Prediction of quality markers of Angong Niuhuang Pills based on LC-MS and pull-down with SPR chips
Elucidating the quality markers(Q-markers)of traditional Chinese medicines is essential for understanding the mechanisms of action and promoting the rational use of traditional Chinese medicines as well as for developing traditional Chinese medicine-derived drugs.Studies have shown that surface plasmon resonance(SPR)is promising in this field.This study proposed a method based on pull-down with SPR chips to predict the Q-markers of Angong Niuhuang pills(AGNHP).Firstly,71 main chemical components of AGNHP were analyzed by UPLC-Q-TOF-MS,and then network pharmacology was employed to predict the potential targets of AGNHP against stroke.Secondly,the STAT3 protein chip was constructed,and the extract of AGNHP was recovered by pull-down of the SPR system for STAT3 ligand.The potential active ingredients were collected,enriched,and identified as coptisine,palmatine,epiberberine,berberine,worenine,demethyleneberberine,jatrorrhizine,tetrahydrocoptisine,baicalein,and baicalin methyl ester.Next,the affinity constants of the 10 active ingredients were determined as 44.7,44,58.1,51.3,39.7,32.1,49.2,69.1,19.7,and 24.9 μmol·L-1,respectively.The molecular docking results showed that the 10 compounds could compete for binding with STAT3.This is the first report that SPR combined with UPLC-Q-TOF-MS is reliable and feasible for determining the active ingredients of AGNHP at the molecular level from complex systems.STAT3 could be used as a potential target for the biological quality evaluation of AGNHP.
万方数据
维普
