Developing a cell-based high throughput screening for USP15 deubiquitinase inhibitor discovery

开发基于细胞的高通量筛选以发现 USP15 去泛素酶抑制剂

基本信息

批准号：
10287750
负责人：
Zhihao Zhuang
金额：
$ 43.47万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10287750
关键词：
Academia Active Sites Biological Biological Assay Biotin Carbidopa Cell Line Cell membrane Cell physiology Cells Chemicals Collaborations Cysteine DNA Repair Deubiquitination Disease Drug Compounding Drug Targeting Economic Burden Ensure Enzymes Event Excision FDA approved Goals Human Impairment Individual Industry Intervention Investigational Drugs Label Lead Levodopa Libraries Malignant Neoplasms Mitochondria Natural Immunity Neurodegenerative Disorders Neurologic Parkinson Disease Pathway interactions Peptides Performance Periodicity Permeability Pharmaceutical Preparations Pharmacology Physiological Play Process Proteins Recombinant Proteins Recombinants Research Personnel Retroviridae Role Running Signal Transduction Societies Speed System Technology Tertiary Protein Structure Testing Therapeutic Translational Research UBD protein Ubiquitin Ubiquitination United States Validation Variant Virus Diseases Work aging population base drug discovery high throughput screening human disease inhibitor/antagonist miniaturize multicatalytic endopeptidase complex nervous system disorder novel polyarginine propargylamine protein complex protein function reduce symptoms robotic system screening small molecule small molecule inhibitor stable cell line success therapy development

项目摘要

Developing a cell-based high throughput screening for USP15 deubiquitinase inhibitor discovery The goal of this project is to develop a ubiquitin probe-based AlphaLISA deubiquitinase HTS assay that enables cell-based and high throughput screening for human deubiquitinase USP15. It is now clear that the human ubiquitin system is fundamentally important to the normal cellular and organismal functions. Deubiquitinases or DUBs as an important class of enzyme have been associated with neurological disorders, particularly Parkinson’s disease (PD). Many disease-relevant and potentially druggable deubiquitinases are currently inaccessible to HTS-based inhibitor discovery due to the difficulty in their purification as homogeneous and active recombinant proteins in quantity large enough for HTS campaign. In this proposal, we aim at developing a cell-based AlphaLISA deubiquitinase assay for high throughput screening against USP15 and other DUBs, and setting the ground work for identifying and developing USP15 inhibitors with a mitophagy-promoting effect and the potential being developed into PD therapeutics.

开发基于细胞的高通量筛选以发现 USP15 去泛素酶抑制剂该项目的目标是开发一种基于泛素探针的 AlphaLISA 去泛素酶 HTS 检测方法，能够对人去泛素酶 USP15 进行基于细胞的高通量筛选。人类泛素系统对于正常的细胞和生物功能至关重要。去泛素酶或 DUB 作为一类重要的酶与神经系统疾病有关，特别是帕金森病 (PD)，许多与疾病相关且可能可药物化的去泛素酶都是如此。由于均质纯化困难，目前无法发现基于 HTS 的抑制剂和活性重组蛋白的数量足以支持 HTS 活动。在本提案中，我们的目标是。开发基于细胞的 AlphaLISA 去泛素酶测定，用于针对 USP15 和其他药物的高通量筛选 DUB，并为识别和开发具有促进线粒体自噬作用的 USP15 抑制剂奠定基础效果和开发成 PD 疗法的潜力。