Comparison of zonal elution and nonlinear chromatography in determination of the interaction between seven drugs and immobilised β2-adrenoceptor

Rapid analysis of interaction between six drugs and β2‐adrenergic receptor by injection amount‐dependent method

“…The affinity rank order of the six drugs to β 2 ‐AR was bambuterol > clenbuterol > tulobuterol > clorprenaline > salbutamol > terbutaline. This order agreed well with the reported affinity rank by frontal analysis (Zhao et al, ), zonal elution and nonlinear chromatography (Li et al, ), even using different batches of β 2 ‐AR column (Table ). Injection amount‐dependent method is believed to be feasible for quantitatively exploring the interaction between ligands and immobilized proteins.…”

“…It is remarkable that high‐performance affinity chromatography (HPAC) (Anguizola et al, ; Moser, White, & Kovacs, ; Schiel, Ohnmacht, & Hage, ) possessing new mathematical tools is relatively rapid for obtaining information on drug–protein interactions. Among the reported mathematical models, frontal analysis (Kamarei, Gritti, Guiochon, & Burchell, ; Ràfols, Zarza, & Bosch, ), zonal elution (Matsuda, Li, Zheng, & Hage, ) and nonlinear chromatography (Li et al, ) are mostly utilized to determine the binding parameters of drug–protein interactions.…”

Confirming therapeutic target of protopine using immobilized β2‐adrenoceptor coupled with site‐directed molecular docking and the target‐drug interaction by frontal analysis and injection amount–dependent method

“…The association constant and the number of binding sites were (1.70 ± 0.44) × 10 4 L/mol and (3.97 ± 0.02) × 10 −7 mol/L. This result was in good line with that determined by zonal elution in literature . Both the association constant of zonal elution and frontal analysis are 10 times lower than that of injection amount–dependent method.…”

“…The hydrogen bonds formed between the hydroxyl side chain of Ser 204 and the metahydroxyl group of the ligand and between the hydroxyl side chain of Ser 207 and the parahydroxyl group of the ligand; the third ligand‐binding site was located at the 2 phenylalanines (Phe), 259 and 290, on the sixth domain . On the basis of these reports and β 2 ‐AR structural difference between this work and the literature, the carboxylate side chain of Asp 113 in the receptor is the high‐affinity site for protopine binding to β 2 ‐AR at low‐drug concentrations presumably. When the high‐affinity binding site is saturated and the drug concentration continuously increased, the weaker interaction at Ser 204 or Ser 207 in β 2 ‐AR becomes an extra force for the drugs binding to β 2 ‐AR.…”

“…This result was in good line with that determined by zonal elution in literature. 46 Both the association constant of zonal elution and frontal analysis are 10 times lower than that of injection amount-dependent method. In particular, the value determined by injection amount-dependent method is more approximated to the data collected by radioligand-binding assay in the reference.…”