Advancing the development of a novel class of small molecules for treating pan-coronavirus infections

推进治疗泛冠状病毒感染的新型小分子的开发

基本信息

批准号：
10672328
负责人：
Shirit Einav
金额：
$ 71.38万
依托单位：
STANFORD UNIVERSITY
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：
2021-08-10 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10672328
关键词：
2019-nCoV Advanced Development Alphavirus Animal Model Antineoplastic Agents Antiviral Agents Binding Biochemistry Biological Assay Biological Markers COVID-19 COVID-19 outbreak COVID-19 treatment CRISPR interference Cells Chemicals Clinical Research Clinical Trials Coronavirus Coronavirus Infections Cultured Cells Data Set Dengue Dengue Virus Development Dimensions Drug Combinations Drug Kinetics Ebola Ebola virus Endosomes Erlotinib Excretory function GAK gene Genetic Genomics Growth Guide RNA Hamsters Human In Vitro Individual Industry Inflammation Inflammatory Inflammatory Response Integration Host Factors Investigational Drugs Laboratories Lead Life Cycle Stages Lung Measures Mediating Membrane Metabolism Modeling Molecular Molecular Target Molecular Virology Monitor Morbidity - disease rate Mus Organoids Pathway interactions Peripheral Blood Mononuclear Cell Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Pharmacology Study Phosphotransferases Production Property Proteomics RNA Viruses Readiness Regimen Reporting Research Resistance Resources Risk Rodent Model Role SARS-CoV-2 entry inhibitor SARS-CoV-2 infection SARS-CoV-2 inhibitor SARS-CoV-2 pathogenesis Safety Series Structure of parenchyma of lung Structure-Activity Relationship Study models Therapeutic Therapeutic Index Toxic effect Toxicology Translating Viral Viral Load result Viral Physiology Virus Virus Diseases Virus Replication Work absorption activity-based protein profiling analog chemical genetics combat cytokine design drug candidate drug development drug repurposing forest future outbreak future pandemic genome-wide in vivo inhibitor insight mortality novel off-label use pandemic coronavirus pharmacologic preclinical safety programs pyridine response side effect small molecule tissue injury tool trafficking treatment response virology virus host interaction

项目摘要

Abstract For the past decade, our laboratory has been studying the role of cellular kinases in intracellular trafficking of RNA viruses and as targets for broad-spectrum antivirals. Furthermore, we have provided a proof of concept for the potential feasibility of the host-targeted broad-spectrum antiviral approach by demonstrating that the inhibition of two cellular kinases, AAK1 and GAK, by novel or the approved anticancer drugs, sunitinib and erlotinib, protects mice from dengue and Ebola viruses with a high barrier to resistance. Since the therapeutic index (TI) of this drug combination is narrower for SARS-CoV-2 infection, here, we focus on an independent class of compounds, the isothiazolo[4,3-b]pyridine-based RMC-113 series, that emerged from our prior work, but does not inhibit AAK1 or GAK. We showed that RMC-113 and 25 related analogs have potent broad- spectrum antiviral activity with a high barrier to resistance. Excitingly, RMC-113 reduces SARS-CoV-2 titer to undetectable levels at non-toxic concentrations and binds PIKFYVE, a cell kinase that regulates endosomal trafficking. We hypothesize that RMC-113 analogs inhibit both multiple distinct steps in the SARS-CoV-2 life cycle and the inflammatory response to this virus, in part by targeting PIKFYVE, thereby offering attractive and safe candidate inhibitors to combat SARS-CoV-2, other pandemic coronaviruses and other emerging viruses. In Aim 1, we will use a multi-dimensional medicinal chemistry approach to optimize the TI and PK profile of lead RMC-113 analogs and define their in vitro therapeutic potential as broad anticoronavirus inhibitors. Aim 2 will determine the effect of prioritized analogs and apilimod, a repurposed drug candidate for COVID-19 that inhibits PIKFYVE, on viral replication, cytokine response and tissue injury in organoids derived from excised normal lung tissue supplemented with PBMCs from 20 human donors and in two rodent models. Aim 3 will generate ADME-toxicity and safety pharmacology datasets to select pre-IND candidates. In Aim 4, we will probe the mechanism of antiviral action of RMC-113. We will validate PIKFYVE as a candidate target and use an unbiased CRISPRi screen to identify RMC-113’s target(s) and profile its chemical-genetic landscape. In parallel, we will design a clickable RMC-113 probe to confirm the molecular target via activity-based protein profiling and to monitor target engagement. Lastly, we will probe functional relevance and specific roles of PIKFYVE and other candidates emerging via these approaches in SARS-CoV-2 infection, and validate them as the molecular target(s) mediating the antiviral effect. The predicted immediate impact is that this project will provide insight into the therapeutic potential and MOA of apilimod, a repurposed drug candidate (beyond the reported effect on viral entry), and will establish a unique human lung organoid model for studying SARS-CoV- 2 pathogenesis and response to treatment under more natural conditions. In the longer term, successful completion of our study will deliver a drug-like small molecule candidate designed to protect against resurge of COVID-19 and to provide readiness for future outbreaks with coronaviruses and other emerging viruses.

