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

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 (K_D) (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 K_D 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 K_D 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.