Exploring the potential mechanism of Jingjie Fangfeng medicine in the treatment of coronavirus pneumonia based on network pharmacology and molecular docking
In December 2019, novel coronavirus pneumonia caused by novel coronavirus (SARS CoV-2) broke out in Wuhan. The main symptoms of patients are fever, fatigue, and dry cough. Severe patients often experience difficulty breathing and/or hypoxemia one week after onset, and in severe cases, they rapidly progress to acute respiratory distress syndrome, septic shock, difficult to correct metabolic acidosis, and coagulation disorders. As of 24:00 on May 26, 2020, China has reported a total of 82993 confirmed cases, with 79 confirmed cases (including 5 severe cases), 78280 cured and discharged cases, and 4634 deaths. At the same time, the incidence of overseas infections continues to rise, with a total of over 5.5 million confirmed cases. This is the third large-scale outbreak of coronavirus in just 20 years of the 21st century, following severe acute respiratory syndrome (SARS) in 2003 and Middle East respiratory syndrome (MERS) in 2015. The coronavirus poses a serious threat to human health every time it spreads on a large scale, bringing severe challenges and heavy burdens to social and economic development. However, there is currently no specific drug for treating pneumonia caused by coronavirus in clinical practice, and supportive treatment is mainly used. Therefore, research on related drugs is urgently needed.

Since the outbreak of the novel coronavirus epidemic, according to the relevant diagnosis and treatment plans and recommendations of provinces, cities and autonomous regions, Chinese medicine has been widely used in the treatment of novel coronavirus, of which many provinces and cities recommend the use of Schizonepeta tenuifolia and Fangfeng in traditional Chinese medicine. Schizonepetae Herba (SH) is the dry aboveground part of Nepeta cataria L., a plant in the family Lamiaceae. It has a pungent taste, a slightly warm nature, and has the effects of relieving external symptoms, dispersing wind, penetrating rashes, and eliminating sores. Saposhnikoviae Radix (SR) is a plant species in the Umbelliferae family, Saposhnikovia divaricata (Trucz.) Schischk Dry roots, with a pungent and sweet taste, and a slightly warm nature. It has the effects of dispelling wind, relieving pain, dispelling dampness, and stopping spasms. Jingjie and Fangfeng are the royal medicines of Fangfeng Tongsheng Wan recorded in the Xuanming Lun Fang. The two are recorded in the Shi Jin Mo Yao Dui as a pair of medicines for dispersing wind and cold, dispelling wind and suppressing dampness. Both Jingjie and Fangfeng have antipyretic, analgesic, sedative, antibacterial, anti-inflammatory, hemostatic and other effects.

Since its proposal in 2008, network pharmacology has become an emerging discipline that systematically reveals the regulatory network effects of traditional Chinese medicine on the body. Due to its holistic and systematic research methods and emphasis on drug interactions, which are consistent with the multi-target and multi pathway characteristics of traditional Chinese medicine, it has become an effective tool for systematically analyzing the multi-target and multi pathway mechanisms of traditional Chinese medicine formulas. Multiple studies have successfully explored the mechanism of action of traditional Chinese medicine using network pharmacology. In summary, this article intends to explore the potential mechanism of the treatment of coronavirus pneumonia by using network pharmacology and molecular docking methods, and provide reference for the clinical application of Jingjie Fangfeng. Firstly, active ingredients and their targets in Schizonepeta tenuifolia were collected by searching TCMSP, ETCM, and BATMAN-TCM databases, and coronavirus pneumonia related targets were collected in GeneCards, OMIM, and NCBI Gene databases. Then, the intersection of the two was analyzed using the STRING database to investigate protein interactions between key targets, and the DAVID database was used for biological function and pathway analysis. Finally, use Autodock software for molecular docking of potential pharmacological substances and key targets.

