COVID-19: Identification and Development of Clinical Candidates to Treat SARS-CoV-2

COVID-19：识别和开发治疗 SARS-CoV-2 的临床候选药物

基本信息

批准号：
10259371
负责人：
Donald Lo
金额：
$ 199.15万
依托单位：
NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
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/10259371
关键词：
2019-nCoV Address Advanced Development Ambulatory Care Antiviral Agents Automobile Driving Autophagocytosis Binding Biological Biological Assay Biological Availability COVID-19 COVID-19 pandemic Cell Surface Receptors Cells Chemistry Clinical Clinical Investigator Clinical Trials Collaborations Collection Communities Coronavirus DNA-Directed RNA Polymerase Data Data Set Development Disease Drug Combinations Drug usage Ebola virus Emergency Situation Enzymes Fluorescence Resonance Energy Transfer Formulation Genome Genomics Glycoproteins HIV Hepatitis C virus Host Defense Mechanism Human In Vitro Infection Influenza Intravenous infusion procedures Investigation Investigational Drugs Investigational New Drug Application Joints Laboratories Lead Libraries Luciferases Mannose Measures Mediating Middle East Respiratory Syndrome Middle East Respiratory Syndrome Coronavirus Names Natural Products Nucleocapsid Proteins Oligosaccharides Patients Peptide Hydrolases Pharmaceutical Preparations Pharmacologic Substance Pharmacology Plasmids Play Positioning Attribute Process Production Proteins Protocols documentation Public Health RNA RNA Polymerase Inhibitor RNA-Directed RNA Polymerase Replicon Reporter Reporter Genes Research Research Activity Research Institute Research Personnel Research Support Resources Role SARS coronavirus Safety Scientist Severe Acute Respiratory Syndrome Signal Transduction Societies System Testing Therapeutic Intervention Time Toxicology Training Transfection Translational Research United States National Institutes of Health Universities Vaccines Viral Viral Antigens Viral Packaging Virus Virus Replication Work atypical pneumonia base biosafety level 3 facility clinical candidate clinical development clinical efficacy clinical investigation coronavirus disease data portal drug candidate drug development drug discovery experience in vivo inhibitor/antagonist microbicide model development novel novel therapeutics open data operation pandemic disease particle pathogen pharmacokinetics and pharmacodynamics phase I trial pre-clinical remdesivir research clinical testing sharing platform small molecule therapeutic candidate therapeutic development viral RNA virtual model

