Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

摘要

Background As the main metabolites of ginsenosides, 20( S , R )-protopanaxadiol [PPD( S , R )] and 20( S , R )-protopanaxatriol [PPT( S , R )] are the structural basis response to a series of pharmacological effects of their parent components. Although the estrogenicity of several ginsenosides has been confirmed, however, the underlying mechanisms of their estrogenic effects are still largely unclear. In this work, PPD( S , R ) and PPT( S , R ) were assessed for their ability to bind and activate human estrogen receptor α (hERα) by a combination of in?vitro and in silico analysis. Methods The recombinant hERα ligand-binding domain (hERα-LBD) was expressed in E.?coli strain. The direct binding interactions of ginsenosides with hERα-LBD and their ERα agonistic potency were investigated by fluorescence polarization and reporter gene assays, respectively. Then, molecular dynamics simulations were carried out to simulate the binding modes between ginsenosides and hERα-LBD to reveal the structural basis for their agonist activities toward receptor. Results Fluorescence polarization assay revealed that PPD( S , R ) and PPT( S , R ) could bind to hERα-LBD with moderate affinities. In the dual luciferase reporter assay using transiently transfected MCF-7 cells, PPD( S , R ) and PPT( S , R ) acted as agonists of hERα. Molecular docking results showed that these ginsenosides adopted an agonist conformation in the flexible hydrophobic ligand-binding pocket. The stereostructure of C-20 hydroxyl group and the presence of C-6 hydroxyl group exerted significant influence on the hydrogen bond network and steric hindrance, respectively. Conclusion This work may provide insight into the chemical and pharmacological screening of novel therapeutic agents from ginsenosides.