Abstract
The growing practice of exploiting noninvasive fluorescence-based techniques to study G protein-coupled receptor pharmacology at the single cell and single molecule level demands the availability of high-quality fluorescent ligands. To this end, this study evaluated a new series of red-emitting ligands for the human β-adrenoceptor family. Upon the basis of the orthosteric ligands propranolol, alprenolol, and pindolol, the synthesized linker-modified congeners were coupled to the commercially available fluorophore BODIPY 630/650-X. This yielded high-affinity β-adrenoceptor fluorescent ligands for both the propranolol and alprenolol derivatives; however, the pindolol-based products displayed lower affinity. A fluorescent diethylene glycol linked propranolol derivative (18a) had the highest affinity (log K(D) of -9.53 and -8.46 as an antagonist of functional β2- and β1-mediated responses, respectively). Imaging studies with this compound further confirmed that it can be employed to selectively label the human β2-adrenoceptor in single living cells, with receptor-associated binding prevented by preincubation with the nonfluorescent β2-selective antagonist 3-(isopropylamino)-1-[(7-methyl-4-indanyl)oxy]butan-2-ol (ICI 118551) ( J. Cardiovasc. Pharmacol.1983, 5, 430-437. ).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adrenergic beta-Agonists / chemical synthesis
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Adrenergic beta-Agonists / chemistry
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Adrenergic beta-Agonists / pharmacology
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Adrenergic beta-Antagonists / chemical synthesis
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Adrenergic beta-Antagonists / chemistry
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Adrenergic beta-Antagonists / pharmacology
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Alprenolol / analogs & derivatives
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Alprenolol / chemical synthesis
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Alprenolol / chemistry
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Alprenolol / pharmacology
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Animals
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Boron Compounds / chemical synthesis*
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Boron Compounds / chemistry
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Boron Compounds / pharmacology
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CHO Cells
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Cricetinae
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Cricetulus
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Drug Partial Agonism
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Fluorescent Dyes / chemical synthesis*
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Fluorescent Dyes / chemistry
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Fluorescent Dyes / pharmacology
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Genes, Reporter
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Humans
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Ligands
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Microscopy, Confocal
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Pindolol / analogs & derivatives
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Pindolol / chemical synthesis
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Pindolol / chemistry
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Pindolol / pharmacology
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Propranolol / analogs & derivatives
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Propranolol / chemical synthesis
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Propranolol / chemistry
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Propranolol / pharmacology
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Radioligand Assay
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Receptors, Adrenergic, beta-1 / metabolism*
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Receptors, Adrenergic, beta-2 / metabolism*
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Receptors, Adrenergic, beta-3 / metabolism*
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Single-Cell Analysis
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Stereoisomerism
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Structure-Activity Relationship
Substances
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Adrenergic beta-Agonists
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Adrenergic beta-Antagonists
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Boron Compounds
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Fluorescent Dyes
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Ligands
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N-(3-hydroxy-1-(naphthalen-1-yl)-1,8,11-trioxa-5-azatridecan-13-yl)-6-(2-(2-(4,4-difluoro-4,4a-dihydro-5-(thiophen-2-yl)-4-bora-3a,4a-diaza-s-indacene-3-yl)vinyl)phenoxyacetamido)hexanamide
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Receptors, Adrenergic, beta-1
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Receptors, Adrenergic, beta-2
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Receptors, Adrenergic, beta-3
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Alprenolol
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Propranolol
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Pindolol