抽象的在过去的十年中，我们的实验室一直在研究细胞激酶在细胞内贩运中的作用 RNA病毒和作为广谱抗病毒药的靶标。此外，我们提供了概念证明为了证明宿主针对宿主的广谱抗病毒方法的潜在可行性通过新颖或认可的抗癌药物抑制两个细胞激酶Aak1和Gak，Sunitinib和 Erlotinib，保护小鼠免受具有高阻力障碍的登革热和埃博拉病毒。自疗法以来 SARS-COV-2感染的这种药物组合的索引（Ti）更窄，在这里，我们专注于独立 Isothiazolo [4,3-B]基于吡啶的RMC-113系列的化合物类别来自我们先前的工作，但不会抑制AAK1或GAK。我们表明，RMC-113和25相关类似物具有潜在的广泛令人兴奋的是，RMC-113将SARS-COV-2滴度降低到在无毒浓度下无法检测到的水平并结合Pikfyve，Pikfyve是一种调节内体的细胞激酶贩运。我们假设RMC-113类似物在SARS-COV-2寿命中抑制了两个不同的步骤循环和对该病毒的炎症反应部分是通过针对pikfyve的，从而提供了吸引力和安全候选抑制剂，以对抗SARS-COV-2，其他大流行冠状病毒和其他新兴病毒。在AIM 1中，我们将使用多维医学化学方法来优化铅RMC-113类似物，并将其体外治疗潜力定义为宽性抗癌病毒抑制剂。目标2 将确定优先类似物和apilimod的效果，Apilimod是Covid-19的重新替代药物的候选药物抑制pikfyve，在病毒复制，细胞因子反应和组织损伤中的细胞动物损伤。正常的肺组织补充了来自20个人类供体的PBMC和两个啮齿动物模型。目标3意志产生毒性和安全药理学数据集，以选择预定候选人。在AIM 4中，我们将探测RMC-113抗病毒作用的机制。我们将验证Pikfyve作为候选目标并使用一个无偏的CRISPRI屏幕，可识别RMC-113的目标并介绍其化学遗传景观。在并行，我们将设计可单击的RMC-113探针，以通过基于活性的蛋白质确认分子靶标分析并监视目标参与。最后，我们将探讨功能相关性和特定作用 Pikfyve和其他通过SARS-COV-2感染中出现的方法出现的候选者，并将其验证为介导抗病毒作用的分子靶标。预测的直接影响是该项目将提供有关重新选择的候选药物的Apilimod的治疗潜力和MOA的见解（超出了报道对病毒进入的影响），并将建立一个独特的人类肺器官模型，用于研究SARS-COV- 2在更自然条件下的发病机理和对治疗的反应。从长远来看，成功我们研究的完成将提供类似药物的小分子候选者，旨在防止重新调节 Covid-19，并为未来的冠状病毒和其他新兴病毒提供准备。