Kinome Drug Bioassay Platform

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.