In vitro activity of RO363, a beta1-adrenoceptor selective agonist.

1 The beta-adrenoceptor stimulant effects of RO363 and (--)-isoprenaline have been compared in a variety of isolated tissue preparations. 2 RO363 is approximately half as potent as (--)-isoprenaline in tissues where actions are due to beta1-receptor activation (guinea-pig atrial and ileal preparations and ventricular strips from the rabbit, rat and guinea-pig. 3 In uterine and lung strip preparations from the guinea-pig, where responses are due to beta2-receptor stimulation. RO363 is 100 to 350 times less active than (--)-isoprenaline and has a low intrinsic activity. 4 In spontaneously contracted tracheal preparations from the guinea-pig, RO363 is a full agonist and is approximately half as potent as (--)-isoprenaline. These effects of RO363 are due to the activation of a population of beta1-receptors in the tissue since RO363 and (--)-isoprenaline have the same relative potencies in trachea, cardiac and ileal preparations. In addition the Kb values for practolol are similar in all these preparations when RO363 is used as the agonist. 5 The results show that RO363 is a potent and highly selective beta1-receptor agonist.     

Human cardiac beta-adrenoceptors: both beta 1- and beta 2-adrenoceptors are functionally coupled to the adenylate cyclase in right atrium

We investigated properties of catecholamine-sensitive adenylate cyclase in membranes from human right atria. Basal adenylate cyclase was activated by Mg2+-ions and guanyl nucleotides [Gpp(NH)p, GTP] in a concentration-dependent manner; guanyl nucleotide activation was strongly dependent on the presence of Mg2+-ions. Catecholamines stimulated adenylate cyclase activity in the following order of potency: (-)-isoprenaline greater than (-)-adrenaline = (-)-noradrenaline greater than phenylephrine, indicating that, in human right atrium, beta 1-adrenoceptors predominate. The beta 1-agonist dobutamine and the beta 2-agonists fenoterol and procaterol activated adenylate cyclase with an intrinsic activity of 0.5-0.7 (isoprenaline = 1.0). Adenylate cyclase activation by dobutamine or procaterol was not additive with the activation induced by isoprenaline. On the contrary, combination of dobutamine (100 microM) and procaterol (10 microM) resulted in activation of adenylate cyclase which was not different from that evoked by saturating concentration of isoprenaline (10 microM), indicating that dobutamine (beta 1) and procaterol (beta 2) produce adenylate cyclase activation through stimulation of different beta-adrenoceptor subtypes. The beta 2-selective antagonist ICI 118,551 was much more potent in inhibiting procaterol-than isoprenaline-stimulated adenylate cyclase activity, whereas the beta 1-selective antagonist betaxolol inhibited isoprenaline-stimulated activity more potently. We conclude that in human right atrium, both beta 1- and beta 2-adrenoceptors are functionally coupled to the adenylate cyclase system.     

A comparative study of beta-adrenoceptors in human and porcine lung parenchyma strip

1. Responses to (+/-)-isoprenaline (Iso), (-)-adrenaline (Adr) and (-)-noradrenaline (NA) were compared in isolated preparations of human and porcine lung parenchyma strip. 2. The order of relaxant potencies of these catecholamines in both human and porcine lung parenchyma was Iso greater than Adr greater than NA (1:0.24:0.01, human; 1:0.21:0.01.pig). These results suggest that beta 2-adrenoceptors predominate in both types of lung parenchyma strip. 3. pA2 values for the beta-adrenoceptor antagonist, propranolol (non-selective), with Iso as the agonist, in human and porcine lung strips were 7.84 and 7.83 respectively and for atenolol were 6.50 and 5.35 respectively. Taken as a whole results indicate the existence of an apparently homogeneous population of beta 2-adrenoceptors in porcine parenchyma strip, while both beta 1 and beta 2-adrenoceptors were revealed in human lung parenchyma.     

