PIKFYVE antagonism as a therapy for C9ORF72-ALS/FTD

PIKFYVE 拮抗剂作为 C9ORF72-ALS/FTD 的治疗方法

基本信息

批准号：
10176613
负责人：
Samuel V Alworth
金额：
$ 53.35万
依托单位：
ACURASTEM, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10176613
关键词：
3-Dimensional Accounting Amyotrophic Lateral Sclerosis Animal Model Antisense Oligonucleotides Autophagocytosis Autopsy Biochemical Biological Assay Blood - brain barrier anatomy C9ORF72 Chemicals Clinic Collection Defect Development Disease model Evaluation Exhibits Genetic Goals Homology Modeling Human Lead Legal patent Lipids Mediating Modeling Modernization Motor Neurons Mus Mutation Nerve Degeneration Northern Europe Patients Phase Phenotype Phosphoric Monoester Hydrolases Phosphotransferases Proteins Safety Specific qualifier value Structure-Activity Relationship Testing Therapeutic Intervention Tissues Toxic effect Validation analog base drug discovery frontotemporal lobar dementia-amyotrophic lateral sclerosis in silico in vitro Assay in vivo inhibitor/antagonist knock-down loss of function mutation motor control new therapeutic target novel pre-clinical programs screening small molecule small molecule inhibitor stem cells targeted treatment therapeutic target trafficking virtual virtual screening

项目摘要

Optimization and validation of PIKFYVE antagonism as a therapy for C9ORF72-ALS/FTD Project Summary / Abstract The C9ORF72 repeat expansion mutation is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), accounting for over 50% of ALS cases in northern Europe and 10% of cases worldwide, making it a critical target for therapeutic intervention. Using patient-specific stem cell-based disease models, animal models, and postmortem tissue analysis, we have identified a new therapeutic target for C9ORF72 ALS/FTD, the lipid kinase PIKFYVE. We find that inhibition of PIKFYVE rescues the endosomal trafficking defects in C9ORF72 motor neurons and restores normal motor neuron survival. PIKFYVE functions in a manner that opposes FIG4, a phosphatase for which a loss-of-function mutation causes ALS. Antisense oligonucleotide-mediated knockdown of PIKFYVE rescues C9-ALS motor neuron survival, without exhibiting any toxicity toward control motor neurons. This combined functional and genetic evidence strongly indicate that small molecule inhibition of PIKFYVE kinase is a viable therapeutic target for C9-ALS/FTD. We have found that Apilimod reverses survival and other functional defects and is an effective PIKFYVE small molecule inhibitor. Apilimod has been tested in the clinic where target engagement without patient toxicity was observed, and we’ve determined it to be well tolerated in mice. We have begun execution of a two-pronged strategy to generate novel inhibitors of PIKFYVE. The first approach focuses on rescaffolding Apilimod to create a small molecule that can cross the BBB and can be patented. As a backup, our second approach employs virtual screening to identify new, patentable chemotypes which inhibit PIKFYVE. To this end, we have constructed 3D homology protein models for human PIKFYVE which we’ve used productively in rescaffolding to generate several Apilimod analogs. We employed the PIKFYVE homology models and the Small Molecule Drug Discovery Suite from Schrodinger and screened over 8 million compounds available virtually from the Icagen and E-molecule electronic compound collections. To develop structure activity relationships (SAR) for this program, we have established a biochemical PIKFYVE kinase assay as the primary assay for all compound evaluations. The goal of this Fast Track project is to identify a potent PIKFYVE inhibitor that is blood-brain-barrier penetrating. In phase I, we will use Apilimod, the 6 analogs from Table 1 and the 3 most promising leads from the in silico screen to test our entire funnel, including in vivo assays. In Phase II we will use these assays to optimize and validate a development candidate. Our specific aims are (Phase I) 1) Validation of primary and secondary in vitro assays; 2) Validation of tertiary assays; 3) Validation of proof of concept assays; (Phase II) 1) Compound optimization in primary and secondary assays; 2) Optimize of compound safety and administration through tertiary assays; 3) Establish in vivo proof of concept in C9ORF72 ALS/FTD.

优化和验证Pikfyve拮抗作用作为C9orf72-Als/ftd的疗法项目摘要 /摘要 C9orf72重复扩张突变是肌萎缩性侧索硬化症（ALS）和额叶痴呆（FTD），占北欧ALS病例的50％以上，占10％的病例全球，使其成为治疗干预的关键目标。使用基于患者的干细胞疾病模型，动物模型和验尸组织分析，我们确定了一个新的治疗靶点 C9orf72 ALS/FTD，脂质激酶pikfyve。我们发现抑制Pikfyve会反应内体 C9ORF72运动神经元的运输缺陷并恢复正常运动神经元存活。 pikfyve功能以反对FIF4的方式，该磷酸酶的功能丧失突变会导致ALS。反义寡核苷酸介导的pikfyve敲除c9-als运动神经元生存，而无需展示对控制运动神经元的任何毒性。这种合并的功能和遗传证据强烈表明小分子抑制pikfyve激酶是C9-ALS/FTD的可行治疗靶标。我们发现，apilimod逆转生存和其他功能缺陷，并且是有效的pikfyve 小分子抑制剂。 apilimod已在诊所进行了测试，在没有患者的情况下，目标参与观察到毒性，并且我们确定它在小鼠中的耐受性很好。我们已经开始执行两种策略生成新型Pikfyve抑制剂。第一种方法侧重于救援 apilimod可以创建一个可以越过BBB并获得专利的小分子。作为备份，我们的第二个方法采用虚拟筛选来识别抑制pikfyve的新的，可专利的化学型。对此最后，我们构建了人类Pikfyve的3D同源蛋白模型，我们在救援以产生几个apilimod类似物。我们采用Pikfyve同源模型和 Schrodinger的小分子药物发现套件，并筛选了超过800万种化合物实际上来自ICAGEN和E-MOLECULE电子化合物集合。发展结构活动该程序的关系（SAR），我们已经建立了生化的pikfyve激酶分析作为主要所有复合评估的测定。这个快速轨道项目的目的是确定潜在的Pikfyve抑制剂，该抑制剂是血脑屏障的穿透。在第一阶段，我们将使用apilimod，表1的6个类似物，以及来自在计算机屏幕上测试我们的整个漏斗，包括体内测定。在第二阶段，我们将使用这些测定优化和验证开发候选人。我们的具体目的是（i）1）验证主要和次要体外测定； 2）验证三级测定； 3）概念证明测定证明；（第二阶段）1）初级和次要测定中的复合优化； 2）优化复合安全和管理通过第三纪念； 3）在C9orf72 ALS/FTD中建立体内概念验证。