项目摘要

TDB scientists have extensive experience in developing novel therapeutic candidates as well as repurposing existing treatments for novel therapeutic purposes. Drug repurposing seeks to explore new uses for existing drugs and therapeutic candidates that already possess detailed pharmacology, formulation, and safety data, which can reduce the time to reach clinical testing for the new indication. TDB rapidly initiated several repurposing campaigns to identify modulators of SARS-CoV-2 activity. Targets under investigation include host defense mechanisms, such as cellular autophagy, and viral entry into cells through interaction of the viral spike protein with human cell surface receptors. TDB scientists also are contributing assay data and protocols to the Open Data Portal, a resource created by NCATS to house a collection of datasets generated from SARS-CoV-2-related assays against all approved drugs in the NCATS Pharmaceutical Collection. This effort aims to share COVID-19 related data in an open data-sharing platform to allow research scientists, clinical investigators and public health officials to prioritize promising compounds and repurposed drugs for further development in treating COVID-19. We have worked in collaboration with experts in and outside of NIH on the development of 8 assays for use in repurposing screens of approved drug collections to identify compounds active against SARS-CoV-2. The compounds identified from such screens have potential for clinical trials as single agents or in drug combinations to treat COVID-19 patients. 1. A SARS-CoV-2 pseudotyped particle (PP) entry assay in collaboration with Gary Whittaker (Cornell University). The PPs were generated with three plasmid transfection system containing MLV gag-pol, CoV spike, and luciferase reporter gene w/ viral packaging signal. In the assay process, the PPs deliver luciferase reporter RNA to host cells via spike-mediated cell entry. This assay has been used for screen the inhibitors of SARS-CoV-2 entry as well as the mechanistic study of other compounds identified from other assays. 2. SARS pseudotyped particle (PP) entry assay and MERS entry assays in collaboration with Gary Whittaker (Cornell University). Both assays have been optimized for 1536-well screens and used for drug repurposing screens of approved drug collection. The identified hits were further evaluated in a cytopathic effect assay with live SARS-CoV-2 infection (Southern Research Institute) to identify anti-SARS-CoV2 compounds that have broad activity against coronaviruses. 3. A SARS-CoV-2 cytopathic effect assay in Vero 6 cells in collaboration with the Southern Research Institute (SRI). Several small collections of approximately 10,000 approved and investigational drugs, preclinical drug candidates, and bioactive compounds were screened using assay ready plates, prepared by NCATS, in the BSL-3 lab at SRI. This assay has also been used to evaluate anti-SARS-CoV-2 compounds identified from NCATSs BSL-2 assays. 4. A SARS-CoV-2 3CL protease (also named main protease) assay. This viral protease plays a critical role in SARS-CoV-2 viral replication. We have developed and optimized this enzyme assay and screened a collection of approximately 10,000 compounds. Currently, a virtual modeling screen of the entire NCATS compound collection is in progress using the data from this focused library screen as a training set. The lead compounds will be used for chemistry optimization for further drug development. 5. A SARS-CoV-2 RNA dependent RNA polymerase (RdRP) assay. This viral RNA polymerase is critical to SARS-CoV-2 viral replication in host cells. Remdesivir, an RdRP inhibitor originally developed for Ebola virus and other viruses, has been authorized by the FDA for emergency use in hospitalized COVID-19 patients. The efficacy and potency of this drug are limited for SARS-CoV-2, as it was not originally developed for this virus. Remdesivir is currently only administered by intravenous infusion that precludes it from outpatient treatment. We are developing a SARS-CoV-2 RdRP enzyme assay and a cell-based RdRP assay to support repurposing screens for new drug development. 6. A SARS-CoV-2 replicon assay for use BSL-2 laboratories. Research involving live SARS-CoV-2 virus requires BSL-3 facility, which greatly limits throughput. We are collaborating with three laboratories to develop a replicon type assay for SARS-CoV-2, similar to our previous work on HCV. This replicon assay will have a SARS-CoV-2 genomic fragment encompassing viral replication machinery, without the viral envelop proteins, and will include a mini-genome reporter gene (luciferase). We will use this assay to screen NCATS compound collections for drug discovery and development. 7. An AlphaLISA assay to detect the SARS-CoV-2 Nucleocapsid (N) protein. Currently, the cytopathic effect (CPE) assay of SARS-CoV-2 allows for the greatest throughput in BSL-3 laboratories, but uses a surrogate readout for host cell infection (cell death). The AlphaLISA N protein assay would allow BSL-3 labs to directly measure viral antigen production upon SARS-CoV-2 infection in host cells. This assay is homogenous and amenable to HTS. It was developed to help increase the throughput of BSL3 labs. 8. A TR-FRET (HTRF) assay to detect the SARS-CoV-2 Nucleocapsid (N) protein. This assay is similar to the AlphaLISA N protein assay, but uses a different readout. This assay could be used as an alternative assay for HTS, or as an orthogonal assay to confirm the hits identified from the above AlphaLISA assay. We also have begun a partnership to rapidly progress a candidate into clinical trials for SARS-CoV-2. Our collaborators recently completed phase I trials of a protein natural product for use as a microbicide against HIV. Its mechanism of action, selectively binding to high-mannose oligosaccharides on viral envelope glycoproteins, is relevant to diverse pathogens, including HIV, Influenza, Ebolavirus, and SARS-CoV. The candidate has shown in vitro and in vivo activity against several coronaviruses, including SARS-CoV and MERS-CoV. Because it binds the spike glycoprotein on SARS-CoV, it is likely to have activity against all coronavirus strains, not only the SARS-CoV-2 driving the current COVID-19 pandemic.