Demonstration of both beta 1- and beta 2-adrenoceptors mediating relaxation of isolated ring preparations of rat pulmonary artery.

1 Cumulative concentration-response (relaxation) curves to three beta-adrenoceptor agonists, fenoterol (beta 2-selective), isoprenaline (non-selective) and noradrenaline (beta 1-selective) were obtained on isolated ring preparations of rat pulmonary artery contracted with 15 mM KCl. alpha-Adrenoceptors and neuronal and extraneuronal uptakes were blocked with phenoxybenzamine. The agonist concentration-response curves were reproducible. 2 Responses to each of the three agonists could be blocked by the beta-adrenoceptor antagonists atenolol (beta 1-selective) or ICI 118,551 (beta 2-selective) confirming the presence of beta-adrenoceptors. 3 The relative potencies of the agonists were isoprenaline : fenoterol : noradrenaline = 100 : 38 : 1.4. This indicated that the predominant beta-adrenoceptor type was beta 2. 4 Schild plots were obtained for atenolol and ICI 118,551 using the three different agonists. For each antagonist the location of the Schild plot varied depending on which agonist was used. This indicated that the beta-adrenoceptor population mediating relaxation responses to beta-adrenoceptor agonists was not homogeneous. 5 Atenolol was most potent when noradrenaline was the agonist and ICI 118,551 was most potent when fenoterol was the agonist. 6 It is concluded that isolated pulmonary artery ring preparations of the rat contain a mixed population of beta 1- and beta 2-adrenoceptors both mediating relaxation.     

An homogeneous population of beta 1-adrenoceptors subserves inhibitory responses in guinea-pig ileal preparations

In transmurally stimulated guinea-pig ileal preparations, the pA2 values obtained with the selective beta 1- and beta 2-adrenoceptor antagonists atenolol and ICI 118,551 were independent of the beta-adrenoceptor agonist used [-)-isoprenaline or fenoterol) and were similar to values reported in isolated cardiac tissue. On the basis of these results, the relaxant responses to beta-adrenoceptor agonists in guinea-pig ileal preparations appear to be mediated by an homogeneous population of beta 1-adrenoceptors.     

The use of the guinea-pig lung parenchyma preparation in studies of the beta-adrenoceptor adenylate cyclase system.

The binding, adenylate cyclase activation, and functional effect of four beta-adrenoceptor agonists were studied in the guinea-pig lung parenchyma preparation and the results were compared with those obtained earlier in guinea-pig left-heart ventricle (beta 1-adrenoceptors) and soleus muscle (beta 2-adrenoceptors) preparations. The pKi-values of the unselective compounds, isoprenaline and orciprenaline, were in good agreement with those obtained in the heart and soleus muscle. The beta 2-adrenoceptor selective compounds KWD 2026 and terbutaline were bound to two sites, one corresponding to the beta 1-adrenoceptors and the other to the beta 2-adrenoceptors. The pKi-value of isoprenaline was in good agreement with its pKact-value indicating that maximum adenylate cyclase activity is obtained when the occupancy of the receptors is maximal. Further, the relative intrinsic efficacy calculated from the functional effect and receptor occupancy agreed well with the relative maximum adenylate cyclase activation by the agonists which was also found earlier for the guinea-pig heart ventricle and soleus muscle preparations. Relative effects were obtained from both functional experiments and from affinity and adenylate cyclase activating studies. There was good agreement between relative effects obtained in these two ways. It is concluded that the guinea-pig lung parenchyma preparation may be useful for the study of the beta-adrenoceptor adenylate cyclase system.     

