Design, Synthesis and Efficacy of New Small Molecule Therapeutics to Impede Myotonic Dystrophy

摘要/摘要 在40多个神经肌肉和神经退行性重复扩张性疾病的不断壮大的家庭中， 包括强直性肌营养不良(DM)，涉及一种具有毒性的强RNA功能获得(GOF)机制 由扩展RNA诱导。在这一机制中，扩展的RNA隔离RNA结合蛋白(RBPs) 导致多条下游RNA加工途径的中断。扩张性的缩减 因此，RNA减轻疾病机制和下游发病机制是一种有吸引力的治疗方法 接近。我们之前已经展示了有希望的小分子功效，包括：(1)放线菌素D 介导的CTG重复序列选择性转录减少；(2)微管抑制物介导 毒性CUG RNA水平的选择性调节；以及(3)二胺介导的毒性RNA的减少。而当 这些结果显示了希望，这些化合物中的许多都是有毒的，并且表现出次优的性质，导致我们 开发一组新的修饰多环化合物(MPC)。这些化合物是基于三个 元素：杂环核心；苯并咪唑侧基；和官能化端基。修改每个 这些元素提供了一大组潜在的化合物，以帮助理解作用机制 并开发新的候选药物，以解决糖尿病迫切的未得到满足的治疗需求。初步数据： 其中两个MPC在纳摩尔范围内的DM1和DM2细胞系中显示了强劲的剪接修复能力 几乎没有相关的毒性或对细胞活性的影响，以及挽救2个独立的DM的错误剪接 老鼠模型。在这项提案中，我们将使用平行的体外和体内设计-模型-测试周期来 系统地修改和评估化合物，重点是替换、测试和提炼 三个MPC元素(核心组、侧组和端组)。这些数据将使您更好地了解其 作用机制及在糖尿病患者来源细胞中的治疗潜力测试 线条和动物模型。该项目的成功完成将提供一种新的治疗方法 小分子，更好地了解它们的作用机制和来自多个动物的体内数据 支持其未来治疗潜力的模型。综上所述，这些信息将针对大型 对糖尿病治疗方法的未得到满足的需求，并为未来的临床研究提供支持数据。

Summary/Abstract Many of the growing family of over 40 neuromuscular and neurodegenerative repeat expansion diseases, including myotonic dystrophy (DM), involve a strong RNA gain-of- function (GOF) mechanism with toxicity induced by expansion RNAs. In this mechanism, the expanded RNAs sequester RNA binding proteins (RBPs) leading to the disruption of multiple downstream RNA processing pathways. The reduction of the expanded RNAs to alleviate disease mechanism and downstream pathogenesis is therefore an attractive therapeutic approach. We have previously demonstrated promising small molecule efficacy including: (1) actinomycin D mediated selective reduction of transcription from expanded CTG repeats; (2) microtubule inhibitors mediated selective modulation of toxic CUG RNA levels; and (3) diamidines mediated reduction of toxic RNAs. While these results show promise, many of these compounds are toxic and display sub-optimal properties leading us to develop a new set of modified polycyclic compounds (MPCs). These compounds are based on three elements: a heterocyclic core; a benzimidazole side group; and functionalized end groups. Modifying each of these elements provides a large panel of potential compounds to aid in understanding mechanism of action and develop new drug candidates to address the urgent unmet therapeutic need in DM. Preliminary data for two of these MPCs shows robust rescue of splicing in both DM1 and DM2 cell lines in the nanomolar range with little associated toxicity or effects on cell viability as well as rescue of mis-splicing in 2 independent DM mouse models. In this proposal, we will use parallel in vitro and in vivo design-model-test cycles to systematically modify and evaluate compounds by focusing on replacement, testing and refinement of the three MPC elements (core, side and end groups). These data will provide a better understanding of their mechanism of action and be followed by testing of their therapeutic potential in DM patient-derived cell lines and animal models. The successful completion of this project will provide a new class of therapeutic small molecules, a better understanding of their mechanism of action and in vivo data from multiple animal models supporting their future therapeutic potential. Taken together this information will address the large unmet need for therapeutic approaches for DM and provide supporting data towards future clinical studies.