Inotropic effects of histamine on developing chick heart: release of transmitters from autonomic nerve terminals

Inotropic effects of histamine were examined in isolated ventricular preparations from late embryonic and hatched chick hearts. In 19 day-old embryonic preparations, histamine had little effect on the contractile force. In preparations from 1 to 2 day-old hatched chick, histamine produced a transient decrease in contractile force followed by a sustained increase. The negative and positive responses were antagonized by atropine and propranolol, respectively, but not by histamine antagonists terfenadine, cimetidine or thioperamide. Acetylcholine produced positive Inotropic responses in the embryo while negative responses were observed after hatching. In myocardium of hatched chicks, compound 48/80, which releases histamine from mast cells, produced a transient decrease in contractile force followed by a sustained increase with a similar magnitude and time course to the case of exogenously applied histamine. The negative and positive responses were inhibited by atropine and propranolol, respectively, but not by terfenadine, cimetidine or thioperamide, which was similar to the case with the responses to histamine. The present results suggest that histamine, either applied exogenously or released from myocardial store sites, produces negative and positive inotropic responses in hatched chick myocardium which are due to release of acetylcholine and norepinephrine, respectively, from autonomic nerve terminals.     

Separate receptors mediating the positive inotropic and chronotropic effect of histamine in guinea-pig atria

The direct positive inotropic effect of histamine was studied on paced left atrial preparations from guinea pigs. Histamine (10^?8 to 10^?4 M) increased the maximum tension developed in left atria incubated at 35°C and driven at 2 Hz. The maximum increase in tension was 60% of that observed with norepinephrine. Metiamide (3 × 10^?5 M; a specific H₂-receptor antagonist) did not alter the inotropic response of left atria to histamine. However, tripelennamine (a typical H₁-receptor antagonist) competitively shifted the histamine inotropic dose—response curve to the right at concentrations from 10^?8 to 10^?7 M. Higher concentrations (3 × 10^?7 and 10^?6 M) caused little further additional shift to the right. The positive chronotropic effect of histamine on spontaneously beating atria was competitively antagonized by metiamide (10^?6 and 3 × 10^?6 M). These results demonstrate that in guinea-pig atria histamine increases myocardial contractility by an interaction with receptors closely related to classical H₁-receptors while its chronotropic effects is mediated by interaction with H₂-receptors.     

Histamine H2 receptor subtypes in the guinea pig right atrium

Inotropic and chronotropic responses to histamine and to selective histamine H2 receptor agonists (4-methyl histamine, dimaprit, impromidine) and antagonists (metiamide and cimetidine) were studied in retrogradely superfused guinea pig right atrium preparations. All agonists tested were acting as chronotropic and, except impromidine, as inotropic agonists; impromidine had a dual inotropic action. Affinities of the antagonists varied, showing approximately 20-fold more activity on chronotropism. The findings suggest the existence of subtypes of histamine H2 receptors in the development of chronotropic and inotropic responses.     

The role of H1 and H2-receptors in the coronary vascular response to histamine of isolated perfused hearts of guinea-pigs and rabbits.

1. The effects of histamine on the isolated perfused hearts of guinea-pigs and rabbits were examined. Records of contractile force, heart rate and coronary perfusion pressure were obtained. 2. Histamine exerted positive inotropic and chronotropic effects which were antagonized by burimamide and attributed to stimulation of H2-receptors. 3. The coronary vascular response to histamine differed between guinea-pigs and rabbits. In guinea-pig hearts, three phases were apparent: (a) An initial vasodilatation preceding any effects on heart force and rate was antagonized by mepyramine and therefore mediated by histamine H1-receptors in the coronary circulation. (b) A secondary vasoconstriction was attributed to the increased myocardial compression during the positive inotropic and chronotropic responses. (c) The final, more predominant, component was a prolonged vasodilatation probably associated with the increased metabolic activity of the heart. 4. The latter two components were abolished together with the myocardial responses by burimamide. The remaining coronary vascular response was biphasic, consisting of a vasodilatation immediately followed by vasoconstriction. Both were antagonized by mepyramine and therefore mediated by H1-receptors. 5. The coronary vascular response of rabbit hearts was similar but no direct vasodilatation was observed and it was concluded that histamine receptors in the coronary vasculature involve only vasoconstriction.     