Effects of full and partial beta-adrenergic agonists and antagonists on human lung adenylate cyclase

The beta-adrenergic stimulation of adenylate cyclase in membranes from human lung was compared to that of adenylate cyclase in membranes with a majority of beta 2-adrenergic receptors (from rat lung) and in membranes with a homogeneous population of beta 2-adrenergic receptors (from rat erythrocytes and reticulocytes). In terms of adenylate cyclase stimulation, three full agonists (isoproterenol, epinephrine and norepinephrine), four partial agonists (procaterol, salbutamol, fenoterol and zinterol), and four antagonists (propranolol, metoprolol, atenolol and practolol) were tested. The potency (Kact or Ki) of the eleven beta-adrenergic agents, and the Hill coefficient (of 1) for the four antagonists tested indicated that the activation of human lung adenylate cyclase occurred through receptors of the beta 2-subtype only. Partial beta-adrenergic agonists were efficiently discriminated by the human lung preparation, as shown by distinct intrinsic activities. The mediocre efficacy and the relatively low potency of all beta-adrenergic agonists on adenylate cyclase suggested a relatively low density of beta 2-adrenergic receptors, as compared to the enzyme density.     

The affinity and efficacy of the selective beta 1-adrenoceptor stimulant RO363 at beta 1- and beta 2-adrenoceptor sites.

(-)-Isoprenaline and the selective beta 1-adrenoceptor agonist RO363 were tested for their inotropic effects in left atrial (beta 1) and relaxant effects in K+-depolarized uterine (beta 2) preparations from the guinea-pig. The drugs had similar activities as positive inotropic agents but RO363 was approximately 400 times less active than (-)-isoprenaline as a uterine relaxant. RO363 had intrinsic activities of 0.8 and 0.25 ((-)-isoprenaline = 1) in atrial and uterine preparations, respectively. Apparent dissociation constants (KD values) determined from the ability of the agonists to displace (-)-[125I]-iodocyanopindolol ([125I]-CYP) bound to membranes prepared from both tissues were used as a measure of affinity. The [125I]-CYP binding sites possessed the characteristics of homogeneous populations of beta 1-adrenoceptors in atrial and beta 2-adrenoceptors in uterine membrane preparations. The pKD values for (-)-isoprenaline were similar in the two tissues (left atria 6.4, uterus 6.0) whilst for RO363 the atrial value (7.8) was considerably greater than that for the uterus (6.0). The latter value is very similar to the pKB value determined from shifts in (-)-isoprenaline curves produced by RO363 in uterine preparations. Graphical plots of the fraction of receptors occupied vs response were constructed. The relative efficacy of (-)-isoprenaline with respect to RO363 was calculated to be 25 in atrial and 2633 in uterine preparations. The selective beta 1-adrenoceptor stimulant actions of RO363 are a reflection of both its greater affinity and efficacy for beta 1- as opposed to beta 2-adrenoceptor sites.(ABSTRACT TRUNCATED AT 250 WORDS)     

Autoradiographic localization of beta-adrenoceptors in pig lung using [125I]-iodocyanopindolol.

The binding of the beta-adrenoceptor radioligand [125I]-iodocyanopindolol (I-CYP) has been studied in pig lung parenchyma and the distribution of binding sites visualised by light microscopic autoradiography. I-CYP binding was saturable (maximum binding capacity Bmax = 51 +/- 3 fmol mg-1 protein), involving sites with high affinity (dissociation constant KD = 73 +/- 10 pM). Specific I-CYP binding was displaceable both by beta-adrenoceptor agonists ((-)-isoprenaline greater than (-)-adrenaline greater than (+/-)-fenoterol greater than (-)-noradrenaline greater than (+)-isoprenaline greater than (+/-)-RO363) and antagonists ((+/-)-propranolol greater than ICI-118551 greater than atenolol), indicating a predominance of beta 2-adrenoceptors. Further analysis showed that displacement data for the beta 1-selective antagonist atenolol and the beta 2-selective antagonist ICI-118551 were fitted best to a 2 binding site model and that both beta 1- and beta 2-adrenoceptors were present in pig lung in the ratio 28:72 respectively. Autoradiographic grains were localized over tissue and were most dense over alveolar walls greater than vascular endothelium greater than vascular smooth muscle greater than bronchial smooth muscle = bronchial epithelium. Atenolol (10(-5) M) caused a 31% reduction in specific grain density over alveolar wall tissue, while a 10 fold lower concentration of ICI-118551 (10(-6) M) caused a 50% decrease. These results are consistent with binding data in pig lung parenchyma demonstrating a mixed population of beta-adrenoceptors with a predominance of the beta 2 subtype.(ABSTRACT TRUNCATED AT 250 WORDS)     

