title: Slow dissociation of long-acting Ca2+ antagonist amlodipine from 3H-PN200-110 binding sites in membranes of rat hearts and brains.
abstract: The dissociation rate of amlodipine ((+/-)-3-ethyl 5-methyl 2-[(2-aminoethoxy)methyl]-4-(o-chlorophenyl)-1,4-dihydro-6-methyl- 3,5- pyridinedicarboxylate benzenesulfonate) from rat heart and brain membranes preincubated with drugs and washed out with buffer was assessed by radioligand binding assay using 3H-PN200-110 as a radioligand. The remaining KCl-induced contraction in rat aortic strips washed out after treatment with this drug and the pKi (inhibition constant) values of the drug were compared with those of nisoldipine, nifedipine, manidipine and benidipine. The inhibition of 3H-PN200-110 binding induced by nifedipine was reversed by washing, whereas that induced by amlodipine, manidipine, and benidipine was not readily reversed under these conditions. When rat aortic strips were pretreated with Ca2+ antagonists, the rank order of the inhibition of contractions induced by 50 mM KCl was manidipine = benidipine > amlodipine > nisoldipine > nifedipine, even though Ca2+ antagonists were not present in the extracellular medium. The pKi values of amlodipine in the heart and brain were 6.86 and 7.41, respectively, and these values were lower than those of the other Ca2+ antagonists. There was a good correlation between the potency of the inhibition of 3H-PN200-110 binding by drugs after the washout of membranes and the inhibition exerted by the drugs in contractions induced by 50 mM KCl after the washout of tissues, although this residual inhibition was not correlated with pKi values. Thus, these results suggest that amlodipine has a very slow rate of dissociation from 3H-PN200-110 binding sites, as do manidipine and benidipine, and this property may explain its long-lasting antihypertensive effect.
authors.affiliation: Department of Pharmacology, Niigata College of Pharmacy, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sugiyama
authors.forename: Y L
authors.initials: YL
authors.lastname: Qu
authors.forename: K
authors.initials: K
authors.lastname: Maruyama
authors.forename: K
authors.initials: K
authors.lastname: Hattori
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: T
authors.initials: T
authors.lastname: Nagatomo
mesh_terms: D017311:Amlodipine / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D017275:Isradipine / Q000378:metabolism*; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013329:Structure-Activity Relationship
chemical_list: D002121:Calcium Channel Blockers; D017311:Amlodipine; D017275:Isradipine