Enzyme Assay Chips for Apoptosis Discovery

描述（由申请人提供）：Reaction Biology公司开发了一种极低成本的纳米升反应微阵列，用于超高通量筛选（uHTS）药物发现、大规模IC50测定和大规模IC50选择性/毒性分析。这些反应比在药物发现中广泛使用的孔板形式小1000到10000倍。目前，在单个微阵列上可以进行3072种药物筛选反应。在第一阶段，Reaction Biology证明：纯化的人caspase在芯片的反应条件下具有强大的活性，可以使用Reaction Biology酶芯片技术进行定量测量。通过第一阶段，RBC已经证明了制造可控且具有成本效益的caspase芯片的可行性。具体目标1涉及使用小型“标准化”文库在微阵列上验证用于IC50分析和uHTS的caspase分析。特异性目标2涉及针对所有10种人类半胱天冬酶筛选两个不同的化学文库（总计175,000种化合物）。

DESCRIPTION (provided by applicant): Reaction Biology Corporation has developed an extremely low cost nanoliter reaction microarrays to serve markets for ultra high throughput screening (uHTS) drug discovery, large scale IC50 determinations, and large scale IC50 selectivity/toxicity profiling. These reactions are 1000 to 10,000-fold smaller than well plate formats used widely in drug discovery. Currently, 3072 drug screening reactions can be carried out on a single microarray. During Phase I, Reaction Biology demonstrated that: purified human caspases have robust activities under the reaction conditions of the chip and can be quantitatively measured using the Reaction Biology enzyme chip technology. Through Phase I, RBC has demonstrated feasibility in creating controllable and cost effective caspase chips. Specific Aim 1 involves the validation of caspase assays on microarrays for IC50 profiling and uHTS using a small "standardization" library. Specific Aim 2 involves the screening of two different chemical libraries (totaling 175,000 compounds) against all 10 human caspases. All hits will be verified by measurement of IC50 on the microarrays. Finally, Specific Aim 3 involves the testing of hits in well plate assays against purified caspases and in a cell-based assay of apoptosis. New inhibitors of caspases may lead to advances in the treatment of neurodegenerative processes during acute injury or in chronic disease.