brl-37344 and ractopamine

Chinese hamster ovary cells expressing the porcine beta2-adrenergic receptor (betaAR) were used to determine the binding kinetics of agonists and antagonists by competitive displacement of the radioligand [125I]iodocyanopindolol. Several purported agonists, including isoproterenol, epinephrine, norepinephine, dobutamine, salbutamol, and terbutaline, exhibited dual-affinity displacement curves, which is characteristic of agonist binding to betaAR. In each case, the addition of guanosine triphosphate (GTP) eliminated the high-affinity state and resulted in a one-site displacement curve. All of the antagonists modeled to only one site in the presence or absence of GTP. Several ligands, including ones used to promote animal growth (clenbuterol, L-644,969, and ractopamine) and the beta3AR-selective agonist BRL 37344 modeled to only one site, suggesting that these ligands would not be full agonists at the porcine beta2AR (pbeta2AR). Most of the tested ligands exhibited binding affinities that were similar to published values for the beta2AR from other species. However, several exceptions were observed. The BRL 37344 ligand bound the pbeta2AR with a 10-fold higher affinity than the human beta2AR, and the Kd of this was similar to Kd values reported for the human and rat beta3AR. The Kd of the pbeta2AR for ICI 118,551 was 50-fold higher than that for the beta2AR from rats and humans. For both BRL 37344 and ICI 118,551 the subtype-selective character of these ligands was different in the pig compared with the human and rat. These data demonstrate the value of using species-specific betaAR for selection of agonists and antagonists. Further, these data support the growing evidence that few ligands are full agonists for pbetaAR and that binding data may be useful for identifying ligands with full agonist potential.

Topics: Animals; Binding, Competitive; CHO Cells; Clenbuterol; Cricetinae; Epinephrine; Ethanolamines; Guanylyl Imidodiphosphate; Humans; Iodocyanopindolol; Kinetics; Ligands; Phenethylamines; Propanolamines; Propranolol; Pyridines; Receptors, Adrenergic, beta-2; Signal Transduction; Swine

The chemical specificity for the beta-adrenoceptor mediated relaxation of rat esophageal smooth muscle was evaluated using selective and non-selective beta-adrenoceptor agonists and antagonists. Pindolol, ICI 89,406, ICI 118551 [erythro-1-(7-methylindan-4-yloxy)-3-(isopropylamine)-but an-2-ol] and the beta-adrenoceptor alkylating agent, pindobind, produced only small rightward shifts in the concentration-response curves of (-)-isoprenaline and (-)-trimetoquinol in this preparation. Rank order potency (pD2 values) of agonists was: (+/-)-trimetoquinol [1-(3',4',5'-trimethoxybenzyl)-6,7-dihydroxy-1,2,3, 4-tetrahydroisoquinoline] (8.34) = (-)-trimetoquinol (8.26) = BRL 37344 [(R* R*)-(+/-)-4-[2'-2-hydroxy 2-(3-chlorophenyl)ethylamino]propyl] phenoxyacetic acid] (8.16) = ICID7114 [(S)-4-(2-hydroxy- 3-phenoxy-propylamino-ethoxy)-N-(2-methoxyethyl) phenoxyacetamide] (8.03) > or = (-)-isoprenaline (7.82) > 3',5'-diiodotrimetoquinol [1-(3',5'-diiodo-4'-methoxybenzyl)-6, 7-dihydroxy-1,2,3,4-tetrahydroisoquinoline] (7.28) > 3'-iodotrimetoquinol [1-(3'-iodo-4',5'-dimethoxybenzyl)-6, 7-dihydroxy-1,2,3,4-tetrahydroisoquinoline] (7.04) > ractopamine (6.84) = 5,8-difluorotrimetoquinol [5,8-difluoro-6,7-dihydroxy-1- (3',4',5'-trimethoxybenzyl)-1,2,3,4-tetrahydroisoquinoline] (6.82) > 8-fluorotrimetoquinol [6,7-dihydroxy-8-fluoro-1-(3',4',5'- trimethoxybenzyl)-1,2,3,4-tetrahydroisoquinoline] (6.56) > or = (-)-noradrenaline (6.46) > or = (-)-adrenaline (6.36) > (+/-)-noradrenaline (6.24) > (+/-)-adrenaline (6.00) > clenbuterol (5.83) > (-)-1-benzyl-6, 7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (5.75). Isomeric activity ratios of trimetoquinol isomers [(-)-(S)- > > (+)-(R)-] in esophageal smooth muscle in the presence and absence of 1 microM pindolol were 1995- and 2951-fold, respectively; and were much greater than those in rat atria (282-fold) and rat trachea (107-fold). The atypical beta/beta 3-adrenoceptor partial agonist, ICI D7114, produced concentration-dependent rightward shifts of the concentration-response curves of (-)-isoprenaline, (-)-trimetoquinol and the reference atypical beta/beta 3-adrenoceptor agonist, BRL 37344. Schild plot analysis of ICI D7114 against trimetoquinol gave slope and pA2 values of 0.91 and of 7.9, respectively. These results clearly demonstrate that the relaxant effects of these agonists in rat esophageal smooth muscle are primarily mediated through the activation of atypical beta/beta 3-adrenoceptors. (-)-Trimetoquinol was as potent as (-)

Topics: Adrenergic beta-1 Receptor Antagonists; Adrenergic beta-2 Receptor Antagonists; Adrenergic beta-Antagonists; Animals; Carbachol; Clenbuterol; Cyclohexane Monoterpenes; Esophagus; Ethanolamines; Female; Heart Atria; In Vitro Techniques; Isoproterenol; Male; Muscle Relaxation; Muscle, Smooth; Phenethylamines; Phenoxyacetates; Phenoxypropanolamines; Pindolol; Propanolamines; Rats; Rats, Sprague-Dawley; Receptors, Adrenergic, beta; Serotonin Antagonists; Trachea; Tretoquinol