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. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Small Business Innovation Research Grants (SBIR) - Phase II (R44)
Project #
5R44CA114995-03
Application #
7246567
Study Section
Special Emphasis Panel (ZRG1-BCMB-L (11))
Program Officer
Song, Min-Kyung H
Project Start
2005-05-01
Project End
2008-06-30
Budget Start
2007-06-01
Budget End
2008-06-30
Support Year
3
Fiscal Year
2007
Total Cost
$472,639
Indirect Cost
Name
Reaction Biology Corporation
Department
Type
DUNS #
611741799
City
Malvern
State
PA
Country
United States
Zip Code
19355
Anastassiadis, Theonie; Deacon, Sean W; Devarajan, Karthik et al. (2011) Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity. Nat Biotechnol 29:1039-45
Horiuchi, Kurumi Y; Ma, Haiching (2009) Fluorescence polarization and time-resolved fluorescence resonance energy transfer techniques for PI3K assays. Methods Mol Biol 572:161-76
Liang, Shuguang; Xu, Wei; Horiuchi, Kurumi Y et al. (2009) Chemical microarrays: a new tool for discovery enzyme inhibitors. Methods Mol Biol 572:149-60
Ma, Haiching; Deacon, Sean; Horiuchi, Kurumi (2008) The challenge of selecting protein kinase assays for lead discovery optimization. Expert Opin Drug Discov 3:607-621
Ma, Haiching; Horiuchi, Kurumi Y (2006) Chemical microarray: a new tool for drug screening and discovery. Drug Discov Today 11:661-8