Using HTS to Identify Inhibitors of R155H-p97/VCP Mutant to treat IBMPFD/ALS

使用 HTS 鉴定 R155H-p97/VCP 突变体抑制剂来治疗 IBMPFD/ALS

基本信息

批准号：
10065879
负责人：
Tsui-Fen Chou
金额：
$ 22.92万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-10-11 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10065879
关键词：
Using HTS Identify Inhibitors R155H

项目摘要

Project Summary Single amino acid mutations in p97 AAA ATPase (also known as VCP, CDC48, TER ATPase) cause autosomal dominant human disorders embracing inclusion body myopathy, Paget's disease of bone, frontotemporal dementia (IBMPFD) and amyotrophic lateral sclerosis (ALS) among others. The most common p97 mutation (R155H) accounts for half the incidence of this rare, progressively debilitating, currently untreatable, fatal disorder. Strong data from IBMPFD/ALS human patients and mouse models suggests that targeting the disease-mutations of p97 will lead to a treatment. Since wild type (WT) p97 and its many ubiquitin-binding cofactors regulate the functions of protein substrates, WT p97’s essential functions must remain intact for human health. Previous studies demonstrated that R155H-p97 has differential sensitivity to inhibitors, higher basal ATPase activity, and is differentially regulated by interacting proteins. This suggests that mutant-selective small molecules can be identified which will inhibit R155H-p97, without effects on wild type. The goal of this project is to identify those mutant-selective small molecule candidates for drug development to treat or prevent IBMPFD/ALS pathology. Three specific aims are proposed to achieve our goal. AIM 1: Identify compounds that inhibit R155H, the most common p97 mutant causing IBMPFD/ALS. Conditions for high-throughput screening were developed and validated during a R155H-p97 pilot screen with the National Center for Advancing Translational Sciences (NCATS) Pharmaceutical Collection of 2,400 approved drugs. This project will carry out a quantitative high-throughput screening (qHTS) of 500,000 diverse compounds for inhibitors of R155H-p97 in the presence of p47 (one of the major cofactors of p97) in collaboration with the NCATS. The compounds will be ranked according to selectivity followed by potency. AIM 2: Test the compounds identified in AIM 1 in a cascade of biochemical and cell-based assays to ensure that the compounds will have the same effect inside as well as outside cells. The compounds will be characterized in established and validated cell-based assays to monitor the expression of seven potential IBMPFD/ALS disease biomarkers. Finally AIM 3: Use the lead compounds that emerge from Aim 2 to evaluate whether they correct disease in fibroblasts obtained from diseased and healthy mice and both human subjects with IBMPFD/ALS and healthy individuals. This data along with evaluation of Blood, Brain Barrier permeability should identify best candidates for further therapy development.

项目概要 p97 AAA ATPase（也称为 VCP、CDC48、TER ATPase）中的单个氨基酸突变导致常染色体显性人类疾病，包括包涵体肌病、佩吉特骨病、其中最常见的是额颞叶痴呆 (IBMPFD) 和肌萎缩侧索硬化症 (ALS)。 p97 突变 (R155H) 占这种罕见的、逐渐使人衰弱的疾病发生率的一半，目前 IBMPFD/ALS 人类患者和小鼠模型的有力数据表明，这是一种无法治愈的致命疾病。针对 p97 的疾病突变将导致治疗，因为野生型 (WT) p97 及其许多。泛素结合辅因子调节蛋白质底物的功能，WT p97 的基本功能必须先前的研究表明，R155H-p97 对人类健康具有不同的敏感性。抑制剂，较高的基础 ATP 酶活性，并且受到相互作用蛋白质的差异调节。可以鉴定突变体选择性小分子，其将抑制 R155H-p97，而不影响野生该项目的目标是确定那些突变选择性小分子候选药物。为了实现我们的目标，我们提出了三个具体目标。目标 1：鉴定抑制 R155H 的化合物，R155H 是导致 IBMPFD/ALS 的最常见 p97 突变体。在 R155H-p97 试验筛选过程中开发并验证了高通量筛选的条件国家转化科学促进中心 (NCATS) 药物收藏 2,400 该项目将对50万种不同药物进行定量高通量筛选（qHTS）。在 p47（p97 的主要辅助因子之一）存在下，用于 R155H-p97 抑制剂的化合物与 NCATS 的合作将根据选择性和功效对化合物进行排名。 2：在一系列生化和细胞检测中测试 AIM 1 中鉴定的化合物，以确保这些化合物在细胞内部和外部都具有相同的效果。在已建立和验证的基于细胞的测定中监测七种潜在 IBMPFD/ALS 的表达最后目标 3：使用目标 2 中出现的先导化合物来评估它们是否存在。纠正从患病小鼠和健康小鼠以及患有此病的人类受试者获得的成纤维细胞中的疾病 IBMPFD/ALS 和健康个体的数据以及血液、脑屏障渗透性的评估。应确定进一步治疗开发的最佳候选者。