Chromatography studies on bio-affinity of nine ligands of α1-adrenoceptor to α1D subtypes overexpres

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.