J. Paul Hieble

In vitro characterization of the α-adrenoceptors in human prostate

The alpha-adrenoceptors of the human prostate gland were characterized in vitro by the use of antagonists selective for alpha 1- and alpha 2-adrenoceptor subtypes. The contractile response induced by norepinephrine in this tissue could be antagonized by prazosin, a selective alpha 1-antagonist, with a receptor dissociation constant (KB) of 4.0 +/- 0.9 nM. The selective alpha 2-antagonists rauwolscine and SK&F 86466 were less potent antagonists of this response, with KB values of 1020 +/- 400 nM and 2400 +/- 800 nM, respectively. The irreversible alpha-antagonists benextramine and phenoxybenzamine, both of which preferentially inactivate the alpha 1-subtype, produced marked depression of norepinephrine-induced contraction. These data would suggest that the alpha-receptors on prostatic smooth muscle are predominantly of the alpha 1-subtype.

GW427353 (Solabegron), a Novel, Selective β3-Adrenergic Receptor Agonist, Evokes Bladder Relaxation and Increases Micturition Reflex Threshold in the Dog

Functional studies have demonstrated that adrenoceptor agonist-evoked relaxation is mediated primarily by β3-adrenergic receptors (ARs) in human bladder. Thus, the use of selective β3-AR agonists in the pharmacological treatment of overactive bladder is being explored. The present studies investigated the effects of a novel selective β3-AR agonist, (R)-3′-[[2-[[2-(3-chlorophenyl)-2-hydroxyethyl]amino]ethyl]amino]-[1,1′-biphenyl]-3-carboxylic acid (GW427353; solabegron) on bladder function in the dog using in vitro and in vivo techniques. GW427353 stimulated cAMP accumulation in Chinese hamster ovary cells expressing the human β3-AR, with an EC50 value of 22 ± 6 nM and an intrinsic activity 90% of isoproterenol. At concentrations of 10,000 nM, GW427353 produced a minimal response in cells expressing either β1-ARs or β2-ARs (maximum response <10% of that to isoproterenol). In dog isolated bladder strips, GW427353 evoked relaxation that was attenuated by the nonselective β-AR antagonist bupranolol and 1-(2-ethylphenoxy)-3-[[(1S)-1,2,3,4-tetrahydro-1-naphthalenyl]amino]-(2S)-2-propanol (SR59230A) (reported to have β3-AR antagonist activity). The relaxation was unaffected by atenolol, a selective β1-AR antagonist, or (±)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl)amino]-2-butanol (ICI 118551), a selective β2-AR antagonist. GW427353 increased the volume required to evoke micturition in the anesthetized dog following acetic acid-evoked bladder irritation, without affecting the ability of the bladder to void. GW427353-evoked effects on bladder parameters in vivo were inhibited by bupranolol. The present study demonstrates that selective activation of β3-AR with GW427353 evokes bladder relaxation and facilitates bladder storage mechanisms in the dog.

Possible structural and functional relationships between imidazoline receptors and alpha 2-adrenoceptors.

Although it is now well established that imidazoline receptors and alpha 2-adrenoceptors are discrete entities with distinct endogenous ligands, the two receptor classes apparently have several common features. While the catecholamines stimulate alpha 2-adrenoceptors but not imidazoline receptors, agmatine, a guanidine analog that may be an endogenous imidazoline receptor ligand, can interact with both I1 and I2 imidazoline receptors as well as alpha 2-adrenoceptors, although, interestingly, other guanidines such as guanabenz are highly selective for alpha 2-adrenoceptors versus I1 receptors. Most I1 receptor agonists such as moxonidine, rilmenidine, and clonidine can also stimulate alpha 2-adrenoceptors, and the same physiological response is produced by activation of central I1 receptors and alpha 2-adrenoceptors, but their anatomical locations differ. The imidazoline idazoxan is an antagonist at I1, I2, and alpha 2-receptors, but minor structural alterations of idazoxan can result in molecules with selectivity for either alpha 2-adrenoceptors or imidazoline receptors. The precise mode of interaction of imidazoline agonists and antagonists with the alpha 2-adrenoceptor is not yet understood, and structures of the imidazoline receptors are still unknown. Nevertheless, the fact that many agents can stimulate or block both receptor classes, combined with the fact that alpha 2-adrenoceptors and I1 receptors can mediate identical physiological responses, suggests that many common structural features may be present.

