Enzyme Assay Chips for Apoptosis Discovery

描述(由申请人提供)：反应生物学公司开发了一种极低成本的纳升反应微阵列，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/毒性分析市场。这些反应比药物发现中广泛使用的孔板形式小1000到10000倍。目前，单个微阵列可以进行3072个药物筛选反应。在第一阶段，反应生物学证明：纯化的人caspase在芯片的反应条件下具有很强的活性，并可以使用反应生物学酶芯片技术进行定量测量。通过第一阶段，RBC已经证明了创造可控和成本效益高的caspase芯片的可行性。具体目标1涉及使用一个小型的“标准化”文库对IC50图谱和uHTS的微阵列上的caspase分析进行验证。具体目标2涉及针对所有10个人类半胱氨酸酶筛选两个不同的化学库(总计175,000个化合物)。 所有的命中都将通过测量微阵列上的IC50来验证。最后，特殊目标3涉及对纯化的半胱氨酸天冬氨酸酶的孔板分析和基于细胞的细胞凋亡分析中的命中测试。新的半胱氨酸氨基转移酶抑制剂可能会在治疗急性损伤或慢性疾病的神经退行性变过程中取得进展。

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.