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

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

展开

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，由严重的急性呼吸综合征引起的冠状病毒疾病Covid-19 冠状病毒2（SARS-COV-2）已感染了全球超过2亿人在美国，有60万人死亡，全球420万人死亡。虽然疫苗已经可用，但已接种疫苗个人仍然可以将疾病传播给未接种且脆弱的人群。作为疫苗的速率美国仍然很低，可能逃避疫苗的新兴变种是一种新的风险。识别小说传统医学的预防剂可能会导致开发易于可用的，具有成本效益的，和对Covid-19的安全饮食干预。在大流行期间，个人转向草药补充剂以防止19岁。有出版在计算机研究和对这些提取物的一些体外研究中，但是支持天然产物的科学（NP）用于预防病毒感染仍然不完整。木制天然产品库（QNPL）是一系列 200,000种植物和真菌提取物，包括美国40种最常用的天然补充剂。病毒进入，在 SARS-COV-2附着于血管紧张素转换酶2（ACE2）细胞表面受体内皮细胞，肺细胞（1型和2型）以及纤毛的支气管上皮细胞，提出了一个有吸引力的选择用于预防效果。我已经从QNPL筛选了2,000提取物，以SARS-COV- 2个假病毒系统，以测试提取抑制病毒进入的病毒系统。哺乳动物细胞毒性测定并行运行测量乳酸脱氢酶（LDH）的形成，以确保抑制不是由于细胞引起的死亡。该建议采用多方面的方法来识别具有直接作用抗病毒特性的药物独一无二的天然产品库。三种具有潜在生物活性的提取物作为直接作用抗病毒剂筛选QNPL后，选择没有明显的毒性。生物测定引导分数耦合到LC- MS/MS分子网络分析将用于鉴定具有直接作用抗病毒的化合物活动。将使用制备的HPLC进一步分离化合物，并在结构上阐明NMR和X- 晶体学以确定病毒抑制剂的绝对结构。馏分和孤立化合物将在相关细胞系和活病毒中的新兴变体中进行测试，以进一步了解活动。此外，我将利用Biolayer干扰和ELISA分析进行行动研究机理。这些研究将导致NP的鉴定，并有可能阻止SARS-COV-2病毒进入相关性人类细胞，增强对植物NP对Covid-19和其他病毒的预防价值的理解。这将实现生物活性饮食补充剂的公式，医学化学的铅的优先级广告系列和识别工具化合物以查询这些途径。