Small Molecule Drug Discovery for CLN3 and CLN6 Disease

项目总结/摘要 CLN 3和CLN 6疾病是更广泛的儿科神经退行性疾病家族的亚型， 神经元蜡样脂褐质病（NCL）或Batten病。NCL影响大约6-8名儿童， 全世界有10万例活产。CLN 3和CLN 6疾病的常见症状包括视力损害， 进展为失明，癫痫发作严重程度增加，认知和运动能力下降进展为 痴呆症，最终过早死亡。对于CLN 3或CLN 6疾病没有治愈或有效的治疗。 知道的 开发治疗CLN 3和CLN 6疾病的新的疾病修饰剂是一个迫切和紧迫的任务。 未满足的医疗需求所有NCL共有的常见表型包括功能失调性自噬 导致储存物质的积累，抗凋亡蛋白Bcl-2的表达减少， 导致神经元凋亡性死亡的神经酰胺产生增加和线粒体功能障碍。 自噬和凋亡是维持细胞内环境稳定的生理过程。功能障碍 一个或在许多情况下，两个过程，是表型在许多神经退行性疾病，此外， 到NCL。虽然单独靶向任一过程都会产生有希望的药理学效果，但不小的 已经鉴定出能够协同调节两者的分子。虽然多目标方法 已经被用于癌症治疗多年，它只是最近才开始被应用于 神经退行性疾病，并且尚未在CLN 3和CLN 6疾病中探索。 通过基于结构的方法，我们已经确定了一个多功能化合物库， 融合自噬激活活性、抗凋亡Bcl-2诱导和神经酰胺合成减少，导致 翻译活性，以保护人诱导多能干细胞（iPSC）衍生的神经元免受外部- 诱导的和表型诱导的细胞凋亡。此外，我们的初步数据表明， 从CLN 3患者获得并衍生为功能性神经元的神经细胞重演了异常的自噬、凋亡 和线粒体功能表型的疾病，并认为这些表型可以挽救选择 铅化合物。此外，我们展示了我们的先导化合物拯救CLN 3疾病的概念验证 在转基因CLN 3小鼠模型中的表型行为缺陷。 该应用程序的目标是优化我们专有的神经保护化合物库， 了解它们的最小药效团，赋予“药物样”特性，识别并消除任何潜在的 毒性，优化药物代谢和药代动力学参数，进一步证实其作用机制， 在另外的患者iPSC衍生的神经元系中起作用并证明概念验证的保护活性， CLN 3和CLN 6转基因小鼠。

PROJECT SUMMARY/ABSTRACT CLN3 and CLN6 disease are subtypes of a wider family of pediatric neurodegenerative diseases called the Neuronal Ceroid Lipofuscinoses (NCLs) or Batten Disease. The NCLs affect approximately 6-8 children per 100,000 live births worldwide. Common symptoms of CLN3 and CLN6 disease include vision impairment which progresses to blindness, seizures which increase in severity, cognitive and motor decline progressing to dementia, and ultimately premature death. No cure or effective treatment for either CLN3 or CLN6 disease is known. The development of new disease-modifying agents to treat CLN3 and CLN6 disease is an urgent and unmet medical need. Common phenotypes that are shared between all NCLs include dysfunctional autophagy leading to accumulation of storage material, reduced expression of the anti-apoptotic protein Bcl-2 and increased ceramide production leading to apoptotic death of neurons, and dysfunctional mitochondria. Autophagy and apoptosis are physiological process that contribute to cellular homeostasis. Dysfunction of one, or in many cases, both processes, is phenotypic across many neurodegenerative diseases in addition to the NCLs. While targeting either process individually results in promising pharmacological effect, no small molecule has been identified that is capable of modulating both synergistically. While a multi-target approach has been used in cancer treatment for many years, it has only recently begun being applied to neurodegenerative diseases and has yet to be explored in CLN3 and CLN6 disease. Through a structure-based approach, we have identified a library of multi-functional compounds that fuse autophagy activation activity, anti-apoptotic Bcl-2 induction and decreased ceramide synthesis resulting in translational activity to protect human induced pluripotent stem cell (iPSC)-derived neurons from externally- induced and phenotype-induced apoptosis. Moreover, we have shown in our preliminary data that iPSCs obtained from CLN3 patients and derived to functional neurons recapitulate the aberrant autophagy, apoptosis and mitochondrial function phenotype of the disease, and that these phenotypes can be rescued by selected lead compounds. Further, we show proof-of-concept that our lead compounds rescue CLN3 disease phenotypic behavioral deficits in a transgenic CLN3 mouse model. The goal of this application is to optimize our proprietary library of neuroprotective compounds to further understand their minimum pharmacophore, confer ‘drug-like’ properties, identify and eliminate any potential toxicity, optimize drug metabolism and pharmacokinetic parameters, further credential their mechanism of action and demonstrate proof-of-concept protective activity in additional patient iPSC-derived neuron lines and CLN3 and CLN6 transgenic mice.