Exploring the Potential of Natural Products to Combat COVID-19

探索天然产品对抗 COVID-19 的潜力

基本信息

批准号：
10696937
负责人：
Caitlin Jaime Risener
金额：
$ 3.64万
依托单位：
EMORY UNIVERSITY
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：
2022-04-01 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10696937
关键词：
2019-nCoV A549 ACE2 Antiviral Agents Binding Biological Assay Biological Sciences Botanicals COVID-19 COVID-19 prevention COVID-19 treatment Cell Death Cell Line Cell Surface Receptors Cell model Cells Cessation of life Chemicals China Chromatography Ciliated Bronchial Epithelial Cell Collaborations Collection Communicable Diseases Contracts Coupled Crystallography Data Development Diet Dietary Intervention Disease Dose Edible Plants Endothelial Cells Ensure Enzyme-Linked Immunosorbent Assay Epithelial Cells Ethnobotany Exhibits Formulation Fractionation Goals Health Care Costs Health Professional Herbal supplement High Pressure Liquid Chromatography Human Immune response In Vitro Indigenous Individual Infection Inflammatory Interferometry Lactate Dehydrogenase Lentivirus Libraries Luciferases Lung Mammalian Cell Measures Medicinal Plants Medicine Modeling Molecular Natural Products Natural Supplement Pathway Analysis Pathway interactions Persons Pharmaceutical Chemistry Pharmacognosy Plants Plaque Assay Population Process Property Province Publishing Recording of previous events Reporter Research Resistance Resolution Resources Risk Running SARS-CoV-2 infection SARS-CoV-2 inhibitor SARS-CoV-2 spike protein SARS-CoV-2 variant Safety Science Serial Passage Severity of illness Standardization Structure Surface System Techniques Testing Therapeutic Toxic effect Traditional Medicine United States Vaccinated Vaccination Vaccinee Vaccines Variant Viral Viral Physiology Virus Virus Diseases Virus Inhibitors Vulnerable Populations X-Ray Crystallography clinically relevant combat coronavirus disease cost effective cytotoxicity dietary dietary supplements efficacy validation high throughput screening in silico insight meter natural product derivative novel pandemic disease pneumocyte prevent receptor receptor binding repository screening secondary metabolite symptom treatment tool unvaccinated vaccine access vaccine hesitancy viral entry inhibitor

项目摘要

ABSTRACT As of August 2021, the coronavirus disease COVID-19, caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has infected more than 200 million people across the globe, causing more than 600,000 deaths in the USA and 4.2 million worldwide. Though vaccines have become available, vaccinated individuals can still spread the disease to unvaccinated and vulnerable populations. As vaccination rates in the United States remain low, emerging variants that may evade the vaccine are a new risk. Identifying novel preventative agents from traditional medicine could lead to the development of a readily available, cost-effective, and safe dietary intervention against COVID-19. During the pandemic, individuals have turned to herbal supplements to prevent COVID-19. There are published in silico studies and a few in vitro studies on these extracts, but the science to support natural products’ (NPs) use to prevent viral infection is still incomplete. The Quave Natural Product Library (QNPL) is a collection of over 2,000 botanical and fungal extracts, including the 40 most used natural supplements in the USA. Viral entry, in which SARS-CoV-2 attaches to the Angiotensin-Converting Enzyme 2 (ACE2) cell surface receptor found on endothelial cells, pneumocytes (type 1 and 2), and ciliated bronchial epithelial cells, presents an attractive option for preventatives. I have screened 2,000 extracts from the QNPL against SARS-CoV- 2 pseudotyped virus system to test which extracts inhibit viral entry. Mammalian cell cytotoxicity assays measuring Lactate Dehydrogenase (LDH) formation were run in parallel to ensure inhibition was not due to cell death. This proposal employs a multi-faceted approach to identify agents with direct-acting antiviral properties using a one-of-a-kind natural product library. Three extracts with potent bioactivity as direct-acting antiviral agents without apparent toxicity were selected after screening the QNPL. Bioassay guided fraction coupled to LC- MS/MS molecular networking analysis will be used for the identification of compounds with direct-acting antiviral activity. Compounds will be further isolated using preparative HPLC and structurally elucidated by NMR and X- crystallography to determine the absolute structure of the viral inhibitor. The fractions and isolated compounds will be tested across emerging variants in relevant cell lines and in live virus to further understand activity. Additionally, I will undertake mechanism of action studies utilizing biolayer interferometry and ELISA assays. These studies will result in the identification of NPs with the potential to block SARS-CoV-2 viral entry in relevant human cells, enhancing understanding of the preventative value of plant NPs for COVID-19 and other viruses. This will enable formulation of a bioactive dietary supplement, prioritization of leads for a medicinal chemistry campaign, and identification of tool compounds to query these pathways.

抽象的截至 2021 年 8 月，由严重急性呼吸系统综合症引起的冠状病毒病 COVID-19 冠状病毒 2 (SARS-CoV-2) 已感染全球 2 亿多人，造成超过美国有 60 万人死亡，全球有 420 万人死亡。随着疫苗接种率的提高，个人仍然可以将疾病传播给未接种疫苗的人群和弱势群体。美国的感染率仍然较低，可能逃避疫苗的新变种是识别新风险的新风险。传统医学的预防药物可能会导致开发一种易于获得、具有成本效益、针对 COVID-19 的安全饮食干预。在大流行期间，人们开始使用草药补充剂来预防 COVID-19。对这些提取物进行了计算机模拟研究和一些体外研究，但支持天然产物（NP）的科学用于预防病毒感染的方法仍然不完整。Quave 天然产物库 (QNPL) 收集了超过 2,000 种植物和真菌提取物，包括美国最常用的 40 种天然补充剂。 SARS-CoV-2 附着在血管紧张素转换酶 2 (ACE2) 细胞表面受体上内皮细胞、肺细胞（1 型和 2 型）和纤毛支气管上皮细胞是一个有吸引力的选择我筛选了 QNPL 中的 2,000 份针对 SARS-CoV 的提取物。 2 假型病毒系统测试其提取物抑制病毒进入哺乳动物细胞的细胞毒性测定。并行进行乳酸脱氢酶 (LDH) 形成的测量，以确保抑制不是由细胞引起的死亡。该提案采用多方面的方法来识别具有直接作用的抗病毒特性的药物独一无二的天然产物库，具有作为直接作用抗病毒剂的有效生物活性的三种提取物。在筛选与 LC- 偶联的生物测定引导级分后，选择了无明显毒性的样品。 MS/MS 分子网络分析将用于鉴定具有直接作用的抗病毒药物的化合物化合物将使用制备型 HPLC 进一步分离，并通过 NMR 和 X- 进行结构阐明。晶体学以确定病毒抑制剂的绝对结构。将在相关细胞系和活病毒中对新出现的变体进行测试，以进一步了解活性。此外，我将利用生物层干涉测量法和 ELISA 测定法进行作用机制研究。这些研究将鉴定出有潜力阻止 SARS-CoV-2 病毒进入相关领域的 NP。人类细胞，增强人们对植物纳米颗粒对 COVID-19 和其他病毒的预防价值的了解。这将使生物活性膳食补充剂的配制成为可能，并优先确定药物化学的先导化合物活动，以及识别工具化合物来查询这些途径。