Frontal analysis of cell-membrane chromatography for determination of drug-alpha(1D) adrenergic receptor affinity

摘要

The aim of the present study was to determine drug-alpha(1D) adrenergic receptor (AR) affinity by frontal analysis of cell-membrane chromatography (CMC). The cell-membrane stationary phase (CMSP) was prepared by immobilizing rat aorta cell membranes on porous silica, and the resulting CMSP was used to determine drug binding affinity to alpha(1D)-AR by frontal analysis. The CMSP of rat aorta was stable and reproducible. Relative binding affinities (dissociation constant, K-d) were determined by frontal chromatography for prazosin (166.13 +/- 18.36 nmol), BMY7378 (537.40 +/- 30.84 nmol), phentolamine (646.92 +/- 23.17 nmol), 5-methylurapidil (725.66 +/- 25.48 nmol), oxymetazoline (910.56 +/- 40.62 nmol) and methoxamine (1299.27 +/- 51.73 nmol). These results were consistent with the affinity rank order and showed a good correlation with the affinity of the same compounds for the cloned alpha(1D)-AR subtype obtained from radioligand-binding assay. The study demonstrates that frontal analysis of CMC may be used for direct determination of drug-receptor binding interactions, and that CMC is an alternative reliable method to quantitatively study ligand-receptor interactions.