Effects of the H1-antagonist promethazine and the H2-antagonist burimamide on chronotropic, inotropic and coronary vascular responses to histamine in isolated perfused guinea-pig hearts.

On guinea-pig atria part of the inotropic response to histamine is attributable to a concomitant increase of the frequency [7]. Since the chronotropic effect of histamine is mediated by a stimulation of H2-receptors a direct interaction of histamine with H1-receptors a direct interaction of histamine with H1-receptors mediating the inotropic response on heart may be overlooked. For this reason the ability of the H1-antagonist promethazine and the H2-antagonist burimamide to inhibit the positive chronotropic, inotropic and coronary vascular responses to histamine was determined in spontaneously beating and electrically driven perfused guinea-pig hearts. (1) Burimamide produced a competitive blockade of the positive chrono- and inotropic responses to histamine. (2) On the other hand, promethazine in concentrations that had no effect on cardiac function by itself, proved to be ineffective against the positive chrono- and inotropic responses produced by histamine on spontaneously beating and electrically driven heart preparations. (3) The predominant coronary vasodilation observed after infusion with histamine was competitively antagonized by promethazine and burimamide. This blockade was not attributable to an interaction with myocardial H2-receptors mediating increases in heart rate and contractility and was, therefore, direct in nature. (4) Based upon the present study and former investigations [7] the following distribution of different histamine receptors in the guinea-pig heart does exist: H1-receptors are present in the atrial muscle and the coronary vascular bed. H2-receptors are located in the sinus node, the ventricular myocardium and the coronary vessels.     

Pharmacological analysis of the tachycardia produced by histamine and specific H1-and H2-receptor agonists in anesthetized dogs.

A study was designed to investigate the mechanism(s) of the tachycardia produced by histamine and specific H₁- and H₂-receptor agonists in anesthetized dogs. I.v. administration of histamine, 2-methylhistamine, and 4-methylhistamine caused a fall in blood pressure and an increase in heart rate. Prior administration of mepyramine, a histamine H₁-receptor antagonist, significantly antagonized the depressor and positive chronotropic effects of histamine and completely abolished the cardiovascular actions of 2-methylhistamine, whereas responses to 4-methylhistamine were not affected. Subsequent administration of metiamide, a specific H₂-receptor antagonist, completely abolished the remaining cardiovascular effects of histamine and the hypotension and tachycardia observed after 4-methylhistamine were also antagonized.In another series of experiments, bilateral vagotomy significantly attenuated the tachycardia and subsequent removal of cardiac sympathetic nerve supply completely abolished it without affecting the depressor effects of these agents. Pharmacological blockade of muscarinic receptors with atropine in another group of animals also significantly attenuated the positive chronotropic effect which was completely prevented by further treatment with propranolol. The hypotension produced by histamine agonists was not affected by atropine and propranolol. These results demonstrate that in pentobarbital-anesthetized dogs, the tachycardia produced by histamine and specific H₁- and H₂-receptor agonists are due to reciprocal alterations in cardiac autonomic activity as a result of the fall in the arterial pressure. Activation of cardiac histamine receptors does not seem to account for the positive chronotropic effects of these compounds in intact dogs.     

Increased sensitivity of neonatal mouse myocardia to autonomic transmitters

1 Chronotropic and inotropic responses to noradrenaline and acetylcholine were examined in isolated right atrial and ventricular preparations from neonatal and adult mice. 2 Noradrenaline and acetylcholine produced positive and negative chronotropic responses, respectively, in the atria from both ages. Noradrenaline produced positive inotropic responses in ventricular preparations from both ages. In all cases, the sensitivity, expressed in terms of pD₂ values, was higher in neonatal preparations. 3 In the ventricle, desipramine produced a leftward shift of the concentration–response curve for noradrenaline in the adult, but no such shift was observed in the neonate. The sensitivity to isoprenaline of ventricular preparations was higher in the neonate than in the adult. 4 Our results demonstrated developmental decreases in sensitivities to autonomic transmitters in mouse myocardia. As for the inotropic response to noradrenaline of ventricular muscle, both pre- and postjunctional mechanisms were responsible for the developmental decrease in sensitivity.     

