Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; School of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230031, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
J Pharm Biomed Anal. 2023 Jan 20;223:115118. doi: 10.1016/j.jpba.2022.115118. Epub 2022 Oct 21.
Coronavirus disease (COVID-19) caused by SARS-COV-2 infection has been widely prevalent in many countries and has become a common challenge facing mankind. Traditional Chinese medicine (TCM) has played a prominent role in this pandemic, and especially TCM with the function of "heat-clearing and detoxifying" has shown an excellent role in anti-virus. Fufang Shuanghua oral liquid (FFSH) has been used to treat the corresponding symptoms of influenza such as fever, nasal congestion, runny nose, sore throat, and upper respiratory tract infections in clinic, which are typical symptoms of COVID-19. The content of chlorogenic acid, andrographolide and dehydrated andrographolide as the quality control components of FFSH is not less than 1.0 mg/mL, 60 μg/mL and 60 μg/mL respectively. In this study, UPLC-Q-TOF-MS/MS was employed to describe the chemical profile of FFSH. Virtual screening and fluorescence resonance energy transfer (FRET) were used to screen the effective components of FFSH acting on SARS-CoV-2 main protease (Mpro). As a result, 214 compounds in FFSH were identified or preliminarily characterized by UPLC-Q-TOF-MS/MS, and 61 active ingredients with potential inhibitory effects on Mpro were selected through receptor-based and ligand-based virtual screening. In particular, quercetin, forsythoside A, and linoleic acid showed a good inhibitory effect on Mpro in FRET evaluation with IC50 values of 26.15 μM, 22.26 μM and 47.09 μM respectively, and had a strong binding affinity with the receptor Mpro (6LU7) in molecular docking. CYS145 and HIS41 were the main amino acid residues affected by small molecules in the protein binding domain. In brief, we characterized, for the first time, 214 chemical components in FFSH, and three of them, including quercetin, forsythoside A and linoleic acid, were screened out to exert beneficial anti-COVID-19 effects through CYS145 and HIS41 sites, which may provide a new research strategy for TCM to develop new therapeutic drugs against COVID-19.
由 SARS-CoV-2 感染引起的冠状病毒病（COVID-19）在许多国家广泛流行，已成为人类共同面临的挑战。传统中药（TCM）在这场大流行中发挥了突出作用，特别是具有“清热解毒”作用的中药在抗病毒方面表现出了优异的作用。复方双花口服液（FFSH）已在临床上用于治疗流感引起的发热、鼻塞、流涕、咽痛和上呼吸道感染等症状，这些都是 COVID-19 的典型症状。FFSH 的质量控制成分绿原酸、穿心莲内酯和脱水穿心莲内酯的含量分别不少于 1.0mg/mL、60μg/mL 和 60μg/mL。在本研究中，采用 UPLC-Q-TOF-MS/MS 描述 FFSH 的化学成分谱。虚拟筛选和荧光共振能量转移（FRET）用于筛选 FFSH 作用于 SARS-CoV-2 主蛋白酶（Mpro）的有效成分。结果，通过 UPLC-Q-TOF-MS/MS 鉴定或初步鉴定了 FFSH 中的 214 种化合物，并通过基于受体和基于配体的虚拟筛选选择了 61 种具有潜在抑制 Mpro 作用的活性成分。特别是槲皮素、连翘酯苷 A 和亚油酸在 FRET 评价中对 Mpro 表现出良好的抑制作用，IC50 值分别为 26.15μM、22.26μM 和 47.09μM，并且与分子对接中受体 Mpro（6LU7）具有很强的结合亲和力。CYS145 和 HIS41 是蛋白质结合域中小分子影响的主要氨基酸残基。总之，我们首次对 FFSH 中的 214 种化学成分进行了特征描述，筛选出其中的槲皮素、连翘酯苷 A 和亚油酸三种成分，通过 CYS145 和 HIS41 位点发挥有益的抗 COVID-19 作用，为 TCM 开发治疗 COVID-19 的新药提供了新的研究策略。
