title: Characterization of a bromoacetylated derivative of pindolol as a high affinity, irreversible beta adrenergic antagonist in cultured cells.
abstract: We have utilized cultured cell lines to test the utility of N8-(bromoacetyl)-N1-[3-(4-indolyoxy)-2-hydroxypropyl]-(Z)-1,8-diam ino- p-menthane, BIM, a recently synthesized, irreversible beta adrenergic antagonist. Previously available irreversible antagonists of beta adrenergic receptors have generally exhibited low affinity (typical IC50 values greater than or equal to 1 microM). By contrast, S49 lymphoma cells incubated with 10 nM BIM for 120 min and then washed extensively showed a 70% loss in beta adrenergic receptors, as measured by [125I]iodocyanopindolol binding. This loss, which could be prevented by propranolol, represented a decrease in receptor number without a change in affinity of the remaining receptors for [125I]iodocyanopindolol. The BIM-induced decrease in binding sites was persistent in membranes incubated for several hours after BIM treatment. BIM did not inactivate alpha-1 adrenergic receptors on Madin Darby canine kidney cells, alpha-2 adrenergic receptors on human erythroleukemia cells, nor did BIM treatment alter guanyl-5'-yl-imidodiphosphate-mediated regulation of agonist binding to the beta adrenergic receptors in S49 cell membranes. BIM treatment decreased cyclic AMP (cAMP) accumulation in S49 cells in response to the beta adrenergic agonist isoproterenol, but increased prostaglandin E1-stimulated cAMP accumulation (P = .09) without altering cAMP production in response to forskolin. The inactivation of beta receptors in S49 cells by BIM (IC50 = 0.30 nM) correlated closely with the loss in beta adrenergic receptor-mediated cAMP accumulation in these cells (IC50 = 0.59 nM), implying the absence of substantial receptor reserve for this response. We conclude that BIM is a potent, irreversible, selective beta adrenergic antagonist for the study of beta adrenergic receptors in cultured cells.
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla.
authors.forename: J R
authors.initials: JR
authors.lastname: Jasper
authors.forename: H J
authors.initials: HJ
authors.lastname: Motulsky
authors.forename: P A
authors.initials: PA
authors.lastname: Insel
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000494:pharmacology*; D000526:Alprenolol / Q000031:analogs & derivatives / Q000494:pharmacology; D000527:Alprostadil / Q000494:pharmacology; D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D000081005:Cyclohexane Monoterpenes; D004305:Dose-Response Relationship, Drug; D020120:Iodocyanopindolol; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D010869:Pindolol / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D011943:Receptors, Adrenergic, beta / Q000032:analysis / Q000187:drug effects / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D000319:Adrenergic beta-Antagonists; D000081005:Cyclohexane Monoterpenes; D008565:Membrane Proteins; D011943:Receptors, Adrenergic, beta; C057316:N(8)-bromoacetyl-N(1)-3'-(4-indolyloxy)-2'-hydroxypropyl-1,8-diamino-4-menthane; C045331:bromoacetylalprenololmenthane; D020120:Iodocyanopindolol; D000526:Alprenolol; D010869:Pindolol; D000242:Cyclic AMP; D000527:Alprostadil