Potential mechanism of 25R-inokosterone in the treatment of osteoporosis: Based on bioinformatics and molecular dynamics model.

25R-inokosterone, a sterone derived from Achyranthes bidentata Bl., has potential anti-osteoporotic effects. However, the underlying mechanism of 25R-inokosterone in the treatment of osteoporosis is unknown. The aim of this study was to investigate the molecular mechanism of 25R-inokosterone's anti-osteoporosis by network pharmacology and molecular docking. First, the structural formula of 25R-inokosterone was obtained by PubChem and potential targets were predicted by SwissTargetPredictive. The genes of osteoporosis (OP) were obtained by GeneCards, OMIM, therapeutic target database and database of gene-disease associations. Protein interactions and functional enrichment of potential targets were analyzed using STRING, gene ontology, and Kyoto encyclopedia of genes and genomes pathway databases. Finally, the hub targets were identified by network pharmacology, and the interaction of their hub targets with 25R-inokosterone was verified by molecular docking and molecular dynamics simulations. The results showed that 43 potential targets were associated with the mechanism of 25R-inokosterone for OP treatment. Enrichment analysis showed that hub genes were mainly associated with prolactin signaling pathway, aldosterone-regulated sodium reabsorption, phosphatidylinositol-3 kinase-Akt signaling pathway, and mTOR signaling pathway. Molecular docking showed that 25R-inokosterone was associated with PIK3CA, MTOR, TNF, MAPK3, CDK2, and NTRK1 with good affinity. Among them, 25R-inokosterone has the highest affinity with PIK3CA/MTOR, which is supported by molecular dynamics simulations. Our findings suggested that 25R-inokosterone may against OP by regulating multiple targets such as PIK3CA, MTOR, TNF, MAPK3, CDK2, and NTRK1 and phosphatidylinositol-3 kinase/AKT/mTOR signaling pathway. These findings provide a theoretical basis for the treatment of OP.