Effects of Ca2+ channel antagonists and ryanodine on H1-receptor mediated electromechanical response to histamine in guinea-pig left atria

Summary Effects of organic Ca²⁺ channel antagonists, Ni²⁺ and ryanodine on the electrophysiological and positive inotropic responses to histamine were examined in isolated guinea-pig left atria. Histamine increased force of contraction, prolonged action potential duration (APD) and hyperpolarized the membrane in a concentration-dependent manner. Histamine at a concentration of 1 mol/l produced a dual-component positive inotropic response composed of an initial increasing phase (initial component) and a second an late developing, greater positive inotropic phase (second component), whereas causing monophasic changes in APD and resting potential. The electrophysiological and dual-component positive inotropic effects induced by histamine were antagonized by chlorpheniramine (1 mol/l) but not by cimetidine (10 mol/l), indicating that both effects are exclusively mediated by H₁-receptors. The positive inotropic response to 1 mol/l histamine was changed by the pretreatment with nifedipine (1 mol/l) and nisoldipine (1 mol/l). In the presence of these dihydropyridines, the second component was almost completely abolished, while the initial component was hardly affected. On the other hand, verapamil (3 mol/l) and diltiazem (10 mol/l) failed to modify the multiphasic inotropic response to histamine. None of the Ca²⁺ channel antagonists affected the histamine-induced APD prolongation. In the presence of Ni²⁺ at a concentration of 0.3 mmol/l, at which it produced no negative inotropic action, the second component of the positive inotropic effect of histamine was specifically suppressed whereas the histamine-induced APD prolongation was unaffected. Preferential attenuation of the second component was also observed in the presence of 30 nmol/l ryanodine. However, the electrophysiological alterations caused by histamine remained unchanged. These results suggest that in guinea-pig left atria the H₁-receptor-mediated prolongation of APD seems unlikely to be due to enhancement of the slow inward Ca²⁺ current. Conversely, the increased Ca²⁺ influx as a result of the APD prolongation may contribute to the second component of the positive inotropic effect of histamine. Dihydropyridine Ca²⁺ channel antagonists, Ni²⁺ and ryanodine are all capable of inhibiting the second component, but do so possibly via different mechanisms, implying the complicated mechanisms underlying the H₁-receptor-mediated positive inotropic effect.Send offprint requests to Y. Hattori     

Differential effects of histamine mediated by histamine H1- and H2-receptors on contractility, spontaneous rate and cyclic nucleotides in the rabbit heart

The effects of histamine on the contractile force, spontaneous rate of contraction, and cyclic AMP and cyclic GMP content were investigated in isolated rabbit cardiac preparations. Histamine had a positive inotropic effect in the left atrium and papillary muscle, and a positive chronotropic effect in the right atrium. Both effects were produced in a concentration-dependent manner. Impromidine also induced the same effect in the left and right atrium as histamine did. The effects produced by histamine and impromidine were antagonized by cimetidine and tiotidine. On the other hand, the positive inotropic response of papillary muscle to histamine was antagonized by mepyramine and chlorpheniramine and was mimicked by 2-(2-pyridyl)ethylamine. Impromidine at a high concentration induced a small increase in the contractile force, an effect which was antagonized by cimetidine. Histamine significantly increased the cyclic AMP levels in both atria but not in papillary muscles. The increase in cyclic AMP was abolished by cimetidine. Histamine also increased cyclic GMP levels in all of the preparations. The increase in cyclic GMP was abolished by chlorpheniramine. The results suggest that both H1- and H2-receptors exist in all parts of the rabbit heart. However, the positive inotropic and chronotropic effects induced by histamine in left and right atrium are mediated predominantly via H2-receptors, whereas the positive inotropic effect in papillary muscle is predominantly mediated via H1 receptors.     

