Discovery of Inositol(1,4,5)trisphosphate Receptor Inhibitors

描述（由申请人提供）：中风已成为工业化国家老年人死亡和发病的重要原因。为了解决日益增加的卫生保健费用负担以及对家庭和受影响个人造成的无法估量的情绪损失，迫切需要新的预防和治疗药物。缺氧和氧化应激导致细胞色素c的释放，这反过来又与控制钙从细胞内储存释放的Ins(1,4,5)P3受体（IP3R）的激活有关。因此，IP3R的小分子拮抗剂将是恢复适当的钙信号通路和防止细胞凋亡的有吸引力的治疗候选者。为了确定该方法的可行性，埃施朗生物科学公司提出合成并鉴定潜在的IP3拮抗剂。具体来说，我们将使用硅建模结合化学文库合成来制备停靠在配体结合位点的无磷酸盐IP3模拟物，以及一系列xestospongin（一种有效的天然IP3R抑制剂）类似物。候选化合物将使用中等通量的细胞内荧光法进行筛选，其中IP3在适当刺激的细胞中释放钙的时间过程在Fluo-3和药物处理的成纤维细胞中进行监测。活性将根据观察到的生物活性通过建模和合成的迭代循环进行优化。

DESCRIPTION (provided by applicant): Stroke has become a significant cause of mortality and morbidity among elderly people in industrialized countries. To address the growing burden of health care costs and the immeasurable emotional toll on families as well as affected individuals, new pharmacological agents for prevention and treatment are desperately needed. Oxygen deprivation and oxidative stress lead to release of cytochrome c, which is related in turn to the activation of Ins(1,4,5)P3 receptors (IP3R) controlling calcium release from intracellular stores. Therefore, small molecule antagonists of IP3R would be attractive therapeutic candidates to restore proper calcium signaling pathways and prevent apoptosis. To establish the feasibility of this approach, Echelon Biosciences proposes to synthesize and identify potential IP3 antagonists. Specifically, we will use in silico modeling in combination with chemical library synthesis to prepare phosphate-free IP3 mimetics that dock in the ligand binding site, as well as a series of xestospongin (a potent natural IP3R inhibitor) analogs. Candidate compounds will be screened using a medium throughput in-cell fluorescence-based assay in which the time course for calcium release by IP3 in appropriately stimulated cells is monitored in Fluo-3 plus drug-treated fibroblasts. Activity will be optimized by iterative cycles of modeling and synthesis based on biological activities observed.