Chromatography studies on bio-affinity of nine ligands of alpha_1-adrenoceptor to alpha_(1D) subtypes overexpressed in cell membrane

摘要

To improve selectivity and specificity of cell membrane Chromatography (CMC), the Chromatography affinities of nine ligands of alpha_1-adrenergic receptor(AR)to alpha_(1D)-AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of alpha_(1D)-AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and alpha_(1D)-AR in CMSP were investigated using CMC. The affinity rank order to alpha_(1D)-AR subtype obtained from CMC for the nine alpha_1-adrenoceceptor ligands is: prazosin, BMY7378, phentolamine, oxymetazoline, 5-methylurapidil, norepinephrine, phenyle-phrine, 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 alpha_(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