Comparison of metoprolol and carvedilol pharmacology and cardioprotection in rabbit ischemia and reperfusion model

Carvedilol, a selective alpha1 and non-selective beta-adrenoceptor antagonist and antioxidant, has been shown to provide significant cardiac protection in animal models of myocardial ischemia. To further explore the mechanisms contributing to carvedilol cardioprotection efficacy, the effects of carvedilol on hemodynamic variables, infarct size and myeloperoxidase activity (an index of neutrophil accumulation) were compared with a beta1-selective adrenoceptor antagonist, metoprolol. Carvedilol (1 mg/kg) or metoprolol (1 mg/kg or 1 mg/kg + 0.5 mg/kg 90 min later) was given intravenously 5 min before reperfusion. In vehicle-treated rabbits, ischemia (60 min) and reperfusion (180 min) resulted in significant increments in left ventricular end diastolic pressure, large infarcts (59+/-2.6% of area-at-risk) and marked increase in myeloperoxidase activity (0.59+/-0.09 U/100 mg tissue). Carvedilol treatment resulted in sustained reduction of pressure-rate-index and significantly smaller infarcts (22.0+/-2.5%, P < 0.01 vs. vehicle) as well as decreased myeloperoxidase activity (0.186+/-0.056 U/100 mg tissue, P < 0.01 vs. vehicle). The highest dose of metoprolol, 1 mg/kg + 0.5 mg/kg, that resulted in pressure-rate-index comparable to that of 1.0 mg/kg carvedilol, failed to reduce myeloperoxidase activity in the ischemic myocardial tissue, and the infarct size (35+/-3.1%) was significantly larger than in carvedilol-treated animals. Taken together, this study suggests that the superior cardioprotection of carvedilol over metoprolol is not a consequence of hemodynamic variances but possibly the result of the additional pharmacological properties of carvedilol such as the antioxidant and anti-neutrophil effects.

Recent advances in the identification of a 1- and a 2-adrenoceptor subtypes: therapeutic implications

The cloning of multiple subtypes of both a1- and a2-adrenoceptors has renewed interest in the therapeutic application of agents interacting with these receptors. Effort has primarily been directed towards the design of uroselective a1-adrenoceptor antagonists for the treatment of benign prostatic hyperplasia (BPH). Evidence is accumulating for the involvement of a novel a1-adrenoceptor, designated as a1L-adrenoceptor, in a1-adrenoceptor-mediated smooth muscle contraction in prostatic and other urogenital tissues. While several antagonists showing a high degree of uroselectivity in animal models have been identified, their clinical superiority over the currently available a1-adrenoceptor antagonists has not yet been demonstrated. It is possible that the interaction with a1-adrenoceptors, as yet uncharacterised subtypes, at non-prostatic sites contributes to the therapeutic activity of this drug class in BPH. The a1-adrenoceptor subtypes involved in the control of vascular tone are currently being evaluated...

Evidence for and against heterogeneity of alpha1-adrenoceptors

Recent experimental evidence has suggested that the alpha 1 adrenoceptor may need to be further subdivided. It can no longer be stated categorically that alpha 1-adrenoceptors are present only at postjunctional sites, in view of several reports of alpha 1-mediated modulation of adrenergic and cholinergic neurotransmission. Furthermore, comparison of the pharmacologic characteristics of the alpha 1-adrenoceptor in different species and/or tissues can show clear differences in sensitivity to selective agonists and antagonists, and differences in the degree of dependence on extracellular calcium. However, in other cases, alpha 1-adrenoceptors at diverse sites have been found to have identical characteristics. Furthermore, the subcategories identified by the various selective agents do not fall into the same discrete groups, in contrast to division of alpha-adrenoceptors into alpha 1 and alpha 2-adrenoceptors. Therefore, at this time it seems premature to subdivide the alpha 1-adrenoceptor further.

