HIGH THROUGHPUT SCREENING FOR MODULATORS OF APOPTOSIS

描述：（改编自申请人的摘要）细胞凋亡是一种细胞自杀机制，与每个细胞的遗传程序紧密相连。促进或阻止细胞凋亡的途径的不适当控制在多种病症如癌症和心脏病的发病机制中是重要的，这些病症在我们的社会中占相当大的发病率和死亡率。选择性诱导或抑制细胞凋亡的化合物的鉴定可能会带来显著的临床益处。然而，可用的评估细胞凋亡的测定不太适合于从大的化合物文库中鉴定这样的促细胞凋亡或抗细胞凋亡剂，因为可以有效进行的那些测定缺乏灵敏度和特异性，并且那些灵敏和特异的测定也是繁琐和劳动密集的。这个I期STTR项目提出验证高通量测定以鉴定在小有机化合物的组合组内启动或防止细胞凋亡的试剂。该项目有两个里程碑：（i）设计、开发和优化灵敏和特异的96孔板形式荧光高通量筛选（HTS）测定，以检测作为细胞凋亡指标的DNA片段化。(ii)验证HTS检测试剂盒的性能和形式，用于多种筛选和检测激活或抑制细胞凋亡的有机化合物。拟议的商业应用：药物筛选-评估化合物库的化合物，调节细胞凋亡。Pro.凋亡化合物（或类似物）可以进一步筛选为癌症或免疫药物治疗疾病的治疗剂。可以筛选抗凋亡化合物用于患有冠状动脉疾病、中风或神经退行性疾病的高风险患者。

DESCRIPTION: (Adapted from the applicant's abstract) Apoptosis is a mechanism of cell suicide hard-wired into the genetic program of every cell. Inappropriate control of the pathways that promote or prevent apoptosis are important in the pathogenesis of diverse conditions such as cancer and heart disease, which account for substantial morbidity and mortality in our society. Tangible clinical benefits could be derived from the identification of compounds that selectively induce or inhibit apoptosis. However, available assays to evaluate apoptosis are not well suited to identify such pro-apoptotic or anti-apoptotic agents from large compound libraries, as those assays that can be performed efficiently lack sensitivity and specificity, and those assays that are sensitive and specific are also tedious and labor intensive. This phase I STTR project proposes to validate a high through-put assay to identify agents that initiate or prevent apoptosis within a combinatorial set of small organic compounds. The project has two milestones: (i) Design, develop and optimize a sensitive and specific 96-well plate format chemiluminescent High Throughput Screening (HTS) assay to detect DNA fragmentation as an indicator of apoptosis. (ii) Validate the performance and format of the HTS assay for multiple screening and detection of organic compounds that activate or inhibit apoptosis. PROPOSED COMMERCIAL APPLICATION: Drug Screening - Evaluating compound libraries for compounds that modulate apoptosis. Pro. apoptotic compounds (or analogues) can be further screened as therapeutic agents for cancer or immune- medicated disease. Anti-apoptotic compounds can be screened for use in high risk patients with coronary disease, stroke, or neurodegenerative disorders.