NOVEL FLUORESCENT ESTROGEN RECEPTOR LIGAND BINDING ASSAY

近年来，雌激素受体(ER)作为一种治疗药物受到越来越多的关注 环境雌激素影响的目标和日益增长的兴趣 对人类健康产生了对改进检测方法的重要需求 确定雌激素受体配体，特别是适合的检测方法 到高通量的筛选格式。 为了满足这一需求，PanVera将研究同质 两种人雌激素的体外荧光偏振分析 受体，ERa和ERbeta。作为实现这一目标的第一步，阶段 I研究将在1)之前开发雌激素受体的荧光配体 雌激素-荧光团偶联物的合成及表征 结构和化学和光物理性质，2)评估 新的偶联物的结合和荧光特性 纯化的ERa和ERbeta，并优化荧光偏振 性能最好的偶联物的测定条件，3) 通过以下方式验证该检测方法用于高通量筛选应用 从一组已知ER配体和测试化合物中筛选 药物化学采集，并比较相对结合力 与使用标准检测方法获得的结果具有亲和力。第二阶段 研究将把这项技术扩展到额外的类固醇激素 感受器。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，以及 医药联盟。 建议的商业应用： 第一阶段SBIR研究将导致开发一种专有的、 用于高通量筛选的均相荧光偏振分析 雌激素受体配体。PanVera将把这些检测商业化 立即通过其国内和国际研究产品 分销网络。第二阶段的研究将产生类似的分析结果。 对于其他类固醇激素受体，包括雄激素受体和 黄体酮受体。在第三阶段，PanVera将继续并扩大 通过全面分销实现每种化验方法的商业化 网络和医药联盟。

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.