权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Evaluation of the sigma-1 receptor as a potential therapeutic target for COVID-19

评估 sigma-1 受体作为 COVID-19 潜在治疗靶点

基本信息

批准号：
10267567
负责人：
Lei Shi
金额：
$ 17.54万
依托单位：
NATIONAL INSTITUTE ON DRUG ABUSE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10267567
关键词：
2019-nCoV Affect Affinity Agonist Antiviral Agents Autophagosome Binding Biological Assay Biological Process Bioluminescence COVID-19 Cardiotoxicity Categories Cells Classification Client Clinical Trials Cocaine Abuse Consensus Coupling Crystallization Dose Endoplasmic Reticulum Energy Transfer Evaluation FDA approved Goals Haloperidol Homo In Vitro Integral Membrane Protein Ligand Binding Ligands Light Lysosomes Membrane Methods Mitochondria Molecular Molecular Conformation Nature Pathologic Pathway Analysis Pentazocine Pharmaceutical Preparations Pharmacology Physiological Property Proteins Protocols documentation Reporting Role Signal Transduction Structure Temperature Therapeutic Therapeutic Effect Therapeutic Intervention Treatment Efficacy Western Blotting base cell type in silico in vitro Assay models and simulation molecular modeling nervous system disorder novel pre-clinical protein protein interaction receptor response scaffold screening sigma-1 receptor stoichiometry targeted treatment therapeutic target virtual screening

项目摘要

Aim 1. Classify antagonists and agonists by ligand-induced 1R oligomerization For several 1R antagonists that have been shown to reduce SARS-CoV-2 infectivity, the original studies in determining their 1R antagonist properties are either vague or do not exist (e.g., those for PB28), not mentioning at in vitro level using 1R specific functional assays as we have developed. Thus, we will use BRET, binding, and western blot assays specifically developed for 1R in my lab, to evaluate whether the ligands that have been found to have either anti-viral or pro-viral activities indeed have opposite functions at 1R, and to assess whether 1R is a valid therapeutic target for COVID-19. If 1R will be found to be an effective therapeutic target for COVID-19 either by us or by other groups, we will seek to develop a BRET assay to characterize ligand-induced effects on the 1R-Nsp6 interaction, similar to our previous characterizations of the protein-protein interactions between 1R and its client proteins. Such an assay will serve to further evaluate the feasibility of targeting the 1R-Nsp6 interface for the therapeutic purposes and to validate the hit compounds from our own virtual screening (see below). Aim 2. Discover and develop the ligands for 1R and against hERG Several 1R antagonists were found to have significant affinities at hERG, which may result in cardiotoxicity when used at high doses. As 1R can be potential drug targets for the treatment of cocaine abuse and COVID-19, we will identify and develop novel high-affinity 1R antagonists that possess low affinity at hERG. Specifically, we will adapt the existing platform that integrated chemoinformatics, molecular modeling, and in vitro assays to i) screen novel 1R ligands that do not possess significant affinity at hERG; and ii) to rationally optimize current scaffolds of the 1R ligands that show promising therapeutic effects. To further enhance the integration, we will use molecular modeling and simulations and network analysis of 1R to investigate the specific binding modes of selected ligands that may induce receptor conformational changes to affect 1R homomerization state. Thus, by in silico characterizations of these changes, we will connect the conformational changes of the 1R (and/or its client proteins) to pharmacological readouts from the BRET and western blot assays we have developed.

目标 1. 通过配体诱导的 1R 寡聚化对拮抗剂和激动剂进行分类对于几种已被证明可以降低 SARS-CoV-2 感染性的 1R 拮抗剂，确定其 1R 拮抗剂特性的原始研究要么含糊不清，要么不存在（例如 PB28 的研究），更没有提到我们开发的在体外水平使用 1R 特异性功能测定法。因此，我们将使用我的实验室专门为 1R 开发的 BRET、结合和蛋白质印迹测定法，来评估已发现具有抗病毒或促病毒活性的配体是否确实在 1R 上具有相反的功能，并评估 1R 是否是 COVID-19 的有效治疗靶点。如果我们或其他团体发现 1R 是 COVID-19 的有效治疗靶点，我们将寻求开发一种 BRET 测定法来表征配体诱导的对 1R-Nsp6 相互作用的影响，类似于我们之前对 1R 与其客户蛋白之间的蛋白质-蛋白质相互作用的表征。这样的测定将有助于进一步评估以 1R-Nsp6 界面为治疗目的的可行性，并验证我们自己的虚拟筛选中的命中化合物（见下文）。目标 2. 发现并开发 1R 和 hERG 的配体研究发现几种 1R 拮抗剂对 hERG 具有显着的亲和力，高剂量使用时可能会导致心脏毒性。由于 1R 可能成为治疗可卡因滥用和 COVID-19 的潜在药物靶点，因此我们将鉴定和开发对 hERG 具有低亲和力的新型高亲和力 1R 拮抗剂。具体来说，我们将采用集成化学信息学、分子建模和体外测定的现有平台来 i) 筛选对 hERG 不具有显着亲和力的新型 1R 配体； ii) 合理优化现有的1R配体支架，显示出有希望的治疗效果。为了进一步增强整合，我们将利用1R的分子建模和模拟以及网络分析来研究可能诱导受体构象变化以影响1R同聚状态的选定配体的特异性结合模式。因此，通过对这些变化进行计算机表征，我们将把 1R（和/或其客户蛋白）的构象变化与我们开发的 BRET 和蛋白质印迹测定的药理学读数联系起来。