Development of Ryanodine Receptor 2 Selective Probes

项目总结 Ryanodine受体2(RyR2)突变导致RyR2介导的失调 钙的释放与多种心血管疾病有关。因此，有几个 小分子探针已被用于心脏疾病的研究和治疗 RyR钙通道的靶向。氟卡胺、丁卡因和丹曲林，最 这些常见的小分子探针，也不是没有问题。例如，氟卡胺是 结构性心脏缺陷或心力衰竭患者的对症应用。此外，选择性 特定的RyR亚型(RyR1、RyR2或RyR3)的特异性很少，而且到目前为止，RyR2选择性探针 并不存在。选择性地调节RyR2的新探针将对两者都有很大的帮助 作为更好地了解RyR2的功能和机制的治疗工具 细胞内钙离子流量。 这项建议的重点是开发一类新的抗心律失常药物，该药物使用 在约翰斯顿实验室发现的Hit化合物ent-verticilide-非天然的对映体 天然产物(-)-垂柳内酯。这种环状低聚体脱脂肽是一种有效的、选择性的 RyR2介导的哺乳动物钙释放的抑制剂，而天然的轮状内酯是无效的。 我们的目标是使用基于结构的设计方法来帮助澄清企业和企业之间的相互作用。 使用RyR2的Verticilide。这种方法可能会导致我们在寻找有效的、 RyR2的选择性调节剂用于心脏病的治疗。我们的放映时间将是 首先在明确定义的RyR2介导的自发钙释放试验中扎根。我们的目标是 为了既包括环大小的同系物的离散集合，又包括“非天然的”对映体，两个 基于天然产品的治疗开发中未被充分开发的变量，以进行筛选 目标(RyR2)，可能导致新的探测。我们还建议使用ent-verticilide 测定ADME的药理学研究，同时促进对ADME的理解 作用机制和RyR2功能。我们的方法非常适合最终的 开发一种有效的、选择性的RyR2抑制剂的目标以及努力实现 明确恩替利特治疗心律失常的作用机制。

PROJECT SUMMARY Ryanodine Receptor 2 (RyR2) mutations that result in dysregulated RyR2-mediated calcium release are associated with a variety of cardiovascular diseases. As a result, several small molecule probes have been used in the study and treatment of cardiac diseases through the targeting of RyR calcium channels. Flecainide, tetracaine, and dantrolene, the most common of these small molecule probes, are not without problems. Flecainide, for example, is contra-indicated in patients with structural heart defects or heart failure. Furthermore, selectivity of specific RyR isoforms (RyR1, RyR2, or RyR3) is rare, and to date, an RyR2-selective probe does not exist. New probes to selectively modulate RyR2 would be of great use both therapeutically and as tools to better understand the function of RyR2 and the mechanism of intracellular calcium flux. This proposal focuses on the development of a new class of antiarrhythmic agents using the hit compound discovered in the Johnston lab, ent-verticilide – the non-natural enantiomer of the natural product (–)-verticilide. This cyclic oligomeric depsipeptide is a potent and selective inhibitor of RyR2-mediated calcium release in mammals while the natural verticilide is inactive. We aim to use a structure-based design approach to help clarify the interaction of ent- verticilide with RyR2. This approach may lead to new findings in our search for potent, selective modulators of RyR2 for the treatment of cardiac disease. Our screening will be grounded first in the well-defined RyR2-mediated spontaneous calcium release assay. We aim to include both discrete collections of ring-size congeners and ‘unnatural’ enantiomers, two underexplored variables in natural-product based therapeutic development, to screen against the target (RyR2), potentially leading to new probes. We also propose to use ent-verticilide in pharmacological studies to determine ADME, while advancing the understanding of the mechanism of action and RyR2 function. Our approach is well suited towards the ultimate goals of both developing a potent, selective inhibitor of RyR2 as well as working towards defining the mechanism of action of ent-verticilide to treat cardiac arrythmias.