Network Pharmacology And Molecular Docking-based Approaches To Investigate The Mechanism Of LianHuaQingWen In Treating COVID-19

BackgroundThe COVID-19 has spread across the globe since its outbreak at the end of 2019 due to its high susceptibility and transmissibility.Patients often accompanied by fever,dry cough,fatigue and other symptoms,and even cause lung inflammation and infiltration,as well as systemic inflammatory cytokine storm.Research on drugs in treating COVID-19 has attracted international attention.Under the situation where there are no antiviral treatments for COVID-19,TCM plays an important role in controlling the spread of inflammation,promoting the absorption of inflammation,preventing development of disease and shortening the treatment cycle.As a commonly used proprietary Chinese medicine for respiratory diseases,LianhuaQingwen（LHQW）was created based on the experience of ancient Chinese doctors in treating acute colds,influenza fever and other respiratory exogenous fevers,and combines with the latest modern pharmacological research.As a result,this Chinese patent medicine has been successfully listed by the National Health Commission of the People’s Republic of China as a recommended drug for the treatment of COVID-19 during the clinical medical observation period.However,the specific mechanism of LHQW in treating COVID-19 is still unclear.Chinese medicine prescriptions have the characteristics of multi-ingredients,multi-targets and overall regulation.Network pharmacology is an effective method to study the mechanism of prescriptions.Molecular docking technology can screen potentially effective Chinese medicine prescriptions or active ingredients by scoring the combination of active ingredients and targets.Molecular docking technology can quickly screen the potential targets,its pharmacological activity and mechanism,and provide a powerful technical means for the modern research of Traditional Chinese medicine.Therefore,this study intends to explore the mechanism of LHQW in treating COVID-19 at molecular and protein levels based on network pharmacology and molecular docking approaches.PurposesIn this study,the molecular mechanism of LHQW in treating COVID-19 were studied through network pharmacology and molecular docking approaches.By screening the potential active ingredients and therapeutic targets of LHQW in treating COVID-19,this study reveals that LHQW inhibiting inflammatory stress storm and regulating the host cell receptor ACE2 of SARS-CoV-2.At the same time,a screening method for other novel co-receptors of the new coronavirus was constructed.Contents and results（1）LHQW regulates the cytokine storm mediated by COVID-19 through multiple targetsIn this study,the pathogenic targets of COVID-19 were screened by using the GeneCards database.The results showed that the core pathogenic target of COVID-19 was TNF,and the secondary core targets were IL6,GAPDH,TP53,IL10,ALB,IL2,and MAPK3.TCMSP and UniProt database were used to screen the main active ingredients and therapeutic targets of LHQW.The results showed that the core therapeutic target of LHQW was AKT1,and the secondary core targets of LHQW were ALB,MAPK3,IL6,TP53,VEGFA,JUN,and EGF.In order to further clarify the common targets,this study mapped the pathogenic targets of COVID-19 and the therapeutic targets of LHQW,and found a total of 45 common targets,among which the core target is IL6,and the secondary core targets were TNF,MAPK8,CXCL8,TP53,MAPK3,IL10,and CASP3.In addition,ACE2 as an important receptor for COVID-19 invades host cell,a PPI network was constructed between ACE2 and the 45 common targets of COVID-19 and LHQW.The results showed that there were 8 targets related to ACE2:ALB,NOS3,IL6,CRP,CAT,TNF,CCL2,and IL1B.Most of these targets are inflammatory factors,which suggests that LHQW may play a role in the prevention and treatment of COVID-19 by regulating the cytokine storm mediated by these inflammatory factors.Functional annotation of 45 common targets was performed by using R language,GO enrichment analysis showed that the response to lipopolysaccharide was the most significant biological process,with a total of 24 targets involved in this process.It contains the core target IL6 and 5 ACE2-related targets（NOS3,IL6,TNF,CCL2,and IL1B）,suggesting that LHQW may inhibit the lipopolysaccharide-mediated inflammatory response and relieve lung damage.KEGG pathway analysis showed that the most significant KEGG pathway was AGE-RAGE signaling pathway,with a total of 19 targets involved in this pathway.It contains the core target IL6 and 5 ACE2related targets（NOS3,IL6,TNF,CCL2,and IL1B）,suggesting that LHQW may relieve tissue damage by regulating the expression of these five targets in the signaling pathway.（2）LHQW regulates core target,secondary core targets and ACE2-related targets through multiple ingredientsIn this study,CDOCKER docking（Discovery Studio）was performed for proteinsmall molecule docking.The results showed that 13 active ingredients（including βcarotene,isovitexin,hyperforin,et al）have good binding activity with core target（IL6）,secondary core targets（TNF,MAPK8,CXCL8,TP53,MAPK3,IL10,CASP3）;ACE2;ACE2-related targets（ALB,NOS3,IL6,CRP,CAT,TNF,CCL2,IL1B）.Therefore,it is suggested that LHQW may play an improtant role in regulating the inflammatory response through multiple active ingredients.（3）LHQW regulates the binding of ACE2 and Spike through multiple ingredientsIn this study,CDOCKER docking（Discovery Studio）was performed for proteinsmall molecule docking,and the results showed that 6 ingredients（β-carotene,wogonin,luteolin,acacia,quercetin and kaempferol）have good binding activity with ACE2Spike（PDB ID:7KJ2）.Moreover,these six active ingredients may affect the binding of ACE2 and Spike through the key amino acids between ACE2 and Spike.（4）Potential co-receptor of ACE2 during the invasion of host cells by SARS-CoVIn this study,based on the biological characteristics of the core target IL6,proteinprotein docking between IL6R/IL6/IL6ST and Spike protein as well as the complex of ACE2-Spike at different stages was performed by HADDOCK rigid docking approach,which could screen the potential novel candidate receptor based on protein-protein affinity.The results showed that IL6ST had a good affinity with Spike protein as well as the complex of ACE2-Spike at different stages.In addition,IL6ST could bind the NTD region of Spike protein even when Spike already bound with ACE2,so the spatial competition between IL6ST and ACE2 was poor.IL6ST may has the possibility to become ACE2 co-receptor.ConclusionThis study revealed that LHQW may regulate the expression of core target,secondary core targets,and ACE2-related targets through active ingredients such as βcarotene,isovitexin,and hyperforin,and has the ability of directly targeting ACE2Spike protein.This study preliminarily confirmed that LHQW could reduce the inflammatory response mediated by COVID-19 at the molecular level.At the same time,the protein-protein docking in this study suggests that IL6ST has the possibility of becoming an ACE2 co-receptor during the process of COVID-19 invading host cells.