Exploring the Potential of Natural Products to Combat COVID-19

探索天然产品对抗 COVID-19 的潜力

• 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
• 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

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.

摘要 截至2021年8月，由严重急性呼吸系统综合征引起的冠状病毒病COVID-19 冠状病毒2（SARS-CoV-2）已感染地球仪的2亿多人， 美国有60万人死亡，全球有420万人死亡。虽然疫苗已经上市，但接种疫苗的 个人仍然可以将疾病传播给未接种疫苗的弱势群体。由于疫苗接种率在 美国仍然很低，可能逃避疫苗的新变种是一个新的风险。鉴定新 传统医学的预防剂可以导致开发一种容易获得的，具有成本效益的， 和安全的饮食干预来预防COVID-19。 在疫情期间，个人已转向草药补充剂以预防COVID-19。已发表 对这些提取物的计算机研究和一些体外研究，但支持天然产品（NP）的科学 用于预防病毒感染的方法仍然不完善。Quave天然产物库（QNPL）是一个超过1000种天然产物的集合。 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。病毒进入， SARS-CoV-2附着在血管紧张素转换酶2（ACE 2）细胞表面受体上， 内皮细胞，肺细胞（1型和2型），纤毛支气管上皮细胞，提出了一个有吸引力的选择 为了预防。我从QNPL中筛选了2,000种提取物， 2假型病毒系统，以测试哪些提取物抑制病毒进入。哺乳动物细胞毒性试验 平行进行测定乳酸脱氢酶（LDH）形成以确保抑制不是由于细胞 死亡 该建议采用多方面的方法，使用 独一无二的天然产物库三种具有直接抗病毒活性的提取物 经QNPL筛选后，选择无明显毒性的动物。生物测定引导部分偶联至LC- MS/MS分子网络分析将用于鉴定具有直接作用的抗病毒化合物 活动化合物将使用制备型HPLC进一步分离，并通过NMR和X-Ray进行结构解析。 晶体学以确定病毒抑制剂的绝对结构。馏分和分离的化合物 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。 这些研究将导致鉴定具有阻断SARS-CoV-2病毒进入相关细胞的潜力的NP。 人类细胞，增强对植物纳米颗粒对COVID-19和其他病毒的预防价值的理解。 这将使生物活性膳食补充剂的配方，药物化学的优先顺序 活动，并确定工具化合物来查询这些途径。