Identification of Therapeutic Small Molecules for Myotonic Dystrophy Type 1

描述（由申请人提供）：肌强直性营养不良症（DM1）是人类最常见的成人发病肌肉营养不良症。目前，没有治愈DM1和相关疾病的药物或FDA批准的药物。DM1是一种常染色体显性遗传病，由DMPK基因3'非翻译区ctg重复序列扩增引起。这种缺陷导致编码扩展CUG重复序列（CUGexp）的突变DMPK rna的表达，形成大的核内rna -蛋白复合物或灶。CUGexp RNA的表达导致异常的RNA剪接，这反过来又与DM1病理的关键特征的发展有关。我们假设降解或分散DM1细胞中CUGexp rna的小分子可以重建正常的剪接模式并逆转DM1病理。为了验证这一假设，我们开发了一种初级HTS和二级命中验证试验，以确定选择性改变DMPK CUGexp rna生物学而不影响正常DMPK转录本的小分子。我们的内部文库是使用强大的机器学习化学信息学平台开发的，由40,000个高度多样化的小分子组成，代表了数百万种化合物的文库。从2500种化合物的筛选中获得的初步结果表明，我们的策略可以快速识别有效的分子，成功逆转患者细胞和DM1小鼠模型中的DM1病理。在共同努力确定一组有效的先导化合物，可以开发作为DM1的治疗鸡尾酒，我们提出以下目标：实施初级HTS和二级命中验证试验，从我们的内部文库中筛选20,000个分子。目标2。测试命中在第三代DM1患者细胞为基础的分析，以确定高效的线索，逆转五种关键的细胞DM1表型。选择性，毒性和铅的协同作用将并行测量。目标3。将对先导化合物的化学结构进行反复细化，以优化药理学性质并建立构效关系。

DESCRIPTION (provided by applicant): Myotonic dystrophy (DM1) is the most common adult onset muscular dystrophy in humans. Currently, there is no cure or an FDA approved drug for DM1 and related diseases. DM1 is an autosomal dominant disorder resulting from the expansion of a CTG-repeat sequence in the 3' untranslated region of the DMPK gene. This defect results in the expression of mutant DMPK RNAs encoding expanded CUG repeats (CUGexp) that form large intra nuclear RNA-protein complexes or foci. Expression of CUGexp RNAs leads to abnormal RNA splicing, which in turn has been linked to the development of key features of DM1 pathology. We hypothesize that small molecules that degrade or disperse CUGexp RNAs in DM1 cells can re-establish normal splice patterns and reverse DM1 pathology. To test this hypothesis, we have developed a primary HTS and a secondary hit validation assay to identify small-molecules that selectively alter the biology of DMPK CUGexp RNAs without affecting the normal DMPK transcripts. Our in house library was developed using a robust machine learning chemoinformatics platform and consists of 40,000 highly diverse small-molecules representing a library of several million compounds. Preliminary results obtained from a screen of 2,500 compounds demonstrate that our strategy allows the rapid identification of potent molecules that successfully reverse DM1 pathology in both patient cells and DM1 mouse models. In a concerted effort to identify a set of potent lead compounds that can be developed as a therapeutic cocktail for DM1 we propose the following Aims: Aim 1. Implement primary HTS and the secondary hit validation assay to screen 20,000 molecules from our in-house library. Aim 2. Test hits in tertiary DM1 patient cell-based assays to identify highly potent leads that reverse five key cellular DM1 phenotypes. Selectivity, toxicity and synergy of leads will be measured in parallel. Aim 3. The chemical structure of lead compounds will be reiteratively refined to optimize pharmacological properties and establish structure-activity relationships. PUBLIC HEALTH RELEVANCE: We have developed a HTS screen to identify compounds that alter the biology of toxic CUGexp RNAs in myotonic dystrophy 1 (DM1). This screen will be used to identify therapeutic compounds that can be used to treat DM1.