Small Molecule Drug Discovery for CLN3 and CLN6 Disease

项目摘要/摘要 CLN3和CLN6疾病是一个更广泛的儿科神经退行性疾病家族的亚型，称为 神经性蜡样脂褐素沉着症(NCLS)或巴顿病。NCLS每年影响大约6-8名儿童 全球有10万名活产婴儿。CLN3和CLN6疾病的常见症状包括视力障碍 进展到失明，癫痫严重程度增加，认知和运动能力下降 痴呆症，并最终过早死亡。对于CLN3或CLN6疾病没有治愈或有效的治疗方法 为人所知。 开发治疗CLN3和CLN6疾病的新的疾病修饰剂是当务之急和 未得到满足的医疗需求。所有NCL之间共有的常见表型包括功能失调的自噬 导致储藏材料堆积，抗凋亡蛋白Bcl2和 神经酰胺的产生增加会导致神经元的凋亡性死亡和线粒体功能障碍。 自噬和细胞凋亡是促进细胞内稳态的生理过程。功能障碍 在许多神经退行性疾病中，一个或在许多情况下，两个过程都是表型 致NCLS。虽然单独针对这两个过程都会产生良好的药理效果，但也不是很小 已经确定了能够协同调节两者的分子。而多目标方法 多年来一直用于癌症治疗，但最近才开始应用于 神经退行性疾病，在CLN3和CLN6病中尚未被探索。 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，该库 融合蛋白自噬激活活性、抗细胞凋亡诱导和神经酰胺合成减少 翻译活性对人诱导多能干细胞(IPSC)来源的神经元的保护作用 诱导和表型诱导的细胞凋亡。此外，我们在初步数据中显示，IPSCs 取自CLN3患者，并衍生为功能神经元，重现异常的自噬、凋亡 和线粒体功能表型的疾病，这些表型可以通过选择拯救 先导化合物。此外，我们还展示了我们的先导化合物拯救CLN3疾病的概念验证 转基因CLN3小鼠模型的表型行为缺陷。 此应用程序的目标是优化我们专有的神经保护化合物库，以进一步 了解它们最低限度的药效基团，赋予“类药物”性质，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 在额外的患者IPSC来源的神经细胞系和 CLN3和CLN6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.