固醇激素受体，包括雄激素受体， 孕酮受体 在第三阶段，PanVera将继续扩大 通过全面分销，实现每种检测试剂的商业化 网络和制药联盟。

The recent increased focus on estrogen receptor (ER) as a therapeutic target and the growing interest in the impact of environmental estrogens on human health has created an important need for improved assay methods to identify estrogen receptor ligands, particularly assay methods suited to high throughput screening formats. To address this need, PanVera will investigate development of homogenous in vitro fluorescence polarization assays for two human estrogen receptors, ERalpha and ERbeta. As a first step towards this goal, Phase I studies will develop fluorescent ligands for estrogen receptors by 1) synthesizing estrogen-fluorophore conjugates and characterizing their structure and chemical and photophysical properties, 2) evaluating the binding and fluorescence characteristics of the new conjugates with purified ERalpha and Erbeta, and optimizing fluorescence polarization assay conditions for those conjugates with the best properties, 3) validating the assay for high throughput screening applications by screening a panel of known ER ligands and test compounds from a pharmaceutical chemical collection, and comparing the relative binding affinities with those obtained using standard assay methods. Phase II studies will extend this technology to additional steroid hormone receptors. In Phase III, PanVera will commercialize this technology through its domestic and international distribution network, and pharmaceutical alliances. PROPOSED COMMERCIAL APPLICATION: The Phase I SBIR studies will result in development of a proprietary, homogenous fluorescence polarization assay for high throughput screening of estrogen receptor ligands. PanVera will commercialize these assays immediately through its domestic and international research products distribution network. Phase II studies will result in similar assays for other steroid hormone receptors, including androgen receptors and progesterone receptors. In Phase III, PanVera will continue and expand commercialization of each assay through its comprehensive distribution network and pharmaceutical alliances.