Effects of ring substitution on the pre- and postjunctional alpha-adrenergic activity of aryliminoimidazolidines

SummaryThe pre- and postjunctional α-adrenergic agonist potency of a series of aryliminoimidazolidines was determined in the isolated rabbit ear artery. This series included clonidine, an antihypertensive agent thought to act by stimulating brainstem α-receptors and known to be a preferentially prejunctional α-adrenergic agonist. Although all of the compounds acted preferentially on the prejunctional α-adrenoceptor, ring substitution had a dramatic effect on both potency and the degree of selectivity. 2-(3,4-Dihydroxyphenylimino) imidazolidine was both the most potent and most selective prejunctional α-agonist in this series.

Adrenoceptor Pharmacology: Urogenital Applications

Although the selective α1-adrenoceptor antagonists were initially developed as antihypertensive drugs, and they are still utilized for this indication, the α1-adrenoceptor blockers are now used extensively for the symptomatic treatment of benign prostatic hyperplasia (BPH). As a result, a number of new drugs in this class have been specifically developed for use in BPH. The utility of α1-adrenoceptor antagonists in BPH derives from the observation, made several decades ago, that the irreversible, α1- adrenoceptor selective antagonist phenoxybenzamine, blocked the contractile activity of norepinephrine in isolated strips of rat or human prostate. Following the further subclassification of α1-adrenoceptors into the α1A-, α1B- and α1D-adrenoceptor subtypes, the relationship between subtype selectivity and efficacy in BPH has been investigated in the hope of developing more selective drugs for the treatment of this disorder. Molecular characterization of the adrenoceptor population in human prostate clearly shows the α1A-adrenoceptor subtype to predominate, and highly selective α1A-adrenoceptor antagonists have been identified and investigated in BPH. However, controversy remains as to whether prostatic smooth muscle contraction is mediated by the α1A-adrenoceptor, or by another novel α1-adrenoceptor subtype (not corresponding to any of the three known recombinant α1-adrenoceptors), or both. α1-Adrenoceptor agonists have been used clinically for the treatment of stress incontinence, acting to increase urethral tone by contracting urethral smooth muscle. Research efforts are ongoing to identify agents of this class having a selective action on urethral versus vascular smooth muscle, in order to produce a greater effect on the urethra without producing dose-limiting increases in blood pressure. Local administration of vascular smooth muscle relaxants, either alone or in combination, has been used for the treatment of erectile dysfunction. An α1-adrenoceptor antagonist is often used as one comportent in such mixtures, which act to relax trabecular smooth muscle. The recent demonstration that a systemically administered drug can produce a sufficiently selective action on cavernosal smooth muscle to allow efficacy without producing limiting systemic side effects has renewed interest in the possibility of systemic administration of α1-adrenoceptor antagonists for this indication.

Functions Mediated by alpha-2 Adrenergic Receptors

This chapter summarizes the diverse physiologic functions that are influenced by alpha-2 adrenergic receptor activation. A great deal of attention has been given to the cardiovascular system, although the physiologic consequences of alpha-2 receptor occupancy are described for other target organs as well. The diverse cellular consequences of alpha-2 adrenergic receptor activation are paralled by the demonstration of several possible biochemical signaling systems that are activated by these receptors, including inhibition of adenylate cyclase, alterations in Ca2+ translocation, and acceleration of Na+/H+ exchange. A similar diversity is known to exist for electrophysiologic consequences of alpha-2 adrenergic receptor occupancy (also explored in Chapter 6). What remains for future experimental data to determine is whether alpha-2 adrenergic receptors all share certain early biochemical and/or electrophysiological events that lead to differing distal effects depending on the particular target cell, or whether distinct signaling mechanisms prevail resulting in the distinct biochemical and physiological changes characteristic of varying target organs.

Demonstration of α2-adrenoceptor-mediated contraction in the isolated canine saphenous artery treated with Bay K 8644

Contractions to B-HT 920 in the plantar branch of the canine saphenous artery in vitro occurred consistently only in the presence of the calcium channel agonist, Bay K 8644 (30 nM). These results indicate that prior activation of the calcium channel may be required to elicit a subsequent response to B-HT 920 in this artery. Furthermore, the contractile response was inhibited by rauwolscine (10 nM) and was unaffected by prazosin (10 nM), indicating that the response is mediated by postjunctional alpha 2-adrenoceptors.