A biophysical assay targeting SARS CoV-2 RNA

针对 SARS CoV-2 RNA 的生物物理检测

基本信息

批准号：
10381446
负责人：
sandra Paige story
金额：
$ 29.99万
依托单位：
NUBAD, LLC
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-27 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10381446
关键词：
2019-nCoV 5&apos Untranslated Regions Address Affect Affinity Amino Sugars Antisense Oligonucleotides Antiviral Agents Awareness Bacterial Genes Binding Biological Assay Biophysics COVID-19 COVID-19 pandemic COVID-19 treatment Caring Cell Culture Techniques Cessation of life Chemicals Chemistry Communication Communities Complex Congestive Coughing Country Development Disease Drug Design Elements Employment Event Exposure to Firefly Luciferases Fluorescence Fluorescent Probes Future Genes Genetic Transcription Genome Government Grant Handwashing Health Health Status Healthcare Systems High Pressure Liquid Chromatography Human In Vitro Individual Infection Infection prevention Interruption Laboratories Lead Legal patent Libraries Life Ligand Binding Ligands Masks Medical Mental Health Messenger RNA Modeling National Institute of Allergy and Infectious Disease Nucleic Acids Persons Phase Population Proteins Proteome Public Health Quarantine RNA RNA Binding RNA Sequences RNA Viruses RNA library RNA replication Rapid screening Reporter Genes Resources Respiratory distress Reverse Transcription Ribonucleic Acid Regulatory Sequences Risk Role SARS coronavirus SARS-CoV-2 infection Schools Scientist Services Severe Acute Respiratory Syndrome Small Business Innovation Research Grant Small Business Technology Transfer Research Social Distance Solid Spanish flu Structure Technology Tennessee Therapeutic Transactivation Transcript Translations Universities Vaccination Vaccines Validation Viral Viral Proteins Virus Virus Replication Work analog assay development biophysical analysis combat coronavirus disease design drug development experimental study genomic RNA high throughput screening improved innovation interest microbial mutant nanomolar new technology novel pandemic coronavirus pandemic disease pandemic influenza phase 2 study phosphorodiamidate morpholino oligomer physical conditioning prevent respiratory screening social stem success symptom management symptom treatment technology development vaccine development viral RNA

项目摘要

PROJECT SUMMARY The world is immersed in a health crisis rivaled only by the Spanish flu pandemic of 1918-1919. The difference between the crisis of today and the crisis a century ago is that we have advanced communication technology greatly so that huge populations of people around the globe are aware of risks and can take appropriate precautions, such as the employment of quarantining, isolation, social distancing, and masks and handwashing. Governments have closed schools and outlawed large social gatherings. Medical care has greatly improved, and those most affected have received symptomatic treatment in the absence of a cure for CoVID-19. The problem must be dealt at multiple fronts, such as vaccine development, drug development and new technologies, assays for mitigating viral effects. The proposed project is significant because it proposes a novel in vitro biophysical screening assay for a unique and yet untapped RNA structure in SARS-CoV-2 virus, that can be used in the future to generate RNA specific antiviral compounds. Nucleic acids are promising avenues for drug design, both as therapeutics and as targets. Here we propose an innovative screening approach for identification of a novel class of ligands that are specific for an RNA element within the viral RNA genome that is vital for replication of the virus, and we propose a biophysical screening assay as a first step for identifying such ligands. First, as outlined in Specific Aim 1, we will characterize a model nucleic acid (RNA) domain that will be synthesized, characterized and used to identify a RNA specific fluorescent probe. The optimized probe will then be used for developing a high-throughput screening assay for discovery of nanomolar binders to this RNA. The RNA specific high-affinity binders will then be combined with sequence-specific RNA binding ligands to validate the assay development and its utility. The mechanism of action will be confirmed using inhibition of firefly luciferase translation in a reporter gene assay (Specific Aim 2). While the focus of this application, as the needed first step, is on the development and validation of the biophysical assay for the underlying SARS-CoV-2 RNA structures, a successful validation of the approach will open the doors for discovery and development of lead compounds for inhibition of SARS-CoV in Phase II studies. NUBAD and its team of scientists and collaborators is uniquely qualified to perform these assays and develop the potential leads in Phase II, in addition to providing a template for the scientific community to use the screening technology for their own discovery platforms. Success of the approach will also allow us to expand the screening technology to other RNA structures in SARS and other RNA viruses, and provide the screening resource as a service to the scientific community.

项目摘要世界沉浸在一次健康危机中，仅与1918 - 1919年西班牙流感大流行有关。区别在一个世纪前的今天的危机和危机之间，我们拥有先进的通信技术很大程度上，全球众多人群都知道风险，并且可以采取适当的态度预防措施，例如使用隔离，孤立，社会疏远以及面具和洗手的预防措施。政府关闭了学校，并取缔了大型社交聚会。医疗已大大改善，在没有治愈的情况下，受影响最大的人接受了症状治疗。这问题必须在多个方面解决，例如疫苗开发，药物开发和新技术，减轻病毒作用的测定法。拟议的项目很重要，因为它提出了小说体外生物物理筛选测定法，用于SARS-COV-2中独特但未开发的RNA结构病毒，将来可以用于产生RNA特异性抗病毒化合物。核酸是药物设计的有希望的途径，无论是治疗剂还是作为靶标。在这里我们提出一个创新的筛选方法，用于鉴定针对RNA的新型配体类别病毒RNA基因组中的元素对于病毒的复制至关重要，我们提出生物物理筛选分析是识别此类配体的第一步。首先，如特定目的所述 1，我们将表征一个模型核酸（RNA）结构域，该结构域将合成，表征和用于确定RNA特异性荧光探针。然后，优化的探针将用于开发高通量筛选用于发现该RNA的纳摩尔粘合剂的测定。然后，RNA特异性高亲和力粘合剂将与序列特异性RNA结合配体结合，以验证测定开发及其效用。这通过抑制记者基因测定中的萤火虫荧光素酶翻译，将确认作用机理（特定目标2）。尽管该应用程序的重点是所需的第一步，但是开发的对基础SARS-COV-2 RNA结构的生物物理测定的验证，成功验证了该方法将打开发现和开发铅化合物以抑制SARS-COV的大门在第二阶段研究中。 Nubad及其科学家和合作者团队具有独特的资格来执行这些除了为科学界提供模板外，测定和开发了第二阶段的潜在潜在客户将筛选技术用于自己的发现平台。方法的成功也将使我们能够将筛选技术扩展到SARS和其他RNA病毒中的其他RNA结构，并提供筛选资源作为科学界的服务。