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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.

项目摘要 p97 AAA ATP酶（也称为VCP，CDC 48，TER ATP酶）中的单个氨基酸突变导致包括包涵体肌病，佩吉特骨病，额颞叶痴呆（IBMPFD）和肌萎缩侧索硬化（ALS）等。最常见的目前，p97突变（R155H）占这种罕见的、进行性衰弱的发病率的一半。无法治愈的致命疾病来自IBMPFD/ALS人类患者和小鼠模型的强有力数据表明，靶向p97的疾病突变将导致治疗。由于野生型（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的基本功能必须为了人类的健康保持完好。先前的研究表明，R155H-p97对抑制剂，更高的基础ATP酶活性，并通过相互作用的蛋白质差异调节。这表明可以鉴定出能抑制R155H-p97，而对野生型无影响的MUR选择性小分子。类型.该项目的目标是鉴定那些多选择性的小分子候选药物，开发用于治疗或预防IBMPFD/ALS病理的药物。为了实现我们的目标，提出了三个具体目标。目的1：鉴定抑制R155 H的化合物，R155 H是引起IBMPFD/ALS的最常见的p97突变体。在R155H-p97中试筛选期间，开发并验证了高通量筛选的条件，国家转化科学推进中心（NCATS）药物收藏2，400件批准的药物。该项目将对50万个不同的基因进行定量高通量筛选（qHTS）。在p47（p97的主要辅因子之一）存在下，与NCATS合作。将根据选择性以及随后的效力对化合物进行排序。目的 2：在一系列生物化学和基于细胞的测定中测试AIM 1中鉴定的化合物，以确保所述化合物在细胞内和细胞外具有相同的效果。这些化合物将被表征在已建立和验证的基于细胞的测定中，监测七种潜在的IBMPFD/ALS的表达，疾病生物标志物。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在从患病和健康小鼠以及两种人类受试者获得的成纤维细胞中纠正疾病， IBMPFD/ALS和健康个体。该数据沿着血液、脑屏障渗透性评价应确定进一步治疗开发的最佳候选药物。