Sustained negative inotropism mediated by alpha-adrenoceptors in adult mouse myocardia: developmental conversion from positive response in the neonate.

1. Inotropic responses to alpha-adrenoceptor stimulation and the effects of antagonists were examined in isolated ventricular preparations from neonatal and adult mice. 2. Phenylephrine, in the presence of propranolol, produced positive inotropic responses in neonates up to 1 week after birth, while it produced negative inotropic responses in mice older than 3 weeks. 3. Both positive and negative responses to phenylephrine in neonates and adults, respectively, were antagonized by prazosin, WB4101 (2-([2,6-dimethoxyphenoxyethyl]aminomethyl)-1,4-benzodioxane) and 5-methylurapidil, but not by atropine, yohimbine or chlorethylclonidine. 4. Noradrenaline (NA) produced positive inotropic responses both in the neonate and adult; the responses were observed in a lower concentration-range in the neonate than in the adult. WB4101 produced a significant leftward shift of the concentration-response curve for noradrenaline in adult preparations while only a slight rightward shift was observed in the neonate. 5. Our results demonstrate the presence of alpha-adrenoceptor-mediated inotropic responses in the mouse ventricular myocardia. The response to phenylephrine changes from a positive to a negative effect during postnatal development. The responses are mediated by alpha 1-adrenoceptors, and modulate the overall inotropic response to NA in the adult.     

Pharmacological characterization of cardiac histamine receptors: Sensitivity to H1- and H2-receptor agonists and antagonists

In the isolated guinea pig heart, histamine H₁-receptor antagonists inhibit the histamine-induced negative dromotropic effect, but not the positive inotropic and chronotropic effects (Levi and Kuye, 1974, European J. Pharmacol. 27, 330). Burimamide, the H₂-receptor antagonist, inhibits the positive chronotropic effect of histamine (Black et al., 1972, Nature 236, 385). In this study, the hypothesis that H₁- and H₂-receptors selectively mediate the various cardiac effects of histamine was tested: (a) by investigating the inhibition of cardiac histamine effects by burimamide; (b) by studying the cardiac effects of 2-methylhistamine (an H₁-agonist) and 4-methylhistamine (an H₂-agonist). Burimamide, as a function of concentration, inhibited the positive inotropic and chronotropic effects of histamine, whereas it attenuated the negative dromotropic effect of histamine at the higher concentration only. 4-Methylhistamine caused dose-dependent positive inotropic and chronotropic but not negative dromotropic effects; conversely, with 2-methylhistamine the negative dromotropic effect prevailed. Sinus tachycardia and slowing of atrioventricular conduction were also obtained in the guinea pig in vivo by the i.v. administration of histamine. Burimamide selectively antagonized the positive chronotropic effect, whereas promethazine (an H₁-antagonist) selectively inhibited the negative dromotropic effect. These results substantiate the hypothesis that H₂-receptors mediate the histamine-induced increase in rate and contractility, whereas H₁-receptors mediate the slowing of atrioventricular conduction.     

Effects of propranolol and a new 'cardioselective' beta-blocker, H 93-26, on responses of isolated rat atria to isoprenaline and noradrenaline

Propranolol and a new ‘cardioselective’ β-adrenergic blocking agent, H 93/26, 1 -isopropylamino-3-[4-(2-meth-oxyethyl)-phenoxyl]-2-propanol, have been studied with regard to their ability to inhibit isoprenaline (IP)- and noradrenaline (NA)-induced chronotropic and inotropic responses of isolated rat atria. NA responses were studied on preparations from hearts which had been ‘chemically sympathectomized’ by 6-hydroxydopamine. This treatment shifted the concentration-effect curves for NA to the left but had no significant influence on the IP curves. Chronotropic and inotropic responses to IP and chronotropic responses to NA were about equally affected by propranolol, no significant differences in K_B being found. H 93/26 also acted as a typical competitive β-blocker with regard to these effector responses but its K_B-values were about 7 times those of propranolol. The results gave no evidence for a differential action of the blockers on the β-adrenoceptors mediating chronotropic and inotropic effects, respectively. With regard to the inotropic responses to NA the plots of log (dose ratio -1) versus log [antagonist] seemed to deviate from the theoretical, linear relationship for both propranolol and H 93/26. Experiments with phenoxybenzamine suggested that this deviation might be due to an α-adrenergic positive inotropic action of NA or to an influence of extraneuronal NA uptake on the responses.     

