β2-adrenergic receptor affinity chromatography with an interaction force analysis model: A method for analysis of active compounds targeting β2-adrenergic receptor

Abstract

Asthma is one of the most prevalent diseases worldwide, and β2-adrenergic receptor (β2AR) agonists have been reported to be highly effective bronchodilators against this disease. In this study, we successfully constructed a novel CHO-β2AR affinity chromatography (CHO-β2AR/AC), which was evaluated by infrared spectroscopic and scanning electron microscope (SEM) analysis. In addition, CHO-β2AR/AC model exhibited good selectivity and reliability with the relative standard deviation smaller than 5.6% after 30 days. Furthermore, an interaction force analysis model was developed based on CHO-β2AR/AC. The results showed that the interaction force analysis model (Φ•E•pKa) exhibited a strong correlation with equilibrium dissociation constant (KD) (r2=0.9284, p=0.002) and a good correlation with logarithm of half-maximum effective concentration (pEC50) values (r2=0.7135, p=0.034). In addition, a pool of clinically approved drugs was screened by this CHO-β2AR/AC model. Codeine wasfound to bind to and activate β2AR with KD value of 4.10 × 10−7 M, leading to increased cyclic adenosine monophosphate (cAMP) production with EC50 of 6.49 × 10−7 M and reduction of intracellular Ca2+ concentration, which in turn relaxes bronchial contraction with EC50 of 2.62 × 10−6 M. Furthermore, the KD value and pEC50 of codeine were within the 95% prediction range of the interaction force analysis model. The results indicate that the CHO-β2AR/AC with interaction force analysis model constructed in this study can be used to effectively and rapidly screen active compounds targeting β2AR.