The Lung-toxin Dispelling Formula No.1 (referred as Harmony Shot, HSh) was established based on the theory of TCM medicinal properties and the classical prescription of TCM. It has demonstrated therapeutic benefits in both disease prevention and treatment against COVID-19. However, the material basis and mechanism of action of HSh are still unclear.

This study integrates network pharmacology, molecular docking, and LC-MS techniques to analyze and clarify the material foundation of HSh. First, the targets were obtained by network pharmacology for pathway analysis, protein-protein interaction (PPI) construction, and prediction of the core active compounds. Then, the binding ability of those active compounds with SARS-CoV-2 3CLpro (6LU7) was evaluated by molecular docking via computational pattern recognition. Finally, the compounds with high 6LU7 inhibitory activity were identified and confirmed with the LC-MS technique in both non-targeted and targeted manners using reference standards.

The global analysis was made and visualized to reveal the intricate network among the compounds, their derived ingredients, the corresponding targets and the various TCM classification and meridian tropism theory. Our results illuminate the effective material basis and possible mechanism of action of HSh prescription, which may be a promising therapeutic TCM prescription for COVID-19 prevention and treatment.