Network Pharmacology and Metabolomics to Uncover the Multi-Target Anti-Non-Small Cell Lung Cancer Mechanism of Baicalin Mediated by Purine Metabolism.

[OBJECTIVE] Baicalin (BA), the primary active component of Scutellaria baicalensis, exhibits anti-tumor potential; however, its multi-target mechanism in the treatment of non-small cell lung cancer (NSCLC) remains poorly understood.

[METHODS] This study systematically elucidated the anti-NSCLC mechanism of BA through an integrated approach that combined network pharmacology, molecular docking, molecular dynamics simulations, in vivo animal models, and untargeted metabolomics using LC-MS. Potential targets were predicted using SwissTargetPrediction and multiple disease databases. A protein-protein interaction (PPI) network was constructed and analyzed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Core targets were validated via molecular docking and simulations. The effects of BA on tumor growth and on the expression of EGFR and TNF-α were assessed in an A549 tumor-bearing nude mouse model. Serum metabolite changes were profiled and linked to associated pathways.

[RESULTS] Sixty overlapping targets were identified, with EGFR, TNF-α, CASP3, PTGS2, EZH2, and IL-2 serving as core nodes. Molecular docking demonstrated strong binding affinity between BA and both EGFR (-9.830 kcal/mol) and PTGS2 (-7.244 kcal/mol). The in vivo xenograft model demonstrated that BA (2.5 mg/kg) significantly inhibited NSCLC tumor growth, with efficacy comparable to paclitaxel. Immunohistochemistry confirmed BA downregulated EGFR and TNF-α expression in tumors. Metabolomics analysis revealed 17 differentially expressed metabolites and four significantly altered metabolic pathways: purine, caffeine, sphingolipid, and pyrimidine metabolism. Purine metabolism exhibited the most pronounced perturbation.

[DISCUSSION] The integrated analysis reveals that BA exerts its anti-NSCLC effects through a multi-target mechanism involving direct interactions with key signaling proteins, such as EGFR and PTGS2, downregulation of oncogenic and inflammatory pathways, and systemic reprogramming of cancer-associated metabolism, with purine metabolism as a central target.

[CONCLUSION] BA exerts its anti-NSCLC effects via multi-target regulation of oncogenic signaling and metabolic reprogramming. This offers preliminary insights that could inform future applications in metabolic-targeted therapies and combination treatments.