Chromatography studies on bioaffinity of nine ligands of α1 adrenoceptor to α1D subtypes overexpressed in cell membrane

To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of α1-adrenergic receptor(AR)to α1D-AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of α1D-AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and α1D-AR in CMSP were investigated using CMC. The affinity rank order to α1D-AR subtype obtained from CMC for the nine α1-adrenoceceptor ligands is: prazosin, BMY7378, phentolamine, oxymetazoline, 5-methylurapidil, norepinephrine, phenylephrine, methoxamine, RS-17053. The affinity rank order is similar and correlates well with that obtained from others’ radioligand binding assays (RBA). CMSP prepared by transfected HEK293 cells with α1D-adrenoceptor cDNA and CMC method could be used to evaluate affinities of drug-receptor and drug-receptor subtypes and to screen drugs selective to α1D-AR.