title: [3H]L-654,284 as a probe of the central alpha 2 adrenoceptor.
abstract: L-654,284 [2R, 12bS)-N-(1,3,4,6,7,12b-hexahydro-2H-benzo[b]-furo[2,3-a] quinolizin-2-yl)-N-methyl-2-hydroxyethanesulfonamide], a potent and selective antagonist of the alpha 2 adrenoceptor, was tritiated to high specific activity. Saturation binding to cell membrane suspensions obtained from calf cerebral cortex revealed a high affinity binding site (0.63 nM). Kinetics of association and dissociation were well represented by single exponential processes, and the equilibrium dissociation constant obtained from the ratio of rate constants agreed well with that found by saturation binding. A direct comparison of saturation binding revealed that the antagonist [3H]L-654,284 had roughly the same affinity for the alpha 2 adrenoceptor as the agonist [3H]clonidine and eight times the affinity of the antagonist [3H]rauwolscine. The maximum receptor densities of these radioligands were not significantly different. Competition assays with a series of compounds of known receptor affinity revealed that [3H]L-654,284 selectively binds to a site with all of the characteristics expected of the alpha 2 adrenoceptor.
authors.affiliation: Department of Medicinal Chemistry, Merck Sharp and Dohme Research Laboratories, West Point, PA 19486.
authors.forename: W C
authors.initials: WC
authors.lastname: Randall
authors.forename: J R
authors.initials: JR
authors.lastname: Huff
authors.forename: J P
authors.initials: JP
authors.lastname: Vacca
authors.forename: J P
authors.initials: JP
authors.lastname: Guare
authors.forename: R
authors.initials: R
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Rosegay
authors.forename: J J
authors.initials: JJ
authors.lastname: Baldwin
mesh_terms: D000317:Adrenergic alpha-Antagonists; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002417:Cattle; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D003000:Clonidine / Q000378:metabolism; D066298:In Vitro Techniques; D011807:Quinolizines; D011869:Radioligand Assay; D011942:Receptors, Adrenergic, alpha / Q000187:drug effects*; D015016:Yohimbine / Q000378:metabolism
chemical_list: D000317:Adrenergic alpha-Antagonists; D011807:Quinolizines; D011942:Receptors, Adrenergic, alpha; D015016:Yohimbine; C047289:N-(1,3,4,6,7,12b-hexahydro-2H-benzo(b)furo(2,3-a)quinolizin-2-yl)-N-methyl-2-hydroxyethanesulfonamide; D003000:Clonidine