A study of beta-adrenoceptors in rat lung parenchymal strip

The aim of the present study was to characterize the beta-adrenoceptor population in rat lung strip. For this purpose, Schild plots were obtained for the beta-adrenoceptor antagonists atenolol (beta 1-selective), butoxamine (beta 2-selective) and propranolol (non-selective), using three different agonists:isoprenaline (non-selective), salbutamol (beta 2-selective) and noradrenaline (beta 1-selective). The slopes of these Schild plots were close to the theoretical value of unity, and pA2 values determined with isoprenaline, salbutamol and noradrenaline as agonists were: for propranolol, 7.86 +/- 0.22, 7.72 +/- 0.15 and 7.89 +/- 0.23; for atenolol, 5.19 +/- 0.05, 5.33 +/- 0.07 and 5.47 +/- 0.22 and for butoxamine, 6.31 +/- 0.11, 6.34 +/- 0.03 and 5.99 +/- 0.23, respectively. These data suggest that pharmacological responses of rat isolated lung strip to beta-adrenoceptor agents are mediated by a homogeneous population of beta 2-adrenoceptors, although the presence of a minor population of beta 1-adrenoceptors undetected by the agonists used cannot be excluded.     

Characterization of the receptor mediating the antianaphylactic effects of beta-adrenoceptor agonists in human lung tissue in vitro.

1 The rank order of potency of six beta-adrenoceptor agonists as inhibitors of the anaphylactic release of histamine from fragments of passively sensitized human lung in vitro was (--)-isoprenaline greater than (--) -adrenaline greater than (+/-)-salbutamol greater than (--)-noradrenaline greater than R0363 greater than H133/22. 2 The beta-adrenoceptor antagonists, propranolol, atenolol and H35/25, blocked the response to both (--)-isoprenaline and (+/-)-salbutamol competitively. Each antagonist gave similar pA2 values with both agonists. pA2 values were consistently at the high end of the range expected for interaction at a beta 2-adrenoceptor. 3 Practolol did not antagonize isoprenaline in a competitive manner but was a competitive antagonist of salbutamol with a pA2 at the high end of the range expected for interaction at a beta 2-adrenoceptor. 4 Data obtained with agonists are consistent with the receptor being of the beta 2-subtype. Data obtained with antagonists indicate a consistently higher affinity for the receptor than observed for the beta 2-subtype in other tissues but do not suggest a novel beta-adrenoceptor subtype on the mast cell of the human lung.     

Biochemical characterization of a new highly cardioselective beta-adrenoceptor antagonist

P0160 (1-phenyl-3-(2-(3-(2-cyanophenoxy)-2-hydroxypropyl)amino) ethylhydantoin HCl) is an aryloxypropanolamine which contains a ureido group as part of the hydantoin ring. This molecule was synthesized to obtain a more cardioselective beta-adrenoceptor blocker. Preliminary data have shown that it is as potent as propranolol and four times more cardioselective than atenolol in pharmacological tests in-vitro and in the conscious rat. In the present study we evaluated the interaction of P0160 with beta-adrenoceptors by radioreceptor binding studies and by measuring adenylate cyclase activity coupled to beta-adrenoceptors. The data indicate that P0160 binds with nanomolar affinity to beta-adrenoceptors labelled with [3H]DHA in the rat heart, but with micromolar affinity in the rat lung. Its binding is stereospecific, the S-(-)isomer being 200 times more active than the R-(+) form. P0160's selectivity between cardiac beta 1- and beta 2-receptors was 1388, about 60 times that for metoprolol. Analysis of the thermodynamic characteristics of P0160's interaction with rat heart beta-adrenoceptors indicated antagonist properties of the same order of magnitude as propranolol, as confirmed by adenylate cyclase studies. These data indicate that P0160 is a potent, specific and selective beta 1-adrenoceptor antagonist, and give a molecular explanation for the cardioselective activity found in pharmacological tests.     

