Sa-Ngiamsuntorn Khanit, Suksatu Ampa, Pewkliang Yongyut, Thongsri Piyanoot, Kanjanasirirat Phongthon, Manopwisedjaroen Suwimon, Charoensutthivarakul Sitthivut, Wongtrakoongate Patompon, Pitiporn Supaporn, Chaopreecha Jarinya, Kongsomros Supasek, Jearawuttanakul Kedchin, Wannalo Warawuth, Khemawoot Phisit, Chutipongtanate Somchai, Borwornpinyo Suparerk, Thitithanyanont Arunee, Hongeng Suradej
Department of Biochemistry, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand.
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
J Nat Prod. 2021 Apr 23;84(4):1261-1270. doi: 10.1021/acs.jnatprod.0c01324. Epub 2021 Apr 12.
The coronaviruses disease 2019 (COVID-19) caused by a novel coronavirus (SARS-CoV-2) has become a major health problem, affecting more than 50 million people with over one million deaths globally. Effective antivirals are still lacking. Here, we optimized a high-content imaging platform and the plaque assay for viral output study using the legitimate model of human lung epithelial cells, Calu-3, to determine the anti-SARS-CoV-2 activity of extract and its major component, andrographolide. SARS-CoV-2 at 25TCID was able to reach the maximal infectivity of 95% in Calu-3 cells. Postinfection treatment of and andrographolide in SARS-CoV-2-infected Calu-3 cells significantly inhibited the production of infectious virions with an IC of 0.036 μg/mL and 0.034 μM, respectively, as determined by the plaque assay. The cytotoxicity profile developed over the cell line representatives of major organs, including liver (HepG2 and imHC), kidney (HK-2), intestine (Caco-2), lung (Calu-3), and brain (SH-SY5Y), showed a CC of >100 μg/mL for extract and 13.2-81.5 μM for andrographolide, respectively, corresponding to a selectivity index of over 380. In conclusion, this study provided experimental evidence in favor of and andrographolide for further development as a monotherapy or in combination with other effective drugs against SARS-CoV-2 infection.
由新型冠状病毒（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）已成为一个重大的健康问题，全球感染人数超过5000万，死亡人数超过100万。目前仍缺乏有效的抗病毒药物。在此，我们优化了一个高内涵成像平台和噬斑测定法，用于使用人肺上皮细胞Calu-3的合法模型进行病毒产量研究，以确定提取物及其主要成分穿心莲内酯对SARS-CoV-2的抗病毒活性。25TCID的SARS-CoV-2在Calu-3细胞中能够达到95%的最大感染性。噬斑测定法显示，在SARS-CoV-2感染的Calu-3细胞中，感染后用提取物和穿心莲内酯处理可显著抑制传染性病毒粒子的产生，其IC50分别为0.036μg/mL和0.034μM。在包括肝脏（HepG2和imHC）、肾脏（HK-2）、肠道（Caco-2）、肺（Calu-3）和大脑（SH-SY5Y）在内的主要器官的细胞系代表上建立的细胞毒性谱显示，提取物的CC50>100μg/mL，穿心莲内酯的CC50为13.2-81.5μM，相应的选择性指数超过380。总之，本研究提供了实验证据，支持提取物和穿心莲内酯作为单一疗法或与其他抗SARS-CoV-2感染的有效药物联合进一步开发。
