Deconvolution of Physicochemical Properties Contributing to Passive Diffusion of Depsipeptides

有助于缩酚肽被动扩散的物理化学性质的反卷积

• 批准号: 10607589
• 负责人: Madelaine Paige Thorpe
• 金额: $ 3.3万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607589
• 关键词: Academic Medical Centers; Accounting; Adult; Affect; Amides; Anti-Arrhythmia Agents; Arrhythmia; Behavior; Biological; Biological Assay; Calcium; Calcium Channel; Cardiac Myocytes; Categories; Cell Membrane Permeability; Cell membrane; Cells; Cellular Membrane; Cessation of life; Characteristics; Chemicals; Collaborations; Coupled; Creativeness; Cyclodepsipeptides; Dantrolene; Data; Depsipeptides; Development; Diffusion; Disease; Drug Kinetics; Elements; Equilibrium; Esters; Exhibits; Extravasation; Feedback; Flecainide; Guidelines; Heart; Hydrogen Bonding; Individual; Laboratories; Lead; Longevity; Measures; Mediating; Methylation; Modification; Molecular Conformation; Molecular Weight; Mutation; Natural Products; Nature; Partition Coefficient; Patients; Penetration; Peptides; Periodicity; Permeability; Pharmaceutical Preparations; Positioning Attribute; Production; Property; Protein Isoforms; Public Health; Quality of life; Rationalization; Research; Route; RyR1; RyR3; Ryanodine Receptor Calcium Release Channel; Sarcoplasmic Reticulum; Scientist; Series; Solubility; Specificity; Structure; Structure-Activity Relationship; System; Testing; Tetracaine; Therapeutic; Toxic effect; Travel; United States; Variant; Ventricular Arrhythmia; Work; analog; design; efficacy study; enantiomer; falls; functional group; improved; in vivo; inhibitor; innovation; interest; lipophilicity; member; membrane activity; novel therapeutics; pharmacologic; pharmacophore; preservation; release of sequestered calcium ion into cytoplasm; single molecule; small molecule; small molecule therapeutics; stereochemistry; success; sudden cardiac death; tool

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.

项目概要 心肌细胞中的钙通过细胞内钙通道兰尼碱受体 2 (RyR2) 释放。在 当 RyR2 发生突变的心脏中，可能会发生自发的 Ca2+ 渗漏，导致心脏衰竭 心律失常。小分子治疗药物如氟卡尼、丁卡因和丹曲林，特异性低、低 膜渗透性、溶解度低、选择性差、效力低或有毒性。之间的治疗选择性 RyR 同种型（RyR1、RyR2、RyR3）的缺乏，增强了我们开发 ent-verticilide 作为抗 心律失常剂。 该提案的重点是发现和开发抗心律失常药物的新疗法。这 拟议的工作是建立在我们发现有效和选择性抑制 RyR2 介导的钙流的基础上 ent-verticilide。通过跨学科合作，发现ent-verticilide是一种选择性的 RyR2 介导的钙释放抑制剂，包括体内功效的初步研究。作为 24 名成员 环缩酚肽，分子量为 853 Da，ent-verticilide 不属于传统的杀虫剂类别 小分子药物。虽然具有体内活性的“超越 5 规则”化合物的数量不断增加，但 对它们的药代动力学 (PK) 的了解已经滞后，因此需要新的化学工具和创造性 推进该领域的策略。我们将通过以下方式研究这种非天然产品及其类似物的渗透性 结构-活性关系概况的开发侧重于渗透性和提高功效。我们 旨在不同程度地系统设计和合成ent-verticilide的结构类似物 N-甲基化。综合之后，我们将研究被动渗透率并收集结构数据 通知 SAR，为具体目标 1 的反馈提供额外的视角。 更改为 ent-verticilide，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研究被动膜的严格工具 使用透化和非透化细胞进行渗透性和基于心肌细胞的功能研究 实现一种整体假设驱动的方法，以发现 ent-verticilide 的类似物如何通过 细胞膜并最终靶向 RyR2。另一个优势是我们准备多样化类似物的定位 包括可能表现出对比渗透性的环链变体。随着了解的加深 作用机制，我们假设我们可以设计具有改进效力的 ent-verticilide 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方法。