Kinome Drug Bioassay Platform

描述(由申请人提供)：反应生物学公司(RBC)开发了一种极低成本的纳升反应微阵列，以满足高通量筛选(HTS)和化合物选择性图谱的各种药物发现需求。这些微阵列可以容纳6000多个单独的反应。每个反应比典型井板形式中使用的反应小1000到10000倍。在癌症、炎症、糖尿病等疾病中，激酶在信号转导和调控过程中发挥着关键作用。然而，寻找选择性的激酶抑制剂仍然是一个挑战。在第一阶段，RBC使用微阵列HTS优化了10种酪氨酸激酶分析和5种丝氨酸/苏氨酸激酶分析。通过与Lopac文库进行筛选，这些微阵列已被验证为具有激酶HTS。此外，在第一阶段开发的几种磷酸酶分析方法已经准备好用于大规模的高温超导运动。在第二阶段，我们建议扩展这项技术，以增加额外的30个激酶(目标1)。针对这些目标的IC50分析将以井板和微阵列格式进行评估，使用来自Lopac图书馆和商业供应商的已知化合物。在目标2中，RBC将开发一种通用的基于放射性同位素33P的激酶分析方法，以实现每10微克纯化的激酶发生100,000个反应。在目标3中，HTS活动使用79,000个不同结构的化合物文库对抗RBC激酶和磷酸酶小组，将建立一个数据库来驱动结构-活性关系(SAR)模型。通过第二阶段资金，RBC将验证这一激酶/磷酸酶HTS平台，以服务于寻求快速分析大型文库或领先系列以对抗从RBC分析集合中选择的众多激酶的客户。使用少量纯化蛋白快速实施HTS也是一个重要的机会。

DESCRIPTION (provided by applicant): Reaction Biology Corporation (RBC) has developed an extremely low cost nanoliter reaction microarray to serve various drug discovery needs for high throughput screening (HTS) and compound selectivity profiling. These microarrays can hold more than 6000 individual reactions. Each reaction is 1000 to 10,000-fold smaller than those used in typical well plate formats. Kinases play a pivotal role in signal transduction and regulation of processes in cancer, inflammation, diabetes etc. However, finding selective kinase inhibitors remains a challenge. In Phase I, RBC optimized over 10 tyrosine kinase assays and 5 serine/threonine kinase assays using microarray HTS. The microarrays have been validated for kinase HTS by screening against the LOPAC library. Additionally, several phosphatase assay methodologies developed during Phase I are ready for large scale HTS campaigns. In Phase II, we propose to expand this technology to add an additional 30 kinases (Aim 1). IC50 profiling against these targets will be evaluated in both well plate and microarray format with known compounds from the LOPAC library and commercial vendors. In Aim 2, RBC will develop a universal radioisotope 33P-based kinase assay to achieve 100,000 reactions per 10 ug of purified kinase. In Aim 3, HTS campaigns using a 79,000 compound library of diverse structures against the RBC panel of kinases and phosphatases will establish a database to drive structure-activity relationship (SAR) models. Through Phase II funding, RBC will validate this kinase/phosphatase HTS platform to serve customers seeking to rapidly profile large libraries or lead series against numerous kinases chosen from the RBC assay set. Rapid HTS implementation for proprietary customer kinases using small amounts of purified protein is also a significant opportunity.