A Gut Microbial Transformation-Integrated Network Pharmacology Approach to Elucidate the Therapeutic Mechanisms of Timosaponin AIII in Diabetes

Objective: Timosaponin AIII, with poorly soluble characteristics, has a potential antidiabetic effect evaluated in vitro and in vivo. The major problem associated with poorly soluble drugs is very low bioavaila-bility. This study aimed to investigate the metabolic profiles and antidiabetic mechanism of Timosaponin AIII. Materials and methods: The metabolic profiles of Timosaponin AIII in intestinal flora were analyzed using LC-MS/MS. Based on mass spectrometry analysis, network pharmacology combined with the GEO database was used to identify potential targets and elucidate the antidiabetic mechanism. Finally, the stability of com-pound-target complexes was further functionally confirmed by molecular docking. Results: As a result, 13 metabolites were identified. After the compound-target network, the genes of its me-tabolites increased by 60 compared to those of Timosaponin AIII. Subsequently, 13 core targets related to antidiabetic efficacy were identified through PPI network analysis. Key genes EGFR, MAPK1, and ICAM1 with strong binding efficiencies with metabolites were identified as crucial targets for the therapeutic effects of Timosaponin AIII. The KEGG analysis indicated that timosaponin AIII combated diabetes through various signaling pathways, including PI3K-Akt, FoxO, and HIF-1 signaling pathways, etc. Conclusions: Taken together, this study clarified the mechanism of Timosaponin AIII against diabetes by identifying additional targets and pathways, and the importance of glycosidic structures. Otherwise, we might provide a solid foundation for the development of clinical applications of Timosaponin AIII. conclusion: It was found that metabolites of timosaponin AIII had strong binding efficiencies with these target genes based on molecular docking results. Taken together, this study not only clarifies the mechanism of timosaponin AIII against diabetes by identifying additional targets and pathways, but also provide an important foundation to explain the pharmacodynamic changes of timosaponin AIII in intestinal flora.