Prenalterol is an agonist at beta 2- as well as at beta 1-adrenoceptors

Prenalterol exerted agonist activity in cat, but not guinea-pig, isolated atria, which contain predominantly beta 1-adrenoceptors. Prenalterol relaxed K+ -contracted rat uterus, but not histamine-contracted cat lung strips; both contain predominantly beta 2-adrenoceptors. The effect of prenalterol in rat uterus was antagonised by the selective beta 2-adrenoceptor antagonist ICI 118551 but not by the selective beta 1-adrenoceptor antagonist atenolol. Thus the ability of prenalterol to exert beta-adrenoceptor activity is tissue-dependent, rather than beta-adrenoceptor subtype-dependent.     

Beta-adrenoceptor selectivity of dobutamine: in vivo and in vitro studies

We compared the beta-adrenoceptor stimulant actions of dobutamine and (-)-isoprenaline in isolated tissue preparations (atria, trachea, and uterus) from the guinea pig and in chloralose-anaesthetized, vagotomized cats (arterial blood pressure, heart rate, hindlimb perfusion pressure, and soleus muscle contractility). The results obtained in these experiments indicate that, on a dose basis, dobutamine shows little selectivity in producing alpha-, beta 1-, and beta 2-adrenoceptor-mediated effects. In phentolamine-treated cats, reductions in arterial pressure and total peripheral resistance produced by infusions of dobutamine were little affected by the beta 2-adrenoceptor-selective antagonist butoxamine, but were antagonized by atenolol. The rise in cardiac output produced by dobutamine involved increases in both heart rate and stroke volume. There was little indication of a selective inotropic action, a feature that confirmed the results obtained in isolated atrial preparations. The increase in cardiac output appeared to involve both alpha- and beta-receptor-mediated actions, because phentolamine reduced the rise in cardiac output by reducing stroke volume.     

Contribution of beta 1- and beta 2-adrenoceptors of human atrium and ventricle to the effects of noradrenaline and adrenaline as assessed with (-)-atenolol.

1. (-)-Atenolol was used as a tool to assess the function of beta 1- and beta 2-adrenoceptors in human heart. Right atrial and left ventricular preparations from patients undergoing open heart surgery were set up to contract isometrically. Membrane particles were prepared for beta-adrenoceptor labelling with [3H]-(-)-bupranolol and adenylate cyclase assays. 2. The positive inotropic effects of (-)-noradrenaline were antagonized to a similar extent by (-)-atenolol in atrial and ventricular preparations. (-)-Atenolol consistently antagonized the effects of (-)-adrenaline to a lesser extent than those of (-)-noradrenaline in atrial preparations. In ventricular preparations (-)-atenolol antagonized the effects of low concentrations of (-)-adrenaline to a lesser extent than those of high concentrations. 3. pKB values (M) of (-)-atenolol, estimated with non-linear analysis from the blockade of the positive inotropic effects of the catecholamines, were 7.4 for beta 1-adrenoceptors and 6.0 for beta 2-adrenoceptors. 4. (-)-Atenolol inhibited the binding of [3H]-(-)-bupranolol to ventricular beta 1-adrenoceptors with a pKD (M) of 5.9 and to ventricular beta 2-adrenoceptors with a pKD of 4.6. 5. (-)-Atenolol inhibited the catecholamine-induced adenylate cyclase stimulation in the atrium and ventricle with pKB values of 5.8-6.4 for beta 1- and pKB values of 4.7-5.7 for beta 2-adrenoceptors. The binding and cyclase assays suggest a partial affinity loss for (-)-atenolol inherent to membrane preparations. 6. beta 1-Adrenoceptors mediate the maximum positive inotropic effects of (-)-noradrenaline in both the atrium and ventricle of man. beta 2-Adrenoceptors appear to be capable of mediating maximal positive inotropic effects of (-)-adrenaline in atrium. In contrast, ventricular beta 2-adrenoceptors mediated only submaximal effects of (-)-adrenaline.     

