Systematic Identification Of Chemical Components In Fufang Shuanghua Oral Liquid And Screening Of Potential Active Components Against SARS-CoV-2 Protease

Objective: Coronavirus pneumonia(COVID-19)caused by novel coronavirus(SARSCoV-2)infection has become widespread in many countries,and has become a common challenge facing mankind.Traditional Chinese medicine(TCM)has played a prominent role in this pandemic,especially with its function of "clearing heat and detoxifying" playing a good antiviral role.Fufang Shuanghua Oral Liquid(FFSH)has been used to treat flu symptoms such as fever,nasal congestion,runny nose,sore throat and upper respiratory tract infection,and is typical of COVID-19.Based on the systematic analysis of FFSH chemical composition spectrum,this study aims to analyze the effects of the main active components of FFSH on the activity of SARS-CoV-2 main protease(Mpro),laying a foundation for the interpretation of the viral action mechanism of FFSH against SARS-CoV-2.Methods: Based on UPLC-Q-TOF-MS and ETCM 2.0 database,qualitative analysis of the chemical constituents of FFSH was conducted,and the mechanism of action of FFSH against SARS-CoV-2 was studied by using network pharmacological methods to provide reference.Furthermore,the influence of FRET on the anti-SARS-CoV-2 Mpro activity of FFSH core components was evaluated by in vitro fluorescence resonance energy transfer method.Results: 1.Combining chemical composition information database in ETCM 2.0database and UPLC-Q-TOF-MS technique,214 chemical components in FFSH were preliminatively identified under positive and negative ion modes,and the classification of chemical components and the identification of medicinal materials were confirmed.2.Based on TCMIP v2.0,99 core targets corresponding to the chemical composition of FFSH and SARS-CoV-2 were finally determined by PPI association analysis and network topology eigenvalue calculation,which were mainly ALB,ACTB,GAPDH,AKT1,TNF and SRC.There are 47 core components,including quercetin,luteolin and diterpene lactones.The main pathways involved include PI3K-Akt signaling pathway,MAPK signaling pathway,TNF signaling pathway,etc.3.The effect of FFSH on Mpro of SARS-CoV-2 was evaluated by FRET method in vitro.The results showed that forsythiate glycoside A,quercetin and linoleic acid had good inhibitory effects on Mpro,and the half-inhibitory concentration values were26.15 μM,22.26 μM and 47.09 μM,respectively.Conclusion: In this study,214 chemical components in FFSH were characterized for the first time,among which three chemical components,quercetin,forsythiate glycoside A and linoleic acid,were screened out to play a beneficial anti-COVID-19 effect through CYS145 and HIS41 sites,which may provide a new research strategy for the development of novel COVID-19 drugs in traditional Chinese medicine.