CaR‐Derived Column Was Prepared to Discover Multiple Active Constituents Simultaneously From Epimedii Folium

The discovery of the bioactive constituents from herbal medicines remains a huge challenge in front of us. Here, a rapid approach was established by immobilizing full-length calcium sensing receptor (CaR) on the solid supports to screen the bioactive constituents from Epimedii Folium. The results indicated that the immobilized CaR can recognize specifically agonist Ca²⁺ as well as allosteric modulators cinacalcet and NPS2143, and the four bioactive compounds including epimedin A, epimedin B, epimedin C, and icariin were simultaneously screened from over 100 constituents in one injection of the Epimedii Folium extract. The chromatographic competitive binding experiment in combination with molecular docking gave the evidence that epimedin B and epimedin C were bound to the CaR's transmembrane domain whereas epimedin A and icariin interacted with the CaR's extracellular domain. These findings contributed to understanding the action mechanism of the four bioactive compounds with the CaR at the molecular level. Our work broke through the conventional way to isolate the interested molecules first and then testify their pharmacological and pharmacodynamic effects, establishing a rapid approach to discover simultaneously multiple active constituents from an herbal medicine.