Selective labelling of beta 1-adrenoceptors in rabbit lung membranes by (-)[3H]bisoprolol

The binding properties of a newly developed, highly selective beta 1-adrenoceptor antagonist radioligand, (-)[3H]bisoprolol (EMD 33512) were investigated in rabbit lung membranes containing a mixture of 80% beta 1-and 20% beta 2-adrenoceptors. The binding of (-)[3H]bisoprolol at 25 degrees C was saturable, of high affinity (KD = 4.7 +/- 0.6 nM, N = 4), rapid and readily reversible. The maximal number of (-)[3H]bisoprolol binding sites (244 +/- 31 fmol bound/mg protein, N = 4), however, was only 80% of the number of sites labelled by the non-selective beta-adrenoceptor radioligand (-)[125I]iodocyanopindolol (299 +/- 36 fmol bound/mg protein, N = 4). beta-Adrenoceptor antagonists (non-selective: propranolol, alprenolol; beta 1-selective: metoprolol, practolol, bisoprolol; beta 2-selective: ICI 118,551) inhibited (-)[3H]bisoprolol binding with monophasic displacement curves and pseudo-Hill coefficients of 1.0 indicating that in rabbit lung membranes (-)[3H]bisoprolol labels a homogeneous class of beta-adrenoceptors. Agonists inhibited binding with an order of potency: (-)-isoprenaline greater than (-)-noradrenaline = (-)-adrenaline, which is a typical one for beta 1-adrenoceptors. It is concluded that in rabbit lung membranes (-)[3H]bisoprolol selectively labels beta 1-adrenoceptors. (-)[3H]Bisoprolol therefore seems to be a suitable ligand for direct determination of the properties of beta 1-adrenoceptors in those tissues where both beta-adrenoceptor subtypes coexist.     

Beta 1-adrenoceptors mediate smooth muscle relaxation in mouse isolated trachea.

1. The relaxant effects to the beta-adrenoceptor agonists isoprenaline, adrenaline, noradrenaline, RO363, procaterol and fenoterol were investigated in carbachol-contracted mouse isolated tracheal preparations. 2. The order of potencies for those beta-adrenoceptor agonists that induced full relaxation of carbachol-contracted mouse tracheal preparations was isoprenaline greater than RO363 greater than noradrenaline = adrenaline greater than fenoterol. The EC50 value of isoprenaline for relaxation was 46 nM. The beta 1-adrenoceptor-selective agonist, RO363 was ten times more potent than the beta 2-adrenoceptor-selective agonist, fenoterol. The highly beta 2-adrenoceptor-selective agonist procaterol was a partial relaxant and induced only 28 +/- 4% relaxation. 3. Relaxations induced by noradrenaline and isoprenaline were not significantly affected by the neuronal uptake inhibitor, cocaine (10 microM) or by the extraneuronal uptake inhibitor, deoxycorticosterone acetate (25 microM) respectively. The alpha-adrenoceptor agonist methoxamine induced no observable elevation of mouse tracheal smooth muscle tone. 4. Schild plots for the beta-adrenoceptor antagonists, atenolol and betaxolol (beta 1-adrenoceptor-selective) and ICI 118,551 (beta 2-adrenoceptor-selective) were linear, with slope values approaching unity. Mean pA2 values derived for atenolol, betaxolol and ICI 118,551 for antagonism of beta-adrenoceptor-mediated relaxation were 7.1, 8.4 and 7.2, respectively. These data were independent of the use of isoprenaline or noradrenaline as the agonist. 5. These findings indicate that beta-adrenoceptor-mediated relaxations of mouse isolated trachea occur predominantly through activation of beta 1-adrenoceptors.     