Structure-activity relationship of imidazolidine derivatives related to clonidine at histamine H2-receptors in guinea-pig isolated atria

1 Cumulative concentration-response relationships for the chronotropic effects of histamine, oxymetazoline, clonidine and thirteen clonidine-like imidazolidine derivatives were examined in isolated spontaneously beating guinea-pig atria.

2 The following compounds induced positive chronotropic effects: histamine, clonidine (2,6-dichloro-phenyliminoimidazolidine) and the 2,6-dibromo, 2,6-dimethyl, 2,6-diethyl, 2,6-dihydroxy, 2,4,6-trimethyl, 3,4-dihydroxy and 2-methyl-5-fluoro analogues of clonidine. These compounds appeared to act as partial agonists on histamine H₂-receptors, with potencies ranging from one tenth to one hundredth and intrinsic activities from approximately 20% to 75% of those of histamine.

3 The following compounds did not induce positive chronotropic effects but rather decreased the atrial rate, usually at high concentrations: oxymetazoline and the 2,3-dichloro, 4-dichloro, 5-dichloro, 2-chloro-4-methyl, 2-methyl-5-chloro, 2,4,6-trichloro analogues of clonidine.

4 The effects of histamine were antagonized by cimetidine, the pA₂ value being 6.68 (s.e. mean = 0.16, n = 3), and also in a concentration-dependent manner by clonidine. Cimetidine antagonized the response to clonidine in a concentration-dependent manner; however, high concentrations of cimetidine depressed the maximal response to clonidine and the slope of the concentration-response curve was no longer parallel to the control curve.

5 The effects of the other compounds which induced positive chronotropic effects were also antagonized by cimetidine (1 μmol/l); however, the effect of the 3,4-dihydroxy derivative was unaffected by cimetidine (1 μmol/l) but was abolished by propranolol (0.3 μmol/l).

6 In general, phenyliminoimidazolidine derivatives with 2,6-substitution on the phenyl ring are active on histamine H₂-receptors, whereas derivatives with 2,3-, 2,4- or 2,5-substitutions are weakly active or inactive. Thus the restriction imposed on the free rotation of the phenyl ring about the carbon-imino nitrogen bond by the introduction of two ortho substituents appears to result in increased agonist activity on the histamine H₂-receptor. The introduction of substituents in the 3, 4 or 5 positions in the phenyl ring may lead to compounds sterically hindered from combining with the histamine H₂-receptor.

7 There is no apparent relationship between the activities of clonidine-like imidazolidine derivatives as agonists on histamine H₂-receptors and their hypotensive activities (as reported in the literature).

     

Actions of mescaline on isolated rat atria.

Mescaline, in concentrations of 5 x 10(-4) and 1 x 10(-3) M, produced negative chronotropic and positive inotropic responses in isolated, spontaneously beating rat atria. In tissues driven at a constant rate, the inotropic response was diminished greatly, indicating that the increment in the force of contraction was secondary to the reduction in rate. These chronotropic and inotropic responses were not altered consistently by pretreatment with the histamine antagonists chlorpheniramine and metiamide.     

The antianaphylactic action of histamine H2-receptor agonists in the guinea-pig isolated heart.