TDB科学家在开发新型治疗候选者以及重新利用现有治疗方法方面具有丰富的经验。药物重新利用试图探索已经拥有详细的药理学，配方和安全数据的现有药物和治疗候选者的新用途，这可以减少为新指示进行临床测试的时间。 TDB迅速发起了几项重新利用的活动，以识别SARS-COV-2活动的调节剂。正在研究的靶标包括通过病毒尖峰蛋白与人类细胞表面受体的相互作用，包括细胞自噬等宿主防御机制，以及病毒进入细胞。 TDB科学家还为开放数据门户提供了分析数据和协议，这是由NCATS创建的资源，用于容纳由SARS-COV-2相关测定的数据集，该数据集针对NCATS Pharmaceutical Collection中的所有批准的药物产生。这项工作旨在在开放数据共享平台中分享相关数据，以允许研究科学家，临床研究人员和公共卫生官员优先考虑有希望的化合物和重新使用的药物，以进一步开发Covid-19。我们已经与NIH内外的专家合作开发了8种测定法，以重新利用认可的药物收集屏幕，以识别有效的SARS-COV-2的化合物。从此类筛选中鉴定出的化合物具有临床试验的潜力，例如单个药物或药物组合，以治疗COVID-19患者。 1。与加里·惠特克（Gary Whittaker）（康奈尔大学）合作，SARS-COV-2伪型粒子（PP）进入测定法。 PPS是用包含MLV GAG-POL，COV SPIKE和LUCIFERASE REPORTER基因带有病毒包装信号的三个质粒转染系统生成的。在测定过程中，PPS通过尖峰介导的细胞进入向宿主细胞传递荧光素酶报告基因RNA。该测定法已用于筛选SARS-COV-2进入的抑制剂，以及从其他测定中鉴定出的其他化合物的机械研究。 2. SARS伪型粒子（PP）进入分析和MERS进入分析与Gary Whittaker（Cornell University）合作。这两种测定法已针对1536孔筛选进行了优化，并用于重新用于批准药物收集的药物。用实时SARS-COV-2感染（南部研究所）中进一步评估了确定的命中率，以鉴定针对冠状病毒活性广泛的抗SARS-COV2化合物。 3。与南方研究所（SRI）合作的Vero 6细胞中的SARS-COV-2细胞病变效应测定。在SRI的BSL-3实验室中，使用NCAT制备的测定板筛选了几种大约10,000名认可药物，研究药物，临床前药物和生物活性化合物。该测定法还用于评估NCATSS BSL-2分析中鉴定的抗SARS-COV-2化合物。 4。SARS-COV-2 3Cl蛋白酶（也称为主蛋白酶）测定法。这种病毒蛋白酶在SARS-COV-2病毒复制中起关键作用。我们已经开发并优化了该酶测定法，并筛选了大约10,000种化合物的集合。当前，正在使用此重点库屏幕中的数据作为培训集进行了整个NCATS化合物集合的虚拟建模屏幕。铅化合物将用于化学优化，以进行进一步的药物开发。 5。SARS-COV-2 RNA依赖性RNA聚合酶（RDRP）测定法。该病毒RNA聚合酶对宿主细胞中的SARS-COV-2病毒复制至关重要。 Remdesivir是一种最初用于埃博拉病毒和其他病毒的RDRP抑制剂，已由FDA授权用于住院的Covid-19患者。 SARS-COV-2的功效和效力受到限制，因为它最初不是针对该病毒开发的。 Remdesivir目前仅通过静脉输注来给药，该输注排除了门诊治疗。我们正在开发SARS-COV-2 RDRP酶测定和基于细胞的RDRP测定法，以支持重新利用新药物开发的筛选。 6。用于使用BSL-2实验室的SARS-COV-2复制子分析。涉及实时SARS-COV-2病毒的研究需要BSL-3设施，这极大地限制了吞吐量。我们正在与三个实验室合作开发SARS-COV-2的复制品类型测定法，类似于我们以前在HCV上的工作。该复制子测定法具有SARS-COV-2基因组片段包含病毒复制机制，而无需病毒包络蛋白，并包括一个小型基因组报告基因（荧光素酶）。我们将使用此测定法筛选NCATS化合物集合进行药物发现和开发。 7。用于检测SARS-COV-2 Nucleocapsid（N）蛋白的α分析。当前，SARS-COV-2的细胞质效应（CPE）测定允许BSL-3实验室中最大的吞吐量，但使用替代读数来进行宿主细胞感染（细胞死亡）。 αN蛋白测定法将允许BSL-3 Labs在宿主细胞中SARS-COV-2感染后直接测量病毒抗原的产生。该测定法是同质的，并且适合HTS。它的开发是有助于增加BSL3实验室的吞吐量。 8。tr-fret（HTRF）测定法以检测SARS-COV-2核素蛋白（N）蛋白。该测定法与αN蛋白测定法相似，但使用了不同的读数。该测定法可以用作HTS的替代测定法，也可以用作正交测定法，以确认从上述alphalisa测定中识别的命中。我们还开始建立合作伙伴关系，以迅速将候选人纳入SARS-COV-2的临床试验。我们的合作者最近完成了对蛋白质天然产品的I期试验，以用作抗艾滋病毒的菌心。它的作用机理在病毒包膜上选择性结合与高臭糖寡糖，与包括HIV，流感，埃博拉病毒和SARS-COV在内的各种病原体有关。该候选者在体外和体内活性表现出针对几种冠状病毒，包括SARS-COV和MERS-COV。由于它在SARS-COV上结合了尖峰糖蛋白，因此它可能具有对所有冠状病毒菌株的活性，而不仅仅是驱动当前Covid-19的大流行的SARS-COV-2。