Jingjie and Fangfeng are common traditional Chinese medicines for dispelling wind, relieving surface heat, clearing heat, and reducing fever. They have pharmacological effects such as anti-inflammatory, antibacterial, antiviral, and antipyretic. Since the outbreak of the novel coronavirus epidemic, Schizonepeta tenuifolia and Saposhnikovia divaricata have been widely used in the treatment of coronavirus pneumonia. They have relatively satisfactory clinical efficacy and improved the clinical cure rate. However, their mechanism of action is not clear, and there is a lack of molecular level experimental research. Therefore, in-depth research and analysis of the mechanism of action of Jingjie Fangfeng in treating coronavirus pneumonia, exploring its potential targets, provides direction for the subsequent molecular mechanism research of Jingjie Fangfeng, and is of great significance for clinical application. This article is based on network pharmacology and molecular docking technology to study the active ingredients, potential targets, and pathways of action of Jingjie Fangfeng in the treatment of coronavirus pneumonia. It was found that Jingjie Fangfeng medicine may act on targets such as IL6, TNF, MAPK1, IL1 β, TP53 through active products such as luteolin, quercetin, wogonin, beta sitosterol, etc., regulating biological processes such as inflammation response, positive regulation of RNA polymerase II promoter transcription, immune response, and negative regulation of apoptosis. It exerts multi-component, multi-target, and multi-channel pharmacological effects against coronavirus pneumonia through TNF signaling pathway, PI3K Akt signaling pathway, cytokine cytokine receptor interaction, MAPK signaling pathway, HIF-1 signaling pathway, Toll like receptor signaling pathway, T cell receptor signaling pathway, etc.

The PPI network parameters show that the degree values of 23 proteins, including IL6, TNF, MAPK1, TP53, IL1 β, RELA, and IL10, are higher than the average, indicating that they are closely related to other proteins and have strong interactions in the PPI network. Among them, inflammatory factor related proteins such as IL6, IL1 β, TNF, and IL10 have high degree values and are located at the center of the network. These target proteins are involved in the immune response and inflammatory process of respiratory diseases, and are closely related to the stimulation of the respiratory tract by the inflammatory response. A recent autopsy study on the remains of patients with coronavirus pneumonia showed significant lung damage and clear inflammatory lesions, indicating that inflammation plays an important role in the development of the disease. Jingjie Fangfeng may exert anti-inflammatory effects through these targets. Kindrachuk et al. found that inhibitors targeting the MAPK and PI3K Akt signaling pathways inhibit coronavirus replication both before and after viral infection. KEGG enrichment results also showed statistical differences in the enrichment of MAPK and PI3K Akt signaling pathways. The HIF-1 signaling pathway also plays an important role in maintaining the homeostasis of alveolar epithelial cells and the pathogenesis of diseases, consistent with the enrichment results of the KEGG pathway. This indicates that the active substances in Jingjie Fangfeng medicine may exert pharmacological effects against coronavirus pneumonia through the MAPK signaling pathway, PI3K Akt signaling pathway, and HIF-1 signaling pathway.

Through network pharmacology screening, we found that luteolin, quercetin, wogonin, and beta sitosterol compounds have moderate values higher than the average in the “component target pathway” network. Therefore, we docked these four components with the top five molecules in the PPI network, and the results showed that the docking energies of these four components with IL6, TNF, MAPK1, and IL1 β were all less than -5 kcal/mol, and the docking energy of beta sitosterol with TP53 was less than -0.5 kcal/mol. This indicates that the active chemical components of Schizonepeta tenuifolia can stably bind to IL6, IL1 β, TNF, and TP53 proteins. Meanwhile, studies have shown that luteolin, quercetin, wogonin, and beta sitosterol all exhibit good anti-inflammatory activity, indicating that these components play an important role in the anti-inflammatory effect of Jingjie Fangfeng medicine. Due to the fact that COVID-19 is a new disease and the disease targets are not fully included, we validated the docking of chemical components with degree values greater than the average with SARS virus ACE2 and COVID-19 main protease, and found that all four chemicals achieved good docking results with ACE2 and COVID-19 main protease. This indicates that the active substance of Jingjie Windproof may bind to these two proteins to exert anti coronavirus pneumonia effects.

In summary, this study used network pharmacology analysis methods to investigate the active substances and targets of Jingjie Fangfeng in the treatment of coronavirus pneumonia, and validated the screened targets through molecular docking technology. At the same time, the active substances were docked with SARS virus receptor ACE2 and COVID-19 main protease, and the potential anti coronavirus potential of Jingjie Fangfeng was discovered. This provides support for the clinical application and development of Jingjie Fangfeng, and lays the foundation for the subsequent screening of active substances and target research of Jingjie Fangfeng by the research group.