Introduction: Diseases associated with Epstein-Barr Virus (EBV) infection present substantial health challenges. While Redujing Oral Liquid (RDJ) has demonstrated efficacy against EBV in preclinical and clinical studies, its active components and mechanisms of action remain unclear. This study aimed to systematically characterize RDJ’s chemical profile and dissect its anti-EBV mechanisms using Ultra-High-Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) and network pharmacology. Method: Ultra-High Performance Liquid Chromatography coupled with Quadrupole-Orbitrap Tandem Mass Spectrometry (UHPLC-Q-Orbitrap MS/MS) was employed to identify chemical components in RDJ. EBV infection-related targets were retrieved from GeneCards and Online Mendelian Inheritance in Man (OMIM) databases to construct a “component-target-disease” network. Protein-Protein Interaction (PPI) networks were built via Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) to screen key targets, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses via Database for Annotation, Visualization, and Integrated Discovery (DAVID) elucidated functional and pathway enrichments. Molecular docking and dynamics simulations validated interactions between active components and core targets. Results: A total of 147 chemical components were identified in RDJ, with flavonoids as the most abundant class. Network pharmacology revealed that RDJ may exert anti-EBV effects through active components (e.g., 7,2-dihydroxy-3',4'-dimethoxyflavone, alpha-methyl-n-butyl shikonin, isobutyl shikonin, norwogonin, baicalein) targeting key proteins, for example, TP53, SRC, STAT3, PIK3CA, and AKT1. Molecular docking and dynamics simulations showed a strong binding affinity between baicalein and PIK3CA, a core target. Discussion: The multi-component, multi-target nature of RDJ suggests its anti-EBV efficacy arises from modulating diverse signaling pathways, with baicalein-PIK3CA interactions as a potential key mechanism. These findings align with RDJ’s clinical utility and provide mechanistic insights for EBV-related disease therapy. Conclusion: This study systematically characterized the active components of RDJ and revealed its anti-EBV mechanisms through an integrated approach combining chemical profiling and network pharmacology. The identified components and pathways, particularly baicalein-mediated regulation of PIK3CA, offer a theoretical foundation for advancing RDJ in clinical and translational research for EBV-associated diseases
Yao et al. (Wed,) studied this question.