The effects of histamine and of H1- and H2-receptor agonists on the response to specific antigen were studied in isolated hearts taken from actively sensitized guinea-pigs. Histamine and H2-receptor agonists (dimaprit, impromidine) dose-dependently decrease the positive chronotropic and inotropic effects, and the severity of arrhythmias evoked by the challenge of sensitized hearts with specific antigen. Nordimaprit and the selective H1-receptor agonist 2-pyridyl-ethyl-amine (2-PEA) did not modify the patterns of cardiac anaphylaxis. The positive inotropic and chronotropic responses of the isolated heart to exogenous histamine appear to be partly reduced in the presence of dimaprit. The H2-receptor agonists decrease the amount of histamine released during cardiac anaphylaxis which is increased by cimetidine, while nordimaprit and PEA were ineffective, indicating an inhibitory function afforded by H2-receptors in cardiac anaphylaxis.     

A study of the actions of histamine on the isolated rat heart

1. The effects of histamine on cardiac force, heart rate and coronary perfusion pressure were studied in the isolated rat heart, using the Langendorff perfused heart preparation. 2. Single injections of histamine induced dose-dependent decreases in contractile amplitude, heart rate and coronary perfusion pressure. 3. Perfusions of metiamide (above 1 × 10^-4 M) had a depressant effect on contractile force and heart rate, whereas diphenhydramine (4 × 10^-6 M) reduced only the heart rate. Both agents caused a fall in coronary perfusion pressure. 4. The negative inotropic and chronotropic effects of histamine on the isolated rat heart were not significantly influenced by either metiamide or diphenhydramine, or a combination of these drugs. However, the fall in coronary perfusion pressure induced by injections of histamine was significantly antagonized by metiamide or diphenhydramine. 5. These results suggest that the effects of histamine on the isolated rat heart may not be due entirely to stimulation of H₁- or H₂-receptors on the cardiac muscle cells. Evidence is presented for the existence of histamine H₁- and H₂-receptors in the coronary vessels.     

Dissociation of phosphoinositide hydrolysis and positive inotropic effect of histamine mediated by H1-receptors in guinea-pig left atria

Summary The present study was undertaken to determine whether the phosphoinositide hydrolysis is responsible for the positive inotropic effect of histamine in guinea-pig left atria. Histamine induced hydrolysis of phosphoinositides and a positive inotropic effect in a concentration-dependent manner. These effects were antagonized by chlorpheniramine (0.1 mol/l) but not by cimetidine (10 mol/l). At a concentration of 1 mol/l histamine produced a dual-component positive inotropic response composed of an initial increasing phase and a second and late developing, greater positive inotropic phase. Histamine (10 mol/l) caused a gradual increase in the formation of [³H]inositol trisphosphate (IP3) and a significant increase in the [³H]IP₃ level was detected 10 min after the stimulation. Thus, the increase in IP₃ did not precede the increase in force of contraction. The phospholipase C inhibitors 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (100 mol/l) and neomycin (100 mol/l) significantly reduced the histamine-induced [³H]inositol monophosphate accumulation. However, pretreatment with the phospholipase C inhibitors did not affect the positive inotropic effect of histamine, either in its extent or in its pattern. The phorbol esters 12-O-tetradecanoylphorbol-13-acetate (TPA) (100 nmol/l) and phorbol-12,13-dibutyrate (PDBu) (100 nmol/l) also significantly inhibited the phosphoinositide hydrolysis induced by histamine. The inhibitory effect of the phorbol esters on the phosphoinositide response was completely abolished in the presence of 10 mol/l 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), a protein kinase C inhibitor. TPA significantly attenuated the positive inotropic effect of histamine without changing the dual-component pattern, whereas PDBu merged two distinct components of the histamine inotropic response into one and potentiated the early part of the positive inotropic effect. However, neither of the changes which the phorbol esters produced in the positive inotropic response to histamine was blocked by H-7. In addition, H-7 itself failed to modify the positive inotropic effect of histamine. These results indicate that histamine induces hydrolysis of phosphoinositides in guinea-pig left atria that is mediated by H₁-receptors, but this biochemical event does not appear to contribute to the H₁-receptor-mediated positive inotropic action. Send offprint requests to Y. Hattori at the above address     

Effects of tetramethylpyrazine on isolated atria of the Guinea-pig.

