Huang Sheng-Teng, Chen Yeh, Chang Wei-Chao, Chen Hsiao-Fan, Lai Hsiang-Chun, Lin Yu-Chun, Wang Wei-Jan, Wang Yu-Chuan, Yang Chia-Shin, Wang Shao-Chun, Hung Mien-Chie
School of Chinese Medicine, China Medical University, Taichung 40402, Taiwan.
Department of Chinese Medicine, China Medical University Hospital, Taichung 40402, Taiwan.
Viruses. 2021 May 2;13(5):826. doi: 10.3390/v13050826.
In late 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic emerged to severely impact the global population, creating an unprecedented need for effective treatments. This study aims to investigate the potential of D. Don (SB) as a treatment for SARS-CoV-2 infection through the inhibition of the proteases playing important functions in the infection by SARS-CoV-2. FRET assay was applied to investigate the inhibitory effects of SB on the two proteases involved in SARS-CoV-2 infection, M and TMPRSS2. Additionally, to measure the potential effectiveness of SB treatment on infection inhibition, cellular models based on the Calu3 and VeroE6 cells and their TMPRSS2- expressing derivatives were assessed by viral pseudoparticles (Vpp) infection assays. The experimental approaches were conjugated with LC/MS analyses of the aqueous extracts of SB to identify the major constituent compounds, followed by a literature review to determine the potential active components of the inhibitory effects on protease activities. Our results showed that SB extracts inhibited the enzyme activities of M and TMPRSS2. Furthermore, SB extracts effectively inhibited SARS-CoV-2 Vpp infection through a TMPRSS2-dependent mechanism. The aqueous extract analysis identified six major constituent compounds present in SB. Some of them have been known associated with inhibitory activities of TMPRSS2 or M. Thus, SB may effectively prevent SARS-CoV-2 infection and replication through inhibiting M and TMPRSS2 protease activities.
2019年末，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）大流行出现，严重影响全球人口，产生了对有效治疗方法前所未有的需求。本研究旨在通过抑制在SARS-CoV-2感染中起重要作用的蛋白酶，研究D. Don（SB）作为SARS-CoV-2感染治疗方法的潜力。采用荧光共振能量转移（FRET）测定法研究SB对SARS-CoV-2感染所涉及的两种蛋白酶M和跨膜丝氨酸蛋白酶2（TMPRSS2）的抑制作用。此外，为了测量SB治疗对感染抑制的潜在有效性，通过病毒假颗粒（Vpp）感染试验评估了基于Calu3和VeroE6细胞及其表达TMPRSS2的衍生物的细胞模型。实验方法与SB水提取物的液相色谱/质谱（LC/MS）分析相结合，以鉴定主要成分化合物，随后进行文献综述，以确定对蛋白酶活性具有抑制作用的潜在活性成分。我们的结果表明，SB提取物抑制了M和TMPRSS2的酶活性。此外，SB提取物通过TMPRSS2依赖性机制有效抑制SARS-CoV-2 Vpp感染。水提取物分析鉴定出SB中存在的六种主要成分化合物。其中一些已知与TMPRSS2或M的抑制活性有关。因此，SB可能通过抑制M和TMPRSS2蛋白酶活性有效预防SARS-CoV-2感染和复制。
