Small molecule neuronal autophagy inducers designed to block neurodegeneration to

描述(由申请人提供)：我们将采用一种新的方法来开发独特的ALS治疗方法，专注于激活自然细胞保护机制--自噬的药物。刺激自噬是治疗导致肌萎缩侧索硬化症的潜在神经变性的一种合乎逻辑的方法。这是神经元清除有毒错误折叠聚集蛋白的主要防御机制，有证据表明ALS运动神经元(MNS)的神经变性是由有毒的错误折叠聚集形式的SOD1、TDP43和FUS积聚引起的。这些蛋白的突变导致家族性ALS(FALS)，这些蛋白的过度表达或异常的翻译后修饰导致散发性ALS(SALS)的MN变性。我们将开发有效刺激ALS MNS自噬的药物，以消除有毒蛋白的积聚，以阻止神经退化和疾病进展，从而治疗SALS患者。我们的合作者芬克贝纳博士此前在ALS的小鼠神经元模型中发现了刺激神经元自噬并阻止疾病进展的药物。他开发了神经元自噬诱导剂(NAI)的药效团模型，将帮助我们合理设计一流的ALS治疗新家族。为了促进这一发现计划，他还开发了来自健康志愿者和FALS和SALS患者的人类MN(I-MN)的ALS创新模型。使用一种名为机器人显微镜(RM)的自动成像和纵向分析系统，他第一个展示了ALS I-MN的疾病表型，因为他们比健康志愿者的I-MN有更大的死亡风险。这种疾病表型为测试治疗肌萎缩侧索硬化症的新药疗效提供了一个强大的、可定义的生理终点。我们将使用这些新的体外检测方法对人类ALS I-MN和最近开发的来自FALS患者的人类I-星形胶质细胞进行分析，以开发一个新的NAI家族。利用药效团模型和迭代的药物化学过程，Nanosyn将完善NAI药效团模型，以确定诱导自噬所需的最小化学结构。初步研究已经确定了这种方法的可行性，确定了具有更高效力和更低安全责任的NAI。Nanosyn将快速合成模型关键化学部分周围的小型化合物库，这些化合物将与来自FALS和SALS患者的人类ALS I-MN模型进行筛选。这项初步筛查将确定在促进人类ALS I-MN存活方面具有更好的效力和有效性的NAI。结构-活性关系研究将进一步完善虚拟高通量筛选的模型，以确定包含CNS特权片段的独特化合物，以选择具有治疗慢性神经退行性疾病所需特性的NAI，例如增加CNS的可用性。在未来的第二阶段SBIR体内研究中，将通过Finkbeiner博士开发的一种新方法在CNS中诱导自噬，并在SOD1-G93A转基因小鼠和过度表达TDP43(A315T)的转基因小鼠中测试Leads在逆转运动性瘫痪、提高存活率和减少运动神经元丢失方面的有效性。

DESCRIPTION (provided by applicant): We will employ a novel approach to develop unique ALS therapeutics by focusing on drugs that activate a natural cellular protective mechanism, autophagy. Stimulating autophagy is a logical approach to treat the underlying neurodegeneration that causes ALS. It is the prime defense mechanism that neurons have for removing toxic misfolded aggregated proteins, and there is evidence that neurodegeneration of motor neurons (MNs) in ALS is caused by the buildup of toxic, misfolded, aggregated forms of SOD1, TDP43 and FUS. Mutations in these proteins cause familial ALS (fALS), and overexpression or abnormal post-translational modification of these proteins leads to MN degeneration in sporadic ALS (sALS). We will develop drugs that effectively stimulate autophagy in ALS MNs to remove the buildup of toxic proteins to block neurodegeneration and disease progression to treat sALS patients. Our collaborator, Dr. Finkbeiner, previously identified drugs that stimulate autophagy in neurons and block disease progression in murine neuronal models of ALS. He developed a pharmacophore model of neuronal autophagy inducers (NAIs) that will help us rationally design a new family of first-in-class ALS therapeutics To facilitate this discovery program, he also developed innovative models of ALS with human MNs (i-MN) derived from healthy volunteers and patients with fALS and sALS. Using an automated imaging and longitudinal analysis system called robotic microscopy (RM) he was the first to show a disease phenotype for ALS i-MNs in that they had a greater risk of death than i-MNs from healthy volunteers. This disease phenotype provides a robust, definable physiological endpoint to test new drugs for efficacy in treating ALS. We will use these novel in vitro assays with human ALS i-MNs and recently developed human i-astrocytes from patients with fALS, to develop a new family of NAIs. Using the pharmacophore model and an iterative medicinal chemistry process, Nanosyn will refine the NAI pharmacophore model to identify minimal chemical structures needed to induce autophagy. Preliminary studies have already established the feasibility of this approach by identify NAI's with increased potency and reduced safety liabilities. Nanosyn will rapidly synthesize small libraries of compounds around key chemical moieties of the model that will be screened against the human ALS i-MN models from patients with fALS and sALS. This primary screening will identify NAIs with improved potency and efficacy in promoting survival of human ALS i-MNs. Structure-activity relationship studies will further refine the model for virtual high-throughput screening to identify unique compounds that incorporate CNS-privileged pieces to select for NAIs with desired properties for treating a chronic neurodegeneration disease, such as increased CNS availability. Leads will be tested in future Phase 2 SBIR studies in vivo for inducing autophagy in the CNS by a novel approach developed by Dr. Finkbeiner that uses GFP-LC3 transgenic mice and for efficacy in SOD1-G93A transgenic mice and transgenic mice over- expressing TDP43 (A315T) in reversing motor paralysis, increasing survival, and reducing motor neuron loss.