UPLC-Q-TOF-MS/MS Combined with Network Pharmacology and Molecular Docking Revealed the Mechanism of Zhi Ke He Ji Oral Solution for the Treatment of Pneumonia.

Abstract

Introduction: This study aimed to clarify the multi-target mechanism of Zhi Ke He Ji Oral Solution (ZKHJ) in anti-pneumonia therapy by integrating UPLC-Q-TOF-MS/MS, network pharmacology, and molecular docking techniques.

Methods: The main chemical constituents of ZKHJ were identified via UPLC-Q-TOF-MS/MS combined with multiple databases. Pneumonia-related targets were retrieved from the OMIM and GeneCards databases, and overlapping targets were screened using a Venn diagram. Cytoscape 3.7.2 was employed to construct a "component-target-pathway" network, while core targets were analyzed for GO functional enrichment and KEGG pathway enrichment. AutoDock- Tools 1.5.6 was used for molecular docking validation to explore potential anti-pneumonia mechanisms, and PyMOL 3.1 was applied to visualize and analyze the resulting complexes.

Results: A total of 114 compounds in ZKHJ were identified by UPLC-Q-TOF-MS/MS, with flavonoids and alkaloids as the main active components. Network pharmacology constructed a "ZKHJ-constituents-targets-pneumonia" network, predicting 223 potential targets; 44 core targets were further screened via topological parameters (Degree, betweenness centrality, closeness centrality) in the PPI network. GO functional enrichment yielded 1,220 significant terms, and KEGG pathway enrichment identified 214 relevant pathways. Molecular docking confirmed that active constituents of ZKHJ could stably bind to core targets (TNF, GAPDH, AKT1, STAT3, NFKB1).

Discussion: An integrated workflow of "literature mining-mass spectrometry-database" was established, constructing a traceable and expandable database of ZKHJ's chemical constituents and corresponding targets. This addresses the gap in standardized chemical profiling of complex herbal formulations and provides a one-stop information source for subsequent studies. Network pharmacology combined with molecular docking revealed that ZKHJ exerts anti-pneumonia effects through a "multi-component, multi-target, multi-pathway" mechanism, linking traditional medicinal efficacy with modern molecular pharmacology. Identifying potential quality markers lays a foundation for ZKHJ standardization, which is an essential prerequisite for its translation into evidencebased medicine. Amid the growing burden of chronic respiratory diseases and drug-resistant bacteria, ZKHJ offers a novel strategy for pneumonia management, and the established workflow provides a replicable model for chemical profiling, mechanism exploration, and standardization of other herbal formulations to facilitate the modernization of traditional herbal medicine.

Conclusion: ZKHJ exhibits potent anti-pneumonia activity, primarily via its flavonoid and alkaloid components that modulate inflammatory and immune pathways. These findings provide a solid theoretical basis for the clinical application of ZKHJ in pneumonia prevention and treatment.