The affinity and efficacy of naturally occurring catecholamines at beta-adrenoceptor subtypes

The contribution of affinity and efficacy to the agonistic actions of the naturally occurring catecholamines, (-)-noradrenaline and (-)-adrenaline at beta-adrenoceptor sites were assessed in guinea-pig driven left atrial (beta 1) and K+-depolarized uterine (beta 2) preparations. The dissociation constants of each agonist, required in these calculations, were calculated using radioligand binding techniques. [125I]Iodocyanopindolol bound to sites in membrane preparations of each tissue which have been shown to represent beta 1-(atria) and beta 2-(uterus) adrenoceptors. It was found that (-)-noradrenaline was approximately 10-fold more selective for the beta 1- as opposed to the beta 2-adrenoceptor in the pharmacological studies. Affinity/efficacy calculations indicated that this selectivity was entirely due to a selective affinity for the beta 1-adrenoceptor subtype. (-)-Noradrenaline, (-)-adrenaline and the reference compound (-)-isoprenaline all had approximately the same efficacy at either beta-adrenoceptor subtype.     

The beta adrenergic receptors of rat corpus luteum membranes..

A comparison has been made between the β receptors of rabbit heart membranes and superovulated rat corpus luteum in terms of adenylate cyclase activation and [³H]dihydroalprenolol binding. Agonist activation and binding agree in rank order, which is isoprenaline > noradrenaline ? adrenaline > salbutamol for heart, and isoprenaline > adrenaline > salbutamol > noradrenaline for corpus luteum. The β₁ selective blockers, practolol, atenolol and ICI 89,406, are all less potent in the corpus luteum than in the heart but the order of potency is not the same in corpus luteum as was previously found in the uterus, and salbutamol. which is a full agonist in uterus, is only a partial agonist in corpus luteum. These findings do not agree with a clear cut separation of receptors into classical β₁ and β₂ types.     

ICI 118,551: an effective ocular hypotensive agent with selectivity for the ciliary process beta 2-adrenoceptor and with minimal cardiac side effects

Prior biochemical studies have shown that the ciliary process epithelium, which is involved in the secretion of aqueous humour, is rich in beta-adrenoceptors with pharmacological characteristics similar to those of the beta 2 subclass. The present experiments demonstrate that the beta-adrenoceptor antagonist, ICI 118,551, is a potent inhibitor of isoprenaline-stimulated adenylate cyclase activity measured in broken cell preparations of rabbit ciliary process. In rabbit cardiac muscle, however, ICI 118,551 is a relatively weak antagonist of isoprenaline-stimulated adenylate cyclase, being approximately 100 fold less potent than the non-selective beta-adrenoceptor antagonist, timolol. ICI 118,551 is also less potent than timolol in inhibiting isoprenaline-sensitive adenylate cyclase of rabbit lung. ICI 118,551 applied topically to eyes of unanaesthetized rabbits causes a dose-dependent decrease in intraocular pressure. Furthermore, in a blind crossover study in rabbits, topically applied ICI 118,551 decreased intraocular pressure for more than 6 h and was more effective than an identical dose of the clinically effective anti-glaucoma agent, timolol. Systemic absorption from topically-applied timolol, but not ICI 118,551, is sufficient to alter cardiac response to subcutaneous administration of isoprenaline. Furthermore, dose-response studies, using direct systemic administration of the two beta-adrenoceptor antagonists, revealed that ICI 118,551 is about 60 times less potent than timolol in blocking isoprenaline-induced cardio-acceleration. ICI 118,551, applied to one eye, causes a decrease in intraocular pressure in the contralateral eye, and systemic administration of ICI 118,551 results in decreased intraocular pressure in both eyes, data indicating that at least part of the ocular hypotensive effect of topical ICI 118,551 is mediated through systemic absorption.(ABSTRACT TRUNCATED AT 250 WORDS)