TDP-43 aggregation inhibitors for the treatment of ALS

 描述(申请人提供)：目前还没有治疗肌萎缩侧索硬化症(ALS)的方法，ALS是一种普遍存在的致命神经退行性疾病， 每一万个人。蛋白质聚集被认为是肌萎缩侧索硬化症和其他多种神经退行性疾病的主要驱动力。TDP-43是ALS中蛋白质聚集体的主要成分，TDP-43的突变足以引起患者的疾病。有些家系TDP-43有常染色体显性遗传突变，导致ALS 100%外显性。这将TDP-43与其他基因定义的突变基因一起置于细胞死亡途径中。TDP-43也是这些基因定义的ALS突变基因中唯一的一个，也是散发性ALS的标志性病理。这就是TDP-43如此重要的原因。TDP-43是一种RNA结合蛋白，在基础条件下是有核的，但在胁迫过程中会转移到细胞质中，在那里它形成被称为应激颗粒(SGS)的细胞质RNA/蛋白质复合体。TDP-43的疾病相关突变增强了TDP-43在体外和动物模型中聚集和形成SGS的能力。在ALS患者的脊髓和脑内，以及在ALS的细胞和动物模型中，胞质TDP-43聚集并与SGS共定位。这些综合观察都表明TDP-43、相关的SGS和ALS的发病机制之间存在着强烈的生物学联系。TPD-43在肌萎缩侧索硬化症的病理生理学中的强大作用表明，TDP-43是一种有效的疾病修改的药理靶点。Aquinnah制药公司已经批准了10种抑制TDP-43和SG聚集的先导化合物，这些化合物是在本杰明·沃洛津博士(波士顿大学医学院)的实验室进行的高通量筛查中发现的。在这个第一阶段的SBIR中，我们建议证明用这些先导化合物靶向TDP-43治疗ALS的可行性。这项建议的目标是确定两种具有最佳脑药代动力学(渗透性和半衰期)的先导化合物，然后使用ALS的TDP-43转基因小鼠模型在体内测试这些化合物改善TDP-43生物标志物的能力。在目标1中，我们将描述每种化合物的一般ADME特征，然后研究其在大脑中的药代动力学，以确定每种先导化合物的脑渗透率和半衰期。在目标2中，我们将选择两个最好的化合物，并与Wolozin博士合作，在ALS的病理TDP-43小鼠模型中测试它们。我们将通过评估潜在疗效的生物标志物，包括TDP-43聚集、磷酸化和泛素化，使用急性剂量的这些化合物来显示生化益处。这一阶段SBIR的成功将导致识别 在ALS急性给药动物模型中，1-2种化合物进入中枢神经系统并抑制TDP-43聚集。这将使更强大的药物开发计划随之而来，包括为长期给药奠定基础，以测试这些化合物在SBIR第二阶段延缓体内疾病进展的能力。

 DESCRIPTION (provided by applicant): There is currently no therapy for Amyotrophic Lateral Sclerosis (ALS), which is a universally fatal neurodegenerative disease that afflicts over 1 out of every 10,000 individuals. Protein aggregation has been implicated as a primary driving force in ALS and multiple other neurodegenerative illnesses. TDP-43 is the principle component of the protein aggregates in ALS, and mutations in TDP-43 are sufficient to cause disease in patients. There are families that have autosomally dominant inherited mutations in TDP-43 that cause ALS with 100% penetrance. This places TDP-43, along with other genetically defined mutant genes, in the cell death pathway. TDP-43 also stands out as the only one of these genetically defined ALS mutant genes that is ALSO the hallmark pathology of sporadic ALS. This is why TDP-43 is so important. TDP-43 is a RNA binding protein that is nuclear under basal conditions but translocates to the cytoplasm during stress where it forms cytoplasmic RNA/protein complexes termed "stress granules" (SGs). Disease-linked mutations in TDP-43 enhance the ability of TDP-43 to aggregate and form SGs in vitro and in animal models. Cytoplasmic TDP-43 aggregates accumulate and also co-localize with SGs in the spinal cord and brain of patients with ALS, as well as in cellular and animal models of ALS. These integrated observations all point to a strong biological connection between TDP-43, the associated SGs, and pathogenesis of ALS. The strong role of TPD-43 in the pathophysiology of ALS points to TDP-43 as a cogent pharmacological target for disease modification. Aquinnah Pharmaceuticals has licensed 10 lead compounds that inhibit TDP-43 and SG aggregation that were identified in a high throughput screen performed in the laboratory of Dr. Benjamin Wolozin (Boston University School of Medicine). In this Phase I SBIR, we propose to demonstrate the feasibility of targeting TDP-43 with these lead compounds for the treatment of ALS. The goal of this proposal is to identify two lead compounds that exhibit the best brain pharmacokinetics (penetration and half-life), and then to test the ability of the compounds to improve TDP-43 biomarkers in vivo using a transgenic mouse model of TDP-43 of ALS. In Aim 1, we will characterize the general ADME characteristics for each compound, and then investigate the pharmacokinetics in the brain to determine brain penetration and half-life for each of the lead compounds. In Aim 2, we will select the two best compounds, and test them in a pathological TDP-43 mouse model of ALS in collaboration with Dr. Wolozin. We will use acute dosing of these compounds to show biochemical benefit by assessing biomarkers of potential efficacy including TDP-43 aggregation, phosphorylation, and ubiquitination. Success in this Phase I SBIR will result in the identification of 1-2 compounds that enter the CNS and inhibit TDP-43 aggregation in an acute dosing animal model of ALS. This will then allow a more robust drug development program to ensue, including setting the stage for chronic dosing to test the ability of these compounds to delay disease progression in vivo in SBIR Phase II.