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Irreversible blockade of β-adrenergic receptors with a bromoacetyl derivative of pindolol

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Summary

A potent irreversible β-adrenergic derivative of pindolol possessing a chemically reactive group (Br-AAM-pindolol) was synthesized. This compound devoid of agonist properties, competed for all (3H)-dihydroalprenolol (3H-DHA) binding sites in C6 glioma cell and rat cerebellum membranes. Pretreatment of C6 glioma cell membranes with Br-AAM-pindolol and subsequent washing resulted in a time- and dose-dependent blockade of β-adrenergic receptors. A 50% blockade was achieved in the presence of 1.6 nM Br-AAM-pindolol.

This blockade occurs specifically at the β-adrenergic receptor level, as: 1) it induced a decrease of maximal isoproterenol stimulated adenylate cyclase activity with no modification of basal and sodium fluoride stimulated activity and 2) decreases of (3H)-DHA binding and stimulation of adenylate cyclase activity by the agonist were suppressed in the presence of isoproterenol, a β-adrenergic agonist. Furthermore, Br-AAM-pindolol treatment did not affect (3H)-diazepam binding in C6 glioma cell membranes.

Pretreatment of C6 glioma cells with Br-AAM-pindolol also reduced the response of adenylate cyclase to isoproterenol and the number of β-adrenergic receptors. The blockade of β-adrenergic receptors of C6 glioma cells by Br-AAM-pindolol was non-competitive, whereas the blockade obtained with AM-pindolol, a derivative of pindolol devoid of alkylating properties, was competitive.

The irreversible blockade of β-adrenergic receptors by Br-AAM-pindolol in rat erythrocyte membranes was substantiated by the demonstration that no recovery of β-adrenergic receptors occurred during long term incubation of the membranes (48 h) following Br-AAM-pindolol treatment and subsequent washing.

Double reciprocal plotting of equiactive isoproterenol concentrations in dose-response curves of adenylate cyclase from membranes of control and Br-AAM-pindolol treated C6 glioma cells permitted calculation of the dissociation constant for isoproterenol from its binding sites (1.5±0.2×10−7 M, n=11). This is very close to the dissociation constant of the agonist derived from binding experiments (1.7±0.5×10−7 M, n=13).

These results suggest that Br-AAM-pindolol is a potent irreversible β-adrenergic antagonist and may be useful for pharmacological and physiological studies of σ-adrenergic receptors.

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Authors and Affiliations

  1. Centre CNRS-INSERM de Pharmacologie-Endocrinologie, Rue de la Cardonille, F-34033, Montpellier, Cedex

    V. Homburger, R. Bouhelal & J. Bockaert

  2. Chare de Neuropharmacologie-INSERM U. 114, Collège de France, 11, Place M. Berthelot, F-75231, Paris, Cedex 05

    H. Gozlan

  3. Chaire de Biochimie Cellulaire-Collège de France, 11, Place M. Berthelot, F-75231, Paris, Cedex 05, France

    M. Lucas

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  1. V. Homburger
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  2. H. Gozlan
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  3. R. Bouhelal
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  4. M. Lucas
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  5. J. Bockaert
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Homburger, V., Gozlan, H., Bouhelal, R. et al. Irreversible blockade of β-adrenergic receptors with a bromoacetyl derivative of pindolol. Naunyn-Schmiedeberg's Arch. Pharmacol. 328, 279–287 (1985). https://doi.org/10.1007/BF00515554

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  • DOI: https://doi.org/10.1007/BF00515554

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