Article date: October 1995
By: P. Chatelain, L. Meysmans, J.R. Mattéazzi, Ph. Beaufort, M. Clinet, in Volume 116, Issue 3, pages 1949-1956
The effects of SR 33589 and amiodarone on the cardiac β‐adrenoceptor were studied in vitro and after chronic treatment by means of [125I]‐(−)−iodocyanopindolol ([125I]‐(−)−CYP) binding and measurement of adenylate cyclase activity.
Binding of [125I]‐(−)−CYP was inhibited in a dose‐dependent manner by SR 33589 (IC50 = 1.8 ± 0.4 μM, nH = 0.93 ± 0.06) and amiodarone (IC50 = 8.7 ± 2.0 μM, nH = 0.92 ± 0.03). Saturation binding experiments indicated a non‐competitive interaction such that SR 33589 (1 and 3 μm) and amiodarone (5 and 10 μm) reduced the Bmax of [125I]‐(−)−CYP binding without any effect on the KD. Kinetic studies showed that the rate of association of [125I]‐(−)−CYP was unchanged while the rate of dissociation was increased both in the presence of SR 33589 (10 μm) and amiodarone (30 μM).
Under the same conditions, the receptor stimulated adenylate cyclase activity was inhibited in a dose‐dependent, but non‐competitive manner, by SR 33589 (isoprenaline‐, glucagon‐ and secretin‐stimulated enzyme inhibited 50% at 6.8 ± 0.6 μM, 31 ± 10 μm and 12 ± 3 μM, respectively) while the basal, GTP‐ and Gpp(NH)p‐stimulated enzyme was inhibited by 5–10% and the NaF and forskolin‐stimulated enzyme by 50% at 500 μM. Amiodarone exhibited a similar pattern of inhibition.
After chronic oral treatment (50, 100, 150 mg kg−1 per day, 14 days), both SR 33589 and amiodarone produced a dose‐dependent decrease in Bmax without any effect on KD as determined from [125I]‐(−)−CYP saturation experiments and a decrease of the isoprenaline‐ and glucagon‐stimulated adenylate cyclase activity without any effect on basal enzyme activity or activity when stimulated by agents acting directly on regulatory catalytic units.
Unlike amiodarone, SR 33589 does not contain iodine substituents. Plasma levels of T3, T4 and rT3 were unchanged after SR 33589 treatment except a decrease in T4 level at the highest dose whilst the T4 T3 ratio and the level of rT3 were dose‐dependently increased by amiodarone treatment.
In vitro, SR 33589 and amiodarone were characterized as non‐competitive β‐adrenoceptor antagonists. Chronic treatment led to a down‐regulation of the β‐adrenoceptor; the down‐regulation cannot be attributed to an indirect effect mediated by the thyroid hormones. To reconcile these opposing observations, we propose that SR 33589 and amiodarone interact with the β‐adrenoceptor at a site close to the intracellular loops which are involved in the coupling with Gs and contain the phosphorylable sites.
The effects of SR 33589 and amiodarone on the cardiac β‐adrenoceptor were studied in vitro and after chronic treatment by means of [125I]‐(−)−iodocyanopindolol ([125I]‐(−)−CYP) binding and measurement of adenylate cyclase activity.
Binding of [125I]‐(−)−CYP was inhibited in a dose‐dependent manner by SR 33589 (IC50 = 1.8 ± 0.4 μM, nH = 0.93 ± 0.06) and amiodarone (IC50 = 8.7 ± 2.0 μM, nH = 0.92 ± 0.03). Saturation binding experiments indicated a non‐competitive interaction such that SR 33589 (1 and 3 μm) and amiodarone (5 and 10 μm) reduced the Bmax of [125I]‐(−)−CYP binding without any effect on the KD. Kinetic studies showed that the rate of association of [125I]‐(−)−CYP was unchanged while the rate of dissociation was increased both in the presence of SR 33589 (10 μm) and amiodarone (30 μM).
Under the same conditions, the receptor stimulated adenylate cyclase activity was inhibited in a dose‐dependent, but non‐competitive manner, by SR 33589 (isoprenaline‐, glucagon‐ and secretin‐stimulated enzyme inhibited 50% at 6.8 ± 0.6 μM, 31 ± 10 μm and 12 ± 3 μM, respectively) while the basal, GTP‐ and Gpp(NH)p‐stimulated enzyme was inhibited by 5–10% and the NaF and forskolin‐stimulated enzyme by 50% at 500 μM. Amiodarone exhibited a similar pattern of inhibition.
After chronic oral treatment (50, 100, 150 mg kg−1 per day, 14 days), both SR 33589 and amiodarone produced a dose‐dependent decrease in Bmax without any effect on KD as determined from [125I]‐(−)−CYP saturation experiments and a decrease of the isoprenaline‐ and glucagon‐stimulated adenylate cyclase activity without any effect on basal enzyme activity or activity when stimulated by agents acting directly on regulatory catalytic units.
Unlike amiodarone, SR 33589 does not contain iodine substituents. Plasma levels of T3, T4 and rT3 were unchanged after SR 33589 treatment except a decrease in T4 level at the highest dose whilst the T4 T3 ratio and the level of rT3 were dose‐dependently increased by amiodarone treatment.
In vitro, SR 33589 and amiodarone were characterized as non‐competitive β‐adrenoceptor antagonists. Chronic treatment led to a down‐regulation of the β‐adrenoceptor; the down‐regulation cannot be attributed to an indirect effect mediated by the thyroid hormones. To reconcile these opposing observations, we propose that SR 33589 and amiodarone interact with the β‐adrenoceptor at a site close to the intracellular loops which are involved in the coupling with Gs and contain the phosphorylable sites.
DOI: 10.1111/j.1476-5381.1995.tb16397.x
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