Deconvolution of Physicochemical Properties Contributing to Passive Diffusion of Depsipeptides

项目摘要 心肌细胞中的钙通过细胞内钙通道Ryanodine受体2（RyR 2）释放。在 RyR 2突变的心脏可能会发生自发性Ca 2+渗漏，导致心脏病 心律不齐小分子治疗剂如氟卡尼、丁卡因和丹曲林具有低特异性、低毒性和低毒性。 膜渗透性、低溶解性、差选择性、低效力或毒性。之间的治疗选择性 RyR亚型（RyR 1，RyR 2，RyR 3）的缺乏，提高了我们开发ent-verticilide作为抗肿瘤药物的兴趣。 麻醉剂。 该提案的重点是发现和开发新的治疗药物作为抗肿瘤药物。的 所提出的工作建立在我们发现RyR 2介导的钙流的有效和选择性抑制的基础上， 对映轮枝芹内酯。通过跨学科的合作，发现ent-verticilide是一种选择性的 RyR 2介导的钙释放的抑制剂，包括体内功效的初步研究。作为一个24人 分子量为853 Da的环状缩酚酸肽，对映轮枝酯福尔斯不属于传统的 小分子药物。虽然具有体内活性的“5倍以上规则”化合物的数量正在增长，但 对它们的药代动力学（PK）的理解已经滞后，因此需要新的化学工具和创造性的方法。 战术来推进战场。我们将研究这种非天然产物及其类似物的渗透性， 开发了一个结构-活性关系曲线，重点是渗透性和增加的功效。我们 目的系统地设计合成不同程度的对映轮叶酯结构类似物 N-甲基化。在综合之后，我们将研究被动渗透率并收集结构数据 为SAR提供信息，为具体目标1的反馈提供额外的视角。按方法论结构 我们将在渗透性，活性，结构， 和构象。这种方法的优点是结合了严格的工具来研究被动膜 渗透性和使用透化和非透化细胞的基于心肌细胞的功能研究 实现一个整体的假设驱动的方法，以发现如何类似物的ent-verticilide旅行通过 细胞膜并最终靶向RyR 2。另一个优势是我们的定位，以制备不同的类似物 其包括可能表现出对比渗透性的环链变体。随着人们对 为了研究其作用机制，我们假设我们可以设计出具有更高效力的恩替维昔利类似物， 选择性，从而提供针对致命性室性心律失常的潜在治疗。

PROJECT SUMMARY Calcium in cardiomyocytes is released via an intracellular calcium channel, ryanodine receptor 2 (RyR2). In hearts where there are mutations in RyR2, spontaneous Ca2+ leakages can occur resulting in cardiac arrhythmias. Small molecule therapeutics such as flecainide, tetracaine, and dantrolene have low specificity, low membrane permeability, low solubility, poor selectivity, low potency, or toxicity. Therapeutic selectivity between RyR isoforms (RyR1, RyR2, RyR3) is lacking, heightening our interest in developing ent-verticilide as an anti- arrhythmic agent. This proposal focuses on the discovery and development of new therapeutics as antiarrhythmic agents. The proposed work is founded on our discovery of potent and selective inhibition of RyR2-mediated calcium flux by ent-verticilide. Through a cross-disciplinary collaboration, it was discovered that ent-verticilide is a selective inhibitor of RyR2-mediated calcium release, including a preliminary study of efficacy in vivo. As a 24 membered cyclic depsipeptide with a molecular weight of 853 Da, ent-verticilide falls outside of the category of a traditional small molecule drug. While “Beyond Rule of 5” compounds with in vivo activity are growing in number, an understanding of their pharmacokinetics (PK) has lagged, thereby requiring new chemical tools and creative tactics to advance the field. We will investigate the permeability of this unnatural product and its analogues by development of a structure-activity relationship profile focused on both permeability and increased efficacy. We aim to systematically design and synthesize structural analogues of ent-verticilide with varying degrees of N-methylation. Following the synthesis, we will study passive permeability and collect structural data to inform SAR, providing additional perspective for feedback to Specific Aim 1. By methodicalstructural change to ent-verticilide, we will create an SAR-based feedback loop between permeability, activity, structure, and conformation. A strength of this approach is the combination of rigorous tools to study passive membrane permeability, and cardiomyocyte-based functional studies using both permeabilized and non-permeabilized cells to achieve an overall hypothesis-driven approach to discover how analogues of ent-verticilide travel through cellular membranes and ultimately target RyR2. Another strength is our positioning to prepare diverse analogues that include ring-chain variants likely to exhibit contrasting permeability. With increased understanding of the mechanism of action, we hypothesize that we can design analogues of ent-verticilide with improved potency and selectivity, thereby providing a potential therapeutic against fatal ventricular arrhythmias.