The chronotropic and inotropic effects of tetramethylpyrazine (TMPZ) obtained from the stem of JATROPHA PODAGRICA H OOK (Euphorbiaceae) have been investigated on guinea-pig isolated atria. TMPZ caused negative chronotropic and inotropic responses on the isolated atria of guinea-pigs. These TMPZ - induced negative chronotropic and inotropic responses were resistant to the action of atropine. Prior administration to the bath fluid of TMPZ also antagonized the positive chronotropic and inotropic effects of isoprenaline, noradrenaline and calcium on the tissue preparations. It is concluded that TMPZ probably exerts a non-specific inhibitory effect on the isolated cardiac muscles examined, and that this non-specific depressant action on the myocardium might have contributed, at least in part, to the observed IN VIVO depressor (blood pressure lowering) effects of the amide alkaloid reported earlier.     

The effect of reserpine treatment on the chronotropic and inotropic sensitivities of the perfused guinea-pig heart to norepinephrine and calcium

The chronotropic and inotropic responses of the isolated perfused guinea-pig heart to norepinephrine and calcium were measured in preparations obtained from untreated and reserpinetreated animals. Dose-response curves were obtained in spontaneously beating as well as electrically paced perfused hearts. The dose-response curve for the chronotropic effect of both norepinephrine and calcium was shifted to the left of control in spontaneously beating preparations from animals pretreated with reserpine (0.1 mg/kg/day) for seven days. However, in these same preparations the dose-response curve for the inotropic effect of only norepinephrine was shifted to the left. This may be reflection of the enhanced sensitivity to the chronotropic action of the agonist since a changing rate is known to influence inotropic response. This conclusion is supported by the finding that, in electrically paced perfused hearts, no postjunctional supersensitivity to the inotropic effects of norepinephrine was demonstrable. Since no supersensitivity was observed to the inotropic effects of calcium, it is concuded that true postjunctional supersensitivity develops to the chronotropic but not to the inotropic effects of drugs.     

Pharmacological characterization of GSK1004723, a novel, long-acting antagonist at histamine H(1) and H(3) receptors

BACKGROUND AND PURPOSE

Preclinical pharmacological characterization of GSK1004723, a novel, dual histamine H₁ and H₃ receptor antagonist.

EXPERIMENTAL APPROACH

GSK1004723 was characterized in vitro and in vivo using methods that included radioligand binding, intracellular calcium mobilization, cAMP production, GTPγS binding, superfused human bronchus and guinea pig whole body plethysmography.

KEY RESULTS

In cell membranes over-expressing human recombinant H₁ and H₃ receptors, GSK1004723 displayed high affinity, competitive binding (H₁ pKi = 10.2; H₃ pKi = 10.6). In addition, GSK1004723 demonstrated slow dissociation from both receptors with a t_1/2 of 1.2 and 1.5 h for H₁ and H₃ respectively. GSK1004723 specifically antagonized H₁ receptor mediated increases in intracellular calcium and H₃ receptor mediated increases in GTPγS binding. The antagonism exerted was retained after cell washing, consistent with slow dissociation from H₁ and H₃ receptors. Duration of action was further evaluated using superfused human bronchus preparations. GSK1004723 (100 nmol·L⁻¹) reversed an established contractile response to histamine. When GSK1004723 was removed from the perfusate, only 20% recovery of the histamine response was observed over 10 h. Moreover, 21 h post-exposure to GSK1004723 there remained almost complete antagonism of responses to histamine. In vivo pharmacology was studied in conscious guinea pigs in which nasal congestion induced by intranasal histamine was measured indirectly (plethysmography). GSK1004723 (0.1 and 1 mg·mL⁻¹ intranasal) antagonized the histamine-induced response with a duration of up to 72 h.

CONCLUSIONS AND IMPLICATIONS

GSK1004723 is a potent and selective histamine H₁ and H₃ receptor antagonist with a long duration of action and represents a potential novel therapy for allergic rhinitis.