Screening of the active compound from Tetradium ruticarpum fruits and analysis of its binding characteristics to the α1A adrenoceptor by high expression α1A adrenoceptor cell membrane chromatography

The dried fruit of the Tetradium ruticarpum (Wu Zhu Yu) tree is commonly used in traditional Chinese medicine, and its decoction can be used for treating conditions such as headaches and hypotension. In the present study, an offline two-dimensional combination method of alpha(1A)/HEK293 cell membrane chromatography (alpha(1A)/CMC) and UHPLC-MS/MS was established to screen and identify the active compound from the Tetradium ruticarpum fruits. The binding characteristics between this active compound and the alpha(1A) receptor were also analyzed by an alpha(1A)/CMC method. By this process, dehydroevodiamine was identified as the potential active compound. Equilibrium dissociation constant (K-d) values between alpha(1A) receptor and dehydroevodiamine, obtained by both stepwise frontal analysis and zonal elution analysis, were (5.18 +/- 0.50) x 10(-6) mol/L and (2.70 +/- 0.74) x 10(-6) mol/L, respectively. Our results indicate that the alpha(1A)/CMC method can not only screen active compounds from complex sample, but can also be used to calculate the binding parameters of the identified compound. (C) 2020 Published by Elsevier B.V.

Automated summary

The study established an α(1A)/CMC method for screening active compounds from Tetradium ruticarpum fruits and analyzing the binding characteristics with the receptor. Dehydroevodiamine was identified as a potential active compound through this method.