Integrative Network Pharmacology Prediction of the Mechanisms of Tri-Ka-Tuk: A Traditional Thai Herbal Formula.

Introduction: Tri-Ka-Tuk, a traditional Thai herbal formula composed of ginger, black pepper, and long pepper, has long been utilized for its therapeutic properties, particularly in promoting digestive health, enhancing metabolism, and reducing inflammation. However, the underlying molecular mechanisms remain poorly understood. This study employs an integrative network pharmacology approach to reveal the bioactive compounds and pathways mediating the formula's therapeutic effects.

Methods: Bioactive compounds of Tri-Ka-Tuk were identified through literature review and database searches. Targets were predicted using SwissTargetPrediction, and gene ontology (GO) enrichment analysis was conducted via ShinyGO. Pathway enrichment analysis utilized KEGG databases to map associated signaling pathways. Protein-protein interaction (PPI) networks were constructed using the STITCH database.

Results: GO analysis revealed that Tri-Ka-Tuk's targets are enriched in biological processes such as protein autophosphorylation and phosphorylation-dependent signaling. Cellular component enrichment indicated involvement in membrane-associated regions critical for signaling. Molecular function analysis highlighted kinase activity and ATP binding as central mechanisms. Pathway enrichment analysis identified significant pathways, including the AGE-RAGE signaling pathway in diabetic complications, the HIF-1 pathway, and cancer-related pathways. PPI network analysis showed that hub proteins such as VEGFA, CDK1, and MAPK1 play key roles in mediating the formula's effects.

Conclusion: The findings suggest that Tri-Ka-Tuk exerts its therapeutic effects by modulating key molecular pathways involved in inflammation, oxidative stress, apoptosis, and metabolism. The integrative network pharmacology approach followed in this research bridges traditional knowledge with modern pharmacological insights, and highlights Tri-Ka-Tuk's potential as a therapeutic agent for managing diabetes, cancer, and metabolic disorders. Experimental validation of these predictions is essential to confirm the efficacy of the formulation and to expand its clinical applications.