Chromatography studies on bioaffinity of nine ligands of α 1 adrenoceptor to α 1D subtypes overexpressed in cell membrane

To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of α 1 -adrenergic receptor(AR)to α 1D -AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of α 1D -AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and α 1D -AR in CMSP were investigated using CMC. The affinity rank order to α 1D -AR subtype obtained from CMC for the nine α 1 -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 α 1D -adrenoceptor cDNA and CMC method could be used to evaluate affinities of drug-receptor and drug-receptor subtypes and to screen